The use of fish parasites as bioindicators of heavy metals in aquatic ecosystems: a review

Parasites are attracting increasing interest from parasite ecologists as potential indicators of environmental quality due to the variety of ways in which they respond to anthropogenic pollution. In environmental impact studies certain organisms provide valuable information about the chemical state of their environment not through their presence or absence but instead through their ability to concentrate environmental toxins within their tissues. Free living invertebrates, notably bivalve molluscs, are commonly employed in this role as `sentinel organisms' to monitor the concentrations of bioavailable metals in aquatic ecosystems. Also certain parasites, particularly intestinal acanthocephalans of fish, can accumulate heavy metals to concentrations orders of magnitude higher than those in the host tissues or the environment. The comparison of metal accumulation capacities between acanthocephalans and established free living sentinel organisms revealed significantly higher concentrations of several elements in Acanthocephalus lucii (Müller) than in the Zebra mussel Dreissena polymorpha (Pallas) which is a commonly used bioindicating organism in Europe. In contrast to the high heavy metal concentrations recorded in adult acanthocephalans, the larval stages in their respective crustacean intermediate hosts show little tendency to accumulate metals. A number of experimental studies demonstrate a clear time dependent accumulation of lead for acanthocephalans in their final hosts. These investigations provide evidence that the extremely high metal concentrations in intestinal acanthocephalans of fish are not the result of a slow process of accumulation but instead a relatively rapid uptake to a steady-state level. Thus, metal concentrations in adult acanthocephalans respond rapidly to changes in environmental exposure of their hosts. The value of parasites for environmental monitoring will be discussed in detail in the present article.     

Biorefining of platinum group metals from model waste solutions into catalytically active bimetallic nanoparticles

Bacteria can fabricate platinum group metal (PGM) catalysts cheaply, a key consideration of industrial processes and waste decontaminations. Biorecovery of PGMs from wastes is promising but PGM leachates made from metallic scraps are acidic. A two‐step biosynthesis ‘pre‐seeds’ metallic deposits onto bacterial cells benignly; chemical reduction of subsequent metal from acidic solution via the seeds makes bioscaffolded nanoparticles (NPs). Cells of Escherichia coli were seeded using Pd(II) or Pt(IV) and exposed to a mixed Pd(II)/Pt(IV) model solution under H₂ to make bimetallic catalyst. Its catalytic activity was assessed in the reduction of Cr(VI), with 2 wt% or 5 wt% preloading of Pd giving the best catalytic activity, while 1 wt% seeds gave a poorer catalyst. Use of Pt seeds gave less effective catalyst in the final bimetallic catalyst, attributed to fewer and larger initial seeds as shown by electron microscopy, which also showed a different pattern of Pd and Pt deposition. Bimetallic catalyst (using cells preloaded with 2 wt% Pd) was used in the hydrogenation of soybean oil which was enhanced by ~fourfold using the bimetallic catalyst made from a model waste solution as compared to 2 wt% Pd preloaded cells alone, with a similar selectivity to cis C18:1 product as found using a Pd‐Al₂O₃ commercial catalyst.     

Palladium exposure of barley: uptake and effects

Motor vehicles are now equipped with exhaust gas catalytic converters containing rare metals, such as palladium (Pd), platinum and rhodium, as catalytic active materials, leading to significantly increased emission of these metals. Compared with platinum and rhodium, low concentrations of Pd have been shown to have more serious effects on cells and organisms. In the present study, uptake of Pd by barley and behaviour of Pd nanoparticles in nutrient solutions used to grow plants were observed in order to develop a model of Pd exposure of plant systems. Pd determination was performed using a selective separation and pre-concentration procedure, which was further developed for this study, and coupled to graphite furnace atomic absorption spectrometry. The results show that uptake of Pd depends on Pd particle diameter. Compared to other toxic metals, like mercury, Pd causes stress effects in leaves at lower concentrations in nutrient solutions. Furthermore, Pd particles are dissolved at different rates, depending on size, in the nutrient solution during plant growth.     

