Identification of antiparasitic drug targets using a multi-omics workflow in the acanthocephalan model.

BACKGROUND: With the expansion of animal production, parasitic helminths are gaining increasing economic importance. However, application of several established deworming agents can harm treated hosts and environment due to their low specificity. Furthermore, the number of parasite strains showing resistance is growing, while hardly any new anthelminthics are being developed. Here, we present a bioinformatics workflow designed to reduce the time and cost in the development of new strategies against parasites. The workflow includes quantitative transcriptomics and proteomics, 3D structure modeling, binding site prediction, and virtual ligand screening. Its use is demonstrated for Acanthocephala (thorny-headed worms) which are an emerging pest in fish aquaculture. We included three acanthocephalans (Pomphorhynchus laevis, Neoechinorhynchus agilis, Neoechinorhynchus buttnerae) from four fish species (common barbel, European eel, thinlip mullet, tambaqui). RESULTS: The workflow led to eleven highly specific candidate targets in acanthocephalans. The candidate targets showed constant and elevated transcript abundances across definitive and accidental hosts, suggestive of constitutive expression and functional importance. Hence, the impairment of the corresponding proteins should enable specific and effective killing of acanthocephalans. Candidate targets were also highly abundant in the acanthocephalan body wall, through which these gutless parasites take up nutrients. Thus, the candidate targets are likely to be accessible to compounds that are orally administered to fish. Virtual ligand screening led to ten compounds, of which five appeared to be especially promising according to ADMET, GHS, and RO5 criteria: tadalafil, pranazepide, piketoprofen, heliomycin, and the nematicide derquantel. CONCLUSIONS: The combination of genomics, transcriptomics, and proteomics led to a broadly applicable procedure for the cost- and time-saving identification of candidate target proteins in parasites. The ligands predicted to bind can now be further evaluated for their suitability in the control of acanthocephalans. The workflow has been deposited at the Galaxy workflow server under the URL tinyurl.com/yx72rda7 .

What contributes to the metal-specific partitioning in the chub-acanthocephalan system?

Physiologically based pharmacokinetic (PBPK) models have been applied to simulate the absorption, distribution, metabolism, and elimination of various toxicants in fish. This approach allows for considering metal accumulation in intestinal parasites. Unlike "semi" physiologically-based models developed for metals, metal accumulation in fish was characterised based on metal-specific parameters (the fraction in blood plasma and the tissue-blood partition coefficient) and physiological characteristics of the fish (the blood flow and the tissue weight) in our PBPK model. In the model, intestinal parasites were considered a sink of metals from the host intestine. The model was calibrated with data for the system of the chub Squalius cephalus and the acanthocephalan Pomphorhynchus tereticolliis. Metal concentrations in this fish-parasite system were monitored in Ag and Co treatments in duplicate during a 48-day exposure phase (Ag and Co were added to tap water at concentrations of 1 and 2 µg/L, respectively) and a 51-day depuration phase. Their concentrations in the gills increased during the exposure phase and decreased in the depuration phase. A similar pattern was observed for Ag concentrations in other chub organs, while a relatively stable pattern for Co indicates regulations in the accumulation of essential metals by chubs. The metals were taken up by the acanthocephalans at similar rate constants. These results indicate that metal availability to parasites, which is determined by the internal distribution and fate, is critical to metal accumulation in the acanthocephalans. The high concentration of Ag in the liver as well as the high rate of Ag excretion from the liver to the intestine might contribute to higher concentrations of metals in the bile complexes in the intestine, which are available to the parasites, but not to the reabsorption by the host intestine. The opposite pattern might explain the lower availability of Co to the acanthocephalans.

The genome, transcriptome, and proteome of the fish parasite Pomphorhynchus laevis (Acanthocephala).

