Toxicity of binary mixtures of Cu, Cr and As to the earthworm Eisenia andrei.

Chromated copper arsenate (CCA) mixtures were used in the past for wood preservation, leading to large scale soil contamination. This study aimed at contributing to the risk assessment of CCA-contaminated soils by assessing the toxicity of binary mixtures of copper, chromium and arsenic to the earthworm Eisenia andrei in OECD artificial soil. Mixture effects were related to reference models of Concentration Addition (CA) and Independent Action (IA) using the MIXTOX model, with effects being related to total and available (H2O and 0.01 M CaCl2 extractable) concentrations in the soil. Since only in mixtures with arsenic dose-related mortality occurred (LC50 92.5 mg/kg dry soil), it was not possible to analyze the mixture effects on earthworm survival with the MIXTOX model. EC50s for effects of Cu, Cr and As on earthworm reproduction, based on total soil concentrations, were 154, 449 and 9.1 mg/kg dry soil, respectively. Effects of mixtures were mainly antagonistic when related to the CA model but additive related to the IA model. This was the case when mixture effects were based on total and H2O-extractable concentrations; when based on CaCl2-extractable concentrations effects mainly were additive related to the CA model except for the Cr-As mixture which acted antagonistically. These results suggest that the CCA components do interact leading to a reduced toxicity when present in a mixture.

The Influence of New and Artificial Aged Microplastic and Leachates on the Germination of Lepidium sativum L.

With the increase in environmental monitoring and assessing, we are gaining insight into the extent of microplastic pollution in our environment. The threat posed by microplastics to biota could come, e.g., from leached substances. As some plastic materials have been decaying in nature for extended periods already, the toxic effects of leaching compounds need to be investigated. It is furthermore essential to understand the adverse effects of new plastic and how these effects differ from the effects elicited by old plastic material. Therefore, in the present study, the effects of exposure to leachates from new and artificial aged polycarbonate as well as new and aged polycarbonate granules on various germination parameters of Lepidium sativum were studied. Germination, root, and shoot length, as well as the calculated germination rate index as a measure for germination speed, was negatively influenced in substrate-free and substrate containing exposures. From an ecological and agricultural point of view, this implies possible yield losses with less germinating seeds, slower plant germination speed, and smaller seedlings in general.

Enchytraeus crypticus Avoid Soil Spiked with Microplastic.

Microplastics (MPs) of varying sizes are widespread pollutants in our environment. The general opinion is that the smaller the size, the more dangerous the MPs are due to enhanced uptake possibilities. It would be of considerably ecological significance to understand the response of biota to microplastic contamination both physically and physiologically. Here, we report on an area choice experiment (avoidance test) using Enchytraeus crypticus, in which we mixed different amounts of high-density polyethylene microplastic particles into the soil. In all experimental scenarios, more Enchytraeids moved to the unspiked sections or chose a lower MP-concentration. Worms in contact with MP exhibited an enhanced oxidative stress status, measured as the induced activity of the antioxidative enzymes catalase and glutathione S-transferase. As plastic polymers per se are nontoxic, the exposure time employed was too short for chemicals to leach from the microplastic, and as the microplastic particles used in these experiments were too large (4 mm) to be consumed by the Enchytraeids, the likely cause for the avoidance and oxidative stress could be linked to altered soil properties.

An uptake and elimination kinetics approach to assess the bioavailability of chromium, copper, and arsenic to earthworms (Eisenia andrei) in contaminated field soils.

The aim of this study was to determine the bioavailability of metals in field soils contaminated with chromated copper arsenate (CCA) mixtures. The uptake and elimination kinetics of chromium, copper, and arsenic were assessed in the earthworm Eisenia andrei exposed to soils from a gradient of CCA wood preservative contamination near Hartola, Finland. In soils contaminated with 1480-1590 mg Cr/kg dry soil, 642-791 mg Cu/kg dry soil, and 850-2810 mg Ag/kg dry soil, uptake and elimination kinetics patterns were similar for Cr and Cu. Both metals were rapidly taken up and rapidly excreted by Eisenia andrei with equilibrium reached within 1 day. The metalloid As, however, showed very slow uptake and elimination in the earthworms and body concentrations did not reach equilibrium within 21 days. Bioaccumulation factors (BAF) were low for Cu and Cr (< 0.1), but high for As at 0.54-1.8. The potential risk of CCA exposure for the terrestrial ecosystem therefore is mainly due to As.

