The factors that influence the composition of marine epiphytic microalgal assemblages are poorly-understood. To address this short-coming, 93 samples were collected from four distinct regions in the Florida Keys National Marine Sanctuary (FKNMS) during winter and summer months to test the model that epiphytic microalgal communities are influenced by environmental gradients related to different sites, seasons, and host macrophyte species. One hundred and eighty-three morphotypes from 13 classes (7 phyla) were identified, dominated by 106 Bacillariophyta (77 identified to species equivalent or below), 37 Cyanophyta (13 identified to species equivalent or below), and 30 Dinophyta (21 identified to species equivalent or below). The largest proportion of variability in epiphytic communities was related to physico-chemical parameters (37%), followed by site location (ocean-versus bayside; 15%), seasonal differences (11%), and host macrophyte species (10%). Four physico-chemical variables were found to be most influential: wave height, temperature, ammonium concentration, and salinity. Only six out of 616 epiphyte - host comparisons exhibited significant differences in individual epiphyte taxon abundance between different host species (within site and season), further demonstrating that host-specificity was not strongly evident in this study. Overall, the results of this (sub)tropical study indicate that changing environmental characteristics between sites and seasons were the primary drivers influencing epiphyte community composition. Similar findings were found in an accompanying study of phytoplankton and other studies from temperate and (sub)polar regions, suggesting that common, underlying processes exist among these disparate environments.

Worldwide, many coral reefs are at risk of shifting to degraded algal-dominated states, due to compromised ecological conditions. Functional diversity of herbivorous fishes maintains coral reef health and promotes reef resilience to disturbances. Given previous evidence, it appears the functional roles of herbivorous fishes differ across geographical locations, indicating a need for further assessment of macroalgal consumption by herbivorous fishes. We assessed functional diversity by examining foraging behavior of herbivorous fish species on macroalgae on a fringing reef in Moorea, French Polynesia. We video-recorded choice experiments containing seven common macroalgae and used Strauss' linear resource selection index to determine macroalgal selectivities. We used cluster analysis to identify any distinct groups within herbivorous fish species, given the macroalgal species they targeted, and fitted generalized linear mixed-effects models to identify factors that best predicted the number of bites taken on macroalgae. Seven species from 3 fish families/tribes took a total of 956 bites. Fish species differed in their selectivity with some species () strongly preferring one or two macroalgal species, while other fish species () were less selective. This resulted in fish species forming two clusters. Only 3 of 7 macroalgae were preferred by any fish species, with two fish species both preferring the same two macroalgae. The limited differences in fish species' preferences for different macroalgae suggests limited functional complementarity. Two models (macroalgal species identity+fish functional group, macroalgal species identity+fish species) best predicted the number of bites taken on macroalgae compared to models incorporating only a single explanatory factor or fish family. In the context of this Moorean fringing reef, there is greater functional redundancy than complementarity of herbivorous fishes consuming macroalgae, and the fishes grouped together according to their relative selectivity. We observed fish species that are not classified as browsers consuming macroalgae, suggesting diets of herbivorous fishes may be broader than previously thought. Finally, we observed macroalgal selectivities and consumption that differed from previous studies for the same fish species. Our results contribute to the understanding of functional diversity of herbivorous fishes across coral reefs, and also highlight the need for additional research to further elucidate the role of context and functional diversity of herbivorous fishes consuming macroalgae on coral reefs.

Herbivory assays are a valuable tool used by ecologists to understand many of the patterns and processes affecting herbivory, a widely recognized driving force in marine communities. However, methods vary substantially among studies in both design and operation, and the effect of these differences has yet to be evaluated. We assessed the effects of several key components of assay design on estimates of herbivory to offer four recommendations. First, we found assays out-planted on sequential days in both predictable and random locations within a 60m site experienced temporal increases in herbivory by an increasingly diverse assemblage of fishes. Thus, we strongly advise against placing herbivory assays in the same site over a series of days. Second, we found while the amount of biomass consumed in assays was density dependent, the percent loss was not. Thus, we recommend researchers report percent consumption because this metric is robust to differences in biomass offered and will facilitate comparisons across studies. Third, we found associational effects, where proximity of species of differing palatabilities impacted estimates of herbivory rate on one or both species, but these impacts were not consistent across species or sites. Thus, we recommend the effect of association be directly tested for multi specie herbivory assays. Fourth, we found no effect of attachment method on estimates of herbivory rate and recommend researchers continue to use the attachment method in which they are most confident. We hope our experimental results prove useful in the future when designing, conducting, and interpreting herbivory assays.