Thorny-headed worms (Acanthocephala) are endoparasites exploiting Mandibulata (Arthropoda) and Gnathostomata (Vertebrata). Despite their world-wide occurrence and economic relevance as a pest, genome and transcriptome assemblies have not been published before. However, such data might hold clues for a sustainable control of acanthocephalans in animal production. For this reason, we present the first draft of an acanthocephalan nuclear genome, besides the mitochondrial one, using the fish parasite Pomphorhynchus laevis (Palaeacanthocephala) as a model. Additionally, we have assembled and annotated the transcriptome of this species and the proteins encoded. A hybrid assembly of long and short reads resulted in a near-complete P. laevis draft genome of ca. 260 Mb, comprising a large repetitive portion of ca. 63%. Numbers of transcripts and translated proteins (35,683) were within the range of other members of the Rotifera-Acanthocephala clade. Our data additionally demonstrate a significant reorganization of the acanthocephalan gene repertoire. Thus, more than 20% of the usually conserved metazoan genes were lacking in P. laevis. Ontology analysis of the retained genes revealed many connections to the incorporation of carotinoids. These are probably taken up via the surface together with lipids, thus accounting for the orange coloration of P. laevis. Furthermore, we found transcripts and protein sequences to be more derived in P. laevis than in rotifers from Monogononta and Bdelloidea. This was especially the case in genes involved in energy metabolism, which might reflect the acanthocephalan ability to use the scarce oxygen in the host intestine for respiration and simultaneously carry out fermentation. Increased plasticity of the gene repertoire through the integration of foreign DNA into the nuclear genome seems to be another underpinning factor of the evolutionary success of acanthocephalans. In any case, energy-related genes and their proteins may be considered as candidate targets for the acanthocephalan control.

Differential mucins secretion by intestinal mucous cells of Chelon ramada in response to an enteric helminth Neoechinorhynchus agilis (Acanthocephala).

Intestinal mucous cells produce and secrete mucins which hydrate, lubricate and protect the intestinal epithelium from mechanical injuries due to the transition of digesta or action of pathogens. Intestinal mucous cells are considered elements of the innate immune system as they secrete lectins, toxins, immunoglobulins, and anti-microbial peptides. Acid mucins can surround and eliminate many pathogenic microorganisms. We performed a quantitative analysis of the density and mucus composition of different intestinal mucous cell types from mullet (Chelon ramada) that were infected solely with Neoechinorhynchus agilis. Most N. agilis were encountered in the middle region of the intestine. Mucous cell types were identified with Alcian Blue (pH2.5) and Periodic acid-Schiff (PAS) histochemistry, and by staining with a panel of seven lectins. Mucus enriched for high viscosity acid mucins was accumulated near points of worm attachment. Parasites were surrounded by an adherent mucus layer or blanket. Ultrastructural examination showed intestinal mucous cells typically possessed an elongated, basally positioned nucleus and numerous electron dense and lucent vesicles in the cytoplasm. The results show both an increase in mucus production and changes in mucin composition in infected mullet in comparison with uninfected conspecifics.

Sclerocollum saudii Al-Jahdali, 2010 (Acanthocephala: Cavisomidae) as a sentinel for heavy-metal pollution in the Red Sea.

Currently, fish helminth parasites, especially cestodes and acanthocephalans, are regarded as sentinel organisms to elucidate metal pollution in aquatic ecosystems. Here, 34 specimens of the fish Siganus rivulatus were collected in the Red Sea, from a seriously polluted, small lagoon named Sharm-Elmaya Bay, at Sharm El-Sheikh, South Sinai, Egypt; 22 (64.7%) were infected by Sclerocollum saudii (Acanthocephala: Cavisomidae). Thus, 22 natural infrapopulations (26-245 individuals) of this parasite were collected from infected fish. Samples of water and sediments from the bay, samples of muscle, intestine and liver from each fish, and samples from the parasite were taken for analysis of heavy metals (cadmium (Cd) and lead (Pb)). Both Cd and Pb concentrations in sediments were higher than those in water. The concentration of these metals were significantly higher in tissues (intestine, liver and muscle) of non-infected fish than those in infected fish, with Pb concentrations consistently higher than those of Cd, and both were drastically decreased in the order: liver > intestine > muscle. Metal concentrations in this acanthocephalan were much higher than those in its fish host. There were strong negative relationships between metal concentrations in tissues (intestine, liver and muscle) of infected fish and infrapopulation size, and between metal concentrations in the acanthocephalan and its infrapopulation size. These relationships strongly suggest competition for these metals between the fish host and its acanthocephalan parasite, and intraspecific competition among acanthocephalan individuals for available metals in the fish intestine. Bioconcentration factors were relatively high, since the mean Cd concentration in S. saudii was 239, 68 and 329 times higher than those in intestine, liver and muscle tissues, respectively, of its fish host. Also, mean Pb concentration was 55, 13 and 289 times higher than those in these tissues, respectively. The host-parasite system described here seems to be promising for biomonitoring of metal pollution in the Red Sea.

