Seasonal variability of antioxidant biomarkers and energy reserves in the freshwater gammarid Gammarus roeseli

In gammarids, behavioural and biochemical biomarkers are commonly used in ecotoxicological studies. In our study, we have investigated seasonal variations of several biochemical biomarkers in Gammarus roeseli, a freshwater species. Organisms were sampled monthly over a 1-year period. Gender was distinguished to measure antioxidant enzyme activities like total glutathione peroxidase (GPxtot), selenium-dependent glutathione peroxidase (SeGPx) and catalase enzymes, lipoperoxidation end-product (malondialdehyde, MDA), and energy reserves with protein and lipid contents. In the same time, usual water physico-chemical parameters were measured at the sampling site. A based-gender difference was observed for parameters related to oxidative stress. Females showed higher antioxidant enzyme activities and lower MDA level than males. Parameters related to oxidative stress and energy reserves appeared correlated with temperature and physiological status of organisms. Females GPx activities were lower in autumn and winter when no breeding occurred. In both gender, MDA levels were correlated with temperature with an increase of lipoperoxidation in summer. Lipid contents were the lowest in summer and the highest in winter, probably due to the reproductive status of organisms and their feeding behaviour. Gender-based differences of biochemical parameters suggest a specific sensitivity of males and females in ecotoxicological experiments. Moreover, organisms could be more vulnerable in summer when MDA levels are high and energy reserves low. Deleterious effect of xenobiotics would be different with gender and season.     

Plant-mediated effects of heavy metal pollution on host choice of a grass miner

We studied the effects of heavy metal exposure on host plant choice and performance of the grass miner Chromatomyia milii (Diptera, Agromyzidae). Cadmium decreased plant growth in a dose-dependent way. C. milii preferred the control to the cadmium-exposed plants for feeding and oviposition. Moreover, preference for the control plants increased with increasing cadmium exposure of the alternative choice. Adult and offspring performance decreased with increasing plant cadmium exposure. This suggests that, at least under our laboratory conditions, host choice of C. milii is adaptive under pollution stress. Foliar cadmium concentration increased and the soluble sugar concentration decreased with increasing cadmium exposure. Regression analysis showed that both latter components might be responsible for the decrease in performance of C. milii on cadmium-exposed plants. The protein and amino acid concentration of the leaves, the amount of structural defenses, and water concentration were not affected by the cadmium treatment.     

The acanthocephalan Paratenuisentis ambiguus as a sensitive indicator of the precious metals Pt and Rh from automobile catalytic converters

Recent studies revealed that intestinal acanthocephalans of fish can accumulate heavy metals to concentrations orders of magnitude higher than those in the host tissues or the aquatic environment. This significant heavy metal accumulation by acanthocephalans, even surpassing that of established free living accumulation bioindicators, encouraged us to study the bioavailability of the platinum-group-metals (PGM) Pt and Rh for parasites. These precious metals are used in catalytic converters of cars for exhaust gas purification in Europe since the early 1980s. In addition to the beneficial effect in reducing the emission of CHx, CO and NOx of cars there is an increasing emission of these metals. However, it still remains unclear if these elements become accumulated in the biosphere and whether they affect the health of organisms. The present study reveals that in European eels (Anguilla anguilla) naturally infected with the eoacanthocephalan parasite Paratenuisentis ambiguus and experimentally exposed to ground catalytic converter material, the parasites take up and accumulate the catalytic active metals Pt and Rh whereas in the examined host tissues we found no metal uptake. Compared with the PGM concentrations in the water the worms contained 1600 times higher Rh and 50 times higher Pt concentrations. Thus, the parasites can be used as sentinel organisms reflecting even very low levels of precious metals.     

Body metal concentrations and glycogen reserves in earthworms (Dendrobaena octaedra) from contaminated and uncontaminated forest soil

Stress originating from toxicants such as heavy metals can induce compensatory changes in the energy metabolism of organisms due to increased energy expenses associated with detoxification and excretion processes. These energy expenses may be reflected in the available energy reserves such as glycogen. In a field study the earthworm, Dendrobaena octaedra, was collected from polluted areas, and from unpolluted reference areas. If present in the environment, cadmium, lead and copper accumulated to high concentrations in D. octaedra. In contrast, other toxic metals such as aluminium, nickel and zinc appeared to be regulated and kept at low internal concentrations compared to soil concentrations. Lead, cadmium and copper accumulation did not correlate with glycogen reserves of individual worms. In contrast, aluminium, nickel and zinc were negatively correlated with glycogen reserves. These results suggest that coping with different metals in earthworms is associated with differential energy demands depending on the associated detoxification strategy.     

