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Contrasting patterns in genetic diversity following multiple invasions of fresh and brackish waters.

Biological invasions may combine the genetic effects of population bottlenecks and selection and thus provide valuable insight into the role of such processes during novel environmental colonizations. However, these processes are also influenced by multiple invasions, the number of individuals introduced and the degree of similarity between source and receiving habitats. The amphipod Gammarus tigrinus provides a useful model to assess these factors, as its invasion history has involved major environmental transitions. This species is native to the northwest Atlantic Ocean, although it invaded both brackish and freshwater habitats in the British Isles after introduction more than 65 years ago. It has also spread to similar habitats in Western Europe and, most recently, to Eastern Europe, the Baltic Sea, and the Laurentian Great Lakes. To examine sources of invasion and patterns of genetic change, we sampled populations from 13 native estuaries and 19 invaded sites and sequenced 542 bp of the mitochondrial COI gene. Strong native phylogeographical structure allowed us to unambiguously identify three allopatrically evolved clades (2.3-3.1% divergent) in invading populations, indicative of multiple introductions. The most divergent clades occurred in the British Isles and mainland Europe and were sourced from the St Lawrence and Chesapeake/Delaware Bay estuaries. A third clade was found in the Great Lakes and sourced to the Hudson River estuary. Despite extensive sampling, G. tigrinus did not occur in freshwater at putative source sites. Some European populations showed reduced genetic diversity consistent with bottlenecks, although selection effects cannot be excluded. The habitat distribution of clades in Europe was congruent with the known invasion history of secondary spread from the British Isles. Differences in salinity tolerance among lineages were suggested by patterns of habitat colonization by different native COI clades. Populations consisting of admixtures of the two invading clades were found principally at recently invaded fresh and brackish water sites in Eastern Europe, and were characterized by higher genetic diversity than putative source populations. Further studies are required to determine if these represent novel genotypes. Our results confirm that biological invasions need not result in diminished genetic diversity, particularly if multiple source populations, each with distinctive genetic composition, contribute to the founding populations.

Adaptation, Physiological↗

Comparative phylogeography of Ponto-Caspian mysid crustaceans: isolation and exchange among dynamic inland sea basins.

The distributions of many endemic Ponto-Caspian brackish-water taxa are subdivided among the Black, Azov and Caspian Sea basins and further among river estuaries. Of the two alternative views to explain the distributions, the relict school has claimed Tertiary fragmentation of the once contiguous range by emerging geographical and salinity barriers, whereas the immigration view has suggested recolonization of the westerly populations from the Caspian Sea after extirpation during Late Pleistocene environmental perturbations. A study of mitochondrial (COI) phylogeography of seven mysid crustacean taxa from the genera Limnomysis and Paramysis showed that both scenarios can be valid for different species. Four taxa had distinct lineages related to the major basin subdivision, but the lineage distributions and depths of divergence were not concordant. The data do not support a hypothesis of Late Miocene (10-5 Myr) vicariance; rather, range subdivisions and dispersal from and to the Caspian Sea seem to have occurred at different times throughout the Pleistocene. For example, in Paramysis lacustris each basin had an endemic clade 2-5% diverged from the others, whereas Paramysis kessleri from the southern Caspian and the western Black Sea were nearly identical. Species-specific ecological characteristics such as vagility and salinity tolerance seem to have played important roles in shaping the phylogeographic patterns. The mitochondrial data also suggested recent, human-mediated cryptic invasions of P. lacustris and Limnomysis benedeni from the Caspian to the Sea of Azov basin via the Volga-Don canal. Cryptic species-level subdivisions were recorded in populations attributed to Paramysis baeri, and possibly in P. lacustris.

Animals↗

Genetic diversity of invasive species in the Great Lakes versus their Eurasian source populations: insights for risk analysis.