Projected increases in ocean CO levels are anticipated to affect calcifying organisms more rapidly and to a greater extent than other marine organisms. The effects of ocean acidification (OA) have been documented in numerous species of corals in laboratory studies, largely tested using flow-through exposure systems. We developed a recirculating ocean acidification exposure system that allows precise CO control using a combination of off-gassing measures including aeration, water retention devices, venturi injectors, and CO scrubbing. We evaluated the recirculating system performance in off-gassing effectiveness and maintenance of target CO levels over an 84-day experiment. The system was used to identify changes in calcification and tissue growth in response to elevated CO (1000 μatm) in three reef-building corals of the Caribbean: , , and . All three species displayed an overall increase in net calcification over the 84-day exposure period regardless of CO level (control +0.28- 1.12 g, elevated CO +0.18- 1.16 g), and the system was effective at both off-gassing acidified water to ambient CO levels, and maintaining target elevated CO levels over the 3-month experiment.

A mesocosm system was developed to simulate estuarine conditions characteristic of short water-residence time ecosystems of the Pacific Coast of North America, and used to evaluate the response of multiple macrophyte metrics to gradients of NO loading and temperature. Replicated experiments found that few responses could be directly attributed to NO loading up to 6 x ambient. Some response metrics exhibited weak relationships with nutrient loading but could not be resolved with available statistical power. While direct nutrient responses were found for some species-specific metrics (e.g. green macroalgal growth and biomass, tissue N%, etc.), many patterns were confounded with temperature. Temperature generally had a larger effect on response metrics than did nutrient load. Experimental macrophyte communities exhibited community shifts consistent with the predicted effects of nutrient loading at 20 °C, but there was no evidence of other eutrophication symptoms (phytoplankton blooms or hypoxia) due to the short system-residence time. The Nutrient Pollution Index (NPI) tracked the NO gradient at 10 °C, but exhibited no response at 20 °C, which may limit the utility of this metric in areas with marked thermal seasonality. Results suggest that teasing apart the influence of temperature and nutrients on the expression of eutrophication symptoms will require complex multi-stressor experiments and the use of indicators that are sensitive across a broad range of conditions.

The ratios of stable isotopes of carbon and nitrogen provide important information on food sources of aquatic organisms and trophic structure of aquatic food webs. For many studies, trophic position and food source are linked to bioaccumulation and trophic transfer of contaminants from prey to predators. In these cases, it is useful to use measurements on whole organisms to make direct comparisons of contaminant bioaccumulation and food web attributes. There is a great deal of variation in methods used for stable isotope analysis, particularly in the selection of tissue type and sample preparation prior to stable isotope analysis. While there have been aquatic studies that examined methodological differences, few have focused on estuarine organisms. In this study, the effects of depuration and tissue dissection on the stable isotope enrichment of common estuarine invertebrates and fish were examined. Homogenized tissues of non-depurated whole organisms were compared to dissected muscle tissue or depurated whole organisms. A 24 h depuration did not change the mean δN and δC values for most species examined. Additionally, as expected, significant differences in carbon and nitrogen signatures were found when muscle tissues were compared to whole organisms. However, differences were small enough that food source as inferred by δC or trophic level as inferred from δN would not be inaccurately represented (differences of <1.9‰ for δC and <1.2‰ for δN). The results of this study suggest that for these common estuarine fish and macroinvertebrates, stable isotopes ratios of samples can be analyzed without depuration in the same way as samples for contaminant analysis, but differences in tissue types must be taken into account when combining data from different sources.

