Acclimation of the intertidal red alga Bangiopsis subsimplex (Stylonematophyceae) to salinity changes

The effect of salinity on growth, photosynthetic performance and osmotic acclimation was investigated in the eulittoral red algal species Bangiopsis subsimplex (Stylonematophyceae). The strain grew in a broad salinity range between 1 and 70 psu showing optimum growth between 10 and 50 psu. The saturation point I_k of the photosynthesis irradiance curves ranged between 153 and 83 μmol photons m^− 2 s^− 1 at all salinities and indicates an adaptation of B. subsimplex to moderate radiation conditions. Adjustments on the photosynthetic level (non-photochemical quenching) were sufficient to prevent damage to the photosynthetic apparatus as F_v/F_m values were constantly high (> 0.7) even when grown at the most hypo- and hypersaline conditions. As main low molecular weight carbohydrates, B. subsimplex contains the heteroside digeneaside and the polyol sorbitol. Digeneaside concentration was low and almost unchanged after hypersaline treatment (< 20 μmol g^− 1 DW), i.e. it did not play a role in osmotic acclimation. By contrast, sorbitol levels increased linearly from 150 to 380 μmol g^− 1 DW with increasing salinities between 5 and 60 psu, indicating its important function as an osmolyte and compatible solute under hypersaline conditions. The data presented are consistent with the natural habitat of B. subsimplex, i.e. the upper eulittoral zone.     

Effects of temperature and salinity on the standard metabolic rate (SMR) of the caridean shrimp Palaemon peringueyi

The standard metabolic rate (SMR) of the caridean shrimp Palaemon peringueyi to changes in temperature (15-30 °C), salinity (0-45‰) and a combination thereof was investigated. The rate of oxygen consumption of the shrimp was determined using a YSI oxygen meter. At a constant salinity of 35‰ the respiration rate of P. peringueyi increased with an increase in temperature and ranged between 0.260 and 0.982 μl O₂ mg wwt^− 1 h^− 1. The Q₁₀ value over the temperature range 15-25 °C was estimated at 3.13. At a constant temperature of 15 °C the respiration rate of P. peringueyi also increased with an increase in salinity and ranged between 0.231 and 0.860 μl O₂ mg wwt^− 1 h^− 1. For combination experiments the absence of any significant difference in the respiration rate of P. peringueyi at the four temperatures over the salinity range 15-35‰ suggests that the shrimp is well adapted to inhabiting environments characterised by variations in salinity and temperature such as those encountered within the middle and lower reaches of permanently open estuaries with substantial freshwater inflow. On the other hand, the total mortality of the shrimp recorded at salinities < 5‰ at all four temperatures suggests that the upper distribution of the shrimp may reflect physiological constraints. Similarly, the increase in the respiration rate of the shrimp at the four temperatures at salinities > 35‰ suggests that the shrimp may experience osmotic stress in freshwater deprived permanently open and intermittently open estuaries where hypersaline conditions may develop.     

Effects of temperature, salinity and irradiance on the growth of the red tide dinoflagellate Gyrodinium instriatum Freudenthal et Lee

The effects of temperature, salinity and irradiance on the growth of the red tide dinoflagellate Gyrodinium instriatum Freudenthal et Lee were examined in the laboratory. Exposed to 45 different combinations of temperature (10–30 °C) and salinity (0–40) under saturating irradiance, G. instriatum exhibited its maximum growth rate of 0.7 divisions/day at a combination of 25 °C and a salinity of 30. Optimum growth rates (>0.5 divisions/day) were observed at temperatures ranging from 20 to 30 °C and at salinities from 10 to 35. The organism could not grow at ≤10 °C. In addition, G. instriatum burst at a salinity of 0 at all temperatures, but grew at a salinity of 5 at temperatures between 20 and 25 °C. It is noteworthy that G. instriatum is a euryhaline organism that can live under extremely low salinity. Factorial analysis revealed that the contributions of temperature and salinity to its growth of the organism were almost equal. The irradiance at the light compensation point (I₀) was 10.6 μmol/(m² s) and the saturated irradiance for growth (I_s) was 70 μmol/(m² s), which was lower than I_s for several other harmful dinoflagellates (90–110 μmol/(m² s)).     