The parasitic nematodes Hysterothylacium sp. type MB larvae as bioindicators of lead and cadmium: a comparative study of parasite and host tissues

Cadmium and lead concentrations were compared in tissues of cutlassfish, Trichiurus lepturus L., its intestinal nematode Hysterothylacium sp. type MB larvae, and in water from the same location in the Sea of Oman. Metal accumulation in hosts, parasites and sea water was measured by ICP-OES. Hysterothylacium larvae from the intestinal lumen and visceral cavity showed much higher metal concentrations than in host tissues or sea water. Statistical analyses revealed no significant differences in metal accumulation between infected and uninfected hosts. Cadmium concentration in the host muscle was lower than in intestine, liver and gonad tissues. The mean concentrations of lead and cadmium in nematodes were 289·03 and 81·5 times higher than in host intestine, 188·4 and 225 times higher than in host muscle, 108·6 and 65·3 times higher than in host gonads, 70·5 and 19·5 times higher than in host liver and 3351 and 148 times higher than in sea water. The results show the value of this and possibly related nematodes as bioindicators of heavy metals and their potential use in environmental studies.     

How parasitism and pollution affect the physiological homeostasis of aquatic hosts

Parasitism poses a serious threat to hosts under certain circumstances, while the well-being of organisms is also negatively affected by environmental pollution. Little information is available on the simultaneous effects of parasites and pollutants on the physiological homeostasis of organisms. The present paper demonstrates that parasites: (i) may influence the metabolism of pollutants in infected hosts, (ii) interact with pollution in synergistic or antagonistic ways, and (iii) may induce physiological reactions in hosts which were thought to be pollutant-induced. Experimental studies on the uptake and accumulation of metals by fish reveal that fish infected with acanthocephalans have lower metal levels than uninfected hosts; e.g. Pomphorhynchus laevis reduces lead levels in fish bile, thereby diminishing or impeding the hepatic intestinal cycling of lead, which may reduce the quantity of metals available for fish. Alterations in pollutant uptake and accumulation in different intermediate and final hosts due to parasites are thus very important in the field of ecotoxicology. In addition to such alterations, there is a close interaction between the effects of pollutants and parasites which seems to be mediated at least partly by the endocrine system, which itself is closely related to the immune system in fish. Laboratory studies on eels experimentally infected with the swimbladder nematode Anguillicola crassus reveal that toxic chemicals such as polychlorinated biphenyls produce immunosuppressive effects which facilitate parasite infection. Similarly, an increase in serum cortisol concentration in eels due to chemical exposure and infection is correlated with decreasing levels of anti-A. crassus antibodies. Furthermore, parasites are able to elicit physiological changes which are attributed to chemicals with endocrine disrupting activity, e.g. the cestode Ligula intestinalis is known to suppress gonad development in roach. The most thoroughly documented examples of endocrine disruption in wild fish are in roach, and it is conceivable that this disruption is not only due to chemical activity but also to parasites such as L. intestinalis or species of the phylum Microspora.     

Heavy Metal Pollution Characteristics of Surface Sediments in Different Aquatic Ecosystems in Eastern China: A Comprehensive Understanding

Aquatic ecosystems in eastern China are suffering threats from heavy metal pollution because of rapid economic development and urbanization. Heavy metals in surface sediments were determined in five different aquatic ecosystems (river, reservoir, estuary, lake, and wetland ecosystems). The average Cd, Cr, Cu, Ni, Pb, and Zn concentrations were 0.716, 118, 37.3, 32.7, 56.6, and 204 mg/kg, respectively, and the higher concentrations were mainly found in sediment samples from river ecosystems. Cd was the most anthropogenically enriched pollutant, followed by Zn and Pb, indicated by enrichment factors >1.5. According to consensus-based sediment quality guidelines, potential ecological risk indices, and risk assessment codes, all five types of aquatic ecosystems were found to be polluted with heavy metals, and the most polluted ecosystems were mainly rivers. Cd was the most serious pollutant in all five aquatic ecosystems, and it was mainly found in the exchangeable fraction (about 30% of the total Cd concentration, on average). The results indicate that heavy metal contamination, especially of Cd, in aquatic ecosystems in eastern China should be taken into account in the development of management strategies for protecting the aquatic environment.     

COMPARISON OF HEAVY METALS IN AQUATIC PLANTS ON CHARITY ISLAND,SAGINAW BAY,LAKE HURON,U.S.A., WITH PLANTS ALONG THE SHORELINE OF SAGINAW BAY

The concentrations of 15 heavy metals in aquatic plants on Charity Island was compared to those in plants on the shoreline of Saginaw Bay, Lake Huron, U.S.A. Heavy metal concentrations were measured by neutron activation analysis. Charity Island was found to have significantly higher levels of nine of fifteen metals investigated. This indicates that distance from known pollution source was not the only factor affecting the heavy metal accumulation of the aquatic plants.     