New England salt marshes are susceptible to degradation and habitat loss as a result of increased periods of inundation as sea levels rise. Increased inundation may exacerbate marsh degradation that can result from crab burrowing and foraging. Most studies to date have focused on how crab burrowing and foraging can impact the dominant low marsh plant species, . Here we used a mesocosm experiment to examine the relationship of foraging and burrowing activity in two dominant New England crab species, and , and the combined effect of inundation, on the dominant high marsh plant species using a 3 × 2 factorial design with three crab treatments (, control) at two levels of inundation (low, high). Plants were labeled with a nitrogen (N) stable isotope tracer to estimate plant consumption by the two crab species. At both levels of inundation, we found that had a significant negative impact on both above- and below-ground biomass by physically clipping and uprooting the plants, whereas had no significant impact. Low inundation treatments for both crab species had significantly greater aboveground biomass than high inundation. Stable N isotope tracer levels were roughly the same for both and tissue, suggesting that the impact of on was not through consumption of the plants. Overall, our results suggest the potential for to negatively impact marsh stability, and that effects of crab foraging behavior may be heightened by increased inundation.

Suspension-feeding porcelain crabs ( spp.) are often the most abundant decapod crustaceans in oyster reef habitat. Analysis of water column and subtidal algal biomass from three Texas estuaries suggests that planktonic food resources are insufficient for porcelain crab growth. Pigment composition of porcelain crab muscle and digestive track contents included the diatom pigment fucoxanthin and cyanobacterial pigment canthaxanthin with digestive track samples containing attached (adnate) benthic diatoms as well as benthic cyanobacteria not found in the water column. Feeding appendages on porcelain crabs include numerous cirri with serrated edges as well as fewer more brush-like longer units. Benthic food resources are in sufficient supply to support porcelain crab biomass.

Carbon isotope fractionation (ε) between the inorganic carbon source and organic matter has been proposed to be a function of CO. To understand the CO-dependency of ε and species-specific differences therein, inorganic carbon fluxes in the four dinoflagellate species and have been measured by means of membrane-inlet mass spectrometry. In-vivo assays were carried out at different CO concentrations, representing a range of CO from 180 to 1200 μatm. The relative bicarbonate contribution (i.e. the ratio of bicarbonate uptake to total inorganic carbon uptake) and leakage (i.e. the ratio of CO efflux to total inorganic carbon uptake) varied from 0.2 to 0.5 and 0.4 to 0.7, respectively, and differed significantly between species. These ratios were fed into a single-compartment model, and ε values were calculated and compared to carbon isotope fractionation measured under the same conditions. For all investigated species, modeled and measured ε values were comparable () and/or showed similar trends with CO (). Offsets are attributed to biases in inorganic flux measurements, an overestimated fractionation factor for the CO-fixing enzyme RubisCO, or the fact that intracellular inorganic carbon fluxes were not taken into account in the model. This study demonstrates that CO-dependency in ε can largely be explained by the inorganic carbon fluxes of the individual dinoflagellates.

To evaluate the effect of elevated pCO(2) exposure on the juvenile growth of the sea urchin Lytechinus variegatus, we reared individuals for three months in one of three target pCO(2) levels: ambient seawater (380 µatm) and two scenarios that are projected to occur by the middle (560 µatm) and end (800 µatm) of this century. At the end of 89 days, urchins reared at ambient pCO(2) weighed 12% more than those reared at 560 µatm and 28% more than those reared at 800 µatm. Skeletons were analyzed using scanning electron miscroscopy, revealing degradation of spines in urchins reared at elevated pCO(2) (800 µatm). Our results indicate that elevated pCO(2) levels projected to occur this century may adversely affect the development of juvenile sea urchins. Acidification-induced changes to juvenile urchin development would likely impair performance and functioning of juvenile stages with implications for adult populations.