Identification of Toxicants from a Highly C10 -C40 -Contaminated Sediment Influenced by the Wood Industry: Petroleum Hydrocarbons or Biogenic Organic Compounds?

Sediment from a log pond located in south Finland contained 15 000 to 50 000 mg/kg dry weight of C10 -C40 hydrocarbons. It was unclear whether they originated from the hydraulic fluid of the log hoist or the wood extractives. In the present study, methods of effect-directed analysis were used for the identification of toxicants. A combination of fractionation, biotesting, and chemical analyses revealed that the key toxicant of log pond sediment was retene, a dialkyl-substituted phenanthrene derived from wood resin acids. In addition, the most toxic fraction included 3 other wood-originated diterpenic compounds. Typical wood extractives such as sesquiterpenes and odd-carbon number alkanes in the range C21 -C33 were identified in the fraction, which showed minor genotoxic potency. The most polar fraction contained triterpenes and showed estrogenic activity. No evidence for the presence of hydraulic fluid in sediment was found. The study also indicated that in cases where the organic matter content of sediment or soil is high, using the results of standard mineral oil analysis in risk management can lead to incorrect actions because standard methods do not differentiate petroleum hydrocarbons from biogenic hydrocarbons. Environ Toxicol Chem 2019;9999:1-11. © 2019 SETAC.

Validation of the OECD reproduction test guideline with the New Zealand mudsnail Potamopyrgus antipodarum using trenbolone and prochloraz.

The Organisation for Economic Cooperation and Development (OECD) provides several standard test methods for the environmental hazard assessment of chemicals, mainly based on primary producers, arthropods, and fish. In April 2016, two new test guidelines with two mollusc species representing different reproductive strategies were approved by OECD member countries. One test guideline describes a 28-day reproduction test with the parthenogenetic New Zealand mudsnail Potamopyrgus antipodarum. The main endpoint of the test is reproduction, reflected by the embryo number in the brood pouch per female. The development of a new OECD test guideline involves several phases including inter-laboratory validation studies to demonstrate the robustness of the proposed test design and the reproducibility of the test results. Therefore, a ring test of the reproduction test with P. antipodarum was conducted including eight laboratories with the test substances trenbolone and prochloraz and results are presented here. Most laboratories could meet test validity criteria, thus demonstrating the robustness of the proposed test protocol. Trenbolone did not have an effect on the reproduction of the snails at the tested concentration range (nominal: 10-1000 ng/L). For prochloraz, laboratories produced similar EC10 and NOEC values, showing the inter-laboratory reproducibility of results. The average EC10 and NOEC values for reproduction (with coefficient of variation) were 26.2 µg/L (61.7%) and 29.7 µg/L (32.9%), respectively. This ring test shows that the mudsnail reproduction test is a well-suited tool for use in the chronic aquatic hazard and risk assessment of chemicals.

NAPL migration and ecotoxicity of conventional and renewable fuels in accidental spill scenarios.

Fuels derived from non-petroleum renewable resources have raised interest due to their potential in replacing petroleum-based fuels, but information on their fate and effects in the terrestrial and aquatic environments in accidental spill scenario is limited. In this study, migration of four fuels (conventional diesel, conventional gasoline, renewable diesel NExBTL, and ethanol-blended gasoline RE85 containing maximum 85% ethanol) as non-aqueous phase liquids (NAPL) in soil was demonstrated in a laboratory-scale experiment. Ecotoxicity data was produced for the same fuels. There was no significant difference in migration of conventional and renewable diesel, but gasoline migrated 1.5 times deeper and 7-9 times faster in sand than diesel. RE85 spread horizontally wider but not as deep (p < 0.05) as conventional gasoline. Conventional gasoline was the most toxic (lethal concentration [LC50] 20 mg/kg total hydrocarbon content [THC]) among the studied fuels in soil toxicity test with earthworm Eisenia fetida followed by ethanol-blended gasoline (LC50 1,643 mg/kg THC) and conventional diesel (LC50 2,432 mg/kg THC), although gasoline evaporated fast from soil. For comparison, the toxicity of the water-accommodated fractions (WAF) of the fuels was tested with water flea Daphnia magna and Vibrio fischeri, also demonstrating groundwater toxicity. The WAF of conventional gasoline and RE85 showed almost similar toxicity to both the aquatic test species. EC50 values of 1:10 (by volume) WAF were 9.9 %WAF (gasoline) and 9.3 %WAF (RE85) to D. magna and 9.3 %WAF (gasoline) and 12.3 %WAF (RE85) to V. fischeri. Low solubility decreased toxicity potential of conventional diesel in aquatic environment, but direct physical effects of oil phase pose a threat to organisms in nature. Renewable diesel NExBTL did not show clear toxicity to any test species.

