Saltwater Exposure Research Articles

Maintenance Challenges in Maritime Environments and the Impact on Urban Mobility: Machico Stayed Bridge

This article investigates the challenges of maintaining the Machico Cable-Stayed Bridge in a marine environment, focusing on its implications for urban mobility. The primary problem addressed is the impact of harsh marine conditions on the structural integrity of the bridge, which poses significant challenges for ongoing maintenance and safety. The research highlights unique aspects such as the effects of saltwater exposure on materials and the interplay between infrastructure and urban transit dynamics. By emphasizing these critical issues, this study aims to provide insights into effective maintenance strategies and contribute to the broader discourse on urban mobility in coastal regions.

Leaf Physiological Responses and Early Senescence Are Linked to Reflectance Spectra in Salt-Sensitive Coastal Tree Species

Salt-sensitive trees in coastal wetlands are dying as forests transition to marsh and open water at a rapid pace. Forested wetlands are experiencing repeated saltwater exposure due to the frequency and severity of climatic events, sea-level rise, and human infrastructure expansion. Understanding the diverse responses of trees to saltwater exposure can help identify taxa that may provide early warning signals of salinity stress in forests at broader scales. To isolate the impacts of saltwater exposure on trees, we performed an experiment to investigate the leaf-level physiology of six tree species when exposed to oligohaline and mesohaline treatments. We found that species exposed to 3–6 parts per thousand (ppt) salinity had idiosyncratic responses of plant performance that were species-specific. Saltwater exposure impacted leaf photochemistry and caused early senescence in Acer rubrum, the most salt-sensitive species tested, but did not cause any impacts on plant water use in treatments with <6 ppt. Interestingly, leaf spectral reflectance was correlated with the operating efficiency of photosystem II (PSII) photochemistry in A. rubrum leaves before leaf physiological processes were impacted by salinity treatments. Our results suggest that the timing and frequency of saltwater intrusion events are likely to be more detrimental to wetland tree performance than salinity concentrations.

COPPER AND MICROPLASTIC EXPOSURE AFFECTS THE GILL GENE EXPRESSION OF COMMON CARP DURING SALTWATER CHALLENGE

The aim of the present study was to assess if pre-exposure to water copper and/or polyvinyl chloride microparticles affects the transcriptional responses of common carp, Cyprinus carpio, to saltwater exposure. Fish were exposed to 0.25 mg/L copper alone (Cu) or in the presence of 0.5 mg/L polyvinyl chloride microparticles (Cu-MPVC) for 14 days, followed by 72 hours of salt water exposure (0 to 13 ppt NaCl). The copper content in the gills and the expression of heat shock protein (hsp70) and cytochrome P450 family 1 (cyp1a) transcripts were examined. The results showed that gill copper levels increased significantly (P = 0.008) in the Cu and Cu-MPVC treatments after 14 days of exposure, compared to the control fish; the Cu and Cu-MPVC treatments had similar gill copper levels. After 14 days of exposure, branchial expression of the hsp70 and cyp1a genes was significantly up-regulated in the Cu and Cu-MPVC treatments. Exposure to salt water led to a significant down-regulation of the gene transcripts in all treatments after 24 h of exposure. At this point, the Cu and Cu-MPVC treatments showed transcripts similar to those of the control fish prior to saltwater exposure. The fish treated with Cu-MPVC showed significantly higher hsp70 expression 72 h after saltwater exposure than the other treatments. At this time point, the control and Cu fish had significantly lower cyp1a expression than before saltwater exposure. In conclusion, the present data suggest that copper exposure induces stress in the fish gills, and the presence of MPCV in the water hampers normal transcriptomic responses of the fish gills to saltwater exposure.

Laser treated novel textures for adhesion performance of aluminum alloy joints exposed to corrosion

The aim of this study was to investigate novel laser-treated structures designed to improve the adhesion performance and corrosion resistance of AA7075-T6 bonded joints. The spacing, position, and surface area parameters of the laser-treated grooves were investigated to understand their role in adhesion performance even under corrosion conditions. Laser-treated surfaces were analyzed using microscopic, topographic, and chemical methods to determine their scientific contribution to adhesion and corrosion performance. Laser-treated adherends were also bonded to mechanically test their bond strength both before and after exposure to corrosion for varying periods of time. Roughness and therefore surface area have a significant effect on adhesion performance; however, it is worth noting that the best adhesion performance both before and after corrosion exposure was achieved when the required surface area was properly designed and, laser treated according to the salt water exposure directions to inhibit its diffusion into the adhesion interface. The performance of the novel laser-treated structures in terms of adhesion performance under corrosive atmosphere was also supported by failure analysis. The mixed failure mode was significantly shifted to the cohesive failure mode, especially for bonded joints with box-type laser-treated structures.