Effects of salinity, temperature and individual variability on the reproduction of Eurytemora affinis (Copepoda; Calanoida) from the Seine estuary: A laboratory study

The brackish water copepod Eurytemora affinis is the most abundant copepod species in the low salinity zone (2-15) of the Seine estuary. Despite its ecological importance, little is known about its population dynamics in the Seine. We studied the effects of temperature (10 °C and 15 °C) and salinity (5, 15 and 25) on reproduction under non-limiting food conditions. We used experiments to determine multiple reproductive parameters for E. affinis. In all experiments, we fed E. affinis a mixture of Rhodomonas marina and Isochrysis galbana. Couples of pre-adult females (C5) and adult males were mated until the female extruded a clutch of eggs and then individual females were observed every 6-12 hours until death to determine (a) embryonic development time, (b) inter clutch time and (c) clutch size throughout their adult lifespan. All reproductive parameters were negatively affected by low temperature (10 °C) and by high salinity (25). At 10 °C and a salinity of 25, mortality during the post-embryonic period was extremely high (85%). Differences in all reproductive parameters between salinities 5 and 15 were minimal. From 15 °C to 10 °C mean latency time (time between hatching of eggs and extrusion of new ones) increased from 0.8 to 2.25 days, the mean embryonic development time from 2.2 to 3.2 days and the mean clutch size decreased from 38 to 22 eggs female^- 1. The mean clutch size decreased when females reached a critical age. The hatching success was high (near 95%) under all conditions except at high salinity. Egg production rates showed no significant differences between salinities 5 and 15 and were significantly higher at 15 °C (13 eggs female^- 1 day^- 1 at salinity 5 and 15) than at 10 °C (4 eggs female^- 1 day^- 1). These values at 15 °C were higher compared to those from other populations of E. affinis in estuaries or lakes. The high reproductive potential of E. affinis from the Seine estuary at 15 °C and low salinities explain its high densities in the low salinity zone during spring and early summer.     

Ion composition and osmolarity of Caspian Sea ctenophore, Mnemiopsis leidyi, in different salinities

The aim was to study osmotic and ionic regulation in alien ctenophore, Mnemiopsis leidyi (total length of 1.5-3 cm), acclimated gradually to artificial sea water of different salinities (8, 13, 18 and 23 ppt) in laboratory conditions (24 ± 2 °C) with the salt composition and proportion of the Caspian Sea. Results showed that this species is hyper-osmoconformer (maintaining internal osmolarity 2-22 mOsmol l^{− 1} above external) in non-lethal salinities ranging from 8 to 23 ppt. The results of ionic regulatory investigations revealed that this species can regulate Ca²⁺ and SO₄²⁻ so that the concentration of Ca²⁺ in the internal fluid and SO₄²⁻ in the external fluid were significantly more in 8 and 13 ppt, respectively. No regulation was observed about other ions such as Cl⁻, Mg²⁺ and K⁺ in the mentioned salinities.     

Salinity induced changes in haemolymph osmolality and total metabolic rate of the mud crab Rhithropanopeus harrisii Gould, 1841 from Baltic coastal waters

The specific metabolic rate (SMR) and haemolymph osmolality (HO) of the mud crab Rhithropanopeus harrisii Gould, 1841 from Baltic brackish waters were measured at a habitat salinity of 7 psu (T = 15 °C, full air saturation) and after step-wise acclimation to a salinity range of 3-27 psu. Values of SMR at 7 psu varied between 0.40 and 3.89 J g^− 1 WW h^− 1 (n = 25, wet weight range 0.051-1.142 g) and were significantly (p < 0.05) related to the specimen's wet weight (WW) according to the power regression SMR = 0.94 WW^− 0.41 (R² = 0.68). The SMR of females did not differ significantly (p > 0.05) from those of males. When exposed to higher salinities, the SMR of R. harrisii decreased significantly (p < 0.05) and reached a minimum value at 23 psu (0.55 ± 0.05 J g^− 1 WW h^− 1, n = 6). Mean haemolymph osmolality at 7 psu amounted to 581 ± 26 mOsm kg^− 1 (n = 5) and was 2.9 times higher than that of the external medium. R. harrisii hyperosmoregulated its body fluids up to 24 psu (727 mOsm kg^− 1) at which salinity the isosmotic point was reached.     

