The influence of salinity on fertilization and larval development of Nereis virens (Polychaeta, Nereidae) from the White Sea

Effect of salinity on fertilization and early development of the polychaeta Nereis virens (Sars) from the White Sea was examined in laboratory experiments. The comparison of salinity resistance of different developmental stages of N. virens showed gradual increase of euryhalinity during ontogenesis—from fertilized eggs to juveniles.Successful fertilization and effective development (≥70-75%) was possible in narrow salinity range 22-34‰. The salinity range of successful development for trochophore and nectochaete larvae reached 14-45‰. This increase of the limits of salinity tolerance in trochophore and nectochaete larvae probably was due to the formation of protonephridium system.Rate of metamorphosis of N. virens was tested under temperature 5, 10, 17 and 23 °C and salinity 22-14‰. The highest rate of metamorphosis was marked at the temperature of 23 °C in salinities higher than 14‰.Our data confirms that N. virens originates from warm seas with oceanic salinity.     

Effects of salinity and water temperature on the ecological performance of Zostera marina

We tested the effects of salinity and water temperature on the ecological performance of eelgrass (Zostera marina L.) in culture-experiments to identify levels that could potentially limit survival and growth and, thus, the spatial distribution of eelgrass in temperate estuaries. The experiments included eight levels of salinity (2.5, 5, 10, 15, 20, 25, 30 and 35‰) and seven water temperatures (5, 10, 15, 20, 25, 27.5 and 30 °C). Low salinity (i.e. 5 and 2.5‰) increased mortality (3–6-fold) and had a strong negative effect on shoot morphology (number of leaves per shoot reduced by 40% and shoot biomass reduced by 30–40%), photosynthetic capacity (P_max—reduced by 30–80%) and growth (production of new leaves reduced by 50–60%, leaf elongation rate reduced by 60–70% and production of side-shoots reduced by 40–60%), whereas eelgrass performed almost equally well at salinities between 10 and 35‰. The optimum salinity for eelgrass was between 10 and 25‰ depending on the response parameter in question. Extreme water temperatures had an overall negative impact on eelgrass, although via different mechanisms. Low water temperatures (5 °C) slowed down photosynthetic rate (by 75%) and growth (production of new leaves by 30% and leaf elongation rate by 80%), but did not affect mortality, whereas high temperatures (25–30 °C) increased mortality (12-fold) and lowered both photosynthetic rate (by 50%) and growth (production of new leaves by 50% and leaf elongation rate by 75%). The optimum water temperature for eelgrass appeared to lie between 10 and 20 °C. These results show that extreme conditions may affect the fitness of eelgrass and, thus, may potentially limit its distribution in coastal and estuarine waters.     

Combined effect of temperature and salinity on the Thermotolerance and osmotic pressure of juvenile white shrimp litopenaeus vannamei (Boone)

Thermotolerance (CTMax) was determined in L. vannamei in three salinities and five acclimation temperatures 20, 23, 26, 29 and 32 °C. In white shrimp, the CTMax was not significantly affected by salinity (P>0.05). A direct relationship was obtained between CTMax and acclimation temperature. The end point of the CTMax in L. vannamei exposed to different combinations of temperature and salinity was defined as the loss of the righting response (LRR). The acclimation response ratio (ARR) for the juveniles of white shrimp ranged from 0.42 to 0.49; values in agreement with other crustaceans from tropical and sub tropical climates. The osmotic pressure of the hemolymph was measured in control organisms and in organisms exposed to CTMax; significant differences were found in organisms maintained in 10 and 40 psu, but there were no significant differences in hemolymph osmotic pressure in those that were acclimated to 26 psu.     

Effect of temperature and salinity on acetylcholinesterase activity, a common pollution biomarker, in Mytilus sp. from the south-western Baltic Sea

