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.     

Response to selection and evolvability of invasive populations

While natural selection might in some cases facilitate invasions into novel habitats, few direct measurements of selection response exist for invasive populations. This study examined selection response to changes in salinity using the copepod Eurytemora affinis. This copepod has invaded fresh water from saline habitats multiple times independently throughout the Northern Hemisphere. Selection response to a constant intermediate salinity (5 PSU) was measured in the laboratory for saline source and freshwater invading populations from the St. Lawrence drainage (North America). These populations were reared under three conditions: (1) native salinities (0 or 15 PSU) for at least two generations, (2) 5 PSU for two generations, and (3) 5 PSU for six generations. Full-sib clutches taken from populations reared under these three conditions were split across four salinities (0, 5, 15, and 25 PSU) to determine reaction norms for survival and development time. Contrasts in survival and development time across the three rearing conditions were treated as the selection response. Selection at 5 PSU resulted in a significant decline in freshwater (0 PSU) tolerance for both the saline and freshwater populations. Yet, evolutionary differences in freshwater tolerance persisted between the saline and freshwater populations. The saline and freshwater populations converged in their high-salinity (25 PSU) tolerance, with an increase in the freshwater population and decline in the saline population. Development time did not shift greatly in response to selection at 5 PSU. For all three rearing conditions, the freshwater population exhibited retarded larval development and accelerated juvenile development relative to the saline population. Results from this study indicate that both the saline and freshwater populations exhibit significant responses to selection for a fitness-related trait critical for invasions into a novel habitat. For the Symposium on “Evolvability and Adaptation of Invasive Species,” Society for the Study of Evolution 2004.     

Evolution of physiological tolerance and performance during freshwater invasions

Invasive species that penetrate habitat boundaries are likelyto experience strong selection and rapid evolution. This studydocuments evolutionary shifts in tolerance and performance followingthe invasion of fresh water by the predominantly estuarine andsalt marsh copepod Eurytemora affinis. Common-garden experimentswere performed on freshwater-invading (Lake Michigan) and ancestralsaline (St. Lawrence marsh) populations to measure shifts inadult survival (at 0, 5, and 25 PSU), and survival during developmentand development time (both using full-sib clutches split across0, 5, 15, and 25 PSU). Results showed clear evidence of heritableshifts in tolerance and performance associated with freshwaterinvasions. The freshwater population exhibited a gain in low-salinitytolerance and a reduction in high-salinity tolerance relativeto the saline population, suggesting tradeoffs. These tradeoffswere supported by negative genetic correlations between survivalat fresh (0 PSU) versus higher salinities. Mortality in responseto salinity occurred primarily before metamorphosis, suggestingthat selection in response to salinity had acted primarily onthe early life-history stages. The freshwater population exhibitedcurious patterns of life-history evolution across salinities,relative to the saline population, of retarded development tometamorphosis but accelerated development from metamorphosisto adulthood. This pattern might reflect tradeoffs between developmentrate and survival in fresh water at the early life-history stages,but some other selective force acting on later life-historystages. Significant effects of clutch (genotype) and clutch-by-salinityinteraction (G x E) on survival and development time in bothpopulations indicated ample genetic variation as substrate fornatural selection. Variation for high-salinity tolerance waspresent in the freshwater population despite negative geneticcorrelations between high- and low-salinity tolerance. Resultsimplicate the importance of natural selection and document theevolution of reaction norms during freshwater invasions.     

Pumping ions: rapid parallel evolution of ionic regulation following habitat invasions

Marine to freshwater colonizations constitute among the most dramatic evolutionary transitions in the history of life. This study examined evolution of ionic regulation following saline-to-freshwater transitions in an invasive species. In recent years, the copepod Eurytemora affinis has invaded freshwater habitats multiple times independently. We found parallel evolutionary shifts in ion-motive enzyme activity (V-type H(+) ATPase, Na(+) /K(+) -ATPase) across independent invasions and in replicate laboratory selection experiments. Freshwater populations exhibited increased V-type H(+) ATPase activity in fresh water (0 PSU) and declines at higher salinity (15 PSU) relative to saline populations. This shift represented marked evolutionary increases in plasticity. In contrast, freshwater populations displayed reduced Na(+) /K(+) -ATPase activity across all salinities. Most notably, modifying salinity alone during laboratory selection experiments recapitulated the evolutionary shifts in V-type H(+) ATPase activity observed in nature. Maternal and embryonic acclimation could not account for the observed shifts in enzyme activity. V-type H(+) ATPase function has been hypothesized to be critical for freshwater and terrestrial adaptations, but evolution of this enzyme function had not been previously demonstrated in the context of habitat transitions. Moreover, the speed of these evolutionary shifts was remarkable, within a few generations in the laboratory and a few decades in the wild.     