Does scope for growth change as a result of salinity stress in the amphipod Gammarus oceanicus?

Physiological performance (feeding, metabolism, growth and excretion) across a broad range of salinity (5-30 psu) were determined for the benthic amphipod Gammarus oceanicus, a species of marine origin inhabiting brackish waters of the southern Baltic Sea. Feeding rates decreased with increasing salinity, whereas the nutritive absorption efficiency increased. Faeces production and ammonia excretion rates decreased strongly from the lowest to the highest salinity by 60% and 58%, respectively. Increasing salinity was accompanied by a reduction in the metabolic rate from 438 J g^− 1 dry wt d^− 1 (5.1 mW g^− 1) at 5 psu to 245 J g^− 1 (2.8 mW g^− 1) at 30 psu. Individuals were able to maintain a positive energy balance at all experimental salinities. The greatest values for scope for growth were recorded at the environmental salinity (7 psu) with a mean of 769 J g^− 1 dry wt d^− 1 (8.7 mW g^− 1).     

Multigenerational analysis of temperature and salinity variability affects on metabolic rate,generation time,and acute thermal and salinity tolerance in Daphnia pulex

Climate change, sea level rise, and human freshwater demands are predicted to result in elevated temperature and salinity variability in upper estuarine ecosystems. Increasing levels of environmental stresses are known to induce the cellular stress response (CSR). Energy for the CSR may be provided by an elevated overall metabolic rate. However, if metabolic rate is constant or lower under elevated stress, energy for the CSR is taken from other physiological processes, such as growth or reproduction. This study investigated the examined energetic responses to the combination of temperature and salinity variability during a multigenerational exposure of partheogenetically reproducing Daphnia pulex. We raised D. pulex in an orthogonal combination of daily fluctuations in temperature (15, 15–25, 15–30 °C) and salinity (0, 0–2, 0–5). Initially metabolic rates were lower under all variable temperature and variable salinity treatments. By the 6th generation there was little metabolic variation among low and intermediate temperature and salinity treatments, but metabolic suppression persisted at the most extreme salinity. When grown in the control condition for the 6th generation, metabolic suppression was only observed in D. pulex from the most extreme condition (15–30 °C, 0–5 salinity). Generation time was influenced by acclimation temperature but not salinity and was quickest in specimens reared at 15–25 °C, likely due to Q₁₀ effects at temperatures closer to the optima for D. pulex, and slowest in specimens reared at 15–30 °C, which may have reflected elevated CSR. Acute tolerance to temperature (LT₅₀) and salinity (LC₅₀) were both highest in D. pulex acclimated to 15–30 °C and salinity 0. LT₅₀ and LC₅₀ increased with increasing salinity in specimens raised at 15 °C and 15–25 °C, but decreased with increasing salinity in specimens raised at 15–30 °C. Thus, increasing temperature confers cross-tolerance to salinity stress, but the directionality of synergistic effects of temperature and salinity depend on the degree of environmental variability. Overall, the results of our study suggest that temperature is a stronger determinant of metabolism, growth, and tolerance thresholds, and assessment of the ecological impacts of environmental change requires explicit information regarding the degree of environmental variability.     

Physiological responses to flooding and light in two tree species native to the Amazonian floodplains

