Developments in Bioremediation of Soils and Sediments Polluted with Metals and Radionuclides. 3. Influence of Chemical Speciation and Bioavailability on Contaminants Immobilization/Mobilization Bio-processes

The biotransformation of metals is an exciting, developing strategy to treat metal contamination, especially in environments that are not accessible to other remediation technologies. However, our ability to benefit from these strategies hinges on our ability to monitor these transformations in the environment. That’s why remediation of contaminated sediments and soil requires detailed in situ characterization of the speciation of the toxic substances and their transformations with respect to time and spatial distribution. The present paper gives an overview of the literature regarding research performed in the laboratory as well as in the field.     

Background and overview of current sediment toxicity identification evaluation procedures

Laboratory bioassays can provide an integrated assessment of the potential toxicity of contaminated sediments to aquatic organisms; however, toxicity as a sole endpoint is not particularly useful in terms of identifying remedial options. To focus possible remediation (e.g., source control), it is essential to know which contaminants are responsible for toxicity. Unfortunately, contaminated sediments can contain literally thousands of potentially toxic compounds. Methods which rely solely on correlation to identify contaminants responsible for toxicity are limited in several aspects: (a) actual compounds causing toxicity might not be measured, (b) concentrations of potentially toxic compounds may covary, (c) it may be difficult to assess the bioavailability of contaminants measured in a sediment, and (d) interactions may not be accounted for among potential toxicants (e.g., additivity). Toxicity identification evaluation (TIE) procedures attempt to circumvent these problems by using toxicity-based fractionation procedures to implicate specific contaminants as causative toxicants. Phase I of TIE characterizes the general physio-chemical nature of sample toxicants. Phase II employs methods to measure toxicants via different analytical methods, and Phase III consists of techniques to confirm that the suspect toxicants identified in Phases I and II of the TIE actually are responsible for toxicity. These TIE procedures have been used to investigate the toxicity of a variety of samples, including sediments. Herein we present a brief conceptual overview of the TIE process, and discuss specific considerations associated with sediment TIE research. Points addressed include: (a) selection and preparation of appropriate test fractions, (b) use of benthic organisms for sediment TIE work, and (c) methods for the identification of common sediment contaminants.     

Assessing the bioaccumulation of contaminants from sediments by fish and other aquatic organisms

Contaminated sediments that are not acutely toxic to aquatic organisms but contain bioaccumulable toxic substances present a common, yet poorly understood problem for regulatory decision makers. In order to recommend options to minimize bioaccumulation of these toxic substances, decision-makers need estimates of 1. which substances are available for accumulation by aquatic organisms; and 2. the potential impacts of such accumulation. The most direct and meaningful approach to estimating bioavailability is measurement of contaminant uptake by aquatic organisms exposed to the sediments of concern. Reasonably reliable methodologies exist for performing such exposures in the laboratory and in situ using marine or freshwater organisms. Such methods can demonstrate short-term potential for bioaccumulation of toxics from the sediments, but not necessarily the biological significance or long-term impact of any accumulated residues in the organisms and transfer of those residues through the food chain. Since most contaminated sediments contain a mixture of toxic substances, determination of the biological significance of their accumulation is not likely in the near future. Thus, the direct measurement of significant bioaccumulation of toxic substances from the sediments remains the most immediately useful index in a decision-making process.Contribution Number 631 of the National Fisheries Center-Great Lakes     

Entrapment of pollutants in Mediterranean sediments and biogeochemical indicators of their impact

The transit and fate of certain pollutants (Cd, Pb, Zn and P) transported to the western Mediterranean sea-floor and a method to assess their impact are described. The spatial distribution of pollutant concentrations in the Rhône prodelta shows that their decline with distance from their source is due both to mixing with unpolluted sediments, release from contaminated particles during transit and release from sediments after deposition. Beyond the continental shelf, metals of anthropogenic origin, mainly incorporated in faecal pellets, sink and become entrapped in deep sea sediments. Because subsaturating concentrations of trace metals are often found in surface pore waters and storage, therefore, still occurs, co-precipitation with other metal ions existing in slightly supersaturated states has also to be considered. Although phosphate tends to precipitate as apatite after entering sea water, its `definitive' storage in sediments is hindered by certain forms of pollution. Low pH and reducing conditions in sediments enhance phosphate release to the overlying water . Eutrophication may then occur in localised areas. The effects of urban waste water contamination on biogeochemical processes in sediments are examined, in particular processes responsible for the transformation of organic nitrogen. Sediment quality appears to be better defined by the effectiveness of diagenetic processes than by pollutant concentration per se. In general, polluted sediments possess weak capacities to transform organic nitrogen relative to the quantities of organic matter that are available. Such sediment characteristics are indicated by the preferential growth of Caulerpa taxifolia over that of Posidonia oceanica.     