Acetylcholinesterase (AChE) activity is a well established biomarker to monitor environmental pollution caused by neurotoxic compounds, such as organophosphorus and carbamate pesticides. The presence of these compounds results in a measurable inhibition of the enzyme. This has been shown for different marine species including blue mussels. Besides pollution, environmental variables may also have a direct or indirect effect on AChE activity, particularly in estuarine and brackish water environments. To assess the impact of abiotic factors on the AChE activity the seasonal course of gill AChE activity was followed in relation to temperature and salinity in Mytilus sp. collected from the south-western Baltic Sea. In addition, the effect of salinity on AChE activity was investigated in an experimental study. The AChE activity showed significant seasonal differences with maximum activities during the summer period and minimum activities in winter. These changes correlate significantly (p<0.001) and positively with water temperature. The experimental exposure of Mytilus sp. to different salinities (5, 7, 16 or 20 psu) resulted in changes in the gill AChE activity. Empirical orthogonal function (EOF) analysis revealed that AChE activity was significantly and negatively correlated with salinity (p<0.01). These results clearly demonstrate the need to consider the potential influence of temperature and salinity on AChE activity when applying this biomarker to monitor exposure to and effect of neurotoxic substances in estuarine and brackish water blue mussels.     

Resistance of larvae of common species of White Sea invertebrates to extreme changes in salinity

This study examines the effect of sharp changes in salinity on pelagic larvae of ten common species of invertebrates from the brackish White Sea (Mollusca, Polychaeta, Echinodermata, Cnidaria, Ascidia). For five species, the low salinity resistance limit was in the range of 8–12‰: for the gastropod Littorina littorea, it was below 8‰; for Dyaphana sp. and the bivalves Hiatella arctica and Heteranomia ovata, it was more than 12‰; and for the ascidian Styela rustica, it was 16‰. About 50% of larvae of four investigated species were able to withstand high salinity and survived at 36–40 and even 50‰ (Littorina). Larvae of littoral-sublittoral species proved to be more euryhaline than larvae of sublittoral species.     

The opercular moult in Spirorbis spirorbis (L.) and S. pusilloides Bush (Polychaeta: Serpulidae)

Opercular moult has been examined in two spirorbids, the tube-incubating Spirorbis spirorbis (L.) and the opercular incubating Spirorbis pusilloides Bush. In both species epithelial ingrowth cuts off the old operculum and a new one differentiates on the proximal side of the constriction. The immediate fate of the severed tissues differs in the two species; in S. spirorbis the epithelia slowly wither as the platelike operculum finally breaks away by rupture of the cuticle while in S. pusilloides there is a cataclysmic autolysis of the epithelial tissues although the cuticle is retained and functions as an incubation chamber which ruptures at the close of incubation to release the larvae and is then shortly discarded.Opercular moult, rare in the immature animal in either species, normally begins with the onset of breeding. In S. pusilloides it is an essential preliminary to brooding and throughout the breeding season opercular moult rigidly alternates with brood incubation to produce a new incubation chamber for each successive brood. In S. spirorbis there is also some evidence of a partial correlation between brooding and opercular moult but the two events overlap and moulting is less frequent than the production of broods. A brief consideration is given to the evolution of the brooding habit and the tentative conclusion reached that opercular incubation has evolved twice in the Spirorbinae.     

Effects of temperature, salinity, and irradiance on the growth of the dinoflagellate Akashiwo sanguinea

The effects of temperature, salinity, and irradiance on the growth of the dinoflagellate Akashiwo sanguinea were examined in the laboratory. The irradiance at the light compensation point (I₀) was 14.40 μmol m^− 2 s^− 1 and the irradiance at growth saturation (I_s) was 114 μmol m^− 2 s^− 1. We exposed A. sanguinea to 48 combinations of temperature (5-30 °C) and salinity (5-40) under saturating irradiance; it exhibited its maximum growth rate of 1.13 divisions/day at a combination of 25 °C and salinity of 20. A. sanguinea was able to grow at temperatures from 10 to 30 °C and salinities from 10 to 40. This study revealed that A. sanguinea was a eurythermal and euryhaline organism; in Japan it should have formed blooms in early summer, when salinity was relatively low. In addition, it was noteworthy that A. sanguinea had markedly cold-durability, retaining the motile form of vegetative cells for more than 50 days at 5 °C and at salinities of 25-30.     