In Amazonian floodplains, plant survival is determined by adaptations and growth strategies to effectively capture sunlight and endure extended periods of waterlogging. By measuring gas exchange, quantum efficiency of photosystem 2 (PSII), and growth parameters, we investigated the combined effects of flooding gradients and light on two common evergreen floodplain tree species, the light-tolerant Cecropia latiloba and the shade-tolerant Pouteria glomerata. Individual plants were subjected to different combinations of light and flooding intensity in short-term and long-term experiments. Plants of C. latiloba lost all their leaves under total submersion treatments (plants flooded to apex and with reduced irradiance) and showed highest maximum assimilation rates (A_max) in not flooded, high light treatments (6.1 μmol CO₂ m⁻² s⁻¹). Individuals of P. glomerata showed similar patterns, with A_max increasing from 1.9 μmol CO₂ m⁻² s⁻¹ under total flooding to 7.1 μmol CO₂ m⁻² s⁻¹ in not flooded, high light treatments. During the long-term flooding experiment, quantum efficiency of PSII (F_v/F_m) of C. latiloba was not affected by partial flooding. In contrast, in P. glomerata F_v/F_m decreased to values below 0.73 after 120 days of total flooding. Moreover, total submergence led P. glomerata to reduce significantly light saturation point (LSP), as compared to C. latiloba. For both species morphological adjustments to long-term flooding, such as the production of adventitious roots, resulted in reduced total biomass, relative growth rate (RGR) and leaf mass ratio (LMR). Growth increase in C. latiloba seemed to be more limited by low-light than by flooding. Therefore, the predominant occurrence of this species is in open areas with high light intensities and high levels of inundation. In P. glomerata flooding induced high reductions of growth and photosynthesis, whereas light was not limiting. This species is more abundant in positions where irradiance is reduced and periods of submergence are slightly modest. We could show that the physiological requirements are directly responsible for the flooding (C. latiloba) and shade (P. glomerata) tolerance of the two species, which explains their local distribution in Amazonian floodplain forests.     

The combined influence of sub-optimal temperature and salinity on the in vitro viability of Perkinsus marinus, a protistan parasite of the eastern oyster Crassostrea virginica

Perkinsus marinus is a major cause of mortality in eastern oysters along the Gulf of Mexico and Atlantic coasts. It is also well documented that temperature and salinity are the primary environmental factors affecting P. marinus viability and proliferation. However, little is known about the effects of combined sub-optimal temperatures and salinities on P. marinus viability. This in vitro study examined those effects by acclimating P. marinus at three salinities (7, 15, 25 ppt) to 10 °C to represent the lowest temperatures generally reached in the Gulf of Mexico, and to 2 °C to represent the lowest temperatures reached along the mid-Atlantic coasts and by measuring changes in cell viability and density on days 1, 30, 60 and 90 following acclimation. Cell viability and density were also measured in 7 ppt cultures acclimated to each temperature and then transferred to 3.5 ppt. The largest decreases in cell viability occurred only with combined low temperature and salinity, indicating that there is clearly a synergistic effect. The largest decreases in cell viability occurred only with both low temperature and salinity after 30 days (3.5 ppt, 2 °C: 0% viability), 60 days (3.5 ppt, 10 °C: 0% viability) and 90 days (7 ppt, 2 °C: 0.6 ± 0.7%; 7 ppt, 10 °C: 0.2 ± 0.2%).     

Salinities, not diets, affect strontium/calcium ratios in otoliths of Anguilla japonica

Although otolith Strontium (Sr)/calcium (Ca) ratios have been widely used to reconstruct the past salinity environmental history of anguillid eels, factors affecting the Sr/Ca ratios in otoliths are incompletely understood. Japanese Eel (Anguilla japonica) elvers (mean length 54.7 ± 2.1 mm) were collected in the estuary during their upstream migration and reared at 5 different salinities (0, 5, 15, 25, and 35 psu) and 3 types of feeding conditions (formulated feed, tubifex, and starvation) for 30 days to evaluate the effects of salinity and diets on otolith Sr/Ca ratios. Ca and Sr concentrations in the ambient water significantly increased with salinity (SAL) as [Ca] _water = 15.50SAL − 5.56, and [Sr] _water = 0.21SAL + 0.03, respectively. Sr/Ca ratios in otoliths increased with salinity (SAL) of the rearing water as [(Sr/Ca) × 1000] _otolith = 0.091SAL + 3.790. In diets, Sr/Ca ratios were 4 times higher in tubifex than in formulated feed. However, in otoliths, ANOVA indicated that Sr/Ca ratios did not differ significantly between groups fed on tubifex or formulated feed (p = 0.118). Otolith Sr/Ca ratios were negatively correlated with fish growth rates while the growth rates differed significantly among rearing conditions with different salinities and diets. Partition coefficients of the Sr/Ca ratios from ambient water to fish tissues and otoliths significantly increased with salinity. The Sr/Ca ratios of Japanese Eel otoliths thus were positively correlated with the ambient salinity and decreased with increasing fish growth rate, but was not affected by fish diet.     