Towards the selection of a produced water enrichment for biological gas hydrate inhibitors

Economic concerns associated with the recovery of non-conventional hydrocarbon reserves include unexpected ice as well as ice-like gas hydrate formation. Antifreeze proteins (AFPs) inhibit ice growth, and experiments with fish, plant, and insect AFPs have shown promise of effective gas hydrate inhibition in lab-scale experiments. If produced on an industrial scale, AFPs could provide a more environmentally friendly alternative to kinetic inhibitors, but a large-scale production of these AFPs is not currently feasible. We believe that these difficulties could be surmounted by the production of microbial AFPs, but to date, only a few such proteins have been identified and purified, and none of these are associated with hydrocarbon reserves. Here, we have used ice-affinity and freeze-thaw stress to select microbes derived from oil and gas formation water, or produced water, as a source of anaerobic microbial communities. Ice-affinity successfully incorporated anaerobic bacteria under aerobic conditions, and the mixed culture had ice-associating properties. Under these conditions, ice-affinity selection does not result in cultivatable isolates, but similar, cultivable microbes were obtained following freeze-thaw selection under anaerobic conditions. Since these mixed cultures inhibited the growth of ice crystals, they also have the potential to inhibit hydrate growth. Overall, freeze-thaw selection provides a promising first step towards the isolation of microbes capable of the inhibition of ice and gas hydrate growth, for possible application for energy exploration and recovery at high-latitudes and in-deep, cold waters.     

In situ and laboratory bioassays to evaluate the impact of effluent discharges on receiving aquatic ecosystems

Effluents are a main source of direct and often continuous input of pollutants into aquatic ecosystems with long-term implications on ecosystem functioning. Therefore, the study of the effects of effluent exposure on organisms, populations or communities within the framework of impact assessment has a high ecological relevance. The aim of this study was to assess the toxicological impact of two effluents, one household wastewater treatment effluent (Effluent 1) and one industrial effluent (Effluent 2), on the receiving aquatic ecosystem using two test species under both in situ and laboratory conditions. Zebra mussel (Dreissena polymorpha) and common carp (Cyprinus carpio) were exposed under laboratory conditions in an online monitoring flow-through system (receiving different concentrations of Effluent 2) and under in situ conditions along the pollution gradient established by these two effluent discharges. Bioassays focussed on growth and condition related endpoints (i.e. condition, growth, lipid budget), since these are key functional processes within organisms and populations. Under laboratory conditions, increasing concentrations of the industrial effluent (Effluent 2) had a negative effect on both zebra mussel and carp energy reserves and condition. Under in situ conditions, the same negative impact of Effluent 2 was observed for zebra mussels, while Effluent 1 had no apparent effect on exposed zebra mussels. Carp growth and condition, on the other hand, were significantly increased at the discharge sites of both effluents when compared to the reference site, probably due to differences in food availability. The results indicate that a combination of in situ and laboratory exposures can illustrate how ecological processes influence bioassay studies. The incorporation of indirect, ecological effects, like changes in food availability, provides considerable benefit in understanding and predicting effects of effluents on selected species under realistic exposure conditions.     

Energy reserves and accumulation of metals in the ground beetle Pterostichus oblongopunctatus from two metal-polluted gradients

Living in an area chronically polluted with metals is usually associated with changes in the energy distribution in organisms due to increased energy expenses associated with detoxification and excretion processes. These expenses may be reflected in the available energy resources, such as lipids, carbohydrates, and proteins. In this context, the energy status of Pterostichus oblongopunctatus (Coleoptera: Carabidae) was studied in two metal pollution gradients near Olkusz and Miateczko ?l?skie in southern Poland. Both regions are rich in metal ores, and the two largest Polish zinc smelters have been operating there since the 1970s. Beetles were collected from five sites at each gradient. Zinc and cadmium concentrations were measured in both the soil and the beetles. The possible reduction in energy reserves as a cost of detoxifying assimilated metals was evaluated biochemically by determining the total lipid, carbohydrates, and protein contents. At the most polluted sites, the Zn concentration in the soil organic layer reached 2,906 mg/kg, and the Cd concentration reached 55 mg/kg. Body Zn and Cd concentrations increased with increasing soil Zn and Cd concentrations (p?=?0.003 and p?=?0.0001, respectively). However, no relationship between pollution level and energetic reserves was found. The results suggest that populations of P. oblongopunctatus inhabiting highly metal-polluted sites are able to survive without any serious impact on their energy reserves, though they obviously have to cope with elevated body metal concentrations.     