Host condition and accumulation of metals by acanthocephalan parasite Echinorhynchus gadi in cod Gadus morhua from the southern Baltic Sea.

In this study, we analyzed the relationship between concentration of metals in the host-parasite system (cod - acanthocephalan Echinorhynchus gadi) and Fulton's condition factor (FCF) of the host. The relationship between metal (Ca, Cd, Cr, Cu, Fe, Hg, K, Mg, Mn, Na, Pb, Sr, Zn) concentrations in E. gadi and cod tissues was expressed as a bioconcentration factor (BCF), the ratio of the concentration in the parasite tissue to that in host tissues. Acanthocephalans accumulated mainly toxic metals (Cd, Pb), as well as Sr, Ca, Na. Cadmium showed the highest bioconcentration in parasites (BCF >200) compared to fish muscle. Significant negative correlation was detected between FCF and the concentration of Cd and Hg in cod liver. In contrast, FCF was positively correlated with the concentration of Hg in acanthocephalan tissues.

Development and Validation of a Biodynamic Model for Mechanistically Predicting Metal Accumulation in Fish-Parasite Systems.

Because of different reported effects of parasitism on the accumulation of metals in fish, it is important to consider parasites while interpreting bioaccumulation data from biomonitoring programmes. Accordingly, the first step is to take parasitism into consideration when simulating metal bioaccumulation in the fish host under laboratory conditions. In the present study, the accumulation of metals in fish-parasite systems was simulated by a one-compartment toxicokinetic model and compared to uninfected conspecifics. As such, metal accumulation in fish was assumed to result from a balance of different uptake and loss processes depending on the infection status. The uptake by parasites was considered an efflux from the fish host, similar to elimination. Physiological rate constants for the uninfected fish were parameterised based on the covalent index and the species weight while the parameterisation for the infected fish was carried out based on the reported effects of parasites on the uptake kinetics of the fish host. The model was then validated for the system of the chub Squalius cephalus and the acanthocephalan Pomphorhynchus tereticollis following 36-day exposure to waterborne Pb. The dissolved concentration of Pb in the exposure tank water fluctuated during the exposure, ranging from 40 to 120 µg/L. Generally, the present study shows that the one-compartment model can be an effective method for simulating the accumulation of metals in fish, taking into account effects of parasitism. In particular, the predicted concentrations of Cu, Fe, Zn, and Pb in the uninfected chub as well as in the infected chub and the acanthocephalans were within one order of magnitude of the measurements. The variation in the absorption efficiency and the elimination rate constant of the uninfected chub resulted in variations of about one order of magnitude in the predicted concentrations of Pb. Inclusion of further assumptions for simulating metal accumulation in the infected chub led to variations of around two orders of magnitude in the predictions. Therefore, further research is required to reduce uncertainty while characterising and parameterising the model for infected fish.

Seasonal profile of metal accumulation in the acanthocephalan Pomphorhynchus laevis: a valuable tool to study infection dynamics and implications for metal monitoring.

