Effect of salinity gradients and heterotrophic microbial activity on biodegradation of nitrilotriacetic acid in laboratory simulations of the estuarine environment

The biodegradation of nitrilotriacetic acid (NTA), a synthetic replacement detergent builder, in the estuarine environment was examined by using a laboratory estuarine simulation. Two interdependent microcosms were used; each of five vessels was equilibrated with a saline gradient between 1.30 and 17.17%, with the final vessel subsequently being increased to a maximum salinity of 31.6%. Each microcosm was seeded simultaneously with heterotrophic bacteria from both fresh and saline sources. Viable counts demonstrated the ability of each microcosm to sustain a mixed heterotrophic bacterial community throughout the range of salinities for 183 days after a stabilization period. Isolation studies demonstrated that both systems contained four bacterial species, representatives of the genera Vibrio and Flavobacterium and members of the coryneform group and the family Enterobacteriaceae. Total bacterial numbers and species diversity decreased with increased salinity. NTA was administered at low and high concentrations, one concentration to each microcosm, initially with the least amount of saline. Removal of both concentrations of NTA occurred and was attributed to biodegradation after a period of bacterial acclimatization. Subsequent dosing of NTA to vessels of higher salinity demonstrated that biodegradation was incomplete at observed mean salinities of greater than 9.18% at low influent NTA concentrations and greater than 5.08% at high influent NTA concentrations. Therefore, acclimatization was dose dependent. It was concluded that NTA acclimatization at the higher salinities ceased because of salinity stress-induced failure of NTA catabolism and not the disappearance of a particular bacterial species.     

Effects of salinity on bacterioplankton: field and microcosm experiments

Bacterial populations in a tidal estuary were monitored with the aim of investigating (i) the relationship between planktonic bacteria and salinity and (ii) the survival of allochthonous bacteria in microcosm experiments at different salinities. With the increase in salinity, bacterial numbers decreased in a curvilinear fashion, rather than monotonically. Maximal abundance for different bacterial groups was found between 7 and 22%₀ S. The salinity alone explained between 19 and 58% of bacterial variability. In microcosm experiments the adverse effect of salinity on the survival of freshwater bacteria and indicators of faecal contamination was particularly important. In freshwater and low salinity experiments grazing by protozoans had similar effects on the survival of allochthonous bacteria.     

Survival of faecal indicator bacteria in tropical estuarine waters (Bangpakong River,Thailand)

AIMS: To investigate the survival of cultivable bacteria in the tropical Bangpakong estuary (Eastern Thailand) under different salinities and light conditions. METHODS AND RESULTS: Dark and light microcosm experiments using membrane diffusion chambers were carried out under three different experimental conditions, namely (i) low salinity, (ii) progressive mixing with brackish water and (iii) fast mixing with high salinity water spiked with raw urban sewage. Faecal coliforms declined faster than faecal enterococci, as shown by survival T90 values ranging from 82.2 +/- 4.2 to 14.5 +/- 0.8 h and 97.5 +/- 0.4-20.6 +/- 1.2 h, respectively. The survival of freshwater heterotrophic bacteria was higher but variable (121.2 +/- 5.0-30.1 +/- 14.3 h), whereas that of heterotrophic marine bacteria was rather stable (81.5 +/- 4.2-44.6 +/- 2.5 h). CONCLUSIONS: Overall survival was higher in low salinities. Light had a further deleterious effect, since it accelerated the decay of faecal indicators, particularly in high salinities. Faecal enterococci had a higher resistance to environmental conditions compared with faecal coliforms. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is relevant to the understanding of the behaviour of different faecal indicator bacteria and the optimization of sewage treatment plants aimed at the reduction and/or elimination of faecal load discharged into estuarine waters submitted to salinity variations.     