Hermit crabs play an important role in the Northern Adriatic Sea due to their abundance, wide range of symbionts, and function in structuring the benthic community. Small-scale (0.25 m(2)) hypoxia and anoxia were experimentally generated on a sublittoral soft bottom in 24 m depth in the Gulf of Trieste. This approach successfully simulates the seasonal low dissolved oxygen (DO) events here and enabled studying the behaviour and mortality of the hermit crab Paguristes eremita. The crabs exhibited a sequence of predictable stress responses and ultimately mortality, which was correlated with five oxygen thresholds. Among the crustaceans, which are a sensitive group to oxygen depletion, P. eremita is relatively tolerant. Initially, at mild hypoxia (2.0 to 1.0 ml l(- 1) DO), hermit crabs showed avoidance by moving onto better oxygenated, elevated substrata. This was accompanied by a series of responses including decreased locomotory activity, increased body movements and extension from the shell. During a moribund phase at severe hypoxia (0.5 to 0.01 ml l(- 1) DO), crabs were mostly immobile in overturned shells and body movements decreased. Anoxia triggered emergence from the shell, with a brief locomotion spurt of shell-less crabs. The activity pattern of normally day-active crabs was altered during hypoxia and anoxia. Atypical interspecific interactions occurred: the crab Pisidia longimana increasingly aggregated on hermit crab shells, and a hermit crab used the emerged infaunal sea urchin Schizaster canaliferus as an elevated substrate. Response patterns varied somewhat according to shell size or symbiont type (the sponge Suberites domuncula). Mortality occurred after extended anoxia (~ 1.5 d) and increased hydrogen sulphide levels (H(2)S ~ 128 μmol). The relative tolerance of crabs and certain symbionts (e.g. the sea anemone Calliactis parasitica) - as potential survivors and recolonizers of affected areas - may influence and promote community recovery after oxygen crises.

Estimating the impacts of global and local threats on coral reefs requires monitoring reef health and measuring coral growth and calcification rates at different time scales. This has traditionally been mostly performed in short-term experimental studies in which coral fragments were grown in the laboratory or in the field but measured ex situ. Practical techniques in which growth and measurements are performed over the long term in situ are rare. Apart from photographic approaches, weight increment measurements have also been applied. Past buoyant weight measurements under water involved a complicated and little-used apparatus. We introduce a new method that combines previous field and laboratory techniques to measure the buoyant weight of entire, transplanted corals under water. This method uses an electronic balance fitted into an acrylic glass underwater housing and placed atop of an acrylic glass cube. Within this cube, corals transplanted onto artificial bases can be attached to the balance and weighed at predetermined intervals while they continue growth in the field. We also provide a set of simple equations for the volume and weight determinations required to calculate net growth rates. The new technique is highly accurate: low error of weight determinations due to variation of coral density (< 0.08%) and low standard error (< 0.01%) for repeated measurements of the same corals. We outline a transplantation technique for properly preparing corals for such long-term in situ experiments and measurements.

Sponges have evolved a variety of chemical and structural defense mechanisms to avoid predation. While chemical defense is well established in sponges, studies on structural defense are rare and with ambiguous results. We used field and laboratory experiments to investigate predation patterns and the anti-predatory defense mechanisms of the sponge Melophlus sarasinorum, a common inhabitant of Indo-pacific coral reefs. Specifically, we aimed to investigate whether M. sarasinorum is chemically or structurally defended against predation and if the defenses are expressed differently in the ectosomal and choanosomal tissue of the sponge. Chemical defense was measured as feeding deterrence, structural defense as feeding deterrence and toughness. Our results demonstrated that chemical defense is evenly distributed throughout the sponge and works in conjunction with a structurally defended ectosome to further reduce predation levels. The choanosome of the sponge contained higher protein levels, but revealed no structural defense. We conclude that the equal distribution of chemical defenses throughout M. sarasinorum is in accordance with Optimal Defense Theory (ODT) in regards to fish predation, while structural defense supports ODT by being restricted to the surface layer which experiences the highest predation risks from mesograzers.