Laboratory experiments on the effects of variable suspended sediment concentrations on the ecophysiology of the porcelain crab Petrolisthes elongatus (Milne Edwards, 1837)

The porcelain crab Petrolisthes elongatus is a particulate suspension feeding species common to coastal areas of New Zealand (NZ). Consistent with the responses of other suspension feeding species, it is likely to be negatively influenced by elevated suspended sediment concentrations. Laboratory experiments were conducted to quantify the effect of temperature (12 °C, 15 °C and 18 °C) and suspended sediment concentration (total particulate matter (TPM): low < 100 mg L^− 1; medium 100-1000 mg L^− 1; high > 1000 mg L^− 1) on the clearance rate (CR in L h^− 1), oxygen uptake rate (VO₂ in mL h⁻¹), net absorption efficiency (AE), and net energy budget (NEB in J h^− 1) of P. elongatus across a range of sizes. Variation in CR and AE was independent of temperature and of body size, but were significantly different (P < 0.05) at low and medium suspended sediment concentrations compared with high suspended sediment concentrations. CR responded in a non-linear manner to changes in TPM, increasing with TPM up to a maximum value at medium-low concentrations (approximately 250 mg L^− 1) and then decreasing thereafter. CR had almost completely shut down at TPM concentrations of > 1000 mg L^− 1 and at particulate organic matter (POM) concentrations of > 250 mg L^− 1. AE was zero at approximate TPM and POM values of 1200 mg L^− 1 and 300 mg L^− 1, respectively. VO₂ was positively correlated with body size and with temperature, but was independent of TPM. NEB values for P. elongatus were low (approx 110 J g^− 1 h^− 1) at low sediment concentrations, were high (approx 320 J g^− 1 h^− 1) at medium sediment concentrations, and were negative (approx − 114 J g^− 1 h^− 1) at high sediment concentrations. These findings indicate that P. elongatus is likely to be food-limited at sediment concentrations of < 100 mg L^− 1, and severely negatively affected at sediment concentrations of > 1000 mg L^− 1, at least for the duration of such events which may persist for 2-3 days in coastal environments where this crab occurs.     

Influence of ammonium sulphate feeding time on fed-batch Arthrospira (Spirulina) platensis cultivation and biomass composition with and without pH control

Previous work demonstrated that a mixture of NH₄Cl and KNO₃ as nitrogen source was beneficial to fed-batch Arthrospira (Spirulina) platensis cultivation, in terms of either lower costs or higher cell concentration. On the basis of those results, this study focused on the use of a cheaper nitrogen source mixture, namely (NH₄)₂SO₄ plus NaNO₃, varying the ammonium feeding time (T = 7-15 days), either controlling the pH by CO₂ addition or not. A. platensis was cultivated in mini-tanks at 30 °C, 156 μmol photons m⁻² s⁻¹, and starting cell concentration of 400 mg L⁻¹, on a modified Schlösser medium. T = 13 days under pH control were selected as optimum conditions, ensuring the best results in terms of biomass production (maximum cell concentration of 2911 mg L⁻¹, cell productivity of 179 mg L⁻¹ d⁻¹ and specific growth rate of 0.77 d⁻¹) and satisfactory protein and lipid contents (around 30% each).     