BACKGROUND: A large number of studies demonstrated that acanthocephalans exhibit a high metal accumulation potential and thus can be used as sensitive accumulation indicators. However, similar to free-living bioindicators, a seasonal variation in metal concentrations in parasites might occur. Accordingly, the influence of seasonality has to be elucidated if parasites should be applied as sentinels. METHODS: In order to assess a possible seasonal profile of element concentrations, the concentrations of As, Cd, Co, Cu, Fe, Mn, Mo, Ni, Pb, V and Zn in the acanthocephalan Pomphorhynchus laevis and in its host barbel (Barbus barbus) were analysed in a seasonal manner (spring, summer and autumn) using inductively coupled plasma mass spectrometry (ICP-MS). RESULTS: Five elements (As, Cd, Cu, Pb and Zn) were detected in significantly higher concentrations in the parasites compared to host muscle, intestine and liver. Their levels in P. laevis showed a clear seasonal pattern, while the concentrations in the fish tissues remained similar during the year. The highest concentrations in the parasites were found in autumn, followed by spring and summer. Evidence from the literature suggests that this profile coincides with the seasonality of acanthocephalan transmission, as their annual concentration profile reflected the mean individual weight pattern during the year. Parasite infrapopulations in autumn consisted mainly of young worms which are characterised by an accelerated metabolism and a higher surface to volume ratio resulting in higher element concentrations when compared to older worms which are assumed to slow down their metabolism and additionally excrete metals with their eggs. CONCLUSIONS: Based on the available data from the present study and literature, a model is suggested, which visualises the accumulation kinetic of several elements under natural conditions. According to the element accumulation data the lifespan of P. laevis in barbel was roughly estimated to range between six and eight months.

Acanthocephalan Parasites (Acanthogyrus sp.) of Nile Tilapia (Oreochromis niloticus) as Biosink of Lead (Pb) Contamination in a Philippine Freshwater Lake.

The potential use of acanthocephalans as bioindicators of Lead (Pb) pollution in Sampaloc Lake, Laguna, Philippines was investigated. Nile tilapias (Oreochromis niloticus) were collected and Pb concentrations were determined in fish tissues and in their acanthocephalan parasites, Acanthogyrus sp. Significantly higher levels of Pb were detected in the parasites relative to the fish host tissues (p = 0.001). Bioaccumulation capacity of the parasites against fish tissues were 102, 119, and 147 times higher than the fish intestine, liver, and muscles, respectively. Pb sensitivity of the parasites was quantified by exact logistic analysis showing higher odds of Pb detection ranging from 18 to 45 folds (p = 0.001-0.009). Interestingly, infected fish showed significantly lower Pb concentration in their tissues compared to uninfected fish (p = 0.001), suggesting parasites were able to sequester Pb and served as active biosinks. The Pb levels in the parasites were also hundred folds higher (988 times) relative to the ambient waters, indicating a potential role of fish parasites as metal biosinks in aquatic ecosystems.

Host-parasite relationships as determinants of heavy metal concentrations in perch (Perca fluviatilis) and its intestinal parasite infection.

The concentrations of As, Cd, Cr, Cu, Hg, Mn, Ni, Pb, and Zn and their bioconcentration factors (BCFs) were determined in two intestinal parasites, an acanthocephalan, Acanthocephalus lucii, a tapeworm, Proteocephalus percae, present in the same host, the European perch (Perca fluviatilis, L.), in the heavily polluted Ruzín reservoir in eastern Slovakia. The bioaccumulation of heavy metals in the fish organs and parasites was studied for acanthocephalan and tapeworm monoinfections or mixed infections by the two parasites and for the size of their parasitic infrapopulations. Bioconcentration factors (c[parasite]/c[muscle tissue]) showed that the concentrations of As, Ni, Pb and Zn were higher in mixed infections than in monoinfections. Negative correlations between heavy metal concentrations in perch organs and the parasites were found. For example, higher concentrations of Ni and Zn in both parasite species corresponded with lower metal concentrations in perch and hard roe. Likewise, significant negative relationships between metal concentrations in fish organs and number of parasites were noticed with lower levels of Pb in fish harbouring higher numbers of tapeworms. Similarly, in both parasite species the concentrations of some essential elements (Cr, Mn) were lower at high infection intensities compared to low intensities. Our study revealed that the differential concentration of heavy metals in perch organs was affected by the type of infection (mono- or mixed-infection), and needs to be considered in field ecotoxicological and parasitological studies as a potentially important factor influencing the pollutant concentrations in fish.