High pressure salt water and low temperature effects on the material performance characteristics of additive manufacturing polymers

The effects of salt water exposure at deep ocean depth pressures when coupled with low temperatures on the material characteristics of three unique additively manufactured polymers has been investigated through a detailed experimental approach. The polymers in the study were manufactured utilizing both Vat Photopolymerization and Material Extrusion printing techniques. The Material Extrusion process was utilized to produce material specimens of Stratasys ULTEM 9085 and Markforged Onyx while the Vat Photopolymerization process was used to produce specimens of Accura ClearVue. The ULTEM 9085 and Markforged Onyx are filament based polymers and the ClearVue is a liquid based resin. The specimens were first submerged in a high pressure, salt water bath of 3.5% NaCl solution at 34.5 MPa (5000 lb/in2) for a total exposure time of 60 days to determine the water absorption characteristics. Subsequent to the salt water exposure at high pressure, the specimens were evaluated to determine changes in tension, compression, flexure, and in-plane fracture properties. To determine the effects of water saturation and low temperature coupling, the mechanical testing was performed at temperatures of 20 °C, 0 °C and −20 °C in both dry and saturated conditions. Additionally, non-destructive testing in the form of TeraHertz and FIRT imaging was conducted to analyze the physical material changes through the thickness of the material due to the saline water absorption. To quantify the change in material storage and loss moduli properties, Dynamic Mechanical Analysis (DMA) characterization was performed on each of the AM polymers in dry and saturated states. The DMA testing also quantified changes in the Glass Transition Temperature because of salt water exposure. In summary, The current study investigates the effects of coupled long term/high pressure salt water exposure with low temperatures on the mechanical and material characteristics of three unique AM polymers by: (1) immersing the materials in a salt water solution at 34.5 MPa for 60 days, (2) Conducting post exposure mechanical testing on the materials at 0 °C and −20 °C with comparisons to 20 °C testing on dry specimens, and quantifies changes in material properties through DMA experiments. The results from all testing in the study show that high pressure salt water exposure when coupled with low temperatures has unique effects on each of the materials considered in the study and careful consideration to each parameter must be given based on the material type when components will be employed in marine operations.

An experimental investigation on mitigating cracks and augmenting the endurance of concrete structures in marine environment by bio-mortar immobilised with halophilic bacteria

In coastal areas, built structures encounter hostile conditions and forces that can cause them to deteriorate over time owing to saltwater exposure, tidal forces, reinforcement corrosion, and freeze–thaw cycles. Early age cracks in such structures accelerate the rate of deterioration, and the current research focuses on alleviating such threats. This paper evaluates the performance of a self-healing mortar made by encapsulating expanded perlite with the bacterium Halobacillus Halophilus MCC2188. Mortar cube specimens of size 70.6 mm × 70.6 mm× 70.6 mm were prepared with cement: fine aggregate in 1:3 ratios. A 10% volume of the fine aggregate fraction was substituted with the expanded perlite immobilised with bacterial spores and nutrients. The expanded perlite aggregates were coated with sodium silicate and cement solution to protect the spores from the nonconducive environment. The specimens were subjected to fully and partially submerged marine water curing. The mechanical properties and self-healing potential were evaluated, and the precipitated polymorphs in completely healed cracks were identified and examined by characterisation techniques such as XRD, FEGSEM, FTIR, and TGA-DTG. The marine bacterium under investigation can tolerate the high salt concentrations commonly found in seawater and saline marshy soil and produce calcite through the metabolism of organic compounds, making it a suitable microorganism for self-healing applications. Crack widths of up to 0.84 mm and 92.79% average strength recovery were achieved in 56 days post-cracking, and the pace of healing was quicker in partially submerged curing conditions. The results showed improved self-healing, strength regain and mechanical strength and proved to be an efficient tool for enhancing the endurance of biomortar in severe marine exposure conditions.