Combining DNA variation data and risk assessment procedures offers important diagnostic and monitoring tools for evaluating the relative success of exotic species invasions. Risk assessment may allow us to understand how the numbers of founding individuals, genetic variants, population sources, and introduction events affect successful establishment and spread. This is particularly important in habitats that are "hotbeds" for invasive species--such as the North American Great Lakes. This study compares genetic variability and its application to risk assessment within and among three Eurasian groups and five species that successfully invaded the Great Lakes during the mid 1980s through early 1990s; including zebra and quagga mussels, round and tubenose gobies, and the ruffe. DNA sequences are compared from exotic and native populations in order to evaluate the role of genetic diversity in invasions. Close relatives are also examined, since they often invade in concert and several are saline tolerant and are likely to spread to North American estuaries. Results show that very high genetic diversity characterizes the invasions of all five species, indicating that they were founded by very large numbers of propagules and underwent no founder effects. Genetic evidence points to multiple invasion sources for both dreissenid and goby species, which appears related to especially rapid spread and widespread colonization success in a variety of habitats. In contrast, results show that the ruffe population in the Great Lakes originated from a single founding population source from the Elbe River drainage. Both the Great Lakes and the Elbe River populations of ruffe have similar genetic diversity levels--showing no founder effect, as in the other invasive species. In conclusion, high genetic variability, large numbers of founders, and multiple founding sources likely significantly contribute to the risk of an exotic species introduction's success and persistence.

Animals↗

Morphology and ultrastructure of Cryptobia eilatica n. sp. (Bodonidae: Kinetoplastida), an ectoparasite from the gills of marine fish.

A marine kinetoplastid flagellate, Cryptobia eilatica n. sp., is described from the gills of cultured gilt-head sea bream Sparus aurata L. and wild black-spot sea bream Diplodus noct (Valenciennes) in the Red Sea. The trophozoite is elongated and lacks a contractile vacuole and undulating membrane. The body averages 13.5 x 4.1 microns, anterior flagellum 9.7 microns and free portion of recurrent flagellum 15.2 microns. The ultrastructural features of the species exhibit great similarity to various previously studied Cryptobiids. Cryptobia eilatica trophozoites feed on bacteria, show a preference for the branchial interlamellar crypts, and attach to the host epithelium by means of the recurrent flagellum. Neither penetration into the epithelial cells, nor any direct damage to host tissue was observed. Cryptobia eliatica inhabits a purely marine habitat, but its trophozoite tolerates salinities as low as 10 ppt.

Animals↗

Molecular and physiological evaluation of subtropical environmental isolates of Acanthamoeba spp., causal agent of Acanthamoeba keratitis.

Previous molecular examination of Acanthamoeba spp. has resulted in the determination of distinct genotypes in this genus (designated T1-T12, T14). Genotype T4 has been responsible for the majority of cases of Acanthamoeba keratitis. Here we examine the relative abundance of environmental T4 isolates on beaches and ask whether they have temperature and salinity tolerances that could enhance pathogenicity. Twenty-four Acanthamoeba strains were isolated from beach sand (n = 20), soil (n = 3), and tap water (n = 1) in south Florida. Phylogenetic analysis identified 19 of 24 isolates as T4, the Acanthamoeba keratitis-associated genotype. The remaining isolates were genotype T5 (4) and T11 (1). Nearly all beach isolates were genotype T4, whereas the tap water and soil isolates were mostly T5. All amoebae grew at 0, 1.0, and 2.0% salt and 19 of 20 beach isolates also grew at 3.2%. No soil or tap-water acanthamoebae reproduced at 3.2%. All isolates grew at 37 degrees C and two (T5) at 42 degrees C. Little correlation existed between beach location, salt-tolerance, and genetic relatedness. Overall, the large majority of environmental isolates obtained were genotype T4, suggesting it may be the most common genotype in this environment and could be a potential source of Acanthamoeba keratitis infections.

Acanthamoeba↗

Adaptive Evolution for Freshwater Adaptation in Coilia nasus by Directional Selection on Osmoregulation Genes.