Parasites and pollution: the effectiveness of tiny organisms in assessing the quality of aquatic ecosystems,with a focus on Africa

The aquatic environment represents the final repository for many human-generated pollutants associated with anthropogenic activities. The quality of natural freshwater systems is easily disrupted by the introduction of pollutants from urban, industrial and agricultural processes. To assess the extent of chemical perturbation and associated environmental degradation, physico-chemical parameters have been monitored in conjunction with biota in numerous biological monitoring protocols. Most studies incorporating organisms into such approaches have focussed on fish and macroinvertebrates. More recently, interest in the ecology of parasites in relation to environmental monitoring has indicated that these organisms are sensitive towards the quality of the macroenvironment. Variable responses towards exposure to pollution have been identified at the population and component community level of a number of parasites. Furthermore, such responses have been found to differ with the type of pollutant and the lifestyle of the parasite. Generally, endoparasite infection levels have been shown to become elevated in relation to poorer water quality conditions, while ectoparasites are more sensitive, and exposure to contaminated environments resulted in a decline in ectoparasite infections. Furthermore, endoparasites have been found to be suitable accumulation indicators for monitoring levels of several trace elements and metals in the environment. The ability of these organisms to accumulate metals has further been observed to be of benefit to the host, resulting in decreased somatic metal levels in infected hosts. These trends have similarly been found for host–parasite models in African freshwater environments, but such analyses are comparatively sparse compared to other countries. Recently, studies on diplozoids from two freshwater systems have indicated that exposure to poorer water quality resulted in decreased infections. In the Vaal River, the poor water quality resulted in the extinction of the parasite from a site below the Vaal River Barrage. Laboratory exposures have further indicated that oncomiracidia of Paradiplozoon ichthyoxanthon are sensitive to exposure to dissolved aluminium. Overall, parasites from African freshwater and marine ecosystems have merit as effect and accumulation indicators; however, more research is required to detail the effects of exposure on sensitive biological processes within these organisms.     

Energy resource reallocation in Daphnia schodleri (Anomopoda: Daphniidae) reproduction induced by exposure to hexavalent chromium

Cladocerans are able to store energy in the form of macromolecules such as proteins, lipids and carbohydrates. However, their ability to do so depends to a large extent on their physiological and reproductive condition as well as their age. These macromolecules constitute the total caloric reserves of the body and may be modified as a result of environmental stress conditions by either physical or nutritional factors or by exposure to toxic compounds. Neonates of the freshwater cladoceran Daphnia schodleri were exposed to 0.0064, 0.032 and 0.064 mg L⁻¹ of hexavalent chromium - Cr(VI) - up to age 5, 7, 14, 21 or 28 d. Survival and fecundity of parthenogenetic females was evaluated for 28 d (maximum exposure period). Body length and maximum width were determined in specimens of all ages. Caloric content was evaluated in progeny and females within each age group. Female survival was significantly lower with exposure to 0.064 mg L⁻¹ of Cr(VI) than with other treatments, and negative effects on reproduction were observed from the lowest concentration on. Largest adult size was attained by control specimens towards the end of the observation period, but no significant differences in neonate size occurred between concentrations. Energy content decreased in adults as toxicant concentration increased, but these females allocated a larger amount of energy reserves to their progeny as a possible compensation aimed at ensuring neonate survival in an adverse environment due to toxicant presence. Sublethal concentrations of Cr(VI) modified energy content in D. schodleri, affecting to varying degrees its survival, growth and reproduction.     