Determination of water quality,and trace metals in endemic Sarotherodon linellii,Pungu maclareni and Clarias maclareni,in crater Lake Barombi Mbo,Cameroon

Considering water pollution as a potential threat to some endemic cichlids of Lake Barombi Mbo, Cameroon, an investigation was done in 2011 to determine trace metals in its water, linking their uptake in gills and liver of fish to water chemistry. ICP-MS and ICP-OES analyses of trace metals based on total concentration of unfiltered lake water samples showed the presence of trace metals. All fish species accumulated Al, Mn and Sr in the highest concentrations in their gills, with Cu, Cd, Co, Cr, Pb and U highest in the liver. Pungu maclareni accumulated Al, Cr, Co, Sr and Pb in the highest concentrations. The highest mean gill Al concentration of 140 µg g^?1 dry weight was measured in P. maclareni gills, this being one of the critically endangered cichlids of the lake. Stable isotope analyses of carbon δ¹³C and nitrogen δ¹⁵N showed that P. maclareni had the highest mean δ¹³C (?30.2‰) and highest concentrations of Cr, Co, Pb and U in liver, probably linking the carbon source to the accumulation of metals. Though trace metal levels in the lake water were low, their presence in fish tissues suggest they are bioavailable, bioaccumulate and may pose a threat to the aquatic biota, and therefore should be monitored.     

The role of a freshwater tidal area with controlled reduced tide as feeding habitat for European eel (Anguilla anguilla,L.)

Implementation of the Controlled Reduced Tide (CRT) technique could increase the total surface of tidal freshwater marshes in Europe and ease implementation of restoration projects in coastal defense and riverine ecosystems. The goal was to determine whether a regularly flooded area connected to a freshwater tidal river could act as an important foraging area for European eel, and if so, to what extent the diet of eels in this flooding area differed from that of eels foraging in the river itself. The stomach contents of eels from the River Schelde were compared with eels from the Lippenbroek, an adjacent CRT area. Prey diversity (H′) of individual eels was about four times higher in the Lippenbroek than in the River Schelde. Moreover, 12 prey categories in eel stomachs from the Lippenbroek were found whereas only three categories were retrieved from eels in the River Schelde. In the Lippenbroek, eels fed on terrestrial organisms (lumbricids, caterpillars and other insects), but also on fish and fish eggs and to a lesser extent on other aquatic prey (Lumbricullidae, chironomids and Hirudinea). In contrast, eels from the main river fed mainly on tubificids, fish, and some gammarids. Consequently, eels in the Schelde estuary are opportunistic feeders, but with a preference for large benthic prey. The number and weight of aquatic organisms ingested by eels in the Lippenbroek is not significantly different from the River Schelde. However, eels foraging in the Lippenbroek area had consumed significantly more terrestrial prey. Furthermore the total caloric value estimated for the ingested prey of eels from the Lippenbroek (derived from the literature) was about twice as high as that for eels from the River Schelde. While the condition index remained inconclusive, an Ancova revealed that eels captured in the Lippenbroek were significantly heavier for a given length than eels captured in the Schelde. The study showed that with a controlled reduced tide to restore lateral connectivity of large tidal rivers with their adjacent floodplains, high quality habitats for the European eel are created. These measures could significantly contribute to the production of eels in better condition, which have better chances to reproduce successfully. Hence, wetland restoration could enhance the recovery of the European eel stocks.     

Comparison between lead accumulation of Pomphorhynchus laevis (Palaeacanthocephala) in the intestine of chub (Leuciscus cephalus) and in the body cavity of goldfish (Carassius auratus auratus)

This experimental study assessed the role of the microhabitat in the uptake of metals by adult acanthocephalans. We examined the accumulation of lead by adult Pomphorhynchus laevis in the intestine of chub (Leuciscus cephalus) and compared it with that in goldfish, Carassius auratus auratus, in which the parasites penetrate the intestinal wall and enter the body cavity. Chub and goldfish experimentally infected with adult Pomphorhynchus laevis were exposed to 0.01 mg l(-1) Pb(2+) over 3 weeks. Lead was rapidly accumulated in the intestinal acanthocephalans reaching a mean concentration of 7.3 microg g(-1). This concentration was significantly greater than in the host muscle, liver and intestine and more than 730 times higher than the exposure concentration. Intraperitoneal P. laevis in goldfish exposed to lead did not accumulate the metal. Thus, it was conclusively shown that metal accumulation in acanthocephalans is associated with the intestinal location and does not occur in the body cavity.     