The molecular mechanisms underlying the adaptation to freshwater habitats in fish of marine origin remain unclear. Grenadier anchovies, such as Coilia nasus, originate from marine environments and include both anadromous and freshwater-resident conspecifics, making them ideal for studying adaptive evolution from marine to freshwater habitats. We conducted a comparative population genomic and transcriptome analysis of two distinct C. nasus lineages, one anadromous and the other freshwater-resident, collected from mainstream and estuarine regions of the Yangtze River, China. By genome-wide genotyping of the anadromous and the freshwater-resident populations, we observed significant divergence in osmoregulation, energy metabolism, and immune response pathways associated with ecological adaptation and energy expenditure for migration. Some ion transport genes such as CAMK1, ATP1α3, KCNJ1 and SLC30A2 were identified that may contribute to freshwater adaptation. Notably, numerous mineralocorticoid signalling genes (e.g., NR3C2, SGK1, ATP1α3, KCNJ1) exhibit dynamic change between the anadromous and freshwater populations, suggesting an important role for the hormone cortisol in regulating salinity acclimation in euryhaline fish. Among these genes, the ion channel ATP1α3 experienced adaptive amino acid substitutions (Val317Ile and Thr329Ser), which appear to be evolutionary hotspots across migratory species based on ortholog comparisons. These variants may facilitate sodium/potassium transport and highlight salinity tolerance as a key driver of divergence in anadromous fish transitioning to freshwater. These results enhance our understanding of the genetic basis underlying freshwater adaptation for an anadromous fish across osmotic boundaries.

Animals↗

Nanomolar levels of dimethylsulfoniopropionate, dimethylsulfonioacetate, and glycine betaine are sufficient to confer osmoprotection to Escherichia coli.

We combined the use of low inoculation titers (300 +/- 100 CFU/ml) and enumeration of culturable cells to measure the osmoprotective potentialities of dimethylsulfoniopropionate (DMSP), dimethylsulfonioacetate (DMSA), and glycine betaine (GB) for salt-stressed cultures of Escherichia coli. Dilute bacterial cultures were grown with osmoprotectant concentrations that encompassed the nanomolar levels of GB and DMSP found in nature and the millimolar levels of osmoprotectants used in standard laboratory osmoprotection bioassays. Nanomolar concentrations of DMSA, DMSP, and GB were sufficient to enhance the salinity tolerance of E. coli cells expressing only the ProU high-affinity general osmoporter. In contrast, nanomolar levels of osmoprotectants were ineffective with a mutant strain (GM50) that expressed only the low-affinity ProP osmoporter. Transport studies showed that DMSA and DMSP, like GB, were taken up via both ProU and ProP. Moreover, ProU displayed higher affinities for the three osmoprotectants than ProP displayed, and ProP, like ProU, displayed much higher affinities for GB and DMSA than for DMSP. Interestingly, ProP did not operate at substrate concentrations of 200 nM or less, whereas ProU operated at concentrations ranging from 1 nM to millimolar levels. Consequently, proU(+) strains of E. coli, but not the proP(+) strain GM50, could also scavenge nanomolar levels of GB, DMSA, and DMSP from oligotrophic seawater. The physiological and ecological implications of these observations are discussed.

Amino Acid Transport Systems↗

Morphology, physiology, and serology of a Pasteurella species pathogenic for white perch. (Roccus americanus).

The Pasteurella species implicated as the etiologic agent of a massive white perch mortality in the Chesapeake Bay and first described by S. F. Snieszko et al. has been characterized further in our laboratory. The general morphology and physiology of this organism is similar to that of the pasteurellae and several known fish pathogens. There are enough dissimilarities, however, to rule out its identification with any established species. The organism is obligately halophilic and grows in a temperature range between 17 and 31 C on ordinary media containing 1% NaCl. It has a relatively narrow range of pH, temperature, and salinity tolerance, and a very short survival time in spent media or brackish water, in contrast to Pasteurella pestis and P. pseudotuberculosis. Serological tests also indicate that this organism is distinct from other species which it resembles. On the basis of classic morphological and physiological criteria, this organism fits best in the genus Pasteurella; the species name piscicida (L. noun piscis, a fish; L.v.L.adj. suffix-cidus, to kill; M.L. noun piscicida, fish killer) is proposed.