Effects of temperature,salinity and their interaction on growth of benthic dinoflagellates Ostreopsis spp. from Thailand

Benthic dinoflagellates Ostreopsis spp. are known as producers of palytoxin and its analogs, resulting occasionally in human health problems worldwide. Although distribution of Ostreopsis spp. along the Thai coasts has been reported, little is known about their growth characteristics. To discuss the bloom dynamics of Ostreopsis spp. in Thailand, first we tested four kinds of media to optimize growth conditions and then clarified the effects of temperature, salinity and temperature–salinity interaction on the growth of strains of the O. cf. ovata Thailand subclade, O. cf. ovata South China Sea subclade, Ostreopsis sp. 6 and Ostreopsis sp. 7. We showed that the f/2 medium was a suitable medium which gave the highest cell yields for all the strains tested. The strains of the O. cf. ovata Thailand subclade, O. cf. ovata South China Sea subclade and Ostreopsis sp. 6 grew in the temperature range 20–32.5 °C, whereas the strain of Ostreopsis sp. 7 grew in 20–30 °C. The semi-optimal temperature ranges (≧80% of the maximal growth rate) for the former three strains were 22.7–27.4 °C, 27.9–30.8 °C and 23.5–26.4 °C, respectively, whereas that of the latter strain was 23–27.2 °C. The optimal temperature for the O. cf. ovata South China Sea subclade was 30 °C, whereas for the others it was 25 °C. All the Ostreopsis strains tested could grow in a salinity range of 20–40. The semi-optimal salinities for the O. cf. ovata Thailand subclade, O. cf. ovata South China Sea subclade Ostreopsis sp. 6 and Ostreopsis sp. 7 were 28.7–35, 23.8–30.8, 29.8–36 and 28–36, respectively. The optimal salinities for the O. cf. ovata Thailand subclade and O. cf. ovata South China Sea subclade were 30 and 25, respectively, whereas for Ostreopsis sp. 6 and Ostreopsis sp. 7 it was 35. In this study, our results suggested that the optimal and tolerable temperature–salinity conditions differ among the Thai Ostreopsis species/clades/subclades. Tolerances of the O. cf. ovata Thailand subclade, O. cf. ovata South China Sea subclade and Ostreopsis sp. 6 to the high temperature of 32.5 °C may allow these organisms to be distributed in the tropical areas, where the water temperature often reaches >30 °C.     

Effects of reduced salinity on the biochemical composition (lipid, protein) of zoea 1 decapod crustacean larvae

Effects of reduced salinities on dry weight (DW) and biochemical composition (total lipid and protein contents) of zoea 1 larvae were evaluated in four decapod crustacean species differing in salinity tolerance (Cancer pagurus, Homarus gammarus, Carcinus maenas, Chasmagnathus granulata). The larvae were exposed to two different reduced salinities (15‰ and 25‰ in C. granulata, 20‰ and 25‰ in the other species) for a long (ca. 50% of the zoea 1 moulting cycle) or a short period (16 h, starting at ca. 40% of the moulting cycle), while a control group was continually maintained in seawater (32‰).In general, the increments in dry weight, lipid and protein content were lower at the reduced salinities than in the control groups. In the zoea 1 of H. gammarus (stenohaline) and C. pagurus (most probably also stenohaline), the lipid and protein contents varied greatly among treatments: larvae exposed to low salinities exhibited very low lipid and protein contents at the end of the experiments compared to the controls. In some cases, there were negative growth increments, i.e. the larvae had, after the experimental exposure, lower lipid and protein contents than at the beginning of the experiment. C. maenas (moderately euryhaline) showed a lower variation in protein and lipid content than the above species. The zoea 1 of C. granulata (fairly euryhaline) showed the lowest variability in dry weight, protein and lipid content. Since salinity tolerance (eury- v. stenohalinity) is associated with the osmoregulatory capacity, our results suggest a relationship between the capability for osmoregulation and the degree of change in the biochemical composition of larvae exposed to variable salinities.Besides larval growth of these species should be affected by natural reductions of salinity occurring in coastal areas at different time scales. These effects may be potentially important for population dynamics since they should influence the number and quality of larvae reaching metamorphosis.     

Effects of temperature on survival, development, growth and feeding of larvae of Yellowtail clownfish Amphiprion clarkii (Pisces: Perciformes)