Biodegradable materials with FDM technology under the aging effect of solar and saltwater exposure

The durability and degradation of polymers is very important for product design in terms of material choice. The degradation behavior of two biodegradable thermoplastic materials manufactured by 3D printing, Enviro ABS and PLA, was studied. The action of the sun and seawater was simulated to find out how they affect the properties of these materials over a period of 8 weeks. The yield strength, maximum elongation, ultimate tensile strength, and microscopy were analyzed, as well as dimensions and mass changes. These biodegradable materials were studied to conclude whether there is an environmentally friendly alternative to traditional ABS, being one of the most widely used petroleum-based plastics in industry and in fused deposition modeling (FDM) or fused filament fabrication (FFF). PLA showed a weight loss and increase in ultimate tensile stress on degradation by sunlight and a prolonged decrease in ultimate tensile stress on degradation by seawater due to humidity absorption. In contrast, Enviro ABS does not show a noticeable difference between the beginning and the end of the test, which leads to the conclusion that Enviro ABS is a good alternative to conventional ABS without forgetting the environmental effects that are currently involved in the manufacture, recycling and composting of this type of material.

Salinity exposure affects lower-canopy specific leaf area of upland trees in a coastal deciduous forest

Sea level rise and increasing storm surges are likely to affect the canopy physiology, ecology, and structure of coastal forests, even well in advance of tree mortality. Laboratory and greenhouse studies have documented that saltwater exposure can trigger changes in leaf-level physiology and morphology, but few in situ studies have examined how tree-specific leaf area (SLA), the ratio of leaf area to mass and a crucial trait and model parameter, is affected by saline soils. We conducted an observational study of SLA in a mid-Atlantic (USA) coastal deciduous forest, taking advantage of a natural gradient in salinity along a tidal creek. Measured SLA of the 239 trees and seven species sampled ranged from Carya glabra (N = 6 trees, mean SLA = 277.9 ± 36.3 cm2/g) to Fagus grandifolia (N = 60, 321.9 ± 62.9 cm2/g); as expected, trees species and canopy position (sun versus shade) significantly affected SLA. For trees (N = 100) directly exposed to the tidal creek, salinity was highly significant after accounting for species (P < 0.001), with trees in the lower reaches of the creek having lower SLA. Leaf area index (LAI), computed from SLA and litter traps, ranged from 4.8 to 15.8 and was inversely related to salinity exposure; the spatial variability in leaf litter production contributed much more to LAI uncertainty than did SLA variability. These in situ results are correlative but consistent with the hypothesis, based on previous greenhouse studies, that the stress of chronic salinity exposure changes species’ leaf morphology. Our findings are useful for understanding the growing effects of saltwater intrusion into upland forests, as well as parameterizing and testing ecosystem-scale models simulating forest stressors and disturbances at the terrestrial-aquatic interface.

Durability Performance of Geopolymer Concrete of Various Strength

Geopolymers, primarily composed of fly ash, have proved an excellent substitute for ordinary portland cement (OPC) in terms of sustainability and productivity. In order to determine the geopolymer concrete's (GPC) resistance to chemical assaults and water permeability, it is necessary to obtain geopolymer concrete (GPC) of varying strengths after normal curing. The objectives of the research was to test the durability performances of the GPC of various strength under normal curing and investigating the optimum strength based on durability testing of the GPC. For this research, different type of cement-to-fly ash ratio was used for various strength data. The appropriate mixture was conducted by using the trial mix method in order to obtain better accuracy of the results data during the mixing design process. To satisfy the varied strength designs, a small proportion of OPC is added to the GPC mixture as part of the mix design. After 28 days of curing, this durability testing is undertaken after the concrete has reached its maximum strength. The compressive strength test and weights were performed and compared to the GPC mix design at 60 °C after heat curing. The 8% OPC replacement has greater resistance to sulfate attack, saltwater exposure, and water permeability compared to the 6% and 7% OPC alternatives. Consequently, the experiment reveals that the GPC's durability and strength increase as the percentage of OPC increases.

Hurricane Kay (September 2022) Impacts Rocky Shores in the Loreto Area of Baja California Sur, Mexico