Toxicity of two types of silver nanoparticles to aquatic crustaceans Daphnia magna and Thamnocephalus platyurus.

Although silver nanoparticles (NPs) are increasingly used in various consumer products and produced in industrial scale, information on harmful effects of nanosilver to environmentally relevant organisms is still scarce. This paper studies the adverse effects of silver NPs to two aquatic crustaceans, Daphnia magna and Thamnocephalus platyurus. For that, silver NPs were synthesized where Ag is covalently attached to poly(vinylpyrrolidone) (PVP). In parallel, the toxicity of collargol (protein-coated nanosilver) and AgNO3 was analyzed. Both types of silver NPs were highly toxic to both crustaceans: the EC50 values in artificial freshwater were 15-17 ppb for D. magna and 20-27 ppb for T. platyurus. The natural water (five different waters with dissolved organic carbon from 5 to 35 mg C/L were studied) mitigated the toxic effect of studied silver compounds up to 8-fold compared with artificial freshwater. The toxicity of silver NPs in all test media was up to 10-fold lower than that of soluble silver salt, AgNO3. The pattern of the toxic response of both crustacean species to the silver compounds was almost similar in artificial freshwater and in natural waters. The chronic 21-day toxicity of silver NPs to D. magna in natural water was at the part-per-billion level, and adult mortality was more sensitive toxicity test endpoint than the reproduction (the number of offspring per adult).

Increasing lake water and sediment oxygen levels using slow release peroxide.

The sediment and hypolimnion of many Finnish lakes suffer from anoxia due to increasing nutrient loading. The aim of this research was to develop a method for increasing the oxygen level using granulated calcium peroxide (CaO2) as a slow oxygen releasing compound. This compound releases oxygen (O2) in a reaction with water during 5 to 7 months. The method was tested in both laboratory and field conditions. In the field test granulated CaO2 were then spread manually from a rowing boat over the whole surface of the test pond. The granules sink onto and into the sediment. No mixing was needed. The dissolved oxygen concentration increased significantly during a laboratory experiment with a CaO2 amendment of 75 g m⁻² and in a pond experiment with a CaO2 amendment of 50 g m⁻². In the pond experiment, the effect was visible for the entire 40-week experiment. In the laboratory, the abundance of aerobic bacteria increased in the sediment after CaO2 addition, while the pond experiment gave more mixed results. The organic matter content of the sediment did not change during the experiment in the control pond, but decreased from 18% to 4% in the pond with the CaO2 amendment. This was possibly due to enhanced microbial activity in the test pond. Although the results show improved oxygen concentrations and effects on the sediment organic matter following CaO2 amendment, the usability of this method in larger lakes remains to be tested.

Selective toxicity at low doses: experiments with three plant species and toxicants.

During the last decade, the paradigm that low toxicant doses often have stimulatory effects on plants has become widely accepted. At the same time, low toxicant doses of metal salts have been observed to inhibit the growth of the most vigorous seedlings of a population in vitro, although mean plant size has remained unaffected. We hypothesized that this kind of selective low-dose toxicity is not restricted to inorganic contaminants. We exposed annual plants (baby's breath Gypsophila elegans, purslane Portulaca oleracea, and duckweed Lemna minor) to 1,3,4,6,7,8-hexahydro-4,6,6,7,8,8-hexamethylcyclopenta-γ-2-benzopyran (HHCB) and 4-tert-octylphenol and lead acetate. As compared to unexposed G. elegans roots, 4-tert-octylphenol did not affect the mean root size of all seedlings, but it reduced the average length of roots longer than the 98(th) percentile. A comparable response was found in case of G. elegans roots treated with lead acetate beyond the 90(th) percentile. The average size of roots beyond the 90(th) percentile was decreased also when L. minor was exposed to lead acetate though the means of all roots were constant. P. oleracea seemed to be insensitive to selective toxicity. We conclude that selective toxicity at low doses should be considered in parallel with hormesis.