Risk Assessment of Heavy Metals in Street Dust Particles to a Stream Network

Runoff with contaminated urban soil has an environmental risk to the aquatic environment. An assessment of heavy metals in street dust particles from a small town and their risks to the township stream network were conducted at Yangtze River delta. This assessment is based on measurement of heavy metal contents in dust particles with different particle sizes, river sediments, and suspended solids of urban runoff. The ranges of heavy metal content were 0.8–4.3, 16–380, 69–240, 9.3–350, 9.6–863 and 67–1170 mg/kg dry street dust, for Cr, Cu, Ni, Pb and Zn, respectively. Approximately 63%-71% of heavy metals were associated with particles less than 250 μ m; this particle size accounted for 40% of the total mass of street dusts. Of the five land use areas, the industrial areas had the highest heavy metal level. The smaller particle size fraction has a higher heavy metal content, low density, high mobility in runoff, and thus is a higher risk to the stream network. The topographical and hydrological features of the landscape also influence the transport of the contaminated street dusts to the aquatic environment.     

Validation of metabolomics for toxic mechanism of action screening with the earthworm Lumbricus rubellus

One of the promises of environmental metabolomics, together with other ecotoxicogenomic approaches, is that it can give information on toxic compound mechanism of action (MOA), by providing a specific response profile or fingerprint. This could then be used either for screening in the context of chemical risk assessment, or potentially in contaminated site assessment for determining what compound classes were causing a toxic effect. However for either of these two ends to be achievable, it is first necessary to know if different compounds do indeed elicit specific and distinct metabolic profile responses. Such a comparative study has not yet been carried out for the earthworm Lumbricus rubellus. We exposed L. rubellus to sub-lethal concentrations of three very different toxicants (CdCl₂, atrazine, and fluoranthene, representing three compound classes with different expected MOA), by semi-chronic exposures in a laboratory test, and used NMR spectroscopy to obtain metabolic profiles. We were able to use simple multivariate pattern-recognition analyses to distinguish different compounds to some degree. In addition, following the ranking of individual spectral bins according to their mutual information with compound concentrations, it was possible to identify both general and specific metabolite responses to different toxic compounds, and to relate these to concentration levels causing reproductive effects in the worms. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Qi Guo and Jasmin K. Sidhu contributed equally to this paper.     

Biodegradation of pyrene in sand,silt and clay fractions of sediment

Microbial degradation is the dominant pathway for natural attenuation of PAHs in environmental compartments such as sediments, which in turn depends on the bioavailability of PAHs. The bioavailability of PAHs has seldom been studied at the sediment particle size scale. We evaluated biodegradation of pyrene by Mycobacterium vanbaalenii PYR-1 as a function of sediment particle sizes, and investigated the relationship between the rate of degradation on sand, silt and clay particles with their individual desorption kinetics measured with the Tenax extraction method. Regression analysis showed that the total organic carbon (TOC), black carbon (BC), and specific surface area (SSA) of the specific particle size fractions, instead of the particle size scale itself, were closely related (P < 0.01) with the mineralization rate. While the fraction in the rapid desorption pool (F _rapid) ranged from 0.11 to 0.38 for the whole sediments and different size groups, the fractions mineralized after 336-h incubation (0.52 to 0.72) greatly surpassed the F _rapid values, suggesting utilization of pyrene in the slow desorption pool (F _slow). A biodegradation model was modified by imbedding a two-phase desorption relationship describing sequential Tenax extractions. Model analysis showed that pyrene sorbed on silt and clay aggregates was directly utilized by the degrading bacteria. The enhanced bioavailability may be attributed to the higher chemical concentration, higher TOC or larger SSA in the silt and clay fractions, which appeared to overcome the reduced bioavailability of pyrene due to sorption, making pyrene on the silt and clay particles readily available to degrading microbes. This conjecture merits further investigation.     

