Copper regulation and homeostasis of Daphnia magna and Pseudokirchneriella subcapitata: influence of acclimation

This study aimed to evaluate (1) the capacity of the green alga Pseudokirchneriella subcapitata and the waterflea Daphnia magna to regulate copper when exposed to environmentally realistic copper concentrations and (2) the influence of multi-generation acclimation to these copper concentrations on copper bioaccumulation and homeostasis. Based on bioconcentration factors, active copper regulation was observed in algae up to 5 microg Cu L(-1) and in daphnids up to 35 mug Cu L(-1). Constant body copper concentrations (13+/-4 microg Cu g DW(-1)) were observed in algae exposed to 1 through 5 microg Cu L(-1) and in daphnids exposed to 1 through 12 microg Cu L(-1). At higher exposure concentrations, there was an increase in internal body copper concentration, while no increase was observed in bioconcentration factors, suggesting the presence of a storage mechanism. At copper concentrations of 100 microg Cu L(-1) (P. subcapitata) and 150 microg Cu L(-1) (D. magna), the significant increases observed in body copper concentrations and in bioconcentration factors may be related to a failure of this regulation mechanism. For both organisms, internal body copper concentrations lower than 13 microg Cu g DW(-1) may result in copper deficiency. For P. subcapitata acclimated to 0.5 and 100 microg Cu L(-1), body copper concentrations ranged (mean+/-standard deviation) between 5+/-2 microg Cu g DW(-1) and 1300+/-197 microg Cu g DW(-1), respectively. For D. magna, this value ranged between 9+/-2 microg Cu g DW(-1) and 175+/-17 microg Cu g DW(-1) for daphnids acclimated to 0.5 and 150 microg Cu L(-1). Multi-generation acclimation to copper concentrations >or =12 microg Cu L(-1) resulted in a decrease (up to 40%) in body copper concentrations for both organisms compared to the body copper concentration of the first generation. It can be concluded that there is an indication that P. subcapitata and D. magna can regulate their whole body copper concentration to maintain copper homeostasis within their optimal copper range and acclimation enhances these mechanisms.     

Effects of pulp mill chlorate on Baltic Sea algae

The long-term effects of pulp mill chlorate on different algal species of the Baltic Sea were studied in land-based model ecosystems simulating the littoral zone. Brown algae (Phaeophyta) exhibited an extraordinarily high sensitivity to chlorate and pulp mill effluents containing chlorate. All brown algal species ceased growth or showed major signs of toxicity at all concentrations tested, down to microgram per litre levels. EC50 levels for growth of Fucus vesiculosus were about 80-100 microg ClO3- litre(-1). Blue-green algae (Cyanophyta) were not deleteriously affected nor were green algae (Chlorophyta). The perennial and annual species of red algae (Rhodophyta) were also unaffected by the effluents. Diatoms did not show any sensitivity and phytoplankton (fresh- and brackish water) were particularly insensitive. A phanerogam, Zostera marina was also unaffected by the treatments.     

Accumulation of lead and cadmium in the marine prosobranch Nerita saxtilis, chemical analysis, light and electron microscopy.

The potential value of the marine prosobranch Nerita saxtilis as an efficient biological monitor to heavy metal pollution in the Red Sea was investigated. Storage ability of lead and cadmium was compared in shell, headfoot and digestive gland of the marine snail N. saxtilis collected from Al-Hamrauin area at El-kuseir (lead, 300.35 +/- 28.53 microg/l, 1,716 +/- 16.14. cadmium 20.01 +/- 1.8 microg/l, 161.72 +/- 21.4 mean +/- S.D. for water and sediment, respectively) relative to that of inhabiting marine water and sediment employing atomic absorption spectrometry to determine the organ with highest capability of heavy metal accumulation. The influence of metal storage on light microscopic structure of that organ was investigated. Also, the ultrastructure localization of storage sites in the same organ was determined employing transmission electron microscopy. The digestive gland was shown to accumulate both metals at conccntrations that are several orders of magnitude higher than those in the surrounding marine water. The bioaccumulation capability of lead and cadmium was ranked in the following order; digestive gland > headfoot > shell for lead and digestive gland > shell > headfoot for cadmium. In spite of its evident highest metal storage capability, no histopathological changes were observed in the digestive gland of that marine prosobranch. Enlarged electron dense vesicles and many granules were observed in ultrathin sections in digestive cells of these snails and are suggested to be the sites of storage of detoxified metals. The results of that finding indicate the possibility of using the marine prosobranch N. saxtilis as biomonitor for heavy metal contaminants in the Red Sea.     