[COMPARATIVE ANALYSIS OF LIPID METABOLISM INDICES IN SOME PARASITES OF THE WHITE CHARR (SALVELINUS ALBUS) FROM THE LAKE KRINOTSKOE].

Comparative study of lipid metabolism indices (total lipids, separate lipid fractions, level of the lipid peroxidation processes, and antioxidant protection) was carried out in three parasite species collected from the white char in the Lake Kronotskoe: Diphyllobothrium ditremum Crepin, 1825 (Cestoda), Philonema oncorhynchi Kuitunen-Ekbaum, 1933 (Nematoda) H Neoechinorhynchus salmonis Ching, 1984 (Acanthocephala). Acanthocephalans possessed significantly greater levels of total lipids, triacylglycerol, and malondialdehyde; nematodes, of cholesterol and sterol esters; and cestodes, in phospholipids and constants of the substrate oxidation. Dependence between lipid metabolism of helminths and their taxonomic affiliation, morpho-functional features, the stage of the life cycle, and the site of infection in the host are discussed.

Acetylcholinesterase activity in the host-parasite system of the cod Gadus morhua and acanthocephalan Echinorhynchus gadi from the southern Baltic Sea.

Acetylcholinesterase (AChE) activity measurement is widely used as a specific biomarker of neurotoxic effects. The aim of this study was to evaluate AChE activity in a host fish (the cod) and its acanthocephalan parasite Echinorhynchus gadi from the southern Baltic. AChE activity in hosts and parasites was inversely related: the highest cod AChE activity corresponded to the lowest E. gadi enzymatic activity and vice versa ("mirror effect"). This is the first report on the simultaneous application of this biomarker in cod and its acanthocephalan parasites. Results obtained for the host-parasite system are complementary and provide comprehensive information about the response of this biomarker. Analysis of the system allows for detection of a greater number of factors influencing AChE activity in the marine environment than separate analysis of the host and parasites. Thus, AChE activity measurement in a host-parasite system may be considered to be a promising tool for biomonitoring.

Comparison of the metal accumulation capacity between the acanthocephalan Pomphorhynchus laevis and larval nematodes of the genus Eustrongylides sp. infecting barbel (Barbus barbus).

BACKGROUND: Metal uptake and accumulation in fish parasites largely depends on the parasite group with acanthocephalans showing the highest accumulation rates. Additionally, developmental stage (larvae or adult) as well as parasite location in the host are suggested to be decisive factors for metal bioconcentration in parasites. By using barbel (Barbus barbus) simultaneously infected with nematode larvae in the body cavity and adult acanthocephalans in the intestine, the relative importance of all of these factors was compared in the same host. METHODS: Eleven elements Arsenic (As), Cadmium (Cd), Cobalt (Co), Copper (Cu), Iron (Fe), Manganese (Mn), Lead (Pb), Selenium (Se), Tin (Sn), Vanadium (V) and Zinc (Zn) were analyzed in barbel tissues (muscle, intestine, liver) as well as in their acanthocephalan parasites Pomphorhynchus laevis and the larval nematode Eustrongylides sp. (L4) using inductively coupled plasma mass spectrometry (ICP-MS). RESULTS: Nine elements were detected in significantly higher levels in the parasites compared to host tissues. The element composition among parasites was found to be strongly dependent on parasite taxa/developmental stage and localization within the host. Intestinal acanthocephalans accumulated mainly toxic elements (As, Cd, Pb), whereas the intraperitoneal nematodes bioconcentrated essential elements (Co, Cu, Fe, Se, Zn). CONCLUSION: Our results suggest that in addition to acanthocephalans, nematodes such as Eustrongylides sp. can also be applied as bioindicators for metal pollution. Using both parasite taxa simultaneously levels of a wide variety of elements (essential and non essential) can easily be obtained. Therefore this host-parasite system can be suggested as an appropriate tool for future metal monitoring studies, if double infected fish hosts are available.

Effect of Acanthocephalus lucii infection on total mercury concentrations in muscle and gonads of fish host (Perca fluviatilis).

