Potentially pathogenic vibrios in brackish waters and mussels

Water and mussel samples were collected from two brackish lakes, used as mussel farms, at different times of the year, for the quantitative analysis of Vibrio spp. and for the isolation of potentially pathogenic species. The isolates underwent cultural and biochemical tests selected for rapid identification. Glucose oxidizing-fermenting and O/129 sensitive strains were distinguished on the basis of the following tests: sucrose and cellobiose utilization, sulphatase activity and polymyxin B resistance performed, respectively, on TCBS, CPC and SPS media. Responses to the presence of beta-galactosidase, salt requirement and growth on triple sugar iron medium were also added. A total of 125 from 152 isolates were referred to the species Vibrio fluvialis (55 strains), V. alginolyticus (40), V. parahaemolyticus (11), V. vulnificus (10) and V. mimicus (9). The remaining 27 isolates were not identified. The isolation of potentially pathogenic vibrios from cultivated mussels is a risk for health of people consuming raw seafood. Therefore, a long-term monitoring programme should also include the search for these bacterial species.     

Ecology of benthic and epiphytic nematodes in brackish waters

Abundances of benthic nematodes from shallow waters in Tvärminne, Finland and in Ringkøbing Fjord, Denmark averaged 1.2–1.5 10⁶ individuals m^?2. Less than 20 species were found. More than 80% of the individuals were made up by Anoplostoma viviparum, Chromadorita fennica, Axonolaimus spinosus, Daptonema trabeculosum and Sabatieria pulchra. S. pulchra is designated as an anoxybiotic species and is attracted to the root tips of young Potamogeton perfoliatus, whilst A. viviparum and C. fennica are oxybiotic species. Abundance of epiphytic nematodes extracted from submerged macrophytes in the Cladophora-Pilayella belt in the outer archipelago of the Finnish archipelago (July 1978) was estimated to reach 5 · 10⁶ individuals m^?2, equivalent to a biomass of 1.2 g org. C m^?2. Abundance and biomass are smaller in the Potamogeton meadows in the inner archipelago, in littoral algal vegetation of the sea zone, and on sublittoral vegetation. Epiphytic nematode species composition is influenced by salinity and food availability and not by physiognomic types or silt contents of the vegetation in the Baltic Sea, European estuaries and lakes. Dominant species are: Chromadorina bioculata, Punctodora ratzeburgensis, Chromadorita tenuis, Adoncholaimus thalassophygas, Theristus acer, Monhystera parva, M. aff. filiformis and Chromadora aff. nudicapitata. A theory is proposed to explain observed seasonal changes in the species composition of epiphytic and benthic nematodes. During winter months, the submerged vegetation is mostly destroyed, being found as detritus on the sea bottom. Sampling benthic nematodes during winter thus includes epiphytic species living in the detritus. In spring the epiphytic nematodes detect the newly growing vegetation, leave the sea bottom, colonize the submerged vegetation and spend their life there.     

DNA barcoding identifies Argentine fishes from marine and brackish waters

Background

DNA barcoding has been advanced as a promising tool to aid species identification and discovery through the use of short, standardized gene targets. Despite extensive taxonomic studies, for a variety of reasons the identification of fishes can be problematic, even for experts. DNA barcoding is proving to be a useful tool in this context. However, its broad application is impeded by the need to construct a comprehensive reference sequence library for all fish species. Here, we make a regional contribution to this grand challenge by calibrating the species discrimination efficiency of barcoding among 125 Argentine fish species, representing nearly one third of the known fauna, and examine the utility of these data to address several key taxonomic uncertainties pertaining to species in this region.

Methodology/Principal Findings

Specimens were collected and morphologically identified during crusies conducted between 2005 and 2008. The standard BARCODE fragment of COI was amplified and bi-directionally sequenced from 577 specimens (mean of 5 specimens/species), and all specimens and sequence data were archived and interrogated using analytical tools available on the Barcode of Life Data System (BOLD; www.barcodinglife.org). Nearly all species exhibited discrete clusters of closely related haplogroups which permitted the discrimination of 95% of the species (i.e. 119/125) examined while cases of shared haplotypes were detected among just three species-pairs. Notably, barcoding aided the identification of a new species of skate, Dipturus argentinensis, permitted the recognition of Genypterus brasiliensis as a valid species and questions the generic assignment of Paralichthys isosceles.