To understand the physiological and ecological responses of marine fishes to the change of water temperature, newly-hatched larvae of Yellowtail clownfish Amphiprion clarkii were reared in captivity at water temperatures of 23, 26 and 29 °C till they completed the metamorphosis to juvenile phase, and larval survival, development, growth and feeding were evaluated during the experimental period. The results showed that water temperature influenced the physiological performance of larvae of A. clarkii significantly. The survival and growth rates of larvae of A. clarkii increased significantly with the increase of water temperature from 23 to 29 °C (P < 0.05). Water temperature also influenced larval development of A. clarkii significantly and larvae reared at 23 °C took longer time for post-larval development and metamorphosis compared to 26 and 29 °C (P < 0.05). Total length and body weight for post-larval development and metamorphosis decreased with the increase of water temperature from 23 to 29 °C (P < 0.05). Q₁₀ in developmental rate was higher than in daily growth rate at the same rearing temperature, indicating that at water temperature had greater influence on larval development than on growth. Water temperature also influenced larval feeding of A. clarkii significantly with feed ration (FR) and feed conversion efficiency (FCE) increased with the increase of water temperature from 23 to 29 °C (P < 0.05). There was a positive correlation between FR and specific growth rate (SGR) (P < 0.05) but not between FCE and SGR (P > 0.05), indicating that FR influenced growth rate significantly in larvae of A. clarkii. This study demonstrated that the physiological responses of larvae of A. clarkii to the change of water temperature and confirmed that water temperature influenced larval survival, development, growth and feeding significantly. This study suggests that the decline of larval survival and growth rates, extension of pelagic larval duration and reduction of larval feeding at lower temperature have ecological impacts on larval dispersal and metamorphosis, juvenile settlement and population replenishment in A. clarkii in the wild.     

Effects of salinity, temperature and pH on the survival of the nemertean Procephalothrix simulus Iwata, 1952

The salinity, temperature and pH tolerance of Procephalothrix simulus Iwata, 1952, were experimentally studied. In hypo-media, the nemerteans could survive 96 h in 3.3‰ solution at 10 °C (median lethal salinity [LS₅₀] was not determined at this temperature), and 96 h LS₅₀ were 7.3‰ and 13.5‰ at 20 °C and 30 °C, respectively. In hyper-media, 96 h LS₅₀ values were 53.9‰, 47.1‰ and 41.4‰ at 10 °C, 20 °C and 30 °C, respectively. The trend of body weight changes in diluted media indicated that this nemertean is a volume regulator. During a 96-h exposure in media at 0 °C, worms were thanatoid but could recover if the temperature was gradually elevated to 20 °C. In thermal tolerance experiments, the nemertean survived 96 h in seawater of 30 °C, and worms suffered high mortalities when the temperature exceeded 32 °C. Present results suggest that the interaction of temperature and salinity on the lethal effects on P. simulus is significant (P < 0.05). Elevated temperature (range 10-30 °C) decreased the worm's solute tolerance, and elevated salinity (range 18-38‰) decreased the worm's thermal tolerance. The survival pH level for this nemertean ranged from 5.00 to 9.20.     

温度和盐度对中华多刺鱼胚胎发育过程的影响

2006年3-5月,在北京怀柔水库入库河流怀九河的西四渡河河段,采集性成熟的中华多刺鱼(Pungitius sinensis),人工干法授精获取受精卵,观察并记录受精卵的胚胎发育历程;设置温度和盐度梯度,观察温度和盐度对中华多刺鱼胚胎发育过程的影响。结果表明:北京地区的中华多刺鱼胚胎发育的起始温度(生物学零度)为4.7℃,适宜的发育水温为14.0℃-16.0℃,与中华多刺鱼繁殖盛期(4月中下旬)自然水体的水温相近;平均水温14.0℃时,中华多刺鱼完成胚胎发育约需183h;水温升至22.5℃时,胚胎的孵化率仅为22.2%;水温升至25.0℃时,受精卵不能孵化,此温度应是中华多刺鱼胚胎发育温度的上限。实验表明,北京地区中华多刺鱼的胚胎对盐度的耐受力较为有限,在与自然水体相同的盐度下(0‰)其孵化率接近100%;随着盐度的升高,孵化率降低,8‰盐度时,仅有56%的受精卵孵化;至20‰时,受精卵全部死亡,此盐度应为中华多刺鱼胚胎发育的盐度上限。     

Effects of temperature,salinity and their interaction on growth of Japanese Gambierdiscus spp. (Dinophyceae)