The samples from 13 perches (Perca fluviatilis) - muscle with skin and bones; fish gonads; and acanthocephalan parasites were analysed for mercury (Hg). Hg concentrations were present in all analysed samples. There were found no statistically significant difference in Hg concentration in fish tissues between perches either with or without infection by the acanthocephalan parasite, Acanthocephalus lucii). In this study there was no evidence that acanthocephalan worms accumulate mercury from hosts. For this reason, A. lucii is not a suitable bioindicator for mercury pollution.

Intestinal parasite Acanthocephalus lucii (Acanthocephala) from European perch (Perca fluviatilis) as a bioindicator for lead pollution in the stream "Jevanský potok" near Prague, Czech Republic.

Lead concentrations in the tissues of perch and its parasites were determined as mg/kg dw. Lead was found at higher concentrations in the acanthocephalans (11.56) than in different tissues (liver, gonads and muscle with skin and bone) of perch. With respect to fish tissues, the highest concentrations of lead were present in the liver (1.24), followed by the gonads (0.57) whereas the lowest concentrations were in the muscle with skin and bone (0.21). The bioconcentration factors for lead indicated that parasites accumulate metals to a higher degree than fish tissues--lead concentrations in acanthocephalans were 9.32, 19.27 and 55.05 higher than in liver, gonads and muscles of host, respectively.

Is metal accumulation in Pomphorhynchus laevis dependent on parasite sex or infrapopulation size?

Concentrations of the elements As, Cd, Co, Cu, Fe, Mn, Mo, Ni, Pb, Sn, V, Zn were analysed by inductively coupled plasma mass spectrometry (ICP-MS) in the acanthocephalan Pomphorhynchus laevis and its fish host Barbus barbus. A total of 27 barbels were collected from the Danube River in autumn 2006 close to the town Kozloduy (685 river kilometer) on the Bulgarian river bank. Fish were divided into 3 groups. According to their P. laevis infrapopulation size hosts were considered as heavily infected (>100 worms per fish) and lightly infected (<20 worms per fish). The third group was used to compare heavy metal concentrations between male and female P. laevis. The 5 elements As, Cd, Cu, Pb and Zn were detected in significantly higher concentrations in parasites compared to host tissues (muscle, intestine, liver). According to the calculated mean bioconcentration factors, 3 more elements (Co, Mn, V) showed usually higher concentrations in P. laevis. Comparisons between heavily and lightly infected fish revealed significant differences only for V with higher concentrations for the heavily infected group. Concerning sex-specific metal accumulation V and Zn showed significant differences (V, at P<0.05; Zn, at P=0.05), with higher levels of both metals in females of P. laevis. Our results suggest that - for the metals analysed - the size of the parasite infrapopulation plays no role in the degree of metal accumulation. Similarly, parasite sex seems not to be a crucial factor for metal accumulation in the parasites. Thus, for metal monitoring purposes there is no need to take these aspects into account, which makes the use of parasites as bioindicators more applicable.

Carotenoid-based colour of acanthocephalan cystacanths plays no role in host manipulation.

Manipulation by parasites is a catchy concept that has been applied to a large range of phenotypic alterations brought about by parasites in their hosts. It has, for instance, been suggested that the carotenoid-based colour of acanthocephalan cystacanths is adaptive through increasing the conspicuousness of infected intermediate hosts and, hence, their vulnerability to appropriate final hosts such as fish predators. We revisited the evidence in favour of adaptive coloration of acanthocephalan parasites in relation to increased trophic transmission using the crustacean amphipod Gammarus pulex and two species of acanthocephalans, Pomphorhynchus laevis and Polymorphus minutus. Both species show carotenoid-based colorations, but rely, respectively, on freshwater fish and aquatic bird species as final hosts. In addition, the two parasites differ in the type of behavioural alteration brought to their common intermediate host. Pomphorhynchus laevis reverses negative phototaxis in G. pulex, whereas P. minutus reverses positive geotaxis. In aquaria, trout showed selective predation for P. laevis-infected gammarids, whereas P. minutus-infected ones did not differ from uninfected controls in their vulnerability to predation. We tested for an effect of parasite coloration on increased trophic transmission by painting a yellow-orange spot on the cuticle of uninfected gammarids and by masking the yellow-orange spot of infected individuals with inconspicuous brown paint. To enhance realism, match of colour between painted mimics and true parasite was carefully checked using a spectrometer. We found no evidence for a role of parasite coloration in the increased vulnerability of gammarids to predation by trout. Painted mimics did not differ from control uninfected gammarids in their vulnerability to predation by trout. In addition, covering the place through which the parasite was visible did not reduce the vulnerability of infected gammarids to predation by trout. We discuss alternative evolutionary explanations for the origin and maintenance of carotenoid-based colorations in acanthocephalan parasites.