Structural changes ofNoetia ponderosa red blood cell membranes during cell volume regulation in reduced salinities: A freeze fracture study

Summary Changes in molluscan blood cell membrane structure coincided with changes in membrane amino acid permeability during cell volume regulation. Blood cells were freeze fractured after the free amino acid permeability of their membranes had been altered by modifying the extracellular Ca²⁺ and intracellular ATP levels and the membrane particles examined for changes in size, number/area and distribution. Test substances that altered the divalent cation or ATP levels also altered membrane particle densities, but not size or distribution, of freeze fractured blood cells. Those test substances (Ca²⁺-free seawater, DNP, low temperature) that inhibited volume regulation and the FAA efflux caused decreased membrane particle density, while those test substances (Co²⁺, Mn²⁺) that potentiated volume regulation and the FAA efflux increased the number of membrane particles/unit area. These changes in membrane particle density appear to result from the changes in surface area due to the treatment effects on cell volume, so that the number of membrane particles per cell remained constant. Therefore, altered membrane FAA permeability is associated with altered membrane particle density, but the effect of this structural alteration on membrane permeability is not clear.Abbreviations FAA free amino acid - DMSO dimethylsulfoxide - DNP dinitrophenol - ASW artificial seawater     

Semiquantitative Confocal Laser Scanning Microscopy Applied to Marine Invertebrate Ecotoxicology

Confocal laser scanning microscopy (CLSM) represents a powerful, but largely unexplored ecotoxicologic tool for rapidly assessing in vivo effects of toxicants on marine invertebrate embryo quality and development. We describe here a new semiquantitative CLSM approach for assessing relative yolk quantity in marine invertebrate embryos (harpacticoid copepods) produced by parents reared from hatching to adult in the polycylic aromatic hydrocarbon chrysene. This method is based on fluorogenic labeling of embryo yolk and subsequent statistical analysis of areal pixel intensities over multiple Z-series using a general linear model (GLM)–nested analysis of variance. The fluorescent yolk-labeling method described here was able to detect statistically significant differences in yolk concentrations in marine copepod (Amphiascus tenuiremis) eggs or embryos from females exposed to ultraviolet light and chrysene-contaminated sediments. Yolk intensities in embryos from females cultured throughout their life cycles in clean sediments were statistically identical with or without UV exposure. In constrast, yolk intensities in embryos of females cultured throughout their life cycle in chrysene-contaminated sediments were significantly higher in the non-UV-exposed treatment with chrysene at 2500 ng/g sediment (65.7% higher) and the UV-exposed treatment with chrysene at 500 ng/g sediment (76.6% higher).     

The role of phytoplankton in the removal of arsenic by sedimentation from surface waters

The role of phytoplankton in the removal of arsenic (As) by particle adsorption and sedimentation was investigated in Moira Lake, Canada. Sampling water and suspended particles over one year illustrated significant variation in As partitioning between particulate and aqueous phases, but failed to establish a correlation between the partition coefficient, K _d, and indicators of phytoplankton biomass. A highly significant inverse logarithmic relationship was noted between K _d and the concentration of suspended particles (log K _d = 5.1 – 1.4 log SS; p = 0.0001) in an apparent demonstration of the particle concentration effect (O' Connor & Connolly, 1980).Particle deposition, measured by means of sediment traps, appeared to include a substantial component of resuspended surficial sediment making sediment trap results unreliable for quantifying the removal of substances from the water column. The As concentration of particles from deep traps deployed during late summer and early fall exceeded the As concentrations of suspended particles and surficial sediment, and may indicate that a highly contaminated nepheloid layer acts as a temporary sink for As.     

Identification of sex-specific DNA methylation changes driven by specific chemicals in cord blood in a Faroese birth cohort