Influence of reduced dietary calcium on the accumulation and effects of lead, cadmium, and aluminum in birds

The influence of low dietary calcium on the accumulation and effects of dietary lead, cadmium and aluminum was examined in zebra finches and ring doves. In zebra finches fed a diet containing 0.3% Ca, the hepatic and renal accumulation of lead was enhanced approximately 400% and of cadmium about 150-200%, compared to birds fed a 3.0% Ca diet. Low dietary Ca also caused bones of female finches to lose an average of about 60% of their normal Ca content. Loss of bone-Ca was also observed in male finches, but was less than in females. In reproductively active ring doves, low (0.4%) dietary Ca enhanced the accumulation of lead and cadmium, but not of aluminum, compared with accumulation in doves consuming a 2.0% Ca diet. Enhanced accumulation of lead and cadmium was accompanied by increased synthesis of the metal-binding protein metallothionein and by greater inhibition of delta-aminolevulinic acid dehydratase activity. These results indicate that, under conditions of reduced dietary Ca availability, such as can occur in acid-impacted environments, wild birds risk increased uptake of certain toxic metals and may accumulate toxic concentrations of these metals more rapidly. Researchers should take account of dietary Ca levels when interpreting results of dosing studies in which metals such as lead and cadmium are administered to birds.     

How the tapeworm <Emphasis Type="Italic">Hymenolepis diminuta</Emphasis> affects zinc and cadmium accumulation in a host fed a hyperaccumulating plant (<Emphasis Type="Italic">Arabidopsis halleri</Emphasis>)

The effects of plant-bound zinc (Zn) and cadmium (Cd) on element uptake and their interactions in a parasite-host system were investigated in a model experiment. Male Wistar rats were divided into four groups (C, P, TC and TP). Groups TC and TP were infected with the rat tapeworm Hymenolepis diminuta. Groups C and TC were fed a standard rodent mixture (ST-1) and received 10.5 mg of Zn per week, while groups P and TP were fed a mixture supplemented with the Zn- and Cd-hyperaccumulating plant Arabidopsis halleri at a dosage of 236 mg Zn/week and 3.0 mg Cd/week. Rats were euthanized after 6 weeks, and Cd and Zn levels were determined in rat and tapeworm tissue. The results indicate that tapeworm presence did have an effect on Cd and Zn concentrations in the host tissue; the majority of tissues in infected rats had statistically significant lower Zn and Cd concentrations than did uninfected rats. Tapeworms accumulated more zinc and cadmium than did the majority of host tissues. This important finding confirms the ability of tapeworms to accumulate certain elements (heavy metals) from the host body to their own body tissues. Thus, tapeworms can decrease heavy metal concentrations in host tissues.     

Species-specific responses to metals in organically enriched river water, with emphasis on effects of humic acids

Invertebrate communities in polluted rivers are often exposed to a wide variety of compounds. Due to complex interactions, 'pollution tolerant' species are not necessarily the most tolerant species to toxicants tested under standard laboratory conditions. It was hypothesized that the distribution of species in polluted rivers is not only dependent on the tolerance of species to toxicants, but also on species-specific capacities to modify or compensate for negative effects of toxicants. To test this hypothesis, species-specific responses to metals in organically enriched river water were studied under controlled conditions. The zebra mussel Dreissena polymorpha and the midge Chironomus riparius were exposed to metal-polluted water from the River Dommel. Additionally, the (interactive) effects of metals and humic acids (HA) on both species were evaluated. In spite of a lower tolerance of Chironomus riparius to metals in laboratory studies, the midge was the most tolerant of the two test species to metal-polluted site water. The results indicated that the sensitivities of the two test species determined in laboratory tests were inversely related to their sensitivities to polluted river water. In accordance with these results, midge larvae were protected from copper (Cu) toxicity by HA, while metal toxicity was not reduced (Cu) or even amplified (cadmium) by HA for the zebra mussel. Thus, the presence of (naturally occurring) HA in site water may partly account for discrepancies between responses of species to bioassays and toxicity tests. It is suggested that these differences in responses to metals in site water are strongly influenced by species-specific preferences for organic compounds (like HA). It is concluded that the response to organic compounds present in site water largely determines whether a species is classified as 'pollution tolerant' or 'pollution sensitive'.     