This project follows a tradition of survey work undertaken to appraise physical and biological damage in the aftermath of hurricane-strength winds and waves at a given locality where conditions were well documented prior to the arrival of a particular storm. The locality is the 12 m limestone terrace at Arroyo Blanco on the eastern shores of Isla del Carmen in Baja California Sur, Mexico. A study undertaken in February 2018 established that the surface of the terrace is covered by a coastal boulder deposit that features large slabs of limestone pealed from the outer edge by strong surf attributed to storms of hurricane intensity but unknown date. The largest slabs tend to be rectilinear in shape vulnerable to dislodgement along horizonal bedding planes and weaknesses in vertical joints. These blocks are sufficiently large and weigh enough that movement by humans without necessary mechanical equipment would be impossible. Hurricane Kay, rated as a Category 2 storm, struck the island on 8 September 2022 and an effort was made to visit the area for reconnaissance and detailed survey work soon afterwards. Although a Category 2 storm lacked the energy to remobilize the largest limestone slabs on the terrace, it was found that the storm was sufficient to disturb the adjacent seabed and redeposit as many as 44 sea fans onto the terrace by overwash; the sea fans belonged to the species Pacifigorgia adamsi. Moreover, a species of land plant common to the limestone terrace is the Gulf Star Violet (Stenotis mucronate); it suffered significant desiccation and death due to saltwater exposure. The occurrence of large potholes on the limestone terrace represents a style of physical erosion previously undocumented at the locality and rarely seen elsewhere on rocky shores.

Juvenile American Alligators (Alligator mississippiensis) Use Behavioral Thermoregulation to Cope with Dehydration and Salt-Loading when Chronically Exposed to 12 ‰ Saltwater

American alligators (Alligator mississippiensis) are native to freshwater habitats in the Southeastern United States. These areas are susceptible to salinity increases due to extreme weather events (e.g., hurricanes) and climate change. The goal of this study was to investigate behaviors in juvenile alligators exposed to 12‰ saltwater for five weeks. Sixteen alligators were maintained in an experimental tank with 90 L of water and a heat lamp overhanging a basking plate. Observations were conducted in freshwater (prior to exposure to saltwater) and over the course of 5 weeks in 12‰ saltwater. This allowed for comparison of behaviors in freshwater and in saltwater in the same group of animals. Behaviors related to thermoregulation (basking and staying in the water) and feeding were recorded via instantaneous sampling, in which sampling intervals of 1 min occurred throughout a 20 min observation period. Saltwater-exposed alligators reduced foraging and feeding within the first week of saltwater exposure and spent significantly less time basking compared with freshwater observations. Furthermore, significantly fewer animals chose to bask over the course of 5 weeks in saltwater. A novel behavior, never seen in freshwater, was the closing of the outer eyelids when alligators were in the saltwater. Collectively, the present study demonstrates that chronic saltwater exposure has significant impact on the thermoregulatory behavior in juvenile alligators concomitantly exposed to dehydration and salt-loading without access to freshwater. As alligator hatchlings emerge during peak hurricane season, the survivability of hatchlings may be compromised if access to freshwater is unattainable.

Responses to saltwater exposure vary across species, populations and life stages in anuran amphibians.

To predict the impacts of environmental change on species, we must first understand the factors that limit the present-day ranges of species. Most anuran amphibians cannot survive at elevated salinities, which may drive their distribution in coastal locations. Previous research showed that coastal Hyla cinerea are locally adapted to brackish habitats in North Carolina, USA. Although Hyla squirella and Hyla chrysoscelis both inhabit coastal wetlands nearby, they have not been observed in saline habitats. We take advantage of naturally occurring microgeographic variation in coastal wetland occupancy exhibited by these congeneric tree frog species to explore how salt exposure affects oviposition site choice, hatching success, early tadpole survival, plasma osmolality and tadpole body condition across coastal and inland locations. We observed higher survival among coastal H. cinerea tadpoles than inland H. cinerea, which corroborates previous findings. But contrary to expectations, coastal H. cinerea had lower survival than H. squirella and H. chrysoscelis, indicating that all three species may be able to persist in saline wetlands. We also observed differences in tadpole plasma osmolality across species, locations and salinities, but these differences were not associated with survival rates in salt water. Instead, coastal occupancy may be affected by stage-specific processes like higher probability of total clutch loss as shown by inland H. chrysoscelis or maladaptive egg deposition patterns as shown by inland H. squirella. Although we expected salt water to be the primary filter driving species distributions along a coastal salinity gradient, it is likely that the factors dictating anuran ranges along the coast involve stage-, species- and location-specific processes that are mediated by ecological processes and life history traits.