Determination of the membrane permeability characteristics of Pacific oyster, Crassostrea gigas, oocytes and development of optimized methods to add and remove ethylene glycol

In order for cryopreservation to become a practical tool for aquaculture, optimized protocols must be developed for each species and cell type. Knowledge of a cell’s osmotic tolerance and membrane permeability characteristics can assist in optimized protocol development. In this study, these characteristics were determined for Pacific oyster oocytes and modified methods for loading and unloading ethylene glycol (EG) were tested. Oocytes were found to behave as ideal osmometers and their osmotically inactive fraction (V_b) was calculated to be 0.48. Oocytes exposed to NaCl solutions of 0.6 to 2.3 Osm fertilized at rates equivalent to oocytes left in seawater. This corresponds to volume changes of +27.3 and −38.1 ± 1.2%. The permeability of the oocytes to water (L_p) was determined to be 3.8 ± 0.4 × 10⁻², 5.7 ± 0.8 × 10⁻², and 13.2 ± 1.3 × 10⁻² μm min⁻¹ atm⁻¹, when measured at temperatures of 5, 10 and 20 °C. The respective EG permeability values (P_s) were 9.5 ± 0.1 × 10⁻⁵, 14.6 ± 1.2 × 10⁻⁵, and 41.7 ± 2.4 × 10⁻⁵ cm min⁻¹. The activation energies for L_p and P_s were determined to be 14.5 and 17.5 kcal mol⁻¹, respectively. Different models for EG loading and unloading from oocytes were developed and tested. Post-thaw fertilization did not differ significantly between a published step addition method and single step addition at 20 °C. This represents a considerable reduction in handling. The results of this study demonstrate that the cryobiological characteristics of a given cell type should be taken into account when developing cryopreservation methods.     

Production of rhizoids by Caulerpa prolifera in culture

Studies on Caulerpa prolifera rhizoids were carried out to determine the possibility of mass culture, because the rhizoids produce a bio-adhesive. Rhizoids can be induced by cutting the base of a blade and floating it in a media or planting it in sand. Measurement of rhizoid production included determination of number, length, and the weight of attached sand grains. The growth experiments were for 1–2 weeks and fronds growth was compared to rhizoid production. Optimal conditions for rhizoid growth included low levels of nitrogen and phosphate (less than 5 and 2 μM, respectively), low irradiance (30 μmol photon m⁻² s⁻¹), moderate temperature (22–28°), continuous shaking, addition of microelements and auxin (1 ppm) and initially detached fronds followed by attachment. Under these optimal conditions maximal weekly growth reached 70–170 rhizoids per blade, 7–11 mm length and maximal attachment to sand grains. Blade growth of C. prolifera responded similarly to rhizoid production and reached a weekly growth rate of 30–130%.     

A cryoprotective and cold-adapted 1,3-β-endoglucanase from cherimoya (Annona cherimola) fruit

A 1,3-β-glucanase with potent cryoprotective activity was purified to homogeneity from the mesocarp of CO₂-treated cherimoya fruit (Annona cherimola Mill.) stored at low temperature using anion exchange and chromatofocusing chromatography. This protein was characterized as a glycosylated endo-1,3-β-glucanase with a M_r of 22.07 kDa and a pI of 5.25. The hydrolase was active and stable in a broad acidic pH range and it exhibited maximum activity at pH 5.0. It had a low optimum temperature of 35 °C and it retained 40% maximum activity at 5 °C. The purified 1,3-β-glucanase was relatively heat unstable and its activity declined progressively at temperatures above 50 °C. Kinetic studies revealed low k_cat (3.10 ± 0.04 s⁻¹) and K_m (0.32 ± 0.03 mg ml⁻¹) values, reflecting the intermediate efficiency of the protein in hydrolyzing laminarin. Moreover, a thermodynamic characterization revealed that the purified enzyme displayed a high k_cat at both 37 and 5 °C, and a low E_a (6.99 kJ mol⁻¹) within this range of temperatures. In vitro functional studies indicated that the purified 1,3-β-glucanase had no inhibitory effects on Botrytis cinerea hyphal growth and no antifreeze activity, as determined by thermal hysteresis analysis using differential scanning calorimetry. However, a strong cryoprotective activity was observed against freeze-thaw inactivation of lactate dehydrogenase. Indeed, the PD₅₀ was 8.7 μg ml⁻¹ (394 nM), 9.2-fold higher (3.1 on a molar basis) than that of the cryoprotective protein BSA. Together with the observed accumulation of glycine-betaine in CO₂-treated cherimoya tissues, these results suggest that 1,3-β-glucanase could be functionally implicated in low temperature-defense mechanism activated by CO₂.     