The effects of salinity,and calcium and potassium variations on the growth of two estuarine red algae

The effects of varying salinity and calcium and potassium concentrations on the growth of two species of estuarine Rhodophyta, Bostrychia radicans Montagne and Caloglossa lepricurii (Montagne) J. Agardh, were examined in unialgal culture. Inocula of settled tetraspores on glass coverslips were incubated in six concentrations of potassium (0. 100, 200, 300, 400 and 500 mg/l) at five salinities (0, 5, 15, 25, and 35%) in the presence of calcium. Growth responses of each alga were determined from the average cell number of 75 tetrasporelings after 4 days in Ott's synthetic sea-water medium. The concentrations of dissolved potassium and calcium in sea water along a salinity gradient in an estuary were determined with atomic absorption spectrophotometry. Four-day-old tetrasporeling of Bostrychia and Caloglossa demonstrated growth over wide ranges of potassium concentrations with growth maxima at 400 500 and 200 400 mg/l, at the optimal salinities for growth of 25 and 15%, respectively. These studies indicate also that the presence of calcium is essential for appreciable growth of both species at each salinity and the effects of variations in potassium are dependent upon the presence of calcium. The abundance of both species in the Mullica River estuary. New Jersey, appears to be a response to a total osmotic effect along the salinity gradient because sufficient levels of the major cations are present. However, the upper limit of both species towards the head of the Mullica River estuary may be determined by combinations of low salinity and low levels of dissolved calcium and potassium rather than by low salinity alone.     

Estuarine microplankton: An experimental approach in combination with field studies

In the natural environment, seasonal climatic changes have the dominant effect on phytoplankton productivity. Salinity effects in estuarine habitats are, however, probably equally important but cannot usually be separated from seasonality. The use of microcosms allows the salinity effect to be studied independent of seasonality.Results of a year-long field study in the Fraser River estuary, British Columbia, Canada, are presented and compared with results of laboratory experiments. In a series of flasks (201 volume), natural low and high salinity waters were mixed in order to give salinities of ? 5, 10, 18, and ? 26%.. The mixed waters were enriched so as to simulate the entrainment of nutrient-rich, saline water in a salt wedge estuary. A distinct pattern of autotrophic and heterotrophic growth developed resembling natural events in the Fraser River estuary during the period between winter and late spring. The salinity values influenced the microplankton ecology with respect to phytoplankton species composition and total biomass. Despite seasonal variability of the source waters, the simulated spring bloom was reproducible under constant laboratory conditions, thus allowing the continued performance of experiments.The microcosms were useful in simulating natural events as well as in testing the impact of natural and man-made perturbations. In the experiments presented, naturally occurring perturbations had a much greater impact on estuarine ecology than addition of anthropogenic pollutants in concentrations much higher than known to occur in moderately polluted estuaries.     

Euryhaline adaptations in the fat sleeper, Dormitator maculatus

Salinity tolerances and plasma osmotic concentrations were determined in the fat sleeper, Dormitator maculatus , a common species in estuarine and coastal fresh waters along the Atlantic and Caribbean coasts of North, Central and South America. Analyses followed sequential laboratory acclimations to a series of ambient salinities at a constant temperature of 20 ± 1° C and photoperiod of 12L: 12D. These fish tolerated a range of ambient salinities from fresh water through a salinity of 75‰. Plasma osmotic concentrations were regulated at an essentially constant level in the salinity range from fresh water through a salinity of 50‰, beyond which plasma concentrations trended upward with increased ambient salinity. We conclude that D. maculatus , while truly euryhaline, docs not show the extreme euryhalme capabilities of several teleost fishes that are 'full-time' estuarine inhabitants.     

Estimation of the contribution of halobacterial to the bacterial biomass and activity in solar salterns by the use of bile salts

Abstract Bile salts (deoxycholate, taurocholate) were used to estimate the contribution of bacteria of the Halobacterium group to bacterial community size and activity at different salinities as found in a multi-pond saltern. Low concentrations of bile salts cause lysis of halophilic archaebacteria of the Halobacterium group, while halophilic eubacteria and halococci remain microscopically intact. Upon addition of bile salts, total bacterial numbers (as estimated microscopically) in saltern ponds at salinities below 250 g/l did not decrease, while above this salinity bacterial numbers decreased by 30–50%. To estimate the contribution of halobacteria to overall heterotrophic activity, the effect of bile salt addition was tested on the incorporation of labelled amino acids. In saltern ponds of a salinity below 250 g/l activity was not greatly inhibited by taurocholate, while at salinity above 300 g/l taurocholate completely abolished incorporation of amino acids.     