Mercury bioaccumulation and decontamination kinetics in the edible cockle Cerastoderma edule

Mercury bioaccumulation and decontamination kinetics in the edible cockle Cerastoderma edule were studied through a mesocosms experiment after a medium-term exposure to the metal.The results revealed that the bivalve presented distinct bioaccumulation kinetics according to the different tissues. While the gills showed a linear accumulation pattern, the digestive gland and the entire organism presented a saturation model, with higher accumulation during the first 7d of exposure and lower during the rest of the time. In addition, the bioaccumulation rate was not proportional to the Hg concentration, since the organisms under lower contamination presented higher bioconcentration factors than the ones under higher contamination. Gills were the tissues with higher mercury accumulation capability.Concerning the decontamination phase, C. edule lost approximately 80% of the mercury after 24 h exposure in clean seawater. Nevertheless, never reached the original condition, showing in the final (20 d detox), Hg levels (>0.5 ppm) higher than those allowed by the legislation regulating human food consumption. This represents a matter of concern for Human health.     

S-metolachlor pulse exposure on the alga Scenedesmus vacuolatus: effects during exposure and the subsequent recovery

In streams and creeks, the aquatic flora is exposed to fluctuating concentrations of herbicides during and following their application. Peak concentrations of herbicides, like the chloroacetanilide S-metolachlor, are usually detected following rain events. In this study, we assessed the effect of S-metolachlor pulse exposure on the algae Scenedesmus vacuolatus. We measured the time-dependency of effects during exposure on algae population and identified the algae development stage most sensitive to S-metolachlor. Furthermore, we assessed the time-to-recovery of the algae following exposure. A 6h pulse exposure at 598mugl(-1) was sufficient to inhibit cell reproduction by 50%. However, the exposure period had to coincide with the cell development stage specifically inhibited by S-metolachlor, which is the end of the cell growth phase. In algae populations composed of cells at all development stages, we initially observed an increase in the size of some algal cells, ultimately leading to an inhibition of the growth rate. In these experimental conditions, effects were observed after 18h of exposure and greatly increased with time. The recovery of algae following exposure to strongly inhibiting S-metolachlor concentrations was delayed and only occurred after 29h. These findings suggest that peak exposure to S-metolachlor may affect the growth of sensitive alga in surface waters, considering that the effects extend beyond the period of exposure.     

Distribution of organotin compounds in tissues of mussels Mytilus edulis and clams Mya arenaria

Concentrations and composition of butyltin (BT) and phenyltin (PT) compounds were compared in tissues of mussels Mytilus edulis and clams Mya arenaria collected from the Mersey Estuary. Tributyltin (TBT) concentrations were consistently highest in digestive gland and low in muscular tissues such as siphons (Mya) and adductor muscle (Mytilus). Lowest concentrations were found in the periostracum surrounding the siphons of Mya. Ratios of monobutyltin (MBT), dibutyltin (DBT) and TBT to total BT content of Mytilus tissues were in the ranges 25-34%, 21-28% and 44-51%, respectively. Much higher proportions of TBT:total BT were characteristic of Mya (>80% in the digestive gland, remaining tissue and gonad) implying that slow degradation rates account for the relatively high levels of TBT in this species. TPT was the only phenyltin compound detected (albeit at relatively low levels), and, like TBT, was present in highest concentrations in the digestive gland of clams--suggesting an important dietary component in the accumulation of OTs in these bivalves.     

A solvent-free method for spiking terrestrial algae (Desmococcus spp.) with pyrene for use in bioassays

Two methods for spiking terrestrial algae (Desmococcus spp., used as a foodstuff in bioassays with Collembola) with pyrene were tested; a "traditional" method that used a carrier solution of pyrene in acetone, and a solvent-free method developed using the principles of partition driven administration (PDA). The PDA method used a pre-spiked, highly-contaminated C18 disk as the source for pyrene, suspended and rotated in an aqueous algal suspension. The pyrene partitioned from the C18 disk into the aqueous phase, and a concentration of 18.4 +/- 0.7 microg pyrene g(-1)dwt. algae (mean +/- sd) was achieved after 18 h exposure by this method, with good reproducibility also observed for lower concentrations achieved over shorter exposure periods. The acetone-spiked algae lost much of its cell integrity and a significant amount of pigment, while cell integrity was maintained after 120 h spiking using the PDA method. Results from a short bioassay with Orchesella cincta (Collembola, Insecta) showed that animals fed acetone-spiked algae had slower growth rates and higher concentrations of pyrene metabolites than animals fed PDA-spiked algae. It is speculated that this was the result of the poor quality of the acetone-spiked food, and that the difference in food quality between treatments stimulated changes in body composition that may have affected the production of pyrene metabolites. The PDA spiking method is expected to be suitable for introducing a range of persistent organic pollutants into other types of sample matrix.     