Animals↗

Lack of osmoregulation in Aplysia brasiliana: correlation with response of neuron R15 to osphradial stimulation.

The exposure of Aplysia brasiliana to dilute seawater (90 and 80%) caused an increase of the relative weight, which returned to the original values after a few hours. Both osmotic and chloride concentrations of the hemolymph decreased on exposure to 80 and 90% dilute seawater, and after 3-h exposure there were no differences between the hemolymph and external media osmotic and chloride concentrations. In contrast to the clear regulatory capabilities reported for A. californica, A. brasiliana cannot maintain the osmolality of its body fluid in dilute media. In A. californica, osphradial receptors and neuron R15 are apparently involved in this regulatory mechanism. Perfusion of osphradium of A. brasiliana with dilute seawater (95-80%) did not affect electrical activity of the bursting neuron R15; perfusion with 70 and 60% seawater caused a transient increase in the duration of the quiescent period. In contrast to the model established for A. californica, in A. brasiliana no relationship was found between exposure of the osphradium to dilute media and electrical activity in neuron R15, which is in accordance with the lack of an osmoregulatory mechanism in this species. Such differences may reflect inherent differences in salinity tolerance between the two species.

Animals↗

Metamorphosis in the summer flounder Paralichthys dentatus: changes in gill mitochondria-rich cells.

Salinity tolerance changes during larval development and metamorphosis in the summer flounder (Paralichthys dentatus) and other teleosts. The physiological mechanisms responsible for osmoregulation during these early stages of development are not well understood. This study characterized changes in ultrastructure, intracellular membranes and immunoreactive Na(+)/K(+)-ATPase of mitochondria-rich cells (MRCs) in the gills of summer flounder during metamorphosis. Gill ultrastructure at the start of metamorphosis revealed only one type of MRC, which had weak reactivity to osmium and lacked a well-defined apical pit. In juveniles, two types of MRCs were observed: light-staining MRCs (LMRCs) with weak reactivity to osmium, and dark-staining MRCs (DMRCs) with strong reactivity to osmium and positioned adjacent to LMRCs. Compared with MRCs at the start of metamorphosis, the mitochondria of juvenile MRCs appeared smaller, with more transverse cristae and electron-dense matrices. Changes in MRCs during metamorphosis were also accompanied by increased immunoreactive Na(+)/K(+)-ATPase. These findings suggest that gill MRCs develop during the metamorphosis of summer flounder as the gill takes on an increasingly important osmoregulatory role.

Journal Article↗

Speciation and dispersal of Tribolodon species (Pisces, Cyprinidae) around the Sea of Japan.

Gene products of 22 protein coding loci from 29 populations of a uniquely sea-going cyprinid genus Tribolodon, T. hakonensis (anadromous and/or river-resident), T. brandtii (anadromous), T. ezoe (freshwater) and T. nakamurai (freshwater), were analyzed in order to infer the genetic divergence and dispersal of Tribolodon species around the Sea of Japan. According to the genetic distance, T. brandtii is located near the center between T. hakonensis and T. ezoe-T. nakamurai cluster. The outgroup root (Pseudaspius leptocephalus, Rhynchocypris lagowskii and Leuciscus waleckii) is attached between T. hakonensis-T. brandtii and the T. ezoe-T. nakamurai clusters. The origin of the genus Tribolodon and its salinity tolerance may date back to the Miocene Sea-of-Japan Lake and the Pliocene Paleo-Sea-of-Japan, respectively, as judged from the divergence time between Tribolodon and the outgroups. The genetic relationships and distribution pattern of Tribolodon species suggest they speciated on the Sakhalin-Japan side, and the continental populations of T. hakonensis and T. brandtii originated from anadromous colonizers. A new faunal element of Japanese freshwater fishes, the Sakhalin-Japan sub-element, is proposed for Tribolodon and some other fishes.