We characterized the planular-zooxanthellae symbiosis of the coral Pocillopora damicornis using criteria that are familiar in studies on corals. Similar to adult corals, planulae exhibited photoacclimation, as changes in symbiont chlorophyll a (chl a); changes in the light-saturation constant for photosynthesis (I(k)); and, at insufficient light, fewer zooxanthellae, decreased respiration, increased weight loss, and increased sensitivity to photoinhibition. Numbers of zooxanthellae in newly-released planulae varied by at least three-fold within broods. Planulae with low versus high numbers of zooxanthellae (termed pale versus dark planulae, respectively) did not differ in symbiont chl-a content, I(k), or biomass-specific rate of dark respiration. Pale planulae had lower rates of photosynthesis, but this difference vanished after three weeks, when zooxanthellar numbers increased by 225% in pale planulae and by 31% in dark planulae. Numbers of zooxanthellae also increased significantly in planulae cultured in ammonium-enriched seawater; ammonium also apparently prevented weight loss and induced settlement. Approximately 70% of photosynthetically-fixed carbon (labeled using (14)C) apparently was translocated from the zooxanthellae to their host. A comparison of planulae cultured at 0.3% versus 11% sunlight suggested that photosynthesis provided ~ 31% of the energy utilized by the latter. Overall, we conclude that the physiology of symbiosis in planulae of P. damicornis is broadly similar to symbiosis physiology in adult corals.

Eutrophication leading to hypoxic water conditions has become a major problem in aquatic systems worldwide. Monitoring the levels and biological effects of lowered oxygen levels in aquatic systems may provide data useful in management of natural aquatic environments. Fishes represent an economically important resource that is subject to hypoxia exposure effects. Due to the extreme diversity of fish species and their habitats, fishes in general have evolved unique capabilities to modulate gene expression patterns in response to hypoxic stress. Recent studies have attempted to document quantitative changes in gene expression patterns induced in various fish species in response to reduced dissolved oxygen levels. From a management perspective, the goal of these studies is to provide a more complete characterization of hypoxia responsive genes in fish, as molecular indicators (biomarkers) of ecosystem hypoxic stress.The molecular genetic response to hypoxia is highly complex and overlaps with other stress responses making it difficult to identify hypoxia specific responses using traditional single gene or low throughput approaches. Therefore, recent approaches have been aimed at developing functional genomic (e.g. high density microarray and real-time PCR) and proteomic (two-dimensional fluorescence difference in gel electrophoresis coupled with mass spectrometry based peptide identification) technologies that employ fish species. Many of the fish species utilized in these studies do not have the advantages of underlying genome resources (i.e., genome or transcriptome sequences). Efforts have attempted to establish correlations between discreet molecular responses elicited by fish in response to hypoxia and changes in the genetic profiles of stressed organs or tissues. Notable progress in these areas has been made using several different versions of either cDNA or oligonucleotide based microarrays to profile changes in gene expression patterns in response to hypoxic stress.Due to these efforts, hundreds of hypoxia responsive genes have been identified both from laboratory reared aquaria fish and from feral fish derived from both fresh and saltwater habitats. Herein, we review these reports and the emergence of hypoxia biomarker development in aquatic species. We also include some of our own recent results using the medaka (Oryzias latipes) as a model to define genetic profiles of hypoxia exposure.

Lepidophthalmus louisianensis burrows deeply into oxygen-limited estuarine sediments and is subjected to extended anoxia at low tides. Large specimens (>2 g) have a lethal time for 50% mortality (LT(50)) of 64 h under anoxia at 25º C. Small specimens (<1 g) have a significantly higher LT(50) of 113 h, which is the longest ever reported for a crustacean. Whole body lactate levels rise dramatically under anoxia and exceed 120 µmol g.f.w.(-1) by 72 h. ATP, ADP, and AMP do not change during 48 h of anoxia, but arginine phosphate declines by over 50%. Thus arginine phosphate may help stabilize the ATP pool. Surprisingly, when compared to the aerobic resting rate, ATP production under anoxia is unchanged during the first 12 h, and drops to only about 50% between 12 and 48 h. Finally, after 48 h of anoxia, a major metabolic depression to less than 5% occurs. Downregulation of metabolism is delayed in L. louisianensis compared to many invertebrates that exhibit facultative anaerobiosis. Bioenergetic constraints as a result of eventual metabolic depression led to ionic disturbances like calcium overload and compromised membrane potential of mitochondria. Because these phenomena trigger apoptosis in mammalian species, we evaluated the susceptibility of ghost shrimp mitochondria to opening of the mitochondrial permeability transition pore (MPTP) and associated damage. Energized mitochondria isolated from hepatopancreas possess a pronounced capacity for calcium uptake. Exogenous calcium does not stimulate opening of the MPTP, which potentially could reduce cell death during prolonged anoxia.