Comparision of the waterborne and dietary routes of exposure on the effects of Benzo(a)pyrene on biotransformation pathways in Nile tilapia (Oreochromis niloticus)

BaP is one of the most studied PAH, due to its ubiquitous presence in aquatic environments and toxicity to aquatic organisms. The main goal of this study was to assess BaP effects in Nile Tilapia after waterborne and dietary exposures, through the evaluation of EROD and GST activities in liver, gills and intestine, and BaP metabolites in bile; and also to evaluate the usefulness of these commonly used biomarkers after two different routes of exposure. Waterborne exposure to BaP led to a significant induction of EROD in all tissues analyzed (644%, 1640% and 2880% in relation to solvent in liver, gill and intestine respectively) while in dietary exposures EROD was induced only in intestine (3143%) after exposure to high BaP concentrations. GST activities with CDNB were slightly induced in liver (40%) and in gill (66%) after water exposure to BaP, and in intestine after dietary exposure to low BaP concentrations (182%). BaP metabolites in bile increased after both exposure routes, and were highly correlated with EROD activity after water exposure. In summary, this work has shown that the effects of BaP on biotransformation pathways depend on the route of exposure. Moreover, barrier tissues like gills and intestine also have an important role in the first-pass metabolism of BaP, reducing the amount of parent compound that reaches the liver to be metabolized. For that reason, EROD activity as a biomarker of exposure should also be applied in extrahepatic organs, like gills and intestine, in monitoring studies. Biliary BaP type metabolites are good reflectors of contamination levels under both exposure routes, while GST activity with CDNB as substrate, as a phase II enzyme, does not seem a reliable biomarker of exposure to BaP regardless the route of exposure.     

Enhanced degradation of benzo[a]pyrene by Mycobacterium sp. in conjunction with green alga

Previous work in this laboratory has confirmed that the bacteria Mycobacterium sp. strain RJGII.135 and Sphingomonas yanoikuyae strain B1 and the green alga Selanastrum capricornutum strain UTEX 1648 degrade benzo[a]pyrene (BaP) to various BaP intermediates. S. capricornutum was first grown with BaP for 4 days. The organic extract of this media was then introduced into separate cultures of strain RJGII.135 and strain B1; separate cultures were grown with BaP for comparison. Cultures grown with BaP and those grown with the algal/BaP extract showed similar mineralization patterns. The quantity of total metabolites formed was greater in bacterial cultures grown with the algal/BaP extract than those grown with BaP alone. For strain RJGII.135, only 27% of the original BaP remained in cultures grown with the algal/BaP extract; 59% remained in cultures grown with BaP. For strain B1, only 6% of the original BaP remained in cultures grown with the algal/BaP extract; 38% remained in cultures grown with BaP. These results indicate that strategies utilizing organisms together may be necessary in being able to degrade large, recalcitrant polycyclic aromatic hydrocarbons (PAHs) such as BaP.     

Elemental concentrations in different species of seaweeds from Loreto Bay, Baja California Sur, Mexico: implications for the geochemical control of metals in algal tissue

Concentration levels of 21 elements were analyzed by instrumental neutron activation analysis (Rb, Cs, Ca, Sr, Ba, Sc, Cr, Fe, Co, Ni, Zn, Se, As, Sb, Th, U, Br, Hf, Ta, Zr, and Ag) in seven different seaweed species (Codium cuneatum, Sargassum sinicola, Padina durvillaei, Laurencia johnstonii, L. papillosa, Gracilaria pachidermatica and Hypnea pannosa), collected in a shallow coastal zone from Bahia de Loreto, Baja California Sur, Mexico. Measured concentrations in algal tissue spanned almost eight orders of magnitude (from 2.0 x 10(-3) microg g(-1) for Hf to 1.2 x 10(5) microg g(-1) for Ca). Ca was consistently the most abundant element in all analyzed seaweeds, followed by Fe and Sr. Brown algae showed a tendency to incorporate higher concentrations of elements than red and green algae. Additionally, there were significant linear correlations (P < 0.05 to P < 0.001) between a total of 76 different pairs of elements, some of them (e.g. Rb-Ni, Rb-Ag, Sc Cr, Sc-Fe, Sc Ni, Sc Hf, Cr Fe, Fe Ni, Fe-Hf and Ni-Th) highly correlated (r2 > 0.900). A significant correlation (r2 = 0.701, n = 18, P < 0.001) exists between our measurements in the tissue of algae and their corresponding average elemental concentrations in oceanic water from the North Pacific Ocean. Hence, overall elemental abundance in algal tissue apparently is controlled by the elemental abundance in oceanic water, whereas metabolic processes as well as environmental factors relevant to each region modify the final concentration of a given element in the body of a macroalgae.     

