Stress levels over time in the introduced ascidian Styela plicata: the effects of temperature and salinity variations on hsp70 gene expression

Species distribution, abundance, and long-term survival are determined by biotic and abiotic regimes. However, little is known about the importance of these factors in species range expansion. Styela plicata is a solitary ascidian introduced all over the world by ship fouling, including salt marsh habitats, where introduced populations must tolerate high seasonal variations in temperature and salinity. To determine the seasonal stress levels in a salt marsh population of S. plicata, we quantified heat shock protein (hsp70) gene expression using quantitative real-time PCR throughout a 2-year cycle. Results showed that hsp70 expression varied over time, with higher stress levels recorded in summer and winter. Periodic conditions of high temperatures, particularly when coupled with low salinities, increased hsp70 gene expression. Mortality events observed every year around June were concurrent with sharp increases in temperature (>6°C), indicating that drastic changes in abiotic factors may overwhelm the observed stress response mechanisms. Determining the ability of introduced species to cope with stress, and the thresholds above which these mechanisms fail, is fundamental to predict the potential expansion range of introduced species and design efficient containment plans.     

Genotype-by-environment interaction for salinity tolerance in the freshwater-invading copepod Eurytemora affinis

This study examined the extent of phenotypic plasticity for salinity tolerance and genetic variation in plasticity in the invasive copepod Eurytemora affinis. Euryemora affinis is a species complex inhabiting brackish to hypersaline environments but has invaded freshwater lakes and reservoirs within the past century. Reaction norm experiments were performed on a relatively euryhaline population collected from a brackish lake with fluctuating salinity. Life history traits (hatching rate, survival, and development time) were measured for 20 full-sib clutches that were split and reared at four salinities (fresh, 5, 10, and 27 practical salinity units [PSU]). On average, higher salinities (10 and 27 PSU) were more favorable for larval growth, yielding greater survival and faster development rate. Clutches differed significantly in their response to salinity, with a significant genotype-by-environment interaction for development time. In addition, genetic (clutch) effects were evident in response to low salinity, given that survival in fresh (lake) water was significantly positively correlated with survival at 5 PSU for individual clutches. Clutches raised in fresh water could not survive beyond metamorphosis, suggesting that acclimation to fresh water could not occur in a single generation. Results suggest the importance of natural selection during freshwater invasion events, given the inability of plasticity to generate a freshwater phenotype, and the presence of genetic variation for plasticity upon which natural selection could act.     

Can sunbleak Leucaspius delineatus or topmouth gudgeon Pseudorasbora parva disperse through saline waters?

In order to determine the potential for the invasive fishes sunbleak Leucaspius delineatus and topmouth gudgeon Pseudorasbora parva to disperse through saline waters their behaviour and physiology were investigated during exposure to salinities of 10·0 and 12·5. Increased salinity caused an increase in whole body cortisol in both species, but sunbleak and topmouth gudgeon showed very different metabolic and behavioural responses to the salinity stress. Sunbleak displayed increased swimming activity in brackish water, which may be important for dispersal through saline waters in the wild, although there were increased metabolic costs associated with this behaviour. Conversely, topmouth gudgeon showed a reduction in both swimming activity and metabolic rate in brackish waters. A pronounced depression in food intake (70–80%) was shown by both species during the salinity exposures. Both sunbleak and topmouth gudgeon, however, showed a full recovery of food intake within 24 h following return to fresh water. Despite the fact that exposure to saline waters is stressful, and affects both physiology and behaviour, rapid recovery of appetite after return to fresh water suggests that short-term use of brackish waters is a feasible dispersal route for sunbleak and topmouth gudgeon in the wild.     

Variation in gene expression along a salinity gradient in wild populations of the euryhaline black-chinned tilapia Sarotherodon melanotheron

This study evaluated variation in expression of 11 genes within and among six wild populations of the black-chinned tilapia Sarotherodon melanotheron distributed along a salinity gradient from 0 to 100. Previous laboratory studies had shown that expression of these genes was sensitive to water salinity; the current study confirmed that a number of them also varied in expression in wild populations along the salinity gradient. Principal component analysis (PCA) first distinguished two, not mutually exclusive, sets of genes: trade-off genes that were highly expressed at one or other extreme of the salinity gradient and stress genes that were up-regulated at the two salinity extremes (i.e. a U-shaped expression pattern). The PCA clearly partitioned the populations into three groups based on their gene expression patterns and their position along the salinity gradient: a freshwater (GL; 0) population, four brackish and seawater (GB, HB, SM, SF; ranging from 20 to 50) populations and a hypersaline (SK, 100) population. Individual variation in gene expression was significantly greater within the populations at the extreme compared to intermediate salinities. These results reveal phenotypically plastic regulation of gene expression in S. melanotheron, and greater osmoregulatory and plasticity costs at extreme salinities, where fitness-related traits are known to be altered.     