Nucleic acid and protein concentrations and their ratios are increasingly used as correlates of nutritional condition and growth in marine species. However, their application in studies of reptile growth has not yet been validated. The green turtle (Chelonia mydas) is an endangered marine reptile for which assessing population health requires knowledge of demographic parameters such as individual growth rates. The purpose of this study was to evaluate a number of biochemical indices ([DNA], [RNA], RNA:DNA ratio, [protein], protein:DNA ratio, and RNA:protein ratio) in liver, heart, and blood as potential predictors of recent growth rate in juvenile green turtles under controlled feeding conditions. Intake of juvenile green turtles was manipulated over twelve weeks to obtain a range of growth rates. With the exception of [RNA](blood), [DNA](heart), and [protein]:[DNA](liver), all biochemical indices demonstrated significant linear relationships with growth rate during the last 1.5 weeks of the study. The best single predictors of recent growth were hepatic [RNA] and [RNA]:[protein], which explained 66% and 49%, respectively, of the variance in growth. Contrary to expectations, these two indices were negatively correlated with growth rate. To investigate the possibility that hepatic [RNA] was higher in slow-growing turtles because of elevated expression of antioxidant genes, we quantified glutathione peroxidase activity and total antioxidant potential. Both measures of antioxidant function were affected by intake and growth histories, but these effects did not explain our results for hepatic RNA and protein concentrations. We developed a model that predicted 68% of the variance in specific growth rate (SGR) with the equation SGR = -0.913(ln[RNA](liver)) + 17.689(Condition Index) + 4.316. In addition, our findings that [DNA] and [RNA]:[DNA] for blood were significantly correlated with SGR demonstrate the potential utility of minimally invasive tissue sampling that could facilitate instantaneous population monitoring.

Aquatic organisms respond to environmental challenges such as thermal stress with the rapid induction of highly conserved polypeptides known as stress proteins or heat shock proteins (Hsps). Solar ultraviolet radiation (UVR, 280-400 nm) is an important environmental stressor in marine ecosystems. Here, we present results of experiments conducted with the marine copepod Acartia tonsa to follow the de novo protein synthesis and measure the level of constitutive and inducible isoforms of the Hsp70 gene family of stress proteins after UV exposure. Animals were collected from Tampa Bay, Florida (USA), and exposed to solar radiation (full spectrum), UV-A (320-400 nm) and PAR (400-700 nm), or PAR only, for periods of 0.5-4 h. Controls were kept in the dark. Protein synthesis was robust under all treatments when the copepods were exposed to low solar radiation intensities. Conversely, high solar radiation intensities (both UV-B and UV-A) caused an overall suppression in the protein synthesis of the copepods with no detectable induction of stress-inducible isoforms of Hsps. Immunochemical assays (western blotting) showed that UVR increased levels (3.5-4-fold increase compared to the dark control) of the constitutively expressed 70 kDa heat-shock (Hsc70) protein in A. tonsa, without indication of inducible isoform upregulation.