Marine toxic dinoflagellates of the genus Gambierdiscus are the causative agents of ciguatera fish poisoning (CFP), a seafood poisoning that is widespread in tropical, subtropical and temperate regions of the world. In the main island of Japan, distributions of Gambierdiscus australes, Gambierdiscus scabrosus and two phylotypes of Gambierdiscus spp. type 2 and type 3, have been reported. To discuss the bloom dynamics of these Japanese species/phylotypes of Gambierdiscus, first we tested six culture media to optimize growth conditions and then clarified the effects of temperature and salinity and temperature–salinity interactions on growth. All strains of the species/phylotypes tested showed the highest cell yields when they were cultivated in IMK/2 medium. G. australes, G. scabrosus and Gambierdiscus sp. type 2 grew in the range 17.5–30 °C, whereas Gambierdiscus sp. type 3 grew in 15–25 °C. The semi-optimal temperature ranges (≥80% of the maximal growth rate) of the former three species/phylotypes were 19–28 °C, 24–31 °C and 21–28 °C, respectively, whereas that of the latter phylotype was 22–25 °C. Hence, Gambierdiscus sp. type 3 may be adapted to relatively lower water temperatures of ≤25 °C. In contrast, G. australes, G. scabrosus and Gambierdiscus sp. type 2 presumably possess adaptability to relatively high water temperatures. The optimal temperature for G. scabrosus was 30 °C, whereas the optimal temperature for the others was 25 °C. G. australes and Gambierdiscus sp. type 3 grew in a salinity range of 25–40 whereas G. scabrosus and Gambierdiscus sp. type 2 grew in salinity 20–40. Furthermore, the semi-optimal salinity range of G. australes, G. scabrosus, Gambierdiscus spp. type 2 and type 3 were salinity 27–38, 24–36, 22–36 and 29–37, respectively. Among the species/phylotypes, G. scabrosus and Gambierdiscus sp. type 2 grew even at salinity 20 where the others did not grow, thus possessing adaptability to low salinity waters. Our results clearly demonstrate that the optimal and tolerable temperature–salinity conditions differ among Japanese Gambierdiscus species/phylotypes. Considering these results, temperature–salinity interactions may play an important role in bloom dynamics and the distribution of the Gambierdiscus species/phylotypes in Japanese coastal waters.     

The effect of temperature and salinity on growth rate and azaspiracid cell quotas in two strains of Azadinium poporum (Dinophyceae) from Puget Sound,Washington State

Azaspiracids (AZA) are novel lipophilic polyether marine biotoxins associated with azaspiracid shellfish poisoning (AZP). Azaspiracid-59 (AZA-59) is a new AZA that was recently detected in strains of Azadinium poporum from Puget Sound, Washington State. In order to understand how environmental factors affect AZA abundances in Puget Sound, a laboratory experiment was conducted with two local strains of A. poporum to estimate the growth rate and AZA-59 (both intra- and extracellular) cell quotas along temperature and salinity gradients. Both strains of A. poporum grew across a wide range of temperatures (6.7 °C to 25.0 °C), and salinities (15 to 35). Growth rates increased with increasing temperature up to 20.0 °C, with a range from 0.10 d⁻¹ to 0.42 d⁻¹. Both strains of A. poporum showed variable growth rates from 0.26 d⁻¹ to 0.38 d⁻¹ at salinities from 15 to 35. The percentage of intracellular AZA-59 in both strains was generally higher in exponential than in stationary phase along temperature and salinity gradients, indicating higher retention of toxin in actively growing cells. Cellular toxin quotas varied by strain in both the temperature and salinity treatments but were highest at the lowest growth rates, especially for the faster growing strain, NWFSC1011.Consistent with laboratory experiments, field investigations in Sequim Bay, WA, during 2016–2018 showed that A. poporum was detected when salinity and temperature became favorable to higher growth rates in June and July. Although current field data of A. poporum in Puget Sound indicate a generally low abundance, the potential of local A. poporum to adapt to and grow in a wide range of temperature and salinity may open future windows for blooms. Although increased temperatures, anticipated for the Puget Sound region over the next decades, will enhance the growth of A. poporum, these higher temperatures will not necessarily support higher toxin cell quotas. Additional sampling and assessment of the total toxicity of AZA-59 will provide the basis for a more accurate estimation of risk for azaspiracid poisoning in Puget Sound shellfish.     