Solution Uptake in Cylindrical Carbon-Fibre-Reinforced Polymer (CFRP) Tendons

Salt water exposure conditions relevant to carbon-fibre-reinforced polymer (CFRP) prestressed concrete structures in marine environments are investigated. The diffusion into relatively small diameter CFRP tendons can be a lengthy process so the prediction of the long-term moisture uptake using short-term experiments on thin films of epoxy would be advantageous. However, the fibre inclusions within a composite introduce complexities, and factors are typically required for correlation with pure epoxy specimens. Diffusion parameters based on moisture uptake result from CFRP tendons exposed to salt water solution at 20°C and 60°C are compared with those obtained using equivalent thin film specimens. The higher temperature is selected to accelerate the moisture uptake. It is found that the measured ratios of tendon and epoxy diffusivity were temperature dependent, and the combination of the higher temperature and salt solution leads to an increased propensity for moisture uptake in the tendon. Existing analytical models to predict the CFRP tendon diffusivity from that of a thin film of epoxy did not appear to capture the observed trends. However, predictions using a unit cell with a fibre interface zone suggest that this may be due to an increased diffusivity in the interphase region.

Year-around survey and manipulation experiments reveal differential sensitivities of soil prokaryotic and fungal communities to saltwater intrusion in Florida Everglades wetlands

Global sea-level rise is transforming coastal ecosystems, especially freshwater wetlands, in part due to increased episodic or chronic saltwater exposure, leading to shifts in biogeochemistry, plant- and microbial communities, as well as ecological services. Yet, it is still difficult to predict how soil microbial communities respond to the saltwater exposure because of poorly understood microbial sensitivity within complex wetland soil microbial communities, as well as the high spatial and temporal heterogeneity of wetland soils and saltwater exposure. To address this, we first conducted a two-year survey of microbial community structure and bottom water chemistry in submerged surface soils from 14 wetland sites across the Florida Everglades. We identified ecosystem-specific microbial biomarker taxa primarily associated with variation in salinity. Bacterial, archaeal and fungal community composition differed between freshwater, mangrove, and marine seagrass meadow sites, irrespective of soil type or season. Especially, methanogens, putative denitrifying methanotrophs and sulfate reducers shifted in relative abundance and/or composition between wetland types. Methanogens and putative denitrifying methanotrophs declined in relative abundance from freshwater to marine wetlands, whereas sulfate reducers showed the opposite trend. A four-year experimental simulation of saltwater intrusion in a pristine freshwater site and a previously saltwater-impacted site corroborated the highest sensitivity and relative increase of sulfate reducers, as well as taxon-specific sensitivity of methanogens, in response to continuously pulsing of saltwater treatment. Collectively, these results suggest that besides increased salinity, saltwater-mediated increased sulfate availability leads to displacement of methanogens by sulfate reducers even at low or temporal salt exposure. These changes of microbial composition could affect organic matter degradation pathways in coastal freshwater wetlands exposed to sea-level rise, with potential consequences, such as loss of stored soil organic carbon.

Microbial Composition of Freshwater Marsh Sediment Responds more Strongly to Microcosm Seawater Addition than Simulated Nitrate or Phosphate Eutrophication.

As sea level rise impacts coastal wetlands, saltmarsh will overtake coastal freshwater marsh in many areas, but changes in the sediment microbiome in response to saltwater intrusion are difficult to predict. Coastal freshwater marsh sediment was exposed to ambient, brackish, and saline conditions as well as to elevated nitrate and phosphate to model the combined stresses of saltwater intrusion and coastal eutrophication. Initially, sediment prokaryotic composition was similar to prior studies of freshwater marsh but diverged over time, reflecting the magnitude of increase in saltwater. There was no observed effect of nutrient amendment, potentially ranking seawater intrusion as a higher-importance compositional driver. Although the previously described loss of methanogenic populations and promotion of sulfate reducers in response to saltwater exposure was observed, taxonomic distribution was not similar to typical meso-polyhaline wetlands. Without colonization by marine taxa, such a community may be short-lived naturally, ultimately equilibrating with more common saltmarsh species. However, the recapitulation of salinity concentration by freshwater sediment microbial composition demonstrates the overwhelming nature of saltwater intrusion relative to other drivers like eutrophication.