Effect of salinity on oxygen consumption in fishes: a review

The effect of salinity on resting oxygen uptake was measured in the perch Perca fluviatilis and available information on oxygen uptake in teleost species at a variety of salinities was reviewed. Trans‐epithelial ion transport against a concentration gradient requires energy and exposure to salinities osmotically different from the body fluids therefore imposes an energetic demand that is expected to be lowest in brackish water compared to fresh and sea water. Across species, there is no clear trend between oxygen uptake and salinity, and estimates of cost of osmotic and ionic regulation vary from a few per cent to >30% of standard metabolism.     

Contrasting patterns of quantitative and neutral genetic variation in locally adapted populations of the natterjack toad, Bufo calamita

The relative importance of natural selection and genetic drift in determining patterns of phenotypic diversity observed in nature is still unclear. The natterjack toad (Bufo calamita) is one of a few amphibian species capable of breeding in saline ponds, even though water salinity represents a considerable stress for them. Results from two common-garden experiments showed a pattern of geographic variation in embryonic salinity tolerance among populations from either fresh or brackish environments, consistent with the hypothesis of local adaptation. Full-sib analysis showed increased variation in survival among sibships within population for all populations as osmotic stress was increased (broad-sense heritability increased as salinity raised). Nevertheless, toads native to the brackish water environment had the highest overall survival under brackish conditions. Levels of population genetic differentiation for salinity tolerance were higher than those of neutral genetic differentiation, the latter obtained through the analysis of eight microsatellite loci. Microsatellite markers also revealed little population differentiation, lack of an isolation-by-distance pattern, and moderate gene flow connecting the populations. Therefore, environmental stress tolerance appears to have evolved in absence of geographic isolation, and consequently we reject the null hypothesis of neutral differentiation.     

Spiniferites cruciformis: a fresh water dinoflagellate cyst?

Palynological studies of cored lacustrine sediments from the late Quaternary of Lake Kastoria, northern Greece, revealed a Late Glacial interval with abundant dinoflagellate cysts. Cyst assemblages include two identifiable species, Spiniferites cruciformis and Gonyaulax apiculata. The presence of the fresh water species G. apiculata is consistent with the lacustrine setting of these deposits, but that of S. cruciformis is anomalous. Previously, this species has only been recorded in abundance from presumed brackish marine sediments from the Black Sea and Marmara Sea sediments where geochemical data clearly record brackish salinities. Therefore, it has been regarded as a low salinity cyst type with a wide range of morphological variation that some workers have suggested to reflect salinity fluctuations. Specimens from Greece display only part of the range of morphological variability previously described from these (brackish) marine settings. Encountered morphological variation includes ellipsoidal/pentameral and cruciform endocyst shapes with rare intermediate shapes, and highly variable septa development. Specimens characterized by extremely reduced ornamentation known from (brackish) marine environments have not been recorded. Our records of S. cruciformis indicate that: (1) it could thrive in fresh water conditions; and (2) that apparently most of the strong morphological variations of the cysts are an intrinsic phenomenon for this taxon, and may only partly be linked to salinity variations as suggested earlier. We suggest that S. cruciformis essentially is a fresh water taxon, and that its records in (brackish) marine environments, with the exception of specimens with strongly reduced ornamentation, may be due to transportation, to short-lived fresh water surface conditions in such environments, or to tolerance of the species to brackish conditions.     