Conclusions/Significance

This study constitutes a significant contribution to the global barcode reference sequence library for fishes and demonstrates the utility of barcoding for regional species identification. As an independent assessment of alpha taxonomy, barcodes provide robust support for most morphologically based taxon concepts and also highlight key areas of taxonomic uncertainty worthy of reappraisal.     

Isolation and evaluation of oil-producing microalgae from subtropical coastal and brackish waters

Microalgae have been widely reported as a promising source of biofuels, mainly based on their high areal productivity of biomass and lipids as triacylglycerides and the possibility for cultivation on non-arable land. The isolation and selection of suitable strains that are robust and display high growth and lipid accumulation rates is an important prerequisite for their successful cultivation as a bioenergy source, a process that can be compared to the initial selection and domestication of agricultural crops. We developed standard protocols for the isolation and cultivation for a range of marine and brackish microalgae. By comparing growth rates and lipid productivity, we assessed the potential of subtropical coastal and brackish microalgae for the production of biodiesel and other oil-based bioproducts. This study identified Nannochloropsis sp., Dunaniella salina and new isolates of Chlorella sp. and Tetraselmis sp. as suitable candidates for a multiple-product algae crop. We conclude that subtropical coastal microalgae display a variety of fatty acid profiles that offer a wide scope for several oil-based bioproducts, including biodiesel and omega-3 fatty acids. A biorefinery approach for microalgae would make economical production more feasible but challenges remain for efficient harvesting and extraction processes for some species.     

Macrofauna distribution in brackish inland waters in relation to chlorinity and other factors

Summary In 1981, 16 brackish inland waters on the Dutch island of Texel were sampled qualitatively for macrofauna. The sampling stations consisted of ditches, small canals and isolated ponds. They were clustered according to their species composition. It appeared that the major fauna differences are closely related to the occurrence of present or past connections of the locality with other water bodies and to the origin and development of a habitat. The distribution of the macrofauna taxa was compared with differences in average chlorinity and variation in chlorinity between the habitats studied. Relations between the number and kind of species and chlorinity were also investigated. It was found that chlorinity cannot be regarded as a master factor affecting the fauna distribution. Hence, it is concluded that, within the range of the mesopolyhalinicum, environmental and historical factors, rather than chlorinity, have to be regarded as the most discriminating abiotic factors determining the species composition in brackish inland waters.     

The distribution of aquatic oligochaetes in brackish inland waters in the SW Netherlands

Macrobenthic fauna was sampled in inland waters with unstable environmental parameters in the SW Netherlands. The distribution of 23 species of aquatic oligochaetes was related to chlorinity. Oligochaetes and other macrofaunal taxa were grouped by cluster analysis. Brackish-marine oligochaetes appeared to have a similarly large tolerance for chlorinity variations, as is known for the better researched euryhaline macrofauna.Communication no. 221.     

The macrofauna distribution in brackish inland waters in relation to chlorinity and other factors

The macrofauna of small isolated brackish waters on a barrier island in the Dutch Wadden Sea was sampled qualitatively. An attempt was made to correlate species composition and species richness to chlorinity characteristics and to data on the water regime of the localities studied.It is shown that the observed differences in the macrofauna cannot be explained by differences in chlorinity only. Present and former connections to other water bodies seem equally important.     

The zooplankton of fresh and brackish waters of the Cape Thompson Area,Northern Alaska

Summary A survey was made of the occurrence of zooplankton in fresh and brackish waters at Cape Thompson, Alaska between June 28, 1960 and August 7, 1961.Fourteen species of Cladocera and 38 species of Copepoda (13 Calanoida, 11 Cyclopoida and 14 Harpacticoida) were distinguished in collections from 10 lakes, 8 lagoons, and 111 pools.Males of Daphnia middendorffiana are illustrated and their geographical significance discussed.Cyclops scutifer had the only hibernal developmental cycle; all others had developmental cycles during early summer or throughout the summer and fall.Relatively complete seasonal developmental cycles are presented for Holopedium gibberum, Daphnia middendorf iana, Heterocope septentrionalis, Eurytemora canadensis, E. gracilicauda, Limnocalanus johanseni, Diaptomus pribilofensis, D. arcticus, Cyclops scutifer, and C. magnus.