Mechanical Properties of Corroded Steel Rebars after 20 Years of Saltwater Exposure

Mechanical Properties of Corroded Steel Rebars after 20 Years of Saltwater Exposure

Environmental Impact on the Behavior of CFRP Sheet Attached to Concrete

Carbon fiber-reinforced polymer (CFRP) has many advantages as a construction/structural-strengthening material. However, there are still concerns regarding the long-term performance of these materials when used with reinforced concrete (RC) structures. Environmental conditions have an adverse effect on the behavior of CFRP and the bond between these sheets and concrete. Therefore, the durability of CFRP used for strengthening RC beams was evaluated under different environmental scenarios, including subjection to immersion in deicing agents, tap water, and saltwater, freeze-and-thaw cycles, and outdoor environmental changes. Laboratory tests were performed to examine the influence of these environmental scenarios on the bonding behavior between CFRP sheets and concrete in terms of deformations and modes of failure. Two types of test setups were performed in this study, namely pull-off shearing and three-point bending. Forty-two concrete prisms with CFRP were prepared and tested by using the pull-off shearing setup. It was observed that as the period of exposure increased, noticeable effects on the CFRP sheet as well as the bond stiffness were observed. Exposure to tap water had a greater impact than saltwater on the CFRP–concrete bond strength as well as the CFRP. In addition, eighteen notched concrete beams strengthened with an external CFRP were tested under three-point bending. The tap water exposure showed a 3.6% increase in the bond strength compared to the control specimen. However, the saltwater exposure showed a 10% increase.

Geomorphological Data from Detonation Craters in the Fehmarn Belt, German Baltic Sea

Military munitions from World War I and II dumped at the seafloor are a threat to the marine environment and its users. Decades of saltwater exposure make the explosives fragile and difficult to dispose of. If required, the munition is blast-in-place. In August 2019, 42 ground mines were detonated in a controlled manner underwater during a NATO maneuver in the German Natura2000 Special Area of Conservation Fehmarn Belt, the Baltic Sea. In June 2020, four detonation craters were investigated with a multibeam echosounder for the first time. This dataset is represented here as maps of bathymetry, slope angle, and height difference to the surrounding. The circular craters were still clearly visible a year after the detonation. The diameter and depth of the structures were between 7.5–12.6 m and 0.7–2.2 m, respectively. In total, about 321 m2 of the seafloor was destroyed along the track line.

Enhanced Hot Salt-Water Corrosion Resistance of NiCoCrAlY-AlSiY Coating by Ion-Beam-Assisted Deposition

A promising ion-beam-assisted deposition (IBAD) method was developed to improve the salt-water corrosion resistance of NiCoCrAlY-AlSiY coating. During hot salt-water exposure, hydrochloric acid (HCl) was produced when chloride salt, water, and metal oxide reacted with each other, while HCl was also produced when chlorine reacted with water. The as-deposited AlSiY layer exhibited a loose texture accompanied by numerous pore defects, which triggered the multi-scale diffusion of HCl, resulting in the large-area corrosion degradation of the coating texture and the rapid diffusion of the NiCoCrAlY bonding layer. By contrast, the ion-beam-assisted AlSiY layer showed a dense texture that effectively inhibited the inner diffusion of HCl and suppressed the corrosion reactions as well as the diffusion of the NiCoCrAlY bonding layer. The current results confirmed the significant potential of IBAD in inhibiting corrosion damage and diffusion of thermal protective coatings.

Molecular mechanisms of local adaptation for salt-tolerance in a treefrog.

Salinization is a global phenomenon affecting ecosystems and forcing freshwater organisms to deal with increasing levels of ionic stress. However, our understanding of mechanisms that permit salt tolerance in amphibians is limited. This study investigates mechanisms of salt tolerance in locally adapted, coastal populations of a treefrog, Hyla cinerea. Using a common garden experiment, we (i) determine the extent that environment (i.e., embryonic and larval saltwater exposure) or genotype (i.e., coastal vs. inland) affects developmental benchmarks and transcriptome expression, and (ii) identify genes that may underpin differences in saltwater tolerance. Differences in gene expression, survival, and plasma osmolality were most strongly associated with genotype. Population genetic analyses on expressed genes also delineated coastal and inland groups based on genetic similarity. Coastal populations differentially expressed osmoregulatory genes including ion transporters (atp1b1, atp6V1g2, slc26a), cellular adhesion components (cdh26, cldn1, gjb3, ocln), and cytoskeletal components (odc1-a, tgm3). Several of these genes are the same genes expressed by euryhaline fish after exposure to freshwater, which is a novel finding for North American amphibians and suggests that these genes may be associated with local salinity adaptation. Coastal populations also highly expressed glycerol-3-phosphate dehydrogenase 1 (gpd1), which indicates they use glycerol as a compatible osmolyte to reduce water loss - another mechanism of saltwater tolerance previously unknown in frogs. These data signify that Hyla cinerea inhabiting coastal, brackish wetlands have evolved a salt-tolerant ecotype, and highlights novel candidate pathways that can lead to salt tolerance in freshwater organisms facing habitat salinization.