Testing the homogenizing effect of low copper sulfate concentrations on the size distribution of Portulaca oleracea seedlings in vitro.

Traditionally, toxicological bioassays rely upon the differences in mean-based statistical tests between the exposed and unexposed plants, and exceptional plant individuals are treated as statistical outliers. Recently, low toxicant concentrations have been observed to affect gene regulation in exposed plant stands and to change the frequency of the largest individuals even if mean plant size remains unchanged. In this paper, we present the results that the latter phenomenon is not restricted to a single toxicant and plant species. Our data analysis consists of two statistical methods that may be of general interest. We utilized the one-tailed Moses extreme reactions test by comparing exposed groups to control plants with and without the trimming of a certain amount of potential outliers from both treatments compared. We also propose that Mann-Whitney U or other tests at ordinal scale can be utilized to analyze if the largest plant individuals in exposed and control treatments come from a single 'survivor' population. We conclude that the results supported the hypothesis that very low toxicant concentrations may have ecological effects on fast-growing plant species. Finally, we discuss the limitations of the statistical methods currently in use.

Ecological risks of an old wood impregnation mill: application of the Triad approach.

Although many studies deal with the distribution and mobility of chromated copper arsenate (CCA) metals in soil, the ecotoxicity of CCA-contaminated soils is rarely studied. The Triad approach was applied to determine the ecological risks posed by a CCA mixture at a decommissioned wood impregnation mill in southern Finland. A combination of (1) chemical analyses; (2) toxicity tests with plants (aquatic: Lemna minor; terrestrial: Lactuca sativa), earthworms (Lumbricus rubellus), and enchytraeids (Enchytraeus albidus) conducted on contaminated soils, their aqueous extracts, and well water collected from the site; and (3) determination of the abundance of enchytraeids and nematodes and the bioaccumulation of metals in plants (horsetail) collected from the field were used to assess the actual risk. Although metal concentrations were low, L. minor growth appeared to be reduced by As contamination of the well water. In soil, metals were heterogeneously distributed with total concentrations of 14.8 to 4360 mg As/kg, 15.2 to 1740 mg Cr/kg, and 4.83 to 790 mg Cu/kg. In several samples, concentrations were above Finnish regulatory guideline values and exceeded the half maximal effective concentration (EC50) or 50% lethal concentration (LC50) values for the toxicity of the individual metals to earthworms and enchytraeids, indicating hazards to the ecosystem. (Bio)availability of metals was high, as indicated by weak electrolyte extractions and body residues in L. rubellus and E. albidus exposed in bioassays. Earthworm survival correlated significantly with body metal concentrations, but not with soil total metal concentrations. Enchytraeid responses in the soil bioassays were less sensitive to CCA metal exposure. Plant growth was affected by CCA pollution, with L. sativa root elongation correlating significantly with total and available As concentrations and L. minor development being significantly reduced in H2O extracts of the most contaminated soil sample. Abundance of enchytraeids and nematodes in the field was much lower than in nonpolluted Finnish soils but did not significantly correlate with CCA contamination. Arsenic accumulation in horsetail did not correlate with As concentrations in soil. Overall, the results of the 3 lines of evidence of the Triad approach indicate possible increased risks to the ecosystem at the most contaminated sites of the CCA treatment area.

Low toxicant concentrations decrease the frequency of fast-growing seedlings at high densities of annual baby's breath (Gypsophila elegans).

Very low toxicant concentrations reduce the frequency of most vital seedlings in dense stands in vitro.

Bioaccumulation of paraquat by Lumbriculus variegatus in the presence of dissolved natural organic matter and impact on energy costs, biotransformation and antioxidative enzymes.