Humic substances

GOALS, SCOPE AND BACKGROUND: Changing environmental conditions and handling stress are well known to cause chronic or acute stress situations in fish with subsequent infections. These requires a therapy by means of antibiotics and chemicals. In contrast to the huge number of pathogens, only a few substances are permitted for application as therapeutics in German aquaculture. Hence, there emerges an urgent need for highly effective and residueless alternatives. MAIN FEATURES: The prophylactic stimulation and training of the defense system of fish by alternative approaches becomes increasingly necessary. One approach is the application of dissolved humic substances (HS) of natural or artificial origin. For example, there exist several reports on the positive effect of HS to fishes. These effects shall be considered in detail. Furthermore, the impact of HS on the constitution of parasites and pathogens will be displayed. The reports on this issue are diverse, if not inconsistent. We try to shed some light on these discrepancies. The last aspect covered by this review is the outdated paradigm that calcium ions act as antidotes. In the presence of HS, even the opposite effect may occur. APPROACH: To overcome old paradigms on HS and their potential interactions with fish and fish parasites, we reviewed recent international literature, as well as 'grey' literature. We also include results from own former and ongoing studies. RESULTS AND DISCUSSION: HS are able to increase the physiological condition of the individuals and to reduce adverse physiological and histological consequences caused by stress; the mechanism behind remains obscure. HS detoxify heavy metals and organic pollutants. Damages caused by several fish pathogens, such as bacteria and parasites, can be repaired more quickly in the presence of HS. Some parasites--mainly fungi--appear to be directly affected by HS. Comparing the fungicidal effects of HS from various sources, evidence is increasing that the aliphatic moiety may be the effective structures. However, further research is necessary to relate more physiological and anti-pathogenic effects to the chemical characteristics of HS. CONCLUSIONS: HS are not real alternatives to strong traditional therapeutics. However, they show different advantages in repairing secondary, stress induced damages in fish. The ecophysiological relevance of HS in either aquatic systems or aquaculture is getting conspicuously. PERSPECTIVES: The lack of therapeutic and antiparasitic substances in aquaculture requires new strategies and ways of thinking. The search for alternatives to the 'traditional' chemical therapeutics calls for the intensive research. Inevitably, this search will lead to an intensive contemplation on HS as 'health promoting substances' and/or even therapeutics. Basic research is needed to detect the functional groups of the HS responsible for the effects observed. Health promoting effects of first investigations made in vitro to affect pathogens via application of HS and several field studies with HS raises hopes for a broader utilisation of HS to reduce stress consequences in fish and fish pathogens residuelessly.     

Interpopulation differences in acute response of Brotia hainanensis (Gastropoda, Prosobranchia) to cadmium: genetic or environmental variance?

Adolescent Brotia hainanensis collected from two separate sites along the same river showed markedly different acute responses to cadmium even after laboratory acclimation for one week. Similar interpopulation differences persisted in the F(1) juveniles (< two-day old) which were descendants of laboratory-cultured snails. Although these results could imply that the inter-site differences in cadmium tolerance for the two populations may have a genetic basis, these differences in metal tolerance in the F(1) juveniles disappeared after the juveniles had been cultured under identical laboratory conditions for one week. Quantitative genetical analyses revealed that variance of initial shell length and survival time of F(1) juveniles exposed to 10 mg/litre cadmium was significantly smaller for juveniles that were offspring of the same female as compared with animals that were descendants of different females. Moreover, the survival time was positively correlated with the initial size of the juveniles. Common environmental and maternal effects have an overwhelming influence on cadmium tolerance in these ovoviviparous snails.     

Effect of gender on physiological and behavioural responses of Gammarus roeseli (Crustacea Amphipoda) to salinity and temperature

The importance of potentially interacting factors in organisms responses to a stress are often ignored or underestimated in ecotoxicology. In laboratory experiments we investigated how gender, temperature and age influence the behaviour and the physiology of the freshwater amphipod Gammarus roeseli under salinity stress. Our results revealed a significant higher sensitivity of females in survival, ventilation and ionoregulation whereas no inter-age differences were reported. Water temperature also exerted a significant effect in survival and ventilation of G. roeseli. Some of those factors appeared to interact significantly. This study provides evidence that gender can affect organisms responses to a stressor and consequently has to be considered while assessing a stress impact. We discussed the potential relationships between biological and behavioural responses.     

Ecological impact assessment of metallurgic effluents using in situ biomarker assays

An ecological impact study was performed based on in situ biomarker assays with the waterflea Daphnia magna. The effects of metallurgic effluents on the energy metabolism, anti-oxidative metabolism and DNA damage were assessed in caged daphnids during a 4-week study. In situ survival and reproduction studies demonstrated a clear impact on these parameters in organisms exposed in the most polluted areas. At the downstream--sublethal--zone the organisms were disturbed within their tolerance limits, resulting in alterations of their energy metabolism. These data suggest an acclimation hypothesis, which was tested through the analysis of the energy metabolism of resident species: isopods and amphipods. These organisms had shifted to a decrease in their overall energy metabolism compared to the upstream region. This change in some biochemical processes suggests a selective advantage to cope with the prevailing environmental conditions. In addition, we found clear genotoxic effects caused by the industrial discharges that might correlate with a reduction in (long-term) survival.     