Reproductive isolation in Chara aspera populations

Possible reproductive isolation between freshwater and brackish water populations of the dioecious charophyte Chara aspera was studied by means of cross-fertilization experiments and AFLP (Amplified Fragment Length Polymorphism). Three Swedish freshwater populations and three (German and Swedish) Baltic Sea populations of C. aspera were sampled. Cross-fertilization experiments were performed in a full combination setup of all populations and with two different salinities (0 and 10 PSU). Both freshwater and brackish water females formed about 70% more gametangia at 0 than at 10 PSU. Male individuals collected from freshwater had higher fertility than brackish water males at both salinities. 57% of all gametangia of females from freshwater developed into oospores compared to only 8% of gametangia of brackish water females. 42% of all oospores were fertilized in crosses between habitats (freshwater–brackish water) compared to 36% in crosses within habitats, the difference was not significant.Oospore and bulbil germination was investigated using propagules from freshwater and brackish water populations and incubation salinities of 0, 5, 10 and 20 PSU. None of the oospores collected from brackish water germinated. Germination of oospores and bulbils from freshwater was higher at 0 and 5 PSU than at higher salinities. Only around 40% of bulbils from brackish water germinated at 20 PSU compared to around 70% at the other three salinities. Germination of all bulbils was delayed at 20 PSU compared to other salinities.Genetic similarities (Jaccard indices of AFLP data) were higher within than between populations, but comparisons within habitat (freshwater–freshwater and brackish water–brackish water) were not different from comparisons between habitats.Our results did not identify any reproductive isolation between freshwater and brackish water populations, but indicate low gene flow between the two habitats. Oospore and bulbil germination success were highest at salinities corresponding to the conditions of their original habitat, suggesting genetic adaptation to their environmental conditions and indicating that propagules transported from freshwater to brackish water or vice versa will hardly develop into fertile plants. Additionally, brackish water plants perform poorer in all aspects of sexual reproduction than freshwater plants. Possibly, successful dispersal of oospores is not subjected to high selective pressure within the Baltic Sea where new sites easily can be colonized by means of vegetative reproduction. We assume that these adaptations will favour speciation within C. aspera and support the idea of the geologically young Baltic Sea as a “cradle of plant evolution”.     

The effect of salinity on larval survival and development in the sea urchin Echinometra lucunter

Summary

Reductions in salinity can have adverse effects on larval development and larval survival in some invertebrate taxa but not others. Salinity tolerance of larvae may be particularly important in echinoderms because they are both poor ion regulators and stenohaline. I examined the effect of six levels of salinity (15, 18, 21, 24, 27 and 33 PSU) on survival and rate of development of larvae in the subtropical sea urchin Echinometra lucunter. In the short-term, mortality rate was significantly lower in 33 PSU than in all other salinities except 27 PSU, and it was significantly greater in 15 and 18 PSU than in all higher salinities. In the long-term, daily and cumulative mortality were significantly greater in 15 PSU than in most other salinities over 11 days of development (except for cumulative mortality in 18 PSU). They were significantly greater in 18 PSU than in 21 PSU or 33 PSU over a period of 13 days. Furthermore, daily mortality was significantly greater in 18 PSU than in 24 PSU or 27 PSU at 13 d after fertilization. Daily and cumulative mortality were significantly lower in 33 PSU than in 21, 24 or 27 PSU over a period of 17 days. Although in the control (33 PSU) 75% of larvae completed development to the 8-arm stage at 35 d, no larvae developed further than the 4-arm stage in 18, 21, 24 or 27 PSU; in 15 PSU, ~60% of larvae did not develop further than swimming blastulae. Since prolonged exposure to salinities as high as 27 PSU (frequently recorded in the adult habitat) can result in great larval losses, adaptive behaviours that prevent larvae from entering water layers of low salinity will enhance their chance for survival.     