Salinity tolerance in Hyalophysa chattoni (Ciliophora, Apostomatida), a symbiont of the estuarine grass shrimp Palaemonetes pugio

The apostome ciliate Hyalophysa chattoni, a symbiont of the estuarine grass shrimp Palaemonetes pugio, was tested for its growth and reproductive ability in a wide range of salinities from 0.1 to 55 ppt. Shrimp, with their attached ciliates, were slowly acclimated to different salinities in order to assess protozoan cell size and division. The trophont and tomont stages of the ciliate life cycle were analyzed. In both stages, cell size increased with salinity from 0.1 to 20 ppt. Cell size leveled in the 20-35 ppt range, and decreased at higher salinities. The number of daughter cells produced per tomont cyst correlated with increased cell size, and also correlated with increased salinity. Additionally, increased salinity correlated with an increase in the percentage of cells able to divide and excyst as tomite stages. These results indicate that H. chattoni is able to grow and divide more effectively at salinities closer to seawater than in the estuarine environment from which they were collected. Though able to survive salinities from 0.1 to 55 ppt, the species is better adapted for an existence in the higher salt concentrations.     

Effects of temperature,ph, salinity,and inorganic nitrogen on the rate of ammonium oxidation by nitrifiers isolated from wetland environments

Ammonium-oxidizing bacteria were examined in two wetland environments, a freshwater marsh and an estuarine bay, during a 2-year period. Two predominant types were consistently isolated, one from each environment. Both isolates were identified as species ofNitrosomonas. Using a closed culture, high cell density assay, the effects of temperature, pH, salinity, Na⁺, K⁺, nitrite, nitrate, and ammonium concentrations on ammonium oxidation were determined. Maximum activity was observed for the freshwater isolate at 35°C, pH 8.5, salinities of 0.3 to 0.5% Na⁺ and K⁺, and ammonium concentrations greater than 0.5 g/l. For the estuarine isolate, maximum activity was observed at 40°C, pH 8.0, salinities of 0.5 to 1.0%, 1.0% Na⁺ and K⁺, and 0.2 g/l ammonium. The estuarine isolate had a Na⁺ requirement which could be partially substituted by the K⁺, suggesting that the organism is a true estuarine bacterium. Nitrite inhibited both isolates at concentrations greater than 5 mg/l, whereas nitrate had no significant effect on either isolate.     

Biodegradation of Phenanthrene by a Halophilic Bacterial Consortium Under Aerobic Conditions

A halophilic bacterial consortium that degraded phenanthrene was developed from oil-contaminated saline soil containing 10% salinity. The biodegradation of phenanthrene occurred at 5%, 10%, and 15% salinity, whereas no biodegradation took place at 0.1% and 20% salinity. A 16S rRNA gene analysis showed that all sequences from the denaturing gradient gel electrophoresis profile were similar to those of halophilic bacteria. This is the first report of a halophilic bacterial consortium capable of degrading phenanthrene under hypersaline conditions.     

Bacterial diversity of a cyanobacterial mat degrading petroleum compounds at elevated salinities and temperatures