Alleles↗

Contrasting physiological responses of dwarf sea-lavender and marguerite to simulated sea aerosol deposition.

Plants of two wild native species from littoral areas, marguerite [Argyranthemum coronopifolium (Willd.) C.J. Humphries] and dwarf sea-lavender [Limonium pectinatum (Aiton) O. Kuntze], grown in an unheated plastic greenhouse, were sprayed 2 to 3 min per day over a 7-d period with different aqueous solutions containing (i) an anionic surfactant (S1); (ii) a solution simulating the composition of sea aerosol (S2); (iii) a solution simulating sea aerosol with anionic surfactant (S3), and (iv) deionized water alone (control). The plant resistance to sea aerosol and the ability to recover from treatments were studied. By the end of the spraying period, marguerite showed a significant reduction in growth compared with control. However, most of the growth parameters were significantly unaffected in dwarf sea-lavender when plants were treated with sea aerosol containing surfactant. Measurements of water relations variables in marguerite showed a slight decrease in leaf turgor potential after spraying with sea aerosol containing surfactant. The surfactant enhanced the foliar absorption of salt in marguerite plants, inducing reductions in leaf stomatal conductance and causing such damage in the photosynthetic apparatus that the level of net photosynthesis decreased and had not recovered by the end of the experiment. The treatments had no effect on leaf stomatal conductance and photosynthesis rate in dwarf sea-lavender plants. The response of the species studied to sea aerosol was related to the degree of salinity tolerance. Although both species are wild native plants from littoral areas, marguerite is not salt tolerant and was the most sensitive to the sea aerosol treatments, while dwarf sea-lavender, a halophyte species, was more efficient at decreasing the toxic salt content of the tissues as its growth and ornamental characteristics were not affected.

Absorption↗

Sodium uptake in Arabidopsis roots is regulated by cyclic nucleotides.

Sodium uptake from the soil is a major cause of salinity toxicity in plants, yet little is known about the mechanisms that underlie Na(+) influx. We have characterized voltage independent channels (VICs) in Arabidopsis roots that are thought to contribute to Na(+) entry. VICs showed no selectivity among monovalent cations, and their gating was found to be voltage independent. However, VIC open probability showed sensitivity to cyclic nucleotides. The presence of micromolar concentrations of cAMP or cGMP at the cytoplasmic side of the plasma membrane evoked a rapid decrease in channel open probability. In accord with predictions from electrophysiological data, our results show that short-term unidirectional Na(+) influx is also reduced in the presence of cyclic nucleotides. Moreover, addition of membrane permeable cyclic nucleotides during growth assays improved plant salinity tolerance, which corresponded with lower levels of Na(+) accumulation in plants. In summary, these data imply that Arabidopsis plants may contain a cyclic nucleotide-based signaling pathway that directly affects Na(+) transport via VICs.

Adaptation, Physiological↗

[Cloning and expression of otsA gene in E. coli].

1.5 kb of otsA gene encoding trehalose synthase has been cloned by PCR amplification. The DNA fragment was ligated to multi-copy vector and transformed to otsBA deleted and otsA deficient strains of E. coli separately. The transformants exhibited growth as well as the otsBA+ wild type on medium containing 0.5 mol/L NaCl. Trehalose was synthesized and accumulated in the transformed cells under osmotic pressure, which was determined by thin layer chromatograph. The results confirmed that otsA gene was functionally expressed in the recipient strains. These studies suggested that engineering otsA gene and trehalose accumulation into crop plants may improve drought and salinity tolerance.