The effects of salinity and temperature on the development and survival of fish parasites

In brackish water the variety of marine and freshwater parasite species is considerably reduced. The distribution in brackish water of most marine endoparasites is restricted by the salinity tolerance of their hosts, most of the parasite species are more tolerant than their hosts. The influence of salinity and temperature on nine species has been examined; first stage larvae of Contracaecum aduncum develop in 0-32‰ salinity; Cryptocotyle lingua proved to be infective at salinities down to 4‰. The greatest resistance was found in Anisakis larvae from herring Clupea harengus , which survived for more than half a year. Parasites in the fish intestines appear to be unaffected by changing water salinities, as the osmolarity in the intestines stays nearly constant. Marine ectoparasites ( Acanthochondria depressa, Lepeophtheirus pectoralis ) survive about three times longer than freshwater species ( Piscicola geometra, Argulus foliaceus ) when salinity is 16‰. High temperature increases the effects of adverse salinities on parasites. There is evidence that none of these ecto-parasitic species can develop within the range of 7-20‰ salinity.     

Notes On The Occurrence Of Clathrus Cancellatus,Tournf., In Argyllshire

Background: Estuaries are characterised by salinity gradients and regular flooding events. These environmental factors form stress gradients, along which species composition changes.

Aims: Analyse and compare patterns of plant species diversity along the estuarine salinity and flooding gradients of the Elbe and Connecticut Rivers.

Methods: Vegetation was sampled at three elevations (low, mid, high) in five sites of each marsh type (fresh, brackish, salt) in both estuaries. Patterns of species density (SD) and evenness (E) along the gradients were analysed and compared between the two estuaries with three-factor ANOVAs.

Results: The regional species pool was 33% higher for the Connecticut than for the Elbe. SD of fresh marshes (19 ± 2.2) was more than twice in the Connecticut than in the Elbe. We found an overall increase in SD from low to high elevation and from salt to freshwater marshes in both estuaries. However, SD and E were strongly depressed at intermediate elevations in the Elbe fresh and brackish marshes.

Conclusions: Although diversity patterns in the two estuaries show overall similarities, patterns of SD and E differ, when particular elevational zones and marsh types are compared. We hypothesise this to be due to evolutionary and historical influences on the regional species pools, shaping the impact of local biotic and abiotic processes.     

Tolerance to salinity and thermal stress by larvae and adults of the serpulid annelid Ficopomatus enigmaticus

The serpulid annelid Ficopomatus enigmaticus is a widely distributed invader of shallow‐water, brackish habitats in subtropical and temperate regions, where it has numerous damaging ecological and economic effects. Its distributional pattern suggests that temperature and salinity play important roles in limiting its distribution, but because other factors often covary with these, drawing strong conclusions from these patterns is difficult. In an effort to more clearly identify the effects of these factors, we examined tolerance to acute thermal (16–28°C) and salinity (0–35 psu) stress by larvae (5‐day exposure, unfed) and adults (14‐day exposure, unfed) of F. enigmaticus in the laboratory experiments. Larvae showed higher mortality at the highest temperature tested 28°C; adult survival was unaffected by temperature. Neither larvae nor adults survived exposure to pure freshwater (0 psu), but survived well at salinities ranging 3.5–35 psu. In addition, high salinity did not slow tube growth in adults. These results suggest that salinity stress, in particular, does not directly limit the distribution of F. enigmaticus to low‐salinity habitats. Experimental work on the distribution of F. enigmaticus is uncommon in the literature, but is likely needed to identify the abiotic or biotic factors that limit the distribution of this frequently invasive species.     

Chronic Salinity Adaptation Modulates Hepatic Heat Shock Protein and Insulin-like Growth Factor I Expression in Black Sea Bream

Black sea bream ( Mylio macrocephalus) hepatic heat shock proteins hsp90, hsp70, and hsp60 were found to be thermally and reversibly inducible as they were elevated 2.0, 3.2, and 2.1 fold, respectively, on acute heat shock and returned to pre-heat-shock levels after a 40-hour recovery period. To establish whether salinity plays a role in regulating heat shock protein (hsp) and insulin-like growth factor-I (IGF-I) expression in a euryhaline marine fish, we adapted groups of juvenile black sea bream to salinities of 50 ppt (hypersaline), 33 ppt (seawater), 12 ppt (isoosmotic), and 6 ppt (hypoosmotic) for 8 months. The lowest levels of hsps were found in fish reared in an isoosmotic salinity and the highest in those adapted to hypersaline and hypoosmotic salinities. Hepatic beta-actin messenger RNA abundance remained unchanged in all groups during salinity adaptation, whereas IGF-I mRNA abundance was highest in isoosmotic adapted black sea bream. This study is the first report of an effect of salinity ranging from hypersaline to hypoosmotic on the expression of different hsp forms and IGF-I in fish, and the possible relationship between environmental salinity, hepatic IGF-I expression, and hsp regulation is discussed.     