Faroe islanders consume marine foods contaminated with methylmercury (MeHg), polychlorinated biphenyls (PCBs), and other toxicants associated with chronic disease risks. Differential DNA methylation at specific CpG sites in cord blood may serve as a surrogate biomarker of health impacts from chemical exposures. We aimed to identify key environmental chemicals in cord blood associated with DNA methylation changes in a population with elevated exposure to chemical mixtures. We studied 72 participants of a Faroese birth cohort recruited between 1986 and 1987 and followed until adulthood. The cord blood DNA methylome was profiled using Infinium HumanMethylation450 BeadChips. We determined the associations of CpG site changes with concentrations of MeHg, major PCBs, other organochlorine compounds [hexachlorobenzene (HCB), p,p’-dichlorodiphenyldichloroethylene (p,p’-DDE) and p,p’-dichlorodiphenyltrichloroethane], and perfluoroalkyl substances. In a combined sex analysis, among the 16 chemicals studied, PCB congener 105 (CB-105) exposure was associated with the majority of differentially methylated CpG sites (214 out of a total of 250). In female-only analysis, only 73 CB-105 associated CpG sites were detected, 44 of which were mapped to genes in the ELAV1-associated cancer network. In males-only, methylation changes were seen for perfluorooctane sulfonate, HCB, and p,p’-DDE in 10,598, 1,238, and 1,473 CpG sites, respectively, 15% of which were enriched in cytobands of the X-chromosome associated with neurological disorders. In this multiple-pollutant and genome-wide study, we identified key epigenetic toxicants. The significant enrichment of specific X-chromosome sites in males implies potential sex-specific epigenome responses to prenatal chemical exposures.     

Contamination effects by a ‘conventional’ and a ‘biodegradable’ lubricant oil on infaunal recruitment to Antarctic sediments: A field experiment

To investigate the impacts of synthetic lubricants on Antarctic infaunal communities, a field experiment was setup near Australia's Casey Station, East Antarctica. Two types of synthetic lubricants were tested: an ‘Unused’ and ‘Used’ conventional synthetic lubricant, and an alternative marketed as ‘biodegradable’. Clean defaunated sediment was contaminated with the lubricants, decanted into trays, and deployed by divers onto the seabed in a randomised block design. Sediments were sampled 5 and 56 weeks after deployment. After 5 weeks, benthic assemblages that had recruited to the lubricant contaminated sediments were significantly different to those in ‘Control’ sediments, and differences were more pronounced after 56 weeks. Total number of individuals did not significantly differ between treatments after 5 weeks. However, after 56 weeks total individuals in the ‘Control’ sediments were significantly greater than in the contaminated sediments. Nototanais antarcticus (tanaid) and to a lesser extent Monoculodes sp. (gammarid), Tanaid sp. IV and Eudorella sp. (cumacean) had significantly higher abundances in the control sediments after 56 weeks compared to the contaminated sediments. Copepods numerically dominated the benthic assemblages at both sampling times; however, their abundance did not significantly differ across treatments. The community recruiting to the contaminated sediments remained different from that in the ‘Control’ sediments for the duration of the experiment (1 year). The ‘biodegradable’ lubricant was just as environmentally harmful to the Antarctic infauna as the ‘conventional’ lubricant currently used at Australia's Antarctic stations. Our results demonstrate that changes to recruitment are one of the potential environmental consequences of a lubricant spill to Antarctic benthic communities, and reinforce the importance of preventative oil spill management and effective clean-up procedures. Further monitoring of this field experiment will provide much needed information about the long-term impacts by synthetic lubricants in the Antarctic marine environment.     

Assessment of the contamination level of bottom sediments of Amursky Bay (Sea of Japan) and their potential toxicity

The contamination of surface bottom sediments of the Amursky Bay (the Sea of Japan) and their potential toxicity were assessed by the computation of contamination indexes and the ecological risk index, as well as by comparison of the concentrations of toxicants in the sediments with the sediment quality guidelines (SQD) values adopted by the US Environmental Protection Agency (EPA). It was shown that sediments from the coastal zone of Amursky Bay adjacent to the Vladivostok City were the most contaminated. The organochlorine pesticide DDT and such heavy metals as zinc, copper, and nickel were major inputs to the potential toxicity of the sediments. Linear regression analysis revealed positive, although not always significant, correlations between the ecological risk index and quotients characterizing the level of pathological changes in the gonads and the number of anomalies in the development of progeny of the sea urchin Strongylocentrotus intermedius. The obtained data array enables us to conclude that contamination of bottom sediments by heavy metals and DDT causes great risk for the reproduction of S. intermedius and other benthic invertebrates in the coastal part of Amursky Bay adjoining the city.     