Dissolved organic matter from natural sources (DNOM) is omnipresent in aquatic ecosystems. Besides affecting bioavailability of substances including xenobiotics, it directly influences physico-chemistry of the habitat and there is increasing evidence for it is interaction with organisms. We investigated direct and interacting effects of DNOM from three sources, Lake Valkea-Kotinen, Svartberget Brook, and Lake Fuchskuhle with the herbicide paraquat on the oligochaete worm Lumbriculus variegatus. Bioavailability of paraquat to L. variegates as well as activities of antioxidative enzymes catalase (CAT) and peroxidase (POD) and biotransformation enzyme soluble glutathione S-transferase (sGST) were assessed without and in the presence of DNOM. Furthermore, metabolic heat dissipation due to the exposure was quantified. Uptake of paraquat into the worms was concentration dependently reduced by DNOM, and with differences concerning the DNOM sources. sGST and CAT responded with increased activities to DNOM (5 and 25 mg C l-1) and paraquat (5.0, 50, and 500 microg l-1) separately. Paraquat at 5.0 microg l-1 and DNOM in combination caused increased activities of sGST, especially at 5 mgC l-1, but inhibition of CAT activities. The latter probably occurred due to saturation of the enzyme. Changes in enzyme activities were independent from the source of DNOM. Increasing DNOM concentrations raised metabolic heat dissipation in L. variegatus with maximum at 3h of exposure. In the combined treatments, metabolic heat dissipation changed more due to the source of DNOM than due to the bioavailability of paraquat.

Body residues as dose for sublethal responses in alevins of landlocked salmon (Salmo salar m. sebago): a direct calorimetry study.

Questions of organism-specific factors, toxicity endpoints, and their relation to mode-of-action all are related to consistency and applicability of body residue-based approaches. To address this issue, direct calorimetry was used to evaluate metabolic responses of alevins of landlocked salmon (Salmo salar m. sebago) to pentachlorophenol (PCP) exposure ranging from 0 to 1.04 microM for 24 h or 24, 48, and 72 h (0, 0.26, and 0.55 microM, respectively). The body residues were used as a dose metric for sublethal responses. The body size, rapid ontogenetic development, and exposure to a specific pollutant all were heat output-modifying factors. The acute exposure (24-72 h) to PCP led to a heat output-enhancing effect, which directly was related to an internal concentration of PCP in the range of 0.01 to 0.15 micromol/g. Within the treatments, body size per level of metabolic rate and magnitude of physiological response were not correlated, thus the alevins with higher mass-specific metabolic rate were not more sensitive to PCP. Primarily, increasing metabolic rate during posthatch development controls PCP toxicity only by affecting bioaccumulation kinetics, not the toxic potency of the chemical. New information of a relationship between observed natural variation in measured physiological trait of fish and PCP-induced response and its body residue-based level is of ecotoxicological importance.

Metabolic rate of Arctic charr eggs depends on their parentage.

The metabolic rate (specific heat output) of individual eyed-stage eggs of Arctic charr Salvelinus alpinus (Linnaeus, 1758) originating from different families was measured with direct microcalorimetry. Metabolic rates varied between 2.3-7.9 microW ind(-1) and 0.06-0.22 microW mg(-1). Absolute heat output was unrelated to egg size, but size-scaled or specific heat output was negatively correlated with egg size, measured as diameter, dry mass or fresh mass. Metabolic rates varied significantly between families, suggesting that genetic and/or maternal effects affect embryonic metabolism in Arctic charr. Heat output increased almost linearly from 3.4 to 16.7 microW ind(-1) (0.09-0.67 microW mg(-1)) during the embryonic development. Although the metabolic rate varied between the families and egg metabolic rate increased during development, there was an unexpected disconnect between metabolic rate and hatching time.

Pentachlorophenol (PCP) bioaccumulation and effect on heat production on salmon eggs at different stages of development.

In this study, pentachlorophenol (PCP) bioaccumulation and its effect on heat dissipation was studied in eggs of the lake salmon (Salmo salar m. sebago). In bioaccumulation studies, the eggs were exposed to low concentrations (0.051-0.056 micromol/l, 13.583-14.915) of waterborne [14C]-labeled PCP at two developmental stages: (1) 3 weeks after fertilization, and (2) just before hatching. The effect of PCP on egg heat dissipation was measured by a microcalorimeter after exposing the eggs to gradual concentrations (0-0.992 micromol/l) of PCP for 48 h. After both the bioaccumulation and heat dissipation experiments, the eggs were dissected and the concentrations of PCP in tissue were determined separately for eggshell, yolk and embryo. The bioaccumulation studies showed that PCP accumulates more in the eggs at the late developmental stage. Bioconcentration factors (BCF) for different tissues were 3-42 times higher for the eggs at the late developmental stage compared with the eggs that were incubated only for 3 weeks. In early developmental stage, the eggshell adsorbs a large portion of the chemical. In late developmental stage, the actual embryo accumulated both proportionately and totally more than other dissected tissues in the beginning of the exposure, but eventually the yolk accumulated highest total amount of the chemical. A probable reason for the higher PCP body burden in the late developmental stage is that the respiration rate and metabolic activity of the embryo increases as it grows. The salmon eggs responded to an exposure to PCP with an elevated rate of heat dissipation. The threshold concentration above which the embryo heat dissipation was amplified was 29.64 micromol/kg embryo wet weight (ww) or 0.28 micromol/l. The highest embryo heat production was measured at the exposure concentration of 0.992 micromol/l. At higher exposure concentrations the heat dissipation decreased. The basic findings of the study are that PCP accumulates in growing embryonic tissue and is able to change the physiology of developing embryo.