Tolerance of Sargassum thunbergii germlings to thermal, osmotic and desiccation stress

The construction of artificial seaweed beds in the intertidal zone is a challenge due to extreme levels of physical stress. In order to provide a basis for the construction using the dispersal of microscopic juveniles, a three-way factorial experimental design was used to evaluate the tolerance of Sargassum thunbergii germlings shortly released from fertile thalli to temperature, salinity and desiccation in this study. Results revealed that temperature, salinity and desiccation significantly affected the growth and survival of germlings. Germlings showed rapid growth with relative growth rate (RGR, % day⁻¹) over 16% when cultured at 25 °C and full immersion in normal seawater. Although growths of germlings subjected to moderate conditions were significantly inhibited, RGRs over 13% were obtained. The RGRs of germlings below 10% were observed only at 35 °C and 9 h desiccation treatments. In comparison to growth, survival was less affected by physical stress. Germlings showed low mortalities below 10% under appropriate conditions (25 °C and 30 °C combined with full immersion), and below 60% under moderate conditions, by the end of experiment. However, the mortality rates increased to over 90% under extreme conditions (9 h desiccation and 35 °C combined with full immersion in salinity of 12). These results showed that S. thunbergii germlings had high tolerance to physical stresses. In addition to the main effects, both two-way and three-way interactions between temperature, salinity and desiccation were significant. Based on the magnitude of effect, desiccation was the predominant factor affecting both growth and survival. According to the results, construction of artificial tanks in natural habitat to minimize desiccation may be an effective strategy for S. thunbergii restoration using germlings.     

Understanding the potential effects of water regime and salinity on recruitment of Melaleuca ericifolia Sm.

Although many emergent wetland plants may readily tolerate rapid changes in flooding and drying under freshwater conditions, their tolerance to dynamic water regimes may be compromised by salinity. Melaleuca-dominated woodlands occur naturally in Australia, south-east Asia and New Caledonia. Coastal wetlands dominated by Swamp paperbark (Melaleuca ericifolia) (Myrtaceae), native to south-east Australia, are commonly degraded as a consequence of altered water regime and salinity. This study simulates the release of M. ericifolia seeds from the aerial canopy under a range of water regime and salinity scenarios to determine conditions limiting sexual recruitment. Plant growth and survival were examined following seed release under two static water regimes (moist and flooded sediment) and two dynamic water regimes (simulated drawdown—“flooded-moist” and simulated re-flooding—“moist-flooded”). All water regimes, excluding the continuously flooded regime, were examined at three salinities: 0.1 dS m⁻¹ (fresh), 8 dS m⁻¹ and 16 dS m⁻¹, over a 50-day period commencing 44 days after the seeds were sown. The flooded treatment was examined at 0.1 dS m⁻¹ only, to confirm that flooding prohibits establishment of M. ericifolia. Seed and seedlings were positively buoyant and establishment was limited to moist soil. Flotation of seedlings in the flooded-moist treatment, however, did not inhibit subsequent establishment upon moist soil, even at the highest salinity of 16 dS m⁻¹. Growth, but not survival, was reduced by salinities of 8 dS m⁻¹ and 16 dS m⁻¹ in the moist treatment. Flotation of seedlings in saline water in the flooded-moist treatment did not reduce growth or survival compared with fresh water. Survival of seedlings in the moist-flooded treatment was lower in the freshwater and 16 dS m⁻¹ treatment compared with the moist treatment, but not at 8 dS m⁻¹. These findings suggest that water regime influences establishment of young M. ericifolia plants more strongly than does salinity, at least up to ∼1/3 seawater and in the short term (<2 months). Seedlings are likely to establish during a drawdown where the soil is exposed at salinities of ≤16 dS m⁻¹. In contrast, premature re-flooding of seedlings, even with fresh water, will compromise survival.     