Cyanobacterial mats of the Arabian Gulf coast of Saudi Arabia experience extreme conditions of temperature and salinity. Because they are exposed to continuous oil pollution, they form ideal models for biodegradation under extreme conditions. We investigated the bacterial diversity of these mats using denaturing gradient gel electrophoresis and 16S rRNA cloning, and tested their potential to degrade petroleum compounds at various salinities (fresh water to 16%) and temperatures (5 to 50 degrees C). Cloning revealed that c. 15% of the obtained sequences were related to unknown, possibly novel bacteria. Bacteria belonging to Beta-, Gamma- and Deltaproteobacteria, Cytophaga-Flavobacterium-Bacteroides group and Spirochetes, were detected. The biodegradation of petroleum compounds at different salinities by mat microorganisms showed that pristine and n-octadecane were optimally degraded at salinities between 5 and 12% (weight per volume NaCl) whereas the optimum degradation of phenanthrene and dibenzothiophene was at 3.5% salinity. The latter compounds were also degradable at 8% salinity. The same compounds were degraded at temperatures between 15 and 40 degrees C but not at 5 and 50 degrees C. The optimum temperature of degradation was 28-40 degrees C for both aliphatics and aromatics. We conclude that the studied microbial mats from Saudi Arabia are rich in novel halotolerant and thermotolerant microorganisms with the potential to degrade petroleum compounds at elevated salinities and temperatures.     

Interaction of salinity and temperature as a mechanism for spatial separation of three co-existing species of Ambassidae (Cuvier) (Teleostei) in estuaries on the south-east coast of Africa

Ambassidae are represented in South African estuaries by three species which are very similar in external morphology, feeding ecology, alimentary system and distribution. The co-existence and spatial separation of these species of Ambassis in the estuaries of southern Africa can be explained by the tolerance of each to salinity and temperature. Investigations of the temperature tolerance ranges of the three species suggest that the osmoregulatory capability of Ambassis productus in reduced salinities (<10%) increases while that of A. gymnocephalus decreases sharply in salinities below 20%. A. natalensis , which is endemic to the south-east coast of Africa, is adapted to a wide range of estuarine conditions. Interaction between salinity and temperature tolerances on the limits within which Ambassis spp. survive is significant with regard to the distribution of the three species in estuaries. Catch data showed that A. productus is restricted to areas of low salinity and seasonal low temperature (upper reaches of estuaries), A. gynmocephalus to high salinities >25% (estuary mouths) and A. natalensis to intermediate salinities associated with the middle reaches of estuaries.     

Ecophysiological adaptability of tropical water organisms to salinity changes]

Physiological response of tropical organisms to salinity changes was studied for some marine, estuarine and freshwater fishes (Astyanax bimaculatus, Petenia karussii, Cyprinodon dearborni, and Oreochromis mossambicus), marine and freshwater crustaceans (Penaeus brasiliensis, Penaeus schmitti and Macrobrachium carcinus), and marine bivalves (Perna perna, Crassostrea rhizophorae, and Arca zebra) collected from Northeast Venezuela. They were acclimated for four weeks at various salinities, and (1) placed at high salinities to determine mean lethal salinity, (2) tested by increasing salinity 5@1000 per day to define upper lethal salinity tolerance limit, or (3) observed in a saline gradient tank to determine salinity preference. Acclimation level was the most significant factor. This phenomenon is important for tropical aquatic organisms in shallow waters, where they can adapt to high salinity during the dry season and cannot lose their acclimation level at low salinity during abrupt rain. For saline adaptation of tropical organisms, this behavior will contribute to their proliferation and distribution in fluctuating salinity environments.     

Hydrocarbon biodegradation in hypersaline environments

When mineral oil, hexadecane, and glutamate were added to natural samples of varying salinity (3.3 to 28.4%) from salt evaporation ponds and Great Salt Lake, Utah, rates of metabolism of these compounds decreased as salinity increased. Rate limitations did not appear to relate to low oxygen levels or to the availability of organic nutrients. Some oxidation of l-[U-C]glutamic acid occurred even at extreme salinities, whereas oxidation of [1-C]hexadecane was too low to be detected. Gas chromatographic examination of hexane-soluble components of tar samples from natural seeps at Rozel Point in Great Salt Lake demonstrated no evidence of biological oxidation of isoprenoid alkanes subject to degradation in normal environments. Some hexane-soluble components of the same tar were altered by incubation in a low-salinity enrichment culture inoculated with garden soil. Attempts to enrich for microorganisms in saline waters able to use mineral oil as a sole source of carbon and energy were successful below, but not above, about 20% salinity. This study strongly suggests a general reduction of metabolic rate at extreme salinities and raises doubt about the biodegradation of hydrocarbons in hypersaline environments.     