Cloning, Molecular↗

Isolation and characterization of an autonomously replicating sequence (ARSD) from the marine yeast Debaryomyces hansenii.

The marine yeast Debaryomyces hansenii is known to tolerate salinities ranging from 0 to 24%. As a first step toward the molecular analysis of halotolerance in this organism, we report the isolation of an autonomously replicating sequence (ARS) and its use in the construction of a shuttle vector. The ARS from D. hansenii (ARSD) is 0.4 kbp long, and the function rests in 0.13 kbp of the sequence. Sequence analysis of ARSD shows strong homology to ARS from other organisms, including a 12-bp consensus sequence common to all ARS functional in Saccharomyces cerevisiae.

Base Sequence↗

[Effects of salinity stress on poplars seedling growth and soil enzyme activity].

A pot culture experiment with two clones of poplars (P1 and P2) showed that soil salinity had a definite inhibitory effect on the establishment and growth of seedlings. The survival rate, seedling height and weight, and root weight were all declined with increasing soil salinity. When the salinity was 0.2%, 0.4%, 0.6% and 0.8%, the shoot growth of P1 and P2 decreased by 23.24%, 48.56%, 70.76% and 83.33%, and 71.77%, 83.25%, 86.28% and 91.39%, respectively, in comparing with the control. Regressive analyses showed that soil salinity had a linear relationship with chlorophyll content, and a binomial relationship with leaf proline content. The activities of soil beta-glucosidase and L-asparaginase were decreased with increasing soil salinity. When the salinity was 0.2%, 0.4%, 0.6% and 0.8%, the beta-glucosidase activity in P1 and P2 soils decreased by 10.96%, 20.07%, 30.96% and 37.44%, and 11.21%, 18.94%, 34.89% and 41.31%, respectively. The salinity tolerance of P1 was better than that of P2.

Asparaginase↗

Distribution of the sibling species of Anopheles farauti in the Cape York Peninsula, northern Queensland, Australia.

The sibling species of Anopheles farauti s.l. were collected in larval and adult surveys from 34 localities on Cape York Peninsula and were identified by isoenzyme electrophoresis. The most common species near the coast was An. farauti 1 which was often found breeding within 100 m of the sea in either brackish or freshwater habitats. Larvae of the other 2 species were not found in brackish water which accords with previous laboratory observations of their lower salinity tolerance. Anopheles farauti 2 appears to have the widest distribution of the 3 sibling species on Cape York Peninsula as it was common in both coastal and inland localities. Anopheles farauti 3 was rarely found near the coast. In one locality at Lockhart River near the east coast of the peninsula larvae of the 3 species were found together in a small muddy creek.

Animals↗

The survivability of the ectoparasitic flagellate Ichthyobodo necator on chum salmon fry (Oncorhynchus keta) in seawater and comparison to Ichthyobodo sp. on Japanese flounder (Paralichthys olivaceus).

Experimental studies revealed that a freshwater ectoparasitic flagellate Ichthyobodo necator (Henneguy, 1883) could survive and reproduce in seawater after infected chum salmon fry, Oncorhynchus keta (Walbaum), were transferred directly from fresh water to 33% seawater. Minor morphological changes (slight reduction in body width, loss of twistlike wrinkles on body surface, and reduction in contractile vacuoles) were observed in the attached form of I. necator following transfer to seawater. The field survey also confirmed that I. necator occurs on chum salmon fry in seawater estuaries (salinity 17-34%) and in freshwater habitats. It was assumed that I. necator acquired salinity tolerance as a result of adapting to the migratory behavior of its anadromous host. Two morphologically similar bodonids, I. necator from chum salmon and Ichthyobodo sp. from marine Japanese flounder, Paralichthys olivaceus (Temminck and Schlegel), were differentiated by cross-infection experiments. Thus, the parasite from marine flounder should be regarded as a separate species from I. necator.

Animals↗