Effect of temperature and salinity on the energetics of juvenile gray snapper (Lutjanus griseus): implications for nursery habitat value

Gray snapper (Lutjanus griseus) encounter a wide range of temperatures and salinities in nearshore and estuarine juvenile habitats. The energetic response of juvenile gray snapper to temperature and salinity was measured in laboratory experiments to determine the influence of these physicochemical factors on the potential value of different juvenile nurseries. Maximum consumption and growth rates of juvenile (25-50 mm SL) gray snapper were determined in 12-day trials at 20 temperature/salinity combinations representing conditions in juvenile habitats. Ad libitum feeding level of individual fish was measured daily. Maximum weight specific feeding rate increased significantly with temperature and salinity; however, the effect of salinity was much less than that of temperature. Linear growth rate and specific growth rate both increased with temperature, and salinity did not have a significant effect on either. Gross growth efficiency (K₁, growth×consumption⁻¹*100) increased with temperature and was significantly lower at high salinities, indicating increased energetic costs. The higher K₁ at lower salinities has several implications for juvenile gray snapper in low salinity habitats: (1) they would need less food to achieve the same somatic growth as juveniles in high salinity habitats; (2) they would have higher growth at limited ration levels as compared to high salinity habitats; and (3) they would have less impact on prey populations than higher salinity habitats assuming similar gray snapper densities.     

The use of metabolism to evaluate the toxicity of dodecil benzen sodium sulfonate (LAS-C12) on the Mugil platanus (mullet) according to the temperature and salinity

The effects of watering exposition to different concentrations of LAS-C12 (1, 2.5 and 5 mg l⁻¹) about the Mugil platanus routine metabolism were evaluated. The metabolic rates were estimated through experiments accomplished in each of the twelve possible combinations of three temperatures (25, 20 and 15 °C) and three salinities (35, 20 and 5). The results show that the oxygen consumption increases according to the LAS-C12 concentration in all temperatures and salinities studied. At the highest concentration employed (5 mg l⁻¹), and the salinity 5 in the temperatures 25 and 20 °C, oxygen consumption increases 80% in relation to the control. In general, the pollutant effects on oxygen consumption were more pronounced at the highest temperatures and salinities 5 and 35.     

In vitro effects of temperature and salinity on fatty acid synthesis in the oyster protozoan parasite Perkinsus marinus

The effects of temperature and salinity on fatty acid synthetic activities in the oyster protozoan parasite, Perkinsus marinus, were tested in vitro at 10, 18 and 28 °C in a salinity of 28 psu and 14, 20 and 28 psu at a temperature of 28 °C using ¹³C sodium acetate as a substrate. Salinity treatments exhibited few treatment effects, but temperature significantly affected cell proliferation, fatty acid content and fatty acid synthesis rates. Fatty acid synthesis rates increased approximately two-fold for every 10 °C increase in temperature; however, the predominant fatty acid synthesized differed between treatments. At 10 °C, the synthesis rate for 18:1(n−9) was not significantly different from the 18 °C treatment and weight percent of 18:1(n−9) was higher at 10 than 18 and 28 °C. In contrast, the synthesis rate for 20:4(n−6) was over five times lower at 10 than at 18 and 28 °C, and the percent fatty acid content of 20:4(n−6) was over two-fold lower at 10 than at 18 and 28 °C. Results suggest that further elongation and desaturation of 18:1(n−9) to 20 carbon polyunsaturated fatty acids may be inhibited at low temperatures. These findings may be relevant to field observations that disease progression and virulence of this parasite are correlated to high water temperatures.     

Effect of salinity, temperature, organic and inorganic nutrients on growth of cultured Fibrocapsa japonica (Raphidophyceae) from the northern Adriatic Sea

The effects of temperature, salinity, inorganic and organic nutrients on the growth and physiological performance of Fibrocapsa japonica have been investigated in strains isolated from the northern Adriatic Sea, where intense and regular blooms have occurred since 1997 in coastal waters during summer. Strains isolated at different times and from different locations appeared homogeneous in terms of both physiological responses and molecular (ITS-5.8S rDNA) characteristics. Growth rates were higher at temperatures between 20 and 26 °C and in a salinity range of 30–35 (0.7 div day⁻¹). The temperature of 16 °C inhibited growth, more markedly at the lowest and highest salinity values, a result also confirmed by a lower photosynthetic efficiency and by an increase in cell volume due to impaired division. Higher cell concentrations were obtained with macronutrients at f/2 levels than in a fivefold diluted medium. Comparing the utilization of 200 μM nitrate to that of different N sources (inorganic and organic), F. japonica showed an efficient growth with equivalent amounts of ammonia, urea and amino acids, such as glycine and tryptophan; glutamate was less effective, while methionine had toxic effects. Organic phosphate, administered as glycerophosphate, could also sustain F. japonica growth, probably on account of an alkaline phosphatase whose activity was enhanced in the presence of the organic form. Vitamins were necessary for growth, though no further stimulation was observed when a surplus of vitamin B12 (3 nM) was added. The addition of 11.7 μM iron instead of 2.3 μM, as well as that of humic acid, with or without macronutrients, did not enhance algal growth either. These results led us to hypothesise that F. japonica blooms became more frequent due to a general seawater temperature increase and to the availability of organic forms which, in coastal anthropized areas, are especially abundant in summer periods.     