Micronutrient seed priming: new insights in ameliorating heavy metal stress

Plants need to survive with changing environmental conditions, be it different accessibility to water or nutrients, or attack by insects or pathogens. Few of these changes, especially heavy metal stress, can become more stressful and needed strong countermeasures to ensure survival of plants. Priming, a pre-sowing hydration treatment, involves pre-exposure of plants to an eliciting component which enhance the plant’s tolerance to later stress events. By considering the role of micronutrients in aiding plants to cope up under adverse conditions, this review addresses various aspects of micronutrient seed priming in attenuating heavy metal stress. Priming using micronutrients is an adaptive strategy that boosts the defensive capacity of the plant by accumulating several active or inactive signaling proteins, which hold considerable importance in signal amplification against the triggered stimulus. Priming induced ‘defence memory’ persists in both present generation and its progeny. Therefore, it is considered a promising approach by seed technologist for commercial seed lots to enhance the vigour in terms of seed germination potential, productivity and strengthening resistance response against metalloid stress. The present review provides an overview regarding the potency of priming with micronutrient to ameliorate harmful effects of heavy metal stress, possible mechanism how attenuation is accomplished, role of priming in enhancing crop productivity and inducing defence memory against the metalloid stress stimulus.

     

Interactive effects of metal contamination and pathogenic organisms on the introduced marine bivalve Ruditapes philippinarum in European populations

In natural environment, marine organisms are concomitantly exposed to pollutants and multiple disease agents resulting in detrimental interactions. The present study evaluated interactive effects of metal contamination (cadmium) and pathogenic organisms (trematode parasites Himasthla elongata and pathogenic bacteria Vibrio tapetis) singularly and in combination on the bivalve Ruditapes philippinarum, an introduced species to Europe, under laboratory controlled conditions. After 7 days, metal bioaccumulation and pathogen load were analyzed as well as metallothionein (MT) response and hemocyte concentrations and activities. Results showed that infection by opportunistic pathogens affects metal accumulation, leading to maximal Cd accumulation in co-infected clams. Among stressors only V. tapetis induced significant effects on immune parameters whereas a particular interaction “trematode-bacteria” was shown on MT responses. Despite low trematode infection in agreement with the resistant status of R. philippinarum to these macroparasites, significant interaction with bacteria and metal occurred. Such results highlight the necessity of taking pathogens into account in ecotoxicological studies.     

Spatial and temporal variations of mercury levels in Okefenokee invertebrates: southeast Georgia

Accumulation of mercury in wetland ecosystems has raised concerns about impacts on wetland food webs. This study measured concentrations of mercury in invertebrates of the Okefenokee Swamp in Georgia, focusing on levels in amphipods, odonates, and crayfish. We collected and analyzed total mercury levels in these invertebrates from 32 sampling stations across commonly occurring sub-habitats. Sampling was conducted in December, May, and August over a two-year period. The highest levels of mercury were detected in amphipods, with total mercury levels often in excess of 20 ppm. Bioaccumulation pathways of mercury in invertebrates of the Okefenokee are probably complex; despite being larger and higher in the food chain, levels in odonates and crayfish were much lower than in amphipods. Mercury levels in invertebrates varied temporally with the highest levels detected in May. There was a lack of spatial variation in mercury levels which is consistent with aerial deposition of mercury.     

Uptake of 203Hg++ and 115 mCd++ by Chlamydomonas reinhardi under various conditions

The uptake of radioactively labeled mercury and cadmium ions by living and dead cells of Chlamydomonas reinhardi WT⁺ was measured at 25 °C under minimal and optimal conditions (dark/nitrogen or light/air, respectively). In each case, incorporation was completely independent of external energy. Living and dead cells incorporated almost the same amount of the added heavy metal ions; after about 4 to 8 hours, saturation was obtained. Furthermore, the distribution of mercury in the system of algea/culture medium/gas phase and the amount of mercury and cadmium adsorbed by cells were studied.