Combined Use of Zero Valent Iron and Magnetic Separation for ex-situ Removal of Bioavailable Metals from Contaminated Sediments

Sandy and organic sediments characterized by different heavy metal binding capacities (HMBC), and contaminated with Copper (Cu), mercury (Hg), or zinc (Zn) were treated ex-situ using a remediation approach consisting of (i) sorption onto oxidized zero-valent iron (ZVI) surfaces and (ii) retrieval of formed metal-ZVI complexes from sediment matrices by magnetic separation. The research focused on the reduction/elimination of the bioavailable fractions of metals, and the efficiency of the method assessed by a combination of a bacterial (MetPLATE?) and an invertebrate (the 48-h Ceriodaphnia dubia acute toxicity test) based bioassays. In sandy sediments, characterized by low HMBC (20.8, 23.5, and 39.6 for Hg, Cu, and Zn, respectively), the determined toxicity units (TU) prior to sediment treatment increased in the order Hg < Cu < Zn, regardless of the bioassay used. The use of ZVI and magnetic separation in these sandy sediments resulted in up to 97% TU reduction. In organic-rich sediments, the affinity of the studied metals for organic matter (OM) resulted in much higher HMBC values (83.9, 108.3, and 136.2 for Cu, Zn, and Hg, respectively) and much lower TU values before sediment treatment with ZVI. The use of MetPLATE? on non-treated sediments resulted in TU values increasing in the order Hg < Cu < Zn, with TU removal efficiencies ranging from 83% to 97% after treatment. The TU values measured with the 48-h C. dubia assay were higher than those obtained with MetPLATE?, and in this case, sediments contaminated with Zn exhibited the lowest percentage of TU removal, with only 81.7% and 80.5% TU removal for sediments with contamination levels of 400 and 800 mg/kg, respectively. For organic sediments contaminated with Cu and Hg, the TU removal exceeded 95%. Overall, this study showed that the proposed remediation method has great potentials with regard to the elimination of the bioavailable metal fractions in contaminated sediments.     

THE EFFECT OF THE ELECTRON-OPAQUE PORE MATERIAL ON EXCHANGES THROUGH THE NUCLEAR ANNULI

To investigate the extent to which the electron-opaque pore material can regulate nucleocytoplasmic exchanges which occur through the nuclear annuli, experiments were performed in which polyvinylpyrrolidone (PVP)-coated colloidal gold particles (25 to 170 A in diameter) were microinjected into the cytoplasm of amebas (Amoeba proteus). The cells were fixed at various times after injection and examined with the electron microscope in order to determine the location of the gold particles. High concentrations of gold were found associated with the pore material at specific points adjacent to and within the pores. It is tentatively suggested that such specific accumulations could be a means of selecting substances from the cytoplasm for transport through the pores. Particles were also scattered throughout the ground cytoplasm and nucleoplasm. A comparison of the diameters of particles located in these two regions showed that the ability of materials to penetrate the nuclear envelope is a function of their size. It was estimated that the maximum size of the particles able to enter the nucleus is approximately 125 to 145 A indiameter. The regulation of exchanges with regard to particle size is thought to be dependent on the specific organization of the electron-opaque pore material.     

Chemical mobility and bioavailabity of sediment-bound heavy metals influenced by salinity

The transfer of metals from contaminated sediments to algal cell walls (Scenedesmus quadricauda) and organisms from various trophic levels (euryhaline osmoconform hydroid Cordylophora caspia and algae Brachiomonas submarina) was studied with a multichamber device. The system consists of a central chamber which contained the mud suspension and six external chambers containing the different biological indicators. The solids in the central and external chambers are separated by 0.45 m-diameter membranes which allow diffusion of the mobilized, dissolved metal compounds. Experiments were performed with dredged sediments at various salinities (0.5, 1.0, 1.5, and 2.0 percent, respectively) and the kinetic of re-adsorption was obtained by taking samples after different time intervals. High enrichment of Cd was found in the living alga Brachiomonas submarina, but on the other side only a weak influence of salinity on re-adsorption could be observed. Model experiments with ionic Cd showed a clear dependency on Cd-sorption on the algae, Cd-concentration in solution, and salinity. These results indicate that the transfer of metals mainly depends on the specific surface properties of the substrates and on the specific chemical form of the dissolved mobilized metal.     