Feeding habitat and silvering stage affect lipid content and fatty acid composition of European eel Anguilla anguilla tissues

Lipids, particularly fatty acids (FAs), are major sources of energy and nutrients in aquatic ecosystems and play key roles during vertebrate development. The European eel Anguilla anguilla goes through major biochemical and physiological changes throughout its lifecycle as it inhabits sea- (SW), and/or brackish- (BW) and/or freshwater (FW) habitats. With the ultimate goal being to understand the reasons for eels adopting a certain life history strategy (FW or SW residency vs. ‘habitat shifting’), we explored differences in lipid content and FA composition of muscle, liver and eyes from eels collected across Norwegian SW, BW and FW habitats, and at different lifecycle stages (yellow to silver). FW and SW eels had a higher lipid content overall compared to BW eels, reflecting differences in food availability and life history strategies. SW eels had higher proportions of certain monounsaturated FAs (MUFAs; 18:1n-9, 20:1n-9), and of the essential polyunsaturated FAs 20:5n-3 (eicosapentaenoic acid, EPA) and 22:6n-3 (docosahexaenoic acid) than FW eels, reflecting a marine-based diet. In contrast, the muscle of FW eels had higher proportions of 18:3n-3, 18:2n-6 and 20:4n-6 (arachidonic acid), as is typical of FW organisms. MUFA proportions increased in later stage eels, consistent with the hypothesis that the eels accumulate energy stores prior to migration. In addition, the decrease of EPA with advancing stage may be associated with the critical role that this FA plays in eel sexual development. Lipid and FA information provided further understanding of the habitat use and overall ecology of this critically endangered species.     

Towards Engineering Nanoporous Platinum Thin Films for Highly Efficient Catalytic Applications

Porous metals attract significant interest for use in diverse electrochemical catalytic applications. However the fabrication of scalable and controlled porous metal structures on the nanoscale, particularly with highly catalytic pure Pt, still remains a significant challenge. We demonstrate highly engineered nanoporous Pt thin films by the dealloying of a Pt‐Si binary alloy system with a predetermined alloy composition. Controlled pore dimensions and nanostructures are obtained by tailoring the Pt‐Si alloy composition followed by selective Si etching. As a result, isotropic open nanopores are formed in continuous Pt ligaments and the porosity becomes larger on increasing the Si/Pt atomic ratio, which leads to the formation of a higher surface area and active catalytic sites. The formed nanoporous Pt film shows a 32‐times‐higher catalytic activity than Pt/C catalysts, with a high current density and low charge‐transfer resistance during methanol electro‐oxidation. The results reported here open up possibilities to develop high‐performance and reliable catalytic electrodes in energy and environmental applications.     

Host-parasite interface of Paratenuisentis ambiguus (Eoacanthocephala) in naturally infected eel and in laboratory-infected sticklebacks and juvenile carp and rainbow trout

Paratenuisentis ambiguus is described from natural infections in adult eels and from laboratory infections in sticklebacks and juvenile carp and rainbow trout. In captured eels, female worms kept reproducing in the laboratory for at least 1 mo. In the 3 small laboratory hosts female worms did not release eggs and longevity did not exceed 3-30 days. It is concluded that the small fishes were unsuitable final hosts. Worm penetration into the intestinal wall of all hosts was shallow. Thus, the small fishes also proved to be unsuitable paratenic hosts. The worms ruptured the intestinal mucosa and the underlying tunica propria and they often seemed to change their sites of attachment. The proboscides carried an osmiophilic surface coat that seemed to be supported by liquid drops from necrotic host tissue and by osmiophilic material apparently discharged from pores in the worm's proboscis hooks. The coat contained lipids, polysaccharides, and/or proteoglycans and likely other substances. Around the hooks the proboscis tegument harbored conspicuous cisternae of rough endoplasmic reticulum as is typical for cells with secretory function. Mostly, the worms were found with semi-invaginated proboscides. The resulting cavity inside the proboscis seemed to collect lipids and other remnants of host cells from the lesions caused by the worms. Whether the apical hollow might function as a gastric cavity is discussed.     