Differences in osmotolerance in freshwater and brackish water populations of Theodoxus fluviatilis (Gastropoda: Neritidae) are associated with differential protein expression

The euryhaline gastropod Theodoxus fluviatilis is found in northern Germany in freshwater or in brackish water habitats in the Baltic Sea. Previous studies have revealed that individuals from both habitats are not distinguishable by morphological characters or by sequence comparison of DNA encoding 16S RNA or cytochrome C. As reported in this study, animals collected in the two habitats differ substantially in their physiological ability to adapt to different salinities. Comparison of accumulation rates of ninhydrin-positive substances (NPS) in foot muscle upon transfer of animals to higher medium salinities revealed that brackish water animals were perfectly able to mobilize NPS, while freshwater animals had only limited ability to do so. In an attempt to explore whether this difference in physiology may be caused by genetic differentiation, we compared protein expression patterns of soluble foot muscle proteins using 2D gel electrophoresis and silver staining. Of the 40 consistently detected protein spots, 27 showed similar levels in protein expression in animals collected from freshwater or brackish water habitats, respectively. In 12 spots, however, protein concentration was higher in brackish water than in freshwater animals. In four of these spots, expression levels followed increases or decreases in medium salinities. In a different set of 4 of these 12 spots, protein levels were always higher in brackish water as compared to freshwater animals, regardless of their physiological situation (14 days in artificial pond water or in medium with a salinity of 16‰). The remaining 4 of the 12 spots had complex expression patterns. Protein levels of the remaining single spot were generally higher in freshwater animals than in brackish water animals. These expression patterns may indicate that freshwater and brackish water animals of T. fluviatilis belong to different locally adapted populations with subtle genetic differentiation.     

Estimating salinity stress via hsp70 expression in the invasive round goby (Neogobius melanostomus): implications for further range expansion

Species invasions often occur on coasts and estuaries where abiotic conditions vary, e.g. salinity, temperature, runoff etc. Successful establishment and dispersal of non-indigenous species in many such systems are poorly understood, partially since the species tend to show genetic and ecological plasticity at population level towards many abiotic conditions, including salinity tolerance. Plasticity may be driven by shifting expression of heat shock proteins such as Hsp70, which is widely recognized as indicator of physical stress. In this study, we developed a qPCR assay for expression of the hsp70 gene in the invasive round goby (Neogobius melanostomus) and tested the expression response of fish collected from a brackish environment in the western Baltic Sea to three different salinities, 0, 10 and 30. hsp70 expression was highest in fresh water, indicating higher stress, and lower at brackish (ambient condition for the sampled population) and oceanic salinities, suggestive of low stress response to salinities above the population’s current distribution. The highest stress in fresh water was surprising since populations in fresh water exist, e.g. large European rivers and Laurentian Great Lakes. The results have implications to predictions for the species’ plasticity potential and possible range expansion of the species into other salinity regimes.

     

Variable salinity tolerance in ascidian larvae is primarily a plastic response to the parental environment

Both phenotypic plasticity and local genetic adaptation may contribute to a species’ ability to inhabit different environmental conditions. While phenotypic plasticity is usually considered costly, local adaptation takes generations to respond to environmental change and may be constrained by strong gene flow. The majority of marine species have complex life-cycles with pelagic stages that might be expected to promote gene flow and plastic responses, and yet several notable examples of local adaptation have been found in species with broadcast larvae. In the ascidian, Ciona intestinalis (Linnaeus, 1767),—a common marine species with broadcast spawning and a short larval stage—previous studies have found marked differences in salinity tolerance of early life-history stages among populations from different salinity regimes. We used common-garden experiments to test whether observed differences in salinity tolerance could be explained by phenotypic plasticity. Adult ascidians from two low salinity populations [2–5 m depth, ~25 practical salinity units (PSU)], and two full salinity populations (25–27 m depth, ~31 PSU) were acclimated for 2–4 weeks at both 25 and 31 PSU. Gametes were fertilized at the acclimation salinities, and the newly formed embryos were transferred to 10 different salinities (21–39 PSU) and cultured to metamorphosis. Adult acclimation salinity had an overriding and significant effect on larval metamorphic success: tolerance norms for larvae almost fully matched the acclimation salinity of the parents, independent of parental origin (deep or shallow). However we also detected minor population differences that could be attributed to either local adaptation or persistent environmental effects. We conclude that differences in salinity tolerance of C. intestinalis larvae from different populations are driven primarily by transgenerational phenotypic plasticity, a strategy that seems particularly favourable for an organism living in coastal waters where salinity is less readily predicted than in the open oceans.     

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.     