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Zusammenfassung Eine Übersicht des Vorkommens vom Zooplankton in der Zeit zwischen dem 28. juni, 1960 and dem 7. August 1961 im Süsswasser und im Brackwasser des Kaps Thompson, Alaska wird hier gegeben.Von den Sammlungen aus 10 Seen, 8 Lagunen and 111 Teichen sind 14 Spezies der Cladoceren and 38 Spezies der Copepoden (darunter 13 der Calanoiden, 11 der Cyclopoiden, und 14 der Harpacticoiden) unterschieden worden.Die männlichen der Daphnia middendorfliana werden hierin illustriert und die geographische Bedeutung erläutert.Cyclops scutifer hatte den einzigen winterlichen Entwicklungszyklus; die anderen hatten Entwicklungszyklen früh in dem Frühling oder während des ganzen Sommers and Herbstes.Jahreszeitliche Entwicklungszyklen, die verhältnismässig vollkommen sind, werden für die folgenden vorgeführt: Holopedium gibberum, Daphnia middendorffiana, Heterocope septentrionalis, Eurytemora canadensis, E. gracilicauda, Limnocalanus johanseni, Diaptomus pribilofensis, D. arcticus, Cyclops scutifer, und C. magnus.

     

Variations in R-plasmid DNA concentrations of Escherichia coli during starvation in sewage and brackish waters

Cell culturability and plasmid stability in Escherichia coli containing plasmids RP1, R388 and pUB824 were studied in raw and treated wastewater, and in brackish water. The E. coli strain survived well in the three samples of water employed. Moreover, the three plasmids were maintained under all conditions studied. Interestingly, plasmid DNA concentration of individual plasmids followed the same evolution as the culturable bacteria in the corresponding selective medium when the bacteria grew in raw or treated wastewater. In contrast, in brackish water, the stress due to the oligotrophic and salinity conditions of the medium produced an initial paradoxical increase in plasmid DNA concentration, followed by a decrease in the number of culturable bacteria in the corresponding selective medium. Maintenance of RP1 (56 kbp) and R388 (33 kbp) was markedly influenced by nutritive conditions, which caused a segregation of the plasmids from cells. The results of the present study suggest that variations in plasmid DNA concentrations in an aquatic environment depend on the quality of the water and also on the molecular weight of the plasmid considered.     

Distribution and antibacterial drug resistance ofAeromonas spp. from fresh and brackish waters in Southern Turkey

The frequency of antibiotic resistance was compared inAeromonas spp. isolated from fresh and brackish water in Southern Turkey. A total of 97Aeromonas spp. strains were isolated from four zones (three from fresh and one from brackish water). Most of the strains isolated from all samples wereAeromonas hydrophila (79.4%), while the amount ofAeromonas sobria andAeromons caviae, were rather lower in the samples examined (17.5% and 3.1% respectively). A high proportion of isolates from all water sources showed resistance to cephalotin (86.6%) and trimethoprim-sulphamethoxazole (69%). On the other hand, a low proportion of bacteria showed resistance to tetracycline (14.4%), chloramphenicol (11.3%), gentamicin (7.2%) and nitrofurantoin (6.8%). Only one strain showing resistance to amikacin was found. Multiple Antibiotic Resistance Index (MARI) to at least two antibiotics was highest in brackish water (zone 4), followed by fresh water (zone 3). MARI values ranging from 0.2 to 0.8 for the bacteria isolated from brackish water. This study suggest that, multiple antibiotic resistantAeromonas spp., especiallyA. hydrophila, can be easily recovered from fresh and brackish water sources in Turkey and these sources may play as a reservoirs responsible for disease pathogen aeromonads.     

Salt adaptation in pseudomonads: characterization of glucosylglycerol-synthesizing isolates from brackish coastal waters and the rhizosphere