The intestinal parasite Pomphorhynchus laevis (Acanthocephala) interferes with the uptake and accumulation of lead (210Pb) in its fish host chub (Leuciscus cephalus)

Uninfected chub as well as fish experimentally infected with the acanthocephalan parasite Pomphorhynchus laevis were exposed to ²¹⁰Pb²⁺ for up to 38 days and the uptake and distribution of lead within different fish organs and the parasites was determined at various time points. Highest metal concentrations were detected in the acanthocephalans, followed by intestine, bile, liver, gill and muscle of the fish host. Infected chub had significantly lower ²¹⁰Pb levels in the gills on day 17 (P≤0.01), in the bile on day 24 (P≤0.05) and in the gills as well as in the intestine on day 38 compared with uninfected fish. A subsequent polynomial regression revealed that lead levels for the infected fish ranged below the levels determined for uninfected fish during most of the exposure period. This is the first proof that P. laevis reduces lead levels in the bile thereby diminishing or even impeding the hepatic intestinal cycling of lead, which may reduce the amount of metals available for the fish organs. This is especially important for ecotoxicological research. For example, organisms used as accumulation indicators may erroneously indicate low levels of pollution if they are infected with parasites which alter their pollutant uptake mechanisms. Additionally, the results gave further experimental evidence for acanthocephalans as accumulation indicators for metals.     

Discrimination of Rhizobium japonicum,Rhizobium lupini,Rhizobium trifolii,Rhizobium leguminosarum and of bacteriods by uptake of 2-ketoglutaric acid,glutamic acid and phosphate

Rhizobium strains (one each of Rh.japonicum, Rh. lupini, Rh. leguminosarum) take up 2-ketoglutaric acid in general much faster and from lower concentrations in the medium than strains of Escherichia coli, Bacillus subtilis and Chromobacterium violaceum. A strain of Enterobacter aerogenes, however, is more similar to some Rhizobium strains. The same strains of Rhizobium take up also phosphate much faster and from lower concentrations than the other bacteria tested. 4 strains of Rh. lupini proved to be significantly different from 4 strains of Rh. trifolii in taking up l-glutamic acid from three to ten times lower concentration within 5 h. A similar difference was noticed between 5 strains of Rh. leguminosarum and 2 strains of Rh. japonicum for the uptake of 2-ketoglutaric acid and of l-glutamic acid. Isolated bacteriods from nodules of Glycine max var. Chippeway have a reduced uptake capacity for glutamic acid and for 2-ketoglutaric acid during the first 10–12 h, but reach the same value after 24 h as free living Rh. japonicum cells. The differences in the uptake kinetics are independent of cell concentration. The group II Rhizobium strains (Rh. japonicum and Rh. lupini, slow growing Rhizobium) are characterized by a rapid uptake of glutamic acid to a lowremaining concentration of 1–3×10^-7 M and an uptake of 2-ketoglutaric acid to a remaining concentration of 2–5×10^-7 M. The group I Rhizobium strains (Rh. trifolii and Rh. leguminosarum, fast growing Rhizobium), can be characterized by a much slower uptake of both substances with a more than ten times higher concentration of both metabolites remaining in the medium after the same time.     

Uptake of antitumor platinum(II)-complexes by cancer cells, assayed by inductively coupled plasma mass spectrometry (ICP-MS)

A systematic study on intracellular Pt uptake and Pt accumulation ratio in breast cancer MCF-7 cell line has been performed on a number of Pt(II)-complexes, namely cisplatin, carboplatin and oxaliplatin, clinically employed as antitumor drugs, trans- and cis-[Pt(Cl)2(pyridine)2] and cis-[Pt(Cl)2(pyridine)(5-SO3H-isoquinoline)] complexes, previously investigated also as potential telomerase inhibitors. In particular, long incubation times have been chosen in order to understand the fate of the complexes in the cells. For this purpose, sub-acute drug concentrations must be employed and, therefore, a very sensitive method of analysis like as inductively coupled plasma mass spectrometry (ICP-MS) superior to the widely employed atomic absorption spectroscopy (AAS) has been adopted. Any relationships among uptake/accumulation and several parameters such as drug structure, lipophilicity, drug concentration and incubation time have been sought and analyzed: the bulk of data point for a passive diffusion mechanism through the cell membrane.     