[The role of parasites in monitoring of environmental pollution]. / Cevre Kirliliginin Izlenmesinde Parazitlerin Rolü

Heavy metals are known for their toxic effects on organisms. The toxic metals such as lead, cadmium and mercury are continuously released into the aquatic environment from different sources. The relationship between parasitism and environmental pollution has been of interest during recent years. Different helminth species, especially Acanthocephala, have been investigated in respect to heavy metal accumulation in their bodies. The importance of parasites for monitoring of environmental pollution was discussed in the present article.

Pollution-induced heat shock protein expression in the amphipod Gammarus roeseli is affected by larvae of Polymorphus minutus (Acanthocephala).

The relationship between the exposure of organisms to chemicals and subsequent alterations in various biochemical processes (commonly referred to as biomarkers) is of growing importance in environmental and ecotoxicological research. However, parasites which also affect the physiological homeostasis of their hosts, and thus may alter biomarker reactions, are usually ignored in environmental research. To address this deficit, we have used the host-parasite system Gammarus roeseli naturally infected with cystacanths of the acanthocephalan Polymorphus minutus to investigate whether infection of gammarids with parasites alters their heat shock protein response following exposure to palladium (Pd). After 24 days of metal exposure relative levels of heat shock protein hsp70 were analysed in the tissues of parasites and intermediate hosts. Simultaneously, the metal accumulation in gammarids and parasites was determined. As none of the infected gammarids showed hsp70 levels at the end of the Pd exposure (either exposed or not), infected and unparasitized G. roeseli were exposed to heat. Again, only uninfected gammarids showed a temperature-dependent increase in hsp70 levels. Interestingly, although the intermediate hosts showed no hsp70 response, exposure to Pd and heat results in increasing hsp70 in the parasites within in the haemocoel of G. roeseli. Heat experiments with isolated cystacanths also showed increasing hsp70 levels in P. minutus with temperature. Concerning uninfected G. roeseli, exposure to Pd and heat causes a hsp70 response. Pd concentrations were found to be higher in the larval parasites than in the gammarids. This result clearly contradicts previous results, as high metal accumulation was so far only described from adult acanthocephalans. Our findings provide experimental evidence that parasites alter the biomarker responses of their host and that the infection status of test animals is extremely important for ecotoxicological studies.

Accumulation of the precious metals platinum, palladium and rhodium from automobile catalytic converters in Paratenuisentis ambiguus as compared with its fish host, Anguilla anguilla.

The platinum group metals (PGM) Pt, Pd and Rh are emitted into the environment mainly by catalytic exhaust gas converters of cars. As PGM accumulate in sediments of aquatic ecosystems, the study was focused on the uptake of the noble metals by European eels, Anguilla anguilla infected with the acanthocephalan Paratenuisentis ambiguus. Eels were exposed to ground catalytic converter material for six weeks. After exposure Pt and Pd were detected in the liver and kidney of the eels and in the parasites. Palladium was also found in fish muscle and intestine. No Rh uptake by the eel tissues and the parasites occurred. Paratenuisentis ambiguus contained the highest levels of both metals with 40 times higher Pt concentrations and four times higher Pd concentrations than the liver of its host. Due to its accumulation capacity for PGM, P. ambiguus can be applied as a sensitive accumulation indicator in field studies to assess the degree of environmental PGM contamination in aquatic ecosystems.