The compatible solute glucosylglycerol (GG) is widespread among cyanobacteria, but, until now, has been reported for only two species of heterotrophic bacteria. About 120 bacterial isolates from coastal regions of the Baltic Sea were screened by HPLC for their ability to synthesize GG. Positive isolates (26) were grouped by SDS-PAGE of whole-cell proteins and representative strains of each group were investigated by sequencing their 16S rRNA genes and phenotypic characterization. All GG-synthesizing isolates were shown to belong to the genus Pseudomonas (sensu stricto) and were assigned to 4 distinct groups, although none of the GG-synthesizing isolates could be unambiguously assigned to described species. The identity of GG was verified by 13C NMR analysis and enzymatic digestion with alpha- and beta-glucosidases. Besides GG, salt adapted cultures of the aquatic isolates accumulated the dipeptide N-acetylglutaminylglutamine amide (NAGGN) and glutamate. The accumulation of noncharged compatible solutes was also tested in previously identified pseudomonads isolated from the rhizosphere of oilseed rape and potato. The majority of these strains were fluorescent species of the genus Pseudomonas and accumulated trehalose and NAGGN when grown under salt stress conditions. However, rhizosphere isolates of Stenotrophomonas maltophilia synthesized GG and trehalose or only trehalose in a strain-dependent manner. These data indicate that the ability to synthesize GG is widely distributed among slightly or moderately halotolerant pseudomonads.     

Lysogens and free viruses in fresh, brackish, and marine waters: a Bayesian analysis

A yearlong study was conducted to determine factors that affect the abundance and distribution of lysogens and free viruses at fresh-, brackish-, and saltwater stations in Newport Bay, CA. The viral and bacterial abundance were highest in the freshwater (average 1.1 × 10⁸ and 1.1 × 10⁷ mL⁻¹, respectively) and lowest in the marine water (average 0.4 × 10⁸ and 0.5 × 10⁷ mL⁻¹, respectively). Bacterial and viral counts were also several times higher during the summer than in winter. Approximately, 35% of the 141 samples were inducible in the presence of mitomycin C. The highest percentage of inducible lysogens was observed in marine waters (42%), while the lowest percentage was observed in the warmer freshwater (23%). A statistical model for the joint occurrence of lysogens and free viruses was formulated and estimated using Bayesian techniques to understand the key environmental determinants of viruses and lysogens. Our results support the existence of significant heterogeneity between the saltwater and freshwater sites. A parsimonious model that combines the two saltwater sites performs best among the specifications that were considered. Bacteria and water temperature were significant determinants of virus counts, whereas lysogen relationships are unclear. Importantly, conditional on the covariates, viruses and lysogen fractions exhibit robust negative correlation.     

Distribution of picophytoplankton communities from brackish to hypersaline waters in a South Australian coastal lagoon

Background

Picophytoplankton (i.e. cyanobacteria and pico-eukaryotes) are abundant and ecologically critical components of the autotrophic communities in the pelagic realm. These micro-organisms colonized a variety of extreme environments including high salinity waters. However, the distribution of these organisms along strong salinity gradient has barely been investigated. The abundance and community structure of cyanobacteria and pico-eukaryotes were investigated along a natural continuous salinity gradient (1.8% to 15.5%) using flow cytometry.

Results

Highest picophytoplankton abundances were recorded under salinity conditions ranging between 8.0% and 11.0% (1.3 × 10⁶ to 1.4 × 10⁶ cells ml^-1). Two populations of picocyanobacteria (likely Synechococcus and Prochlorococcus) and 5 distinct populations of pico-eukaryotes were identified along the salinity gradient. The picophytoplankton cytometric-richness decreased with salinity and the most cytometrically diversified community (4 to 7 populations) was observed in the brackish-marine part of the lagoon (i.e. salinity below 3.5%). One population of pico-eukaryote dominated the community throughout the salinity gradient and was responsible for the bloom observed between 8.0% and 11.0%. Finally only this halotolerant population and Prochlorococcus-like picocyanobacteria were identified in hypersaline waters (i.e. above 14.0%). Salinity was identified as the main factor structuring the distribution of picophytoplankton along the lagoon. However, nutritive conditions, viral lysis and microzooplankton grazing are also suggested as potentially important players in controlling the abundance and diversity of picophytoplankton along the lagoon.

Conclusions

The complex patterns described here represent the first observation of picophytoplankton dynamics along a continuous gradient where salinity increases from 1.8% to 15.5%. This result provides new insight into the distribution of pico-autotrophic organisms along strong salinity gradients and allows for a better understanding of the overall pelagic functioning in saline systems which is critical for the management of these precious and climatically-stress ecosystems.     