Effects of salinity and temperature on the survival and byssal attachment of the lion's paw scallop Nodipecten nodosus at its southern distribution limit

The lion's paw scallop, Nodipecten nodosus, is subject to wide temperature variations on seasonal and short-term scales, and may be exposed to low-salinity events, caused by oceanographic and meteorological processes at its southern distribution limit (Santa Catarina State, Brazil). Such variations may have important implications on the distribution and on aquaculture site selection. The upper and lower temperature tolerances and the percentage of byssal attachment at different temperatures (11 to 35 °C) were studied for spat, juvenile and adult scallops. The lethal and sublethal effects of reduced salinity (13‰ to 33‰) on spat, juvenile and adult scallops were studied at ambient temperature (23.5 °C) and on spat also at low (16 °C) and high (28 °C) temperatures during 96-h bioassays. In addition, the influences of short exposure (1 h) to low salinity (13‰ and 17‰) at different temperatures (16 and 28 °C), and the effects of exposure (2 and 4 h) to high temperature (33 °C) at ambient salinity (33‰) were studied. N. nodosus is a moderately eurythermal but stenohaline tropical species, adults having lower tolerance to high temperature and low salinity than spat. Lethal temperatures for a 48-h exposure (LT₅₀) were 29.8 °C for adult and juveniles, and 31.8 °C for spat. Maximum rate of byssal attachment occurred in a narrower temperature range for juveniles and adults (23 to 27 °C) than for spat (19 to 27 °C), which are suggested as the optimum ranges of temperatures for growth. Lethal salinities (LC₅₀) for a 48-h exposure at ambient temperature were 23.2‰, 23.6‰ and 20.1‰ for adults, juveniles and spat, respectively, but the percent byssal attachment was significantly reduced below salinities of 29‰ indicating that scallops were physiologically stressed. A 1-h exposure to 17‰ was lethal to spat at 28 °C, but at 16 °C there was a 28.5% survival, 96 h after the exposure. Temperatures and salinity in coastal areas of southern Brazil can reach levels leading to sublethal effects, and in some sites, it may surpass the limits of tolerance for the survival of the species.     

Biomarker responses of the estuarine brown shrimp Crangon crangon L. to non-toxic stressors: Temperature, salinity and handling stress effects

Biochemical biomarkers in common estuarine species, such as the brown shrimp Crangon crangon, have the potential to provide early warning of contaminant exposure from field collected samples and through the development of in situ tests. The biomarkers acetylcholinesterase (AChE), lactate dehydrogenase (LDH) and glutathione S-transferase (GST) have been shown to provide evidence of exposure to contaminants in a number of species and field situations. As they may naturally respond to the marked physicochemical changes found in estuaries (thus confounding contaminant-induced effects), this work aims to determine the effects of salinity, temperature and handling stressors on these biomarkers in C. crangon.AChE recovery in field-collected shrimp transplanted to clean laboratory conditions suggests the presence of inhibiting factors at the sampling site (River Minho estuary). Maintenance time in stock tanks had effects that led to the choice of a minimum 15-day maintenance period of C. crangon in the laboratory before subsequent use of the enzymes as effect criterions in toxicity assays. Field levels of biomarker activity were unaffected following field-laboratory transportation of C. crangon, making this factor unlikely to jeopardize detection of contaminant associated effects. LDH levels were significantly lower under conditions that mimic a diurnal salinity fluctuation, increasing under low salinity conditions; this potentially indicates increased energy costs associated with raised osmoregulatory demands. It is recommended that a lower limit to field exposure of in situ tests should be based on salinity. Higher temperatures led to higher AChE activities and this is in agreement with the existing evidence of increases of endogenous AChE levels as a function of temperature (within a certain range). To avoid misinterpretation of biomarker responses, studies such as this are an important contribution to the establishment of reference activity levels against which biomarker changes can be estimated and are therefore essential preliminary steps in the development of in situ bioassays using biomarkers.