Impacts of the swimbladder nematode Anguillicola crassus on Anguilla anguilla: variations in liver and spleen masses

Variations in the liver and spleen masses of the eel Anguilla anguilla were analysed in relation to the parasite load of Anguillicola crassus at autopsy (current infection by swimbladder lumen worms) and in relation to the severity of damage observed in the swimbladder (a way of assessing the intensity of past infections). None of these measures of parasite pressure were shown to account for variation in the relative liver mass, either when controlling for somatic mass or eel age. In marked contrast, a significant increase in spleen size was revealed in eels harbouring many lumen worms and also in eels with severe damage in the swimbladder. Splenic enlargement was nearly two‐fold higher among severely affected eels (harbouring more than seven lumen parasites and showing severe damage in the swimbladder) than among infection‐free eels (no lumen parasites and no pathological signs in the swimbladder). Several possible hypotheses are reviewed before arguing for an adaptive host response involving the haematological and immunological functions of the spleen. Indeed, among eels with no pathological signs in the swimbladder, the relative spleen mass was positively associated with the mass of lumen parasites, which suggests a hyper‐synthesis of blood cells by the spleen in response to the bloodsucking activity of lumen worms. Nevertheless, among eels with no lumen parasites at autopsy, there was still an increase in spleen size in relation to the severity of the swimbladder damage, which also suggests a hyper‐synthesis of splenic immune cells (lymphocytes and macrophages) in reaction to damaged tissues and particularly to larvae in the swimbladder wall.     

Metallobacteriochlorophylls as potential dual agents for photodynamic therapy and chemotherapy

A theoretical analysis of bacteriochlorophyll a containing its non-native divalent metal ions: Co, Ni, Cu, Zn, Ru, Rh, Pd, and Pt, has been carried out by means of density functional theory (DFT) calculations. The main stress was put on the derivatives with metals, which already found applications as coordination compounds in anti-tumor therapy (Ru, Pt, Pd, and Rh). The idea was to combine their cytotoxic properties with the known suitability of bacteriochlorophylls macrocycle for photodynamic therapy. The geometries of the studied systems are compared and reveal a number of similarities. The cores of the modified bacteriochlorophylls are flat, and the introduced metal ions lie in plane of the macrocycle, showing its large ability to accommodate metal ions of different sizes. However, four metal–nitrogen bonds, linking the central ions with the macrocycle ligand, are not equivalent. Metals are the strongest attached to nitrogens, which come from the pyrrole, which is fused with isocyclic ring. Based on the known spectroscopic data, the absorption properties of the proposed systems are predicted. Finally, it is found that all studied metal–macrocycle adducts are stable in aqueous media. The only exceptions are Mg-BChla (the finding is reflected by experimental facts) and Zn-BChla. The predicted high stability of Ru-, Rh-, Pt- and Pd-bacteriochlorophylls might turn out beneficial for therapeutic purposes.     

Element concentrations in the archiacanthocephalan Macracanthorhynchus hirudinaceus compared with those in the porcine definitive host from a slaughterhouse in La Paz, Bolivia

Concentrations of lead and cadmium, determined by electrothermal atomic absorption spectrometry, and concentrations of the elements barium, cadmium, copper, iron, magnesium, manganese, nickel, lead, selenium and strontium, determined by inductively coupled plasma mass spectrometry, in the acanthocephalan Macracanthorhynchus hirudinaceus and its porcine final host, sampled at a slaughterhouse in La Paz, Bolivia, were compared. Inductively coupled plasma mass spectrometry analysis revealed that most of the elements were found at higher concentrations in the acanthocephalan than in different tissues of its host. The bioconcentration of elements in M. hirudinaceus compared with the host intestine, listed in order of decreasing values, was as follows: Cd > Pb > Ni > Sr = Cu > Mg > Se > Fe = Mn = Ba. Analysis by electrothermal atomic absorption spectrometry showed that M. hirudinaceus contained 85, 85, 56 and 24 times higher lead levels compared with hosts muscle, liver, kidney and intestine, respectively. The mean cadmium concentration of the parasite was 32 times higher than that of the liver and five times higher compared with porcine kidney. The metal distribution within the body of M. hirudinaceus was as follows: cement gland > testes > lemnisci > eggs = tegument for lead and lemnisci > testes > cement gland > tegument > eggs for cadmium. Therefore, the hypothesis that parasites excrete toxic metals with the shells of their eggs seems not to be valid for M. hirudinaceus. It is concluded, that not only eoacanthocephalans and palaeacanthocephalans parasitising fish, but also archiacanthocephalans from mammalian hosts, are able to bioaccumulate metals.     