Sublethal energetic responses by Pisidium amnicum (Bivalvia) exposed to pentachlorophenol at two temperatures.

Sublethal effects of pentachlorophenol (PCP) were studied in the freshwater clam Pisidium amnicum by examining the cause-effect relationship between PCP body burden and the rate of the whole animal heat output. In addition, the modifying effects of temperature (7 and 15 degrees C) and trematode parasites as natural stress factors were assessed. Results showed that even a low PCP body burden was sufficient to increase the heat output during the periods of aerobic metabolism and the magnitude of responses increased linearly with increasing body burden. During a valve-closure period, however, PCP had no effect on anaerobic heat output. The rise of the maximum heat output in response to PCP was more pronounced at the higher temperature. The response in heat output was similar in both infected and uninfected clams, but the general level of heat output was lower in infected clams. The increasing PCP body burden also increased the duration of valve-closure time. The results of this study suggested that microcalorimetry could be a useful tool in assessing sublethal responses by the clam, which offers several calorimetrically measurable parameters such as aerobic and anaerobic heat output and behavior.

Physiological energetics of a midge, Chironomus riparius Meigen (Insecta, Diptera): normoxic heat output over the whole life cycle and response of larva to hypoxia and anoxia.

The rate of metabolism of laboratory reared Chironomus riparius was monitored by direct calorimetry over the entire life cycle from egg to adult stage. The metabolic response of the fourth instar larva to decreasing oxygen concentrations and anoxia was also measured. Normoxic measurements were carried out at 20°C and the hypoxic-anoxic experiments at 10°C. In larvae with body sizes ranging from 0.0028 to 0.645 mg ash-free dry mass (afdm), the rate of heat dissipation was related to body mass by a power function, with a mass exponent of 0.71±0.02 corresponding to an exponent of -0.29 for the relationship between mass-specific metabolic rate and body mass. However, the allometric equations applicable to larvae would not predict the metabolic rates of eggs, pupae and adults. Single egg batches used in the experiments consisted of 354±90 eggs, the individual egg with a mass of 0.99±0.01 µg (mean±SD). The mass-specific rate of heat dissipation of the egg (13.7±1.8 µ W mg-1 afdm) was considerably lower than that of the first and second instar larvae (44-53 µ W mg-1) but equal to that of fourth instar larvae (13.1±3.9 µ W mg-1). Heat dissipation by a pupa shortly before adult emergence was high (14.8±1.8 µ W mg-1), probably due to high metabolism during metamorphosis. Emergence of the adult in the calorimeter was indicated by a short but intense burst of heat. The newly emerged imago had a ca. 20-35% higher metabolic rate than the pupa. In response to reduced O2 partial pressure the fourth instar larva of C. riparius displayed metabolic regulation. In continuously declining oxygen partial pressure, the fourth instar larva maintained its aerobic energy metabolism (4.2 µ W mg-1) with only a small decrease down to 0.8 kPa, corresponding to an oxygen concentration of 0.42 mg O2l-1 H2O. Below this critical oxygen concentration (Pc), the rate of heat dissipation decreased rapidly down to the anoxic level which was only 14-17% of the normoxic level. The high relative reduction of metabolic rate under anoxia gives a wrong impression of short-term tolerance of C. riparius to anoxia. The absolute energetic costs of C. riparius associated with anaerobic energy metabolism (0.64±0.11 µ W mg-1) are almost 6 times higher than those of more anoxia tolerant invertebrates such as sphaeriid bivalves.