Impact of rhizobacterial inoculants on plant growth and enzyme activities in soil treated with contaminated bottom sediments

The impact of contaminated bottom sediments on plant growth and soil enzyme activities was evaluated in a greenhouse pot study. The sediments were moderately contaminated with zinc and heavily contaminated with polycyclic aromatic hydrocarbons and polychlorinated dibenzo-p-dioxins and furans. The sediments were mixed with soil and planted with either Festuca arundinacea or Tagetes patula. The capacity of two rhizobacterial strains (Massilia niastensis P87 and Streptomyces costaricanus RP92), previously isolated from contaminated soils, to improve plant growth under the chemical stress was tested. Application of sediments to soil was severely phytotoxic to T. patula and mildly to F. arundinacea. On the other hand, the addition of sediments enhanced the soil enzymatic activity. Inoculation with both bacterial strains significantly increased shoot (up to 2.4-fold) and root (up to 3.4-fold) biomass of T. patula. The study revealed that the selected plant growth-promoting bacterial strains were able to alleviate phytotoxicity of bottom sediments to T. patula resulting from the complex character of the contamination.     

Real-time analysis of protein and protein mixture interaction with lipid bilayers

Artificial lipid bilayers in the form of planar supported or vesicular bilayers are commonly used as models for studying interaction of biological membranes with different substances such as proteins and small molecule pharmaceutical compounds. Lipid membranes are typically regarded as inert and passive scaffolds for membrane proteins, but both non-specific and specific interactions between biomolecules and lipid membranes are indeed ubiquitous; dynamic exchange of proteins from the environment at the membrane interface can strongly influence the function of biological membranes. Such exchanges would either be of a superficial (peripheral) or integrative (penetrating) nature. In the context of viral membranes (termed envelopes), this could contribute to the emergence of zoonotic infections as well as change the virulence and/or pathogenicity of viral diseases. In this study, we analyze adsorption/desorption patterns upon challenging tethered liposomes and enveloped virus particles with proteins – or protein mixtures - such as bovine serum albumin, glycosylphosphatidylinositol anchored proteins and serum, chosen for their different lipid-interaction capabilities. We employed quartz crystal microbalance and dual polarization interferometry measurements to measure protein/membrane interaction in real time. We identified differences in mass uptake between the challenges, as well as differences between variants of lipid bilayers. Tethered viral particles showed a similar adsorption/desorption behavior to liposomes, underlining their value as model system. We believe that this methodology may be developed into a new approach in virology and membrane research by enabling the combination of biophysical and biochemical information.     

Microbial biomass and activities associated with subsurface environments contaminated with chlorinated hydrocarbons

Soil microcosms and enrichment cultures from subsurface sediments and groundwaters contaminated with trichloroethylene (TCE) were examined. Total lipids, [I‐‘⁴C]acetate incorporation into lipids, and [Me‐³H]thymidine incorporation into DNA were determined in these subsurface environments. In heavily TCE‐contam‐inated zones (greater than 500 mg/L) radioisotopes were not incorporated into lipids or DNA. Radioisotope incorporation occurred in sediments both above and below the TCE plume. Phospholipid fatty acids (PLFA) were not detected, i.e., less than 0.5 pmol/L in heavily contaminated groundwater samples. In less contaminated waters, extracted PLFA concentrations were greater than 100 pmollL and microbial isolates were readily obtained. Degradation of 30–100 mg/L TCE was observed when sediments were amended with a variety of energy sources. Microorganisms in these subsurface sediments have adapted to degrade TCE at concentrations greater than 50 mg/L.     

Behavioural adaptations of the fiddler crabs Uca vocans borealis (crane) and Uca lactea lactea (De Haan) for coexistence on an intertidal shore

The distribution and coexistence of the fiddler crabs U. vocans borealis and U. lactea lactea was investigated in the upper shore of a sandy beach with respect to particle size, water content and total organic carbon content of the sediments. A clear segregation of habitats between the two species was apparent. U. lactea lactea were only found in sediments with significantly lower total organic carbon content. U. lactea lactea had a higher ratio for the size of third maxilliped to body size than U. vocans borealis. There was no statistical difference in the median sediment particle size of the habitats where both species were found on the study shore. U. lactea lactea was larger on shores where U. vocans borealis were absent. Smaller U. vocans borealis individuals occupied sediments with higher water content and finer particles. Smaller individuals of U. vocans borealis also possessed fewer spoon-tipped setae on second maxilliped than their larger conspecifics, which were found mostly on coarser sediments. U. lactea lactea spent significantly more time on the surface than in the burrow during low tide when compared with U. vocans borealis. Both Uca species exhibited similar desiccation resistance. U. lactea lactea tends to keep its frontal region in close contact with sediments to maintain moisture when being exposed to air.