Comparison of low salinity tolerance in Callinectes sapidus Rathbun and Callinectes similis Williams postlarvae upon entry into an estuary

Planktonic larvae of estuarine crabs are commonly exported to the continental shelf for development and then return to coastal and estuarine areas as postlarvae (megalopae). Megalopae returning to estuaries must be adapted to survive in brackish water whereas those of coastally distributed species should not need such adaptations. We investigated 1) whether megalopae of the estuarine crab Callinectes sapidus and the coastal crab Callinectes similis undergo changes in salinity tolerance upon entry into an estuary and 2) what factors induce those changes. Megalopae were collected at a coastal site and a nearby estuarine site and exposed to a range of salinities (5, 10, 15, 20 and 30) for 6 h. Percent survival was determined after 24 h reintroduction to the collection site water. We also investigated 1) whether increased salinity tolerance was induced by reduced salinity or estuarine chemical cues, 2) the time to acclimation and 3) the salinity necessary for acclimation. C. sapidus megalopae from the estuarine site were more likely to survive exposure to low salinities than those from the coastal site. C. sapidus megalopae from the coastal site exhibited increased survival after acclimation to salinities of 27 and 23 for 12 h. Estuarine chemical cues had no effect on salinity tolerance. C. similis megalopae were less likely to survive at low salinities and did not exhibit an acclimation response upon exposure to reduced salinities. These results suggest that megalopae of C. sapidus are physiologically adapted to recruit to estuaries whereas megalopae of C. similis are unable to acclimate to low salinity conditions.     

Ecophysiological behavior of Caquetaia kraussii (Pisces: Cichlidae) exposed to different temperatures and salinities.

Tropical river sardine, Caquetaia kraussii, captured from La Aguá lagoon (Sucre State, Venezuela) were acclimatized for four weeks at 22, 24, 30 and 32 degrees C and at 0, 5, 10, 15 and 17@1000 salinity. To evaluate effects of thermal response to acclimatization level, the fish were transferred suddenly from lower temperatures (22 and 24 degrees C) to higher ones (32 and 30 degrees C) respectively. Then thermal resistance time was measured at the lethal temperature of 40.9 degrees C for 30 days. We considered that acclimatization process completed when resistance time was stabilized at the new temperature regime. For the saline effect, the concentrations of sodium and potassium were measured in the tissues at each treatment: gills, white muscle, gut and heart. The results showed that thermal tolerance increased rapidly in 3 h with a 6 degrees C rise in temperature (from 24 to 30 degrees C) and in 24 h with a 10 degrees C rise (22 to 32 degrees C). With decreasing temperatures, the acclimatization level reached its lowest in 11 days with a 6 degrees C decreases (from 30 to 24 degrees C) and in 14 days with a 10 degrees C decrease (32 to 22 degrees C). Caquetaia kraussii regulates as much sodium as potassium in gills and white muscle tissues at all salinity levels tested; however, gut and heart tissues showed significantly different regulations among salinities examined.     

Distribution of heterotrophic flagellates at the littoral of estuary of Chernaya River (Kandalaksha Bay, White Sea)

Species composition, distribution, and the character of structural changes in the heterotrophic flagellate community were studied along environmental gradients in the Chernaya River estuary. There were 99 species and forms of heterotrophic flagellates, subdivided into three groups: prevalently marine species and euryhaline species preferring biotopes either of higher or decreased salinity. The heterotrophic flagellate community of the estuary was continuously divided into two distinct variants: (1) cenosis of halophilic species, prevalently of sea forms and euryhaline species preferring biotopes of increased salinity; (2) cenosis of halophobic species with prevalence of euryhaline forms gravitating to fresh biotopes. The arbitrary and indistinct boundary between the variants of the community ran at a salinity of 9–10‰. The response of estuarine communities of heterotrophic flagellates and infusorians to variation of abiotic factors was similar and differed from response of communities of microphyto-, meiozoo-and macrozoobenthos; this implied similarity of the response mechanism to environmental factors in organisms of one level of organization.     