Sexual and asexual reproductive strategies of invasive Eurasian milfoil (Myriophyllum spicatum) in estuarine environments

The proliferation and domination of many of the world’s ecosystems by invasive species is thought to have numerous deleterious impacts on biodiversity and ecosystem services. As a first step toward improving our understanding of factors controlling the reproductive potential of one such species, we report on the impacts of varying salinity on reproductive strategies of Eurasian milfoil, an invasive species of submerged vegetation that has persisted in Gulf of Mexico estuaries for decades. We present results from a series of experiments designed to determine relative incidences of sexual (flowering) and asexual (fragmentation) reproduction at salinities commonly encountered in these estuaries (0, 5, 15 psu). Because fragmentation is thought to be responsible for milfoil spread in freshwaters, we also investigate the effects of salinity on shoot breaking strength and fragment root production. Experimental results indicate that production of fragments and plant breaking strength are not significantly impacted by increased salinity. In contrast, flowering and fragment root production are decreased at elevated salinities. These results suggest that milfoil’s invasiveness may be diminished in brackish water. As such, the removal of impediments to brackish water intrusion may aid in reducing milfoil’s proliferation via disruption of physiological processes such flowering and fragment root generation.     

Effects of developmental acclimation on adult salinity tolerance in the freshwater-invading copepod Eurytemora affinis

Invasive species are commonly thought to have broad tolerances that enable them to colonize new habitats, but this assumption has rarely been tested. In particular, the relative importance of acclimation (plasticity) and adaptation for invasion success are poorly understood. This study examined effects of short-term and developmental acclimation on adult salinity tolerance in the copepod Eurytemora affinis. This microcrustacean occurs in estuarine and salt marsh habitats but has invaded freshwater habitats within the past century. Effects of short-term acclimation were determined by comparing adult survival in response to acute versus gradual salinity change to low salinity (fresh water). Effects of developmental acclimation on adult tolerance were determined using a split-brood 4 x 2 factorial experimental design for one brackish-water population from Edgartown Great Pond, Massachusetts. Twenty full-sib clutches were split and reared at four salinities (fresh, 5, 10, and 27 practical salinity units [PSU]). On reaching adulthood, clutches from three of the salinity treatments (no survivors at fresh) were split into low- (fresh) and high- (40 PSU) salinity stress treatments, at which survival was measured for 24 h. Short-term acclimation of adults did not appear to have a long-term affect on low-salinity tolerance, given that gradual transfers to fresh water enhanced survival relative to acute transfers in the short term (after 7 h) but not over a longer period of 8 d. Developmental acclimation had contrasting effects on low- versus high-salinity tolerance. Namely, rearing salinity had a significant effect on tolerance of high-salinity (40 PSU) stress but no significant effect on tolerance of low-salinity (freshwater) stress. In addition, there was a significant effect of clutch on survival under freshwater conditions, indicating a genetic component to low-salinity tolerance but no significant clutch effect in response to high salinity. While developmental acclimation might enhance survival at higher salinities, the minimal effect of acclimation and significant effect of clutch on low-salinity tolerance suggest the importance of natural selection during freshwater invasion events.     

Laboratory simulation of the effects of environmental salinity on wild‐caught juveniles of European sea bass Dicentrarchus labrax and gilthead seabream Sparus aurata

Gilthead seabream Sparus aurata and European sea bass Dicentrarchus labrax , are two important species in Mediterranean aquaculture. In the wild, their juveniles occur in brackish areas such as lagoons and river deltas. Even though the juveniles seem to favour brackish environments, low salinity incurs an energy cost for osmoregulation. This paper presents the results of a series of laboratory experiments exploring the effects of salinity on growth, feeding, food conversion, survival and maintenance energy requirements of wild‐caught juveniles. The fish were kept in the laboratory, divided in groups of 20 in small tanks of 50 l each, and supplied with biologically filtered seawater of four salinity levels (8, 18, 28‰ and natural seawater) and fixed temperature (20 ± 1·4° C). The fish were fed pelleted feed throughout the experiment. Both species showed great similarity in their responses to lower salinities. Satiation time for both species increased with decreasing salinity, while maintenance requirements (required daily ration and energy) increased as with increasing salinity. Growth and feed conversion is highest for salinities around 28‰ and lower for salinities above and below. Both species share common physiological features, and intermediate salinities are optimal for their performance in nature and in captivity.     