Rapid evolution of body fluid regulation following independent invasions into freshwater habitats

Colonizations from marine to freshwater environments constitute among the most dramatic evolutionary transitions in the history of life. Colonizing dilute environments poses great challenges for acquiring essential ions against steep concentration gradients. This study explored the evolution of body fluid regulation following freshwater invasions by the copepod Eurytemora affinis. The goals of this study were to determine (1) whether invasions from saline to freshwater habitats were accompanied by evolutionary shifts in body fluid regulation (hemolymph osmolality) and (2) whether parallel shifts occurred during independent invasions. We measured hemolymph osmolality for ancestral saline and freshwater invading populations reared across a range of common-garden salinities (0.2-25 PSU). Our results revealed the evolution of increased hemolymph osmolality (by 16-31%) at lower salinities in freshwater populations of E. affinis relative to their saline ancestors. Moreover, we observed the same evolutionary shifts across two independent freshwater invasions. Such increases in hemolymph osmolality are consistent with evidence of increased ion uptake in freshwater populations at low salinity, found in a previous study, and are likely to entail increased energetic costs upon invading freshwater habitats. Our findings are consistent with the evolution of increased physiological regulation accompanying transitions into stressful environments.     

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.     

Seasonal plant water uptake patterns in the saline southeast Everglades ecotone

The purpose of this study was to determine the seasonal water use patterns of dominant macrophytes coexisting in the coastal Everglades ecotone. We measured the stable isotope signatures in plant xylem water of Rhizophora mangle, Cladium jamaicense, and Sesuvium portulacastrum during the dry (DS) and wet (WS) seasons in the estuarine ecotone along Taylor River in Everglades National Park, FL, USA. Shallow soilwater and deeper groundwater salinity was also measured to extrapolate the salinity encountered by plants at their rooting zone. Average soil water oxygen isotope ratios (δ ¹⁸O) was enriched (4.8 ± 0.2‰) in the DS relative to the WS (0.0 ± 0.1‰), but groundwater δ ¹⁸O remained constant between seasons (DS: 2.2 ± 0.4‰; WS: 2.1 ± 0.1‰). There was an inversion in interstitial salinity patterns across the soil profile between seasons. In the DS, shallow water was euhaline [i.e., 43 practical salinity units (PSU)] while groundwater was less saline (18 PSU). In the WS, however, shallow water was fresh (i.e., 0 PSU) but groundwater remained brackish (14 PSU). All plants utilized 100% (shallow) freshwater during the WS, but in the DS R. mangle switched to a soil–groundwater mix (δ 55% groundwater) while C. jamaicense and S. portulacastrum continued to use euhaline shallow water. In the DS, based on δ ¹⁸O data, the roots of R. mangle roots were exposed to salinities of 25.4 ± 1.4 PSU, less saline than either C. jamaicense (39.1 ± 2.2 PSU) or S. portulacastrum (38.6 ± 2.5 PSU). Although the salinity tolerance of C. jamaicense is not known, it is unlikely that long-term exposure to high salinity is conducive to the persistence of this freshwater marsh sedge. This study increases our ecological understanding of how water uptake patterns of individual plants can contribute to ecosystem levels changes, not only in the southeast saline Everglades, but also in estuaries in general in response to global sea level rise and human-induced changes in freshwater flows.     

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.     

Elongation and mat formation of <Emphasis Type="Italic">Chara aspera</Emphasis> under different light and salinity conditions

Former laboratory results indicate that shoot elongation at low light intensities of Chara aspera is absent already at 10 psu which is within the physiologically optimal salinity range for brackish water populations. To investigate if similar restrictions occur in the field, density and morphology of C. aspera were compared between three freshwater and three brackish water sites along its depth range. The lower depth limit of C. aspera varied considerably among sites (30–600 cm) related to turbidity. Light availability at the lower depth limit corresponded to about 15% of surface irradiance in freshwater and brackish water with lower salinity (3.4 psu). Total length increased and fresh weight:length ratio decreased with depth at these sites indicating shoot elongation related to lower light availability. Due to shoot elongation, light availability was far higher at the upper parts of the shoot than at the bottom in the turbid sites. Light availability at the lower depth limit was higher (about 40%) at two sites with higher salinity (7–8 psu), where no shoot elongation was observed at the lower depth limit. Instead, the plants were stunted and often covered with filamentous algae or shaded by other rooted submerged macrophytes indicating competitive disadvantages of C. aspera at higher salinities. As growth in high densities (mat formation) exposes the plants to severe self-shading, it is suggested that shoot elongation is a prerequisite to mat formation. Dense vegetation of C. aspera was found only in freshwater and brackish water with lower salinity. Single, richly branched plants occurred in clearwater sites with higher salinity. C. aspera was not found in “double stress” environments with both high turbidity and high salinity: We asume that the species is a poor competitor under these conditions. Our results indicate that morphological differences between freshwater and brackish water populations of C. aspera are at least partly explained by salinity rather than genetic differences.     