Different reproductive success at low salinity determines the estuarine distribution of two Palaemon prawn species

The two prawn species Palaemon adspersus Rathke and P. elegans Rathke differ in their distribution patterns in estuaries: P. adspersus occurs at lower salinities and also extends further into the Baltic than P. elegans . Yet, at low salinities adult survival does not differ between the two species. Reproductive success was, however, substantially reduced in P. elegans at low salinity, but not in P. adspersus . Berried P. elegans females from the Swedish west coast hatched significantly fewer clutches at 10%‰ than did P. adspersus females from the same locality. Furthermore, larval survival in P. elegans was significantly lower at 5 and 7.5‰ than in P. adspersus . At higher salinities (10 and 24.5‰) no interspecific differences in larval survival were found, except in one experiment where P. elegans larvae had a lower mortality. It is concluded that the different estuarine distributions of the two palaemonid prawn species result from these interspecific differences in reproductive success at low salinity.     

Biodegradation of crude oil across a wide range of salinities by an extremely halotolerant bacterial consortium MPD-M,immobilized onto polypropylene fibers

The bacterial consortium MPD-M, isolated from sediment associated with Colombian mangrove roots, was effective in the treatment of hydrocarbons in water with salinities varying from 0 to 180 g L(-1). Where the salinity of the culture medium surpassed 20 g L(-1), its effectiveness increased when the cells were immobilized on polypropylene fibers. Over the range of salinity evaluated, the immobilized cells significantly enhanced the biodegradation rate of crude oil compared with free-living cells, especially with increasing salinity in the culture medium. Contrary to that observed in free cell systems, the bacterial consortium MPD-M was highly stable in immobilized systems and it was not greatly affected by increments in salinity. Biodegradation was evident even at the highest salinity evaluated (180 g L(-1)), where biodegradation was between 4 and 7 times higher with immobilized cells compared to free cells. The biodegradation of pristane (PR) and phytane (PH) and of the aromatic fraction was also increased using cells immobilized on polypropylene fibers.     

Responses of the saltmarsh rush Juncus kraussii to salinity and waterlogging

Juncus kraussii Hochst., an important saltmarsh macrophyte, is intensively harvested for many commercially orientated products and current populations are under threat of overexploitation. In saline, intertidal mud banks, this species occurs on higher ground, suggesting that it is adapted to lower salinities and less frequent inundation. The objectives of this study were to determine biomass accumulation, as well as morphological and physiological adaptations of J. kraussii to salinity and waterlogging stresses. Plants collected from the field were subjected to 0.2, 10, 30, 50 and 70% seawater under drained or flooded conditions for three months. Measurements were made of biomass accumulation, CO₂ exchange, chlorophyll fluorescence, ion and water relations. Furthermore, seed germination responses to a range of salinities were investigated. Total dry biomass accumulation, as well as the number and height of culms, decreased with increase in salinity under both flooded and drained conditions. Generally, CO₂ exchange, stomatal conductance, Photosystem II (PSII) quantum yield and electron transport rate (ETR) through PSII declined with increase in salinity in both the flooded and drained treatments. Predawn and midday ψ in culms decreased with increase in salinity, being lower under drained than flooded conditions. Inorganic solute concentrations in culms increased with increase in salinity, with Na⁺ and Cl⁻ being the predominant ions. Na⁺/K⁺ ratios in culms increased significantly with increase in salinity. Proline concentrations in roots and culms, which increased with salinity, were considerably higher under drained than flooded conditions. Germination was best at salinities less than 20% seawater and decreased significantly with further increase in salinity to 110% seawater. Transfer of ungerminated salt-treated seeds to distilled water stimulated germination. This study has demonstrated that J. kraussii is a highly salt and flood tolerant species, being able to grow and survive in salinities up to 70% seawater, under both drained and flooded conditions. Maximal growth occurred at low salinities (<10% seawater) under flooded condition.