Physiological responses to salinity changes of the isopod Idotea chelipes from the Baltic brackish waters

Survival, haemolymph osmolality and the specific metabolic rate of the isopod Idotea chelipes from the brackish waters of the southern Baltic Sea (salinity 7 psu) were investigated after acclimation to different salinities in the range of 3-30 psu keeping other parameters constant (T=15 degrees C, full air saturation). Haemolymph osmolality of I. chelipes increased from 379+/-33 mmol kg(-1) at 3 psu to 935+/-12 mmol kg(-1) at 30 psu. In the salinity range of 3-11 psu I. chelipes exhibited a hyperosmotic pattern of haemolymph regulation, whereas from 14 to 25 it behaved like an osmoconformer. Isoosmosis occurred at 28.5 psu (920 mmol kg(-1)). The specific metabolic rate of I. chelipes was significantly (P<0.05) related to salinity, with maximum (2.4+/-0.8 J g(-1) wm h(-1)) and minimum values (0.6+/-0.3 J g(-1) wm h(-1)) at 7 and 20 psu, respectively. The changes in the specific metabolic rate and in the difference between the osmolality of haemolymph and external medium were significantly correlated for I. chelipes (P<0.05). Rates of change per salinity unit were, however, unequal varying according to salinity transition. These differences indicate that osmotic adjustment may be a more or less costly in terms of energy according to salinity.     

Microfossils in surface sediments of brackish waters on the west coast of South Africa and their palaeoecological implications

A faunistic survey covering 25 sites in estuaries, coastal lakes and ponds along the west coast of South Africa between the Cape of Good Hope in the south and the Olifants River in the north was carried out in May 2014. The study aimed to establish a dataset with ecological and distribution data of ostracods and foraminifers for later palaeoenvironmental reconstructions. Canonical correspondence analysis showed that the distribution of the 19 foraminifer and 32 ostracod taxa was controlled mainly by habitat structure, but that specific conductivity (salinity) was best and solely described by the second axis, highlighting the usefulness of the two microfossil groups for salinity reconstructions. Habitat structure was demonstrated by the foraminifer species Trochamminita irregularis’ preference for stillwater areas of lakes. Sarscypridopsis aculeata and Sarscypridopsis glabrata were the dominant ostracod species in coastal ponds and lakes. However, their living distributions excluded each other, with S. aculeata preferring areas of lower salinity and S. glabrata dominating areas of higher salinity up to hyperhaline conditions in small, closed water bodies.     

Using the salinity preferences of benthic macroinvertebrates to classify running waters in brackish marshes in Germany

Marshland streams along European coastlines are typically located in the transition zone from limnic to brackish conditions. They constitute a distinct and important stream type within the German stream typology, which is predominantly characterised by tidal influence and salinity. According to the Water Framework Directive (WFD) the basis for the abiotic classification of European transitional and coastal waters is the ‘Venice System’ (Weber, M., 2005. Ergebnisse der Bestandsaufnahme 2004 zur Umsetzung der europäischen Wasserrahmenrichtlinie (WRRL) in den Küstengewässern Mecklenburg-Vorpommerns. Rostocker Meeresbiol. Beitr. 14, 7–15). However, the extreme spatial and temporal variability in salinity and tidal influence determines and controls the macroinvertebrate communities of marshland river and stream ecosystems and hampers the application of the ‘Venice System’. Earlier, purely biotic classifications of brackish waters typically classify different brackish water zones, each with a specific macroinvertebrate community, and often yield unsatisfactory results due to the extensively dynamic abiotic factors found in marshland streams. In practice, the permanent impact of tidal charge means that stable salinity zones do not exist. This effect can than be intensified by freshwater discharges after heavy rainfall or high tides in combination with strong winds that hinder discharge into the open sea. To overcome the restricted applicability of abiotic and biotic classification systems, we present a scoring system for marshland streams that combines the abiotic salinity classes of the ‘Venice System’ with the salinity preferences of the macroinvertebrate community. This proposal is based on literature data for macroinvertebrate salinity tolerances and preferences. According to the 137 references that were evaluated, 151 macroinvertebrate taxa were classified into six salinity groups. In a second step, we applied this scoring scheme to empirical data from two official quality assessment surveys. These surveys had been carried out in ten estuary tributaries of German marshland areas in Lower Saxony and Schleswig-Holstein in order to implement the WFD. Applying the scoring system to these data revealed a clear (and expected) gradient that was not expressed in the abiotic data. Thus we conclude that the biotic salinity preferences of benthic macroinvertebrates are a useful baseline metric for the ecological assessment of marshland streams and other transitional waters. Regional revision and adaptation of taxonomic salinity preferences would broaden the applicability of this assessment to transitional waters worldwide.