Effects of salinity on the growth and morphology of the invasive,euryhaline hydroid Cordylophora (Phylum Cnidaria,Class Hydrozoa)

The invasive, euryhaline hydroid Cordylophora sp. is a colonial cnidarian present in both freshwater and brackish water habitats. Individuals contend with osmotic stress at the tissue and cellular level. It has been suggested that this hydroid's ability to expand its range of distribution by invading new habitats is due in large part to an ability to acclimate to new salinities. The purpose of this study was to assess colony growth and morphological changes at various salinities in freshwater and brackish genotypes of Cordylophora sp. Single genotypes from a known freshwater clade (0.5 psu; Des Plaines River) and a known brackish clade (16 psu; Napa River) were cultured and gradually transitioned to 12 different salinities ranging 0.5–22 psu, and we characterized the growth rates and hydranth morphological features at each salinity. Colony growth was optimal at 0.5 psu for the freshwater genotype and 10 psu for the brackish genotype. Changes in hydranth morphology in the freshwater genotype were primarily observed at higher salinities, while morphological changes in the brackish genotype primarily occurred at lower salinities. Our results for the brackish genotype generally concur with previous work, but this study is the first to document the response of a freshwater genotype of Cordylophora sp. to various salinities. Differences in growth between these two genotypes strongly support the previously proposed existence of multiple cryptic species. Furthermore, because this hydroid is quite prevalent in freshwater and brackish systems as a fouling organism, understanding the effects of various salinities on the successful establishment of Cordylophora sp. is an important contribution to the understanding of the ecophysiology and management of this invasive hydroid.     

First description of an euryoecious acanthoecid choanoflagellate species,Enibas tolerabilis gen. et sp. nov. from a salar in the Chilean Andes based on morphological and transcriptomic data

In general, acanthoecid choanoflagellates have been described to occur exclusively in brackish water to marine habitats. Only recently, two studies have proven their existence in inland waters. One of them has shown, that an acanthoecid species from a small lake (near Apia on the island of Upolu, Samoa) is strictly freshwater adapted, not able to tolerate even brackish water. In this study, we present the first euryoecious acanthoecid species, able to live and reproduce in freshwater as well as under hypersaline conditions. The new species, Enibas tolerabilis gen. et sp. nov. was isolated in 2017 from the Salar de Ascotán in the Altiplano at 3750 m a. s. l., Northern Chile. The salinity at the time of sampling was 6 PSU. A series of autecological experiments have revealed a salinity tolerance from freshwater up to 70 PSU. In our phylogenetic analysis, E. tolerabilis gen. et sp. nov. clustered within the family of Acanthoecidae, forming a well-supported sister clade together with two other, environmental choanoflagellate sequences. We erected a new genus, Enibas gen. nov. and described the morphology, molecular biology and autecology for E. tolerabilis gen. et sp. nov. which has a stephanoecid-like lorica morphology. We emphasize that the definition of the genus Stephanoeca, being polyphyletic, is in urgent need of revision as we showed that this morphology is present in both acanthoecid families.     

Salinity tolerance of laboratory-reared larvae of the California killifish, Fundulus parvipinnis Girard

Newly hatched larvae of the California killifish ( Fundulus parvipinnis ) reared in the laboratory, were tolerant of salinities from fresh water to 70‰. Their salinity tolerance was influenced by incubation salinity; larvae hatched in lower incubation salinities exhibited greater freshwater tolerance than those hatched in higher salinities. In gradual acclimation tests, the upper median lethal salinity for the larvae was 130‰. Freshwater tolerance of the larvae decreased with age; yolk sac larvae were completely tolerant of fresh water while larvae more than 15 days old were least resistant.     

A Morphometric Study of Euryhalinity in Marine Populations of the Ciliate Genus Euplotes

ABSTRACT. Twenty different clonal strains of marine and brackish Euplotes , representing four morphotypes, were tested for hyposalinity tolerance by a method which gradually acclimated the cells to lower salinity medium. The lowest salinities in which the strains could thrive ranged from 60% of normal seawater to complete freshwater. The morphological effects of culture medium salinity were also examined for two strains of a small "Euplotes charon" morphotype, as well as for two mating compatible "Euplotes vannus" strains and several of their exconjugates. There were no differences between the euryhaline strains grown in fresh or saltwater, except for a slight increase in overall cell size in one strain when cultured in freshwater medium. E. vannus strains increased in overall cell size with decreased salinity; also, the dorsal surface of the cells can become disorganized when the cells are cultured in 30% normal seawater.