The role of disturbance in community ecology has been studied extensively and is thought to free resources and reset successional sequences at the local scale and create heterogeneity at the regional scale. Most studies have investigated effects on either the disturbed patch or on the entire community, but have generally ignored any effect of or on the community surrounding disturbed patches. We used marine fouling communities to examine the effect of a surrounding community on species abundance within a disturbed patch and the effect of a disturbance on species abudance in the surrounding community. We varied both the magnitude and pattern of disturbance on experimental settlement plates. Settlement plates were dominated by a non-native bryozoan, which may have established because of the large amount of initial space available on plates. Percent cover of each species within the patch were affected by the surrounding community, confirming previous studies' predictions about edge effects from the surrounding community on dynamics within a patch. Disturbance resulted in lower percent cover in the surrounding community, but there were no differences between magnitudes or spatial patterns of disturbance. Disturbance lowered population growth rates in the surrounding community, potentially by altering the abiotic environment or species interactions. Following disturbance, the recovery of species within a patch may be affected by species in the surrounding community, but the effects of a disturbance can extend beyond the patch and alter abundances in the surrounding community. The dependence of patch dynamics on the surrounding community and the extended effects of disturbance on the surrounding community, suggest an important feedback of disturbance on patch dynamics indirectly via the surrounding community.

Two common tropical estuarine pufferfishes were used in this study. The main species was Sphoeroides testudineus Linnaeus, 1758, a very abundant species in the estuaries of Paranaguá Bay (Paraná, Brazil), found in waters of salinity between 0 per thousand (tidal creeks) and 34 per thousand (tidal plains). The second species was S. greeleyi Gilbert, 1900, a species limited in distribution to an area of higher salinity ( approximately 30 per thousand) than S. testudineus. The present work thus aimed at evaluating the capacity of ionic regulation of both species of pufferfishes when submitted to salinity decrease, relating the results with both species' distribution in nature. Ion regulation curves for sodium (Na(+)), chloride (Cl(-)), and magnesium (Mg(2+)) ions after 6 h and 15 days of exposure of the abundant S. testudineus to the salinities of 30 per thousand, 20 per thousand, 10 per thousand, and 5 per thousand were elaborated, as well as for Cl(-) and Mg(2+) after 6 h and 15 days of exposure of both species to the extreme salinities of 35 per thousand and 5 per thousand. Both species kept their plasma Cl(-) ( approximately 120-160 mM), and Mg(2+) ( approximately 1.3 mM) concentrations stable, as did S. testudineus for Na(+) ( approximately 130 mM). Na(+) (measured only for S. testudineus) and Cl(-) were either hyper-regulated (in 5 per thousand) or kept iso-ionic ( approximately 7-10 per thousand), but more often hypo-regulated (20-35 per thousand). In contrast, Mg(2+) was strongly hypo-regulated in all salinities. According to their distribution records in nature, S. greeleyi was less able to tolerate strong sea water dilution, showing mortality after 5 days in 5 per thousand water. These estuarine pufferfishes are thus efficient regulators of plasma ionic concentrations in diluted sea water, as expected from their occupation of estuaries. The experiments have supported the distribution records of both species in the estuarine complex and resident estuarine species were thus characterized with respect to their osmoregulatory capacities.

Salt marshes have the potential to intercept nitrogen that could otherwise impact coastal water quality. Salt marsh plants play a central role in nutrient interception by retaining N in above- and belowground tissues. We examine N uptake and allocation in two dominant salt marsh plants, short-form and . Nitrogen uptake was measured using N tracer experiments conducted over a four-week period, supplemented with stem-level growth rates, primary production, and microbial denitrification assays. By varying experiment duration, we identify the importance of a rarely-measured aspect of experimental design in N tracer studies. Experiment duration had a greater impact on quantitative N uptake estimates than primary production or stem-level relative growth rates. Rapid initial scavenging of added N caused apparent nitrogen uptake rates to decline by a factor of two as experiment duration increased from one week to one month, although each experiment shared the qualitative conclusion that roots scavenged N approximately twice as rapidly as . We estimate total N uptake into above- and belowground tissues as 154 and 277 mg N·m·d for and , respectively. Driving this pattern were higher N content in leaves and belowground tissue and strong differences in primary production; and produced 8.8 and 14.7 g biomass·m·d. Denitrification potentials were similar in sediment associated with both species, but the strong species-specific difference in N uptake suggests that -dominated marshes are likely to intercept more N from coastal waters than are short-form marshes. The data and source code for this manuscript are available as an R package from https://github.com/troyhill/NitrogenUptake2016.