The role of dissolved organic matter, particularly free amino acids and humic substances, in freshwater ecosystems

1. Although the mass of dissolved organic matter (DOM) often exceeds that of living organisms in freshwaters, little is known about the roles of its constituent molecules as sources of energy and information for aquatic organisms. In the present review attention is focused on free amino acids (FAA) and humic substances (HS) as examples of labile and refractory components within DOM. 2. The following questions are addressed. (i) What are spatiotemporal patterns in the distribution of DOM, HS and FAA? (ii) What are the origins of the components of DOM and how are their concentrations regulated? (iii) What is the significance of the spatial and temporal distributional patterns of DOM, HS and FAA to detritivorous invertebrates and other organisms associated with them? (iv) What is the relevance of DOM to the food web concept and to the biochemical ecology of freshwater ecosystems? 3. Concentrations of DOM, FAA and HS within lentic ecosystems are ranked as follows: Sediment pore water > Air–water interface > Midwater column. Comparisons between water bodies show that the concentrations of labile constituents of DOM, such as FAA, are usually positively correlated with base cations, nutrients and biological activity. In contrast, HS concentrations are negatively correlated with base cations or nutrients but positively correlated with the rate of biological degradation (the maximum values occurring in the autumn). The FAA : HS ratios might serve therefore as an indicator of the potential productivity of a water body. 4. External sources of DOM in general, and FAA and HS in particular, include rainwater, windborne material, surface flow and groundwater. The relative importance of these allochthonous sources of DOM decreases along the length of lotic ecosystems and also with increase in size of lentic ecosystems. Internal sources of FAA and HS include synthesis or polymerization from existing organic matter, degradation of organic matter and release from both living and dead organisms. The net accumulation of DOM released by living bacteria, phytoplankton, epilithon, macrophytes and invertebrates is much reduced due to heterotrophic uptake. Hence, most of the allochthonous DOM in freshwater originates from dead organic matter deposited on the sediment. Phytoplankton-dominated ecosystems may, however, differ, as most of their DOM may be recycled within the water column. 5. The factors that determine the external concentrations of DOM, FAA and HS are discussed. Evidence is cited in support of the following testable hypotheses. (i) The rates of production of DOM components will be favoured by increasing base cation and nutrient concentrations. (ii) Colloidal clay, base cations, biopolymers and living organisms, particularly bacteria, facilitate the removal of HS. Consequently, base-rich eutrophic waters tend to have lower HS concentrations than oligotrophic, base-deficient waters. (iii) As a result of higher productivity and selective removal of FAA, eutrophic waters tend to have higher FAA concentrations than those that are oligotrophic. 6. Labile DOM components, such as FAA, act as sources of information for aquatic organisms. More research is needed in this field. There is a consensus that DOM acts as an important source of energy for aquatic bacteria, thus forming the microbial loop. However, higher eukaryotic organisms also utilize DOM, including components released by bacteria and plants as metabolic end-products and photoassimilates, respectively. As a result, these DOM components may be more important as food for macrodecomposers than the microdecomposers themselves. HS may also benefit aquatic organisms by promoting their growth and protecting them from inimical forces. Conversely, the removal of photons and the release of toxins by HS may be detrimental to aquatic organisms. 7. It is concluded that the central dogma of the foodweb, and its implicit assumption that the energy flow in aquatic ecosystems can be quantified solely by measuring rates of photosynthesis, ingestion of solid food and its digestion by higher organisms, is invalid. To extend our understanding of the role of DOM as a source of nutrition and information to aquatic organisms it is suggested that the subject should be studied within the context of ‘modules’ which have the following properties: (i) the components have co-evolved; (ii) the more vulnerable components will have protective mechanisms; (iii) the components will derive mutual benefits from co-existence; (iv) sedentary components will release kairomonal attractants or developmental primers; (v) living components will exchange energy and information; (vi) the module will collapse following the removal of strongly interactive keystone species. An example of a three-component, three-subset module, is provided by tubificid worms, epilithic bacteria and algae. A more complex module consisting of pulmonate snails, associated macrophytes, their epiphytic bacteria and algae has four components and six subsets. The elucidation of the interactive mechanisms within such modules demands an interdisciplinary approach, involving microbiology, biochemistry and behavioural biology.