Effect of salinity on the distribution of Ponto-Caspian gammarids in a non-native area – environmental and experimental study

The native area of gammarids from the so-called ‘Caspian complex’, Pontogammarus robustoides (G.O. Sars, 1894), Obesogammarus crassus (G.O. Sars, 1894), Dikerogammarus haemobaphes (Eichwald, 1841) and D. villosus (Sowinsky, 1894), is associated with brackish waters. Over the last several decades they have colonized the European inland waters and part of the brackish Baltic Sea. It is believed that anthropogenic increase in the salinity of inland waters facilitated their expansion. However, the influence of salinity on the dispersal of gammarid species outside their native area is not fully understood. We tested the hypothesis that salinity was a major factor in determining distribution, based on the abundance of Gammaridae in three coastal areas of low salinity (brackish Baltic), i.e. 0.3, 3.4 and 7.3 PSU, successfully inhabited by them. Additionally, for the first time, the effect of water salinity on the osmoregulatory capacity of O. crassus was examined under laboratory conditions, for the salinities given above. The experiments showed that similarly as in the case of other Caspian complex species, salinity values of about 7 PSU create better conditions for osmoregulation in O. crassus than lower salinities (i.e. 0.3 and 3.4 PSU). In the environmental part of the study, we observed that only D. villosus achieved a significantly higher abundance in the area of 7.3 PSU. Thus, we concluded that in the range of 0.3–7.3 PSU, salinity is not a key factor governing the distribution of Ponto-Caspian gammarids.     

Local adaptation of an anuran amphibian to osmotically stressful environments

Abstract. Water salinity is an intense physiological stress for amphibians. However, some species, such as Bufo calamita, breed in both brackish and freshwater environments. Because selection under environmentally stressful conditions can promote local adaptation of populations, we examined the existence of geographic variation in water salinity tolerance among B. calamita populations from either fresh or brackish water ponds in Southern Spain. Comparisons were made throughout various ontogenetic stages. A combination of field transplant and common garden experiments showed that water salinity decreased survival probability of individuals in all populations, prolonged their larval period, and reduced their mass at metamorphosis. However, significant population X salinity interactions indicated that the population native to brackish water (Saline) had a higher salinity tolerance than the freshwater populations, suggesting local adaptation. Saline individuals transplanted to freshwater environments showed similar survival probabilities, length of larval period, and mass at metamorphosis than those native to freshwater. This indicates that increased tolerance to osmotic stress does not imply a loss of performance in freshwater, at least during the larval and juvenile phases. Despite the adaptive process apparently undergone by Saline, all populations still shared the same upper limit of embryonic stress tolerance (around 10 g/l), defining a window of salinity range within which selection can act. Significant differences in embryonic and larval survival in brackish water among sibships for all populations suggest the existence of a genetic basis for the osmotic tolerance.     

Hydromineral regulation in the saline water corixid Trichocorixa reticulata (Hemiptera: Corixidae)

The aquatic corixid Trichocorixa reticulata (Guerin-Meneville) inhabits coastal marshes, brackish water ponds and salt ponds of high salinity, suggesting the presence of well developed mechanisms for hydromineral regulation.Groups of corixids acclimated in salinities ranging from fresh water to just above 300% sea water (100‰) were analyzed for total body water content, haemolymph ionic and osmotic levels, and haemolymph free amino acids.Results indicate an excellent ability to maintain haemolymph Na⁺, Cl^?, Mg²⁺ and K⁺ hyperosmotic to the medium at low salinities and hyposmotic at high salinities. Calcium appears to conform closely to changes in external medium, becoming hyposmotic at very high salinities (80‰).Total haemolymph osmotic pressure was well regulated, the freezing point depression varying from 0.75°C in distilled water to 1.15°C in salinities of 100‰. Total body water was maintained at approx. 75% of the total animal wet weight at all salinities tested.Free amino acids were maintained between 40–60 mM in all tests and did not appear to change with salinity.