Short-term exposure to waterborne free silver has acute effects on membrane current of Xenopus oocytes

Waterborne free silver can cause osmo- and ionoregulatory disturbances in freshwater organisms. The effects of a short-term exposure to extracellular Ag⁺ ions on membrane currents were investigated in voltage-clamped defolliculated Xenopus oocytes. At a holding potential of − 60 mV, ionic silver (1 μM Ag⁺) increased inward currents (=I_Ag) from − 8 ± 2 nA to − 665 ± 41 nA (n = 74; N = 27). I_Ag activated within 2 min of silver exposure and then rose impetuously. This current was largely reversible by washout and repeatable. I_Ag reversed around − 30 mV and rectified slightly at more positive potentials. Na⁺-free bath conditions reduced the silver-induced current to a smaller but sustained current. The response to silver was abolished by the Cl⁻ channel blockers DIDS and SITS, whereas niflumic acid strongly potentiated I_Ag. Intraoocyte injection of AgNO₃ to about 1 mM [Ag]_i strongly potentiated I_Ag. Extracellular application of either dithiothreitol (DTT), a compound known to reduce disulfide bridges, or l-cysteine abolished Ag⁺-activated increase of membrane current. In contrast, n-ethylmaleimide (NEM) which oxidizes SH-groups potentiated I_Ag. Hypoosmotic bath solution significantly increased I_Ag whereas hyperosmolar conditions attenuated I_Ag. The activation of I_Ag was largely preserved after chelation of cytosolic Ca²⁺ ions with BAPTA/AM. Taken together, these data suggest that Xenopus oocytes are sensitive to short-term exposure to waterborne Ag⁺ ions and that the elicited membrane currents result from extra- and intracellular action of Ag⁺ ions on peptide moieties at the oocyte membrane but may also affect conductances after internalization.     

Hypoxia and sulphide influence gamete production in Ulva sp.

Gamete production after exposure to hypoxia or sulphide was studied in the marine macroalga Ulva sp. collected in the Sacca di Goro, Italy. Experiments were carried out on discs (12 mm diameter) of thalli cultured in artificial sea water in laboratory at 20 ± 1 °C, 152 μmol m⁻² s⁻¹, 16 h photoperiod and 30‰ salinity. Dehydration of thallus was used as inducer of gametogenesis and growth and gamete release during recovery after 10, 20, 30 or 40 min dehydration (20 ± 1 °C, 25% humidity) were analysed. Unlike non-dehydrated thalli the dehydrated ones produced gametes. Thallus discs, non-dehydrated or subjected to 30 min dehydration, were exposed to hypoxia (1.78–4.02 μmol O₂ L⁻¹) or sulphide (1 mM) for 3, 5, or 7 days at 20 °C in the dark. Non-dehydrated and dehydrated thalli maintained in normoxic conditions in the dark were the controls. Gamete density was checked by counting at the end of the incubation period and during the subsequent 7 days of recovery under 16 h photoperiod in normoxic conditions. Non-dehydrated thalli maintained in normoxic conditions in the dark released gametes when returned to light suggesting that dark constitutes a stimulus to gamete production. The presence of gametes at the end of 3 days incubation of dehydrated thalli in normoxia demonstrated that gametogenesis can occur even in the dark. However, gametes were not present at the end of incubation in hypoxic and sulphidic conditions. Actually, during hypoxic incubation oxygen consumption in D-thalli was very low, only 0.117 × 10⁻³ μmol O₂ mg⁻¹ h⁻¹ compared to 5.93 × 10⁻³ μmol O₂ mg⁻¹ h⁻¹ in normoxia, denoting a reduction of the metabolic rate that could not sustain gametogenesis. During recovery after incubation in normoxic, hypoxic or sulphidic conditions densities of gametes from dehydrated thalli showed significant differences and resulted after hypoxia > after normoxia > after sulphide. Differences in non-dehydrated thalli were not significant. Dehydrated thalli, still green at the end of the incubation period, underwent blanching in the course of recovery in parallel to gamete production, while non-dehydrated thalli maintained their green colour even after exposure to sulphide. Our findings suggest that macroalga Ulva sp. can survive exposure to darkness, severe hypoxia and high sulphide levels and can maintain gamete production even when the exposure to these stress conditions is joined to dehydration.