Toxicity of imidacloprid to the terrestrial isopod Porcellio scaber (Isopoda, Crustacea)

Imidacloprid is a neonicotinoid insecticide with neurotoxic action that, as a possible alternative for commonly used organophosphorus pesticides, has gained registration in about 120 countries for use in over 140 agricultural crops. Only few data are available on its toxicity for soil invertebrates. We therefore assessed the effects of imidacloprid on survival, weight gain, feeding rate, total protein content, glutathione S-transferase activity (GST), and digestive gland epithelial thickness in juveniles and adults of the terrestrial isopod Porcellio scaber. After two weeks of feeding on imidacloprid-dosed food, weight gain (NOEC 5 microg/g dry food) and feeding rate (NOEC 10 microg/g) in juveniles, and feeding rate (NOEC<10 microg/g) and digestive gland epithelial thickness (NOEC<10 microg/g) in adults were most affected. In juveniles induction of GST activity and increase of total protein content per wet animal weight was detected at 5 microg/g dry food, whereas in adults a reduction of GST was observed at 25 microg/g (NOEC 10 microg/g). An estimate of actual intake rates suggests that imidacloprid affects isopods at similar exposure concentrations as insects. The toxicity of imidacloprid was similar to that of the organophosphorus pesticide diazinon, tested earlier using the same methods [Stanek, K., Drobne, D., Trebse, P., 2006. Linkage of biomarkers along levels of biological complexity in juvenile and adult diazinon fed terrestrial isopod (Porcellio scaber, Isopoda, Crustacea). Chemosphere 64, 1745-1752]. At actual environmental concentrations, diazinon poses a higher risk to P. scaber. Due to its increasing use in crop protection and higher persistence in soil, imidacloprid might however, be potentially more dangerous after long-term application. We conclude that toxicity testing with P. scaber provides relevant, repeatable, reproducible and comparable toxicity data that is useful for the risk assessment of pesticides in the terrestrial environment.     

Bioaccumulation studies of organochlorinated pesticides in tissues of Cyprinus carpio

Freshwater fish Cyprinus carpio was selected for the study of bioaccumulation of organochlorinated pesticides in tissues like gills, muscle, intestine, kidney, and liver in a continuous fed system. The pesticides used were Aldrin, Dieldrin, BHC, and DDT. The bioaccumulation of Dieldrin was maximum of 85.0 microg g(-1) wet weight in liver tissue while minimum of 7.30 microg g(-1) wet weight for DDT at 30 days exposure time. Bioconcentration factor (BCF) has followed the same trend in liver tissue for Dieldrin and DDT. The rate of bioaccumulation was found to be maximum of 4.3879 microg g(-1) wet weight in liver tissue and minimum of 0.0021 microg g(-1) wet weight in gill tissue for 30 days exposure. As evidenced by the increasing values of BCF, pesticide uptake also showed increased trend with the increase in exposure time. A high correlation coefficient ranging between 0.7247 and 0.9616 between the pesticide concentration and exposure time was observed. Based on actual BCF values, log Kow were calculated and the values are well within the reported values of 6.5 indicating efficient relationship between BCF and log Kow because beyond the 6.5 the bioconcentration levels off.     

Macro, minor and toxic elemental uptake and distribution in Hypoxis hemerocallidea, "the African Potato"--an edible medicinal plant

The elemental uptake and distribution, in various parts of the admired herbal plant, Hypoxis hemerocallidea, the 'African potato' and its ability to accumulate elements in response to the growth soil quality are investigated. The total and exchangeable concentrations of twelve elements in the growth soils and their distribution in the roots, potato bulb and leaves of the plants grown under four different settings were compared. The typical concentrations of the twelve selected elements, in the bulb and leaves of the plant grown in a nursery pot (site 2) were (in microg g(-1)dry weight) Ca (8430 and 27075), Mg (2113 and 1566), Fe (66 and 150), Al (10 and 368), Zn (105 and 6.1), Mn (42 and 51), Cu (7.2 and 20.8), Ba (0.23 and 4.44), Co (0.20 and 0.42), As (2.05 and 24.56), Hg (0.92 and 1.82) and Cr (0.13 and 0.33). Except for Ca, Mg, Zn and Mn, the exchangeable cation concentrations in all the growth soils were low. Ca, Mg, Mn, Zn and As had bioaccumulation factors >1. Fe, Al and Co concentrations were high in the roots with little in the rest of the plant. High concentration of arsenic (approximately 13 microg g(-1) dry weight) with bioaccumulation factors of 7 and 20 were observed in the roots and leaves of the plant respectively (site 2), but the concentration of mercury in bulb was very low (0.92 microg g(-1) dry weight).     

Bioaccumulation of aluminium in the freshwater snail Lymnaea stagnalis at neutral pH

This study examined the accumulation of aluminium (Al) by the freshwater snail Lymnaea stagnalis at neutral pH, when most Al would be predicted to be in an insoluble form (Al(OH)(3)). Snails were exposed to a range of Al concentrations (38-285 microg l(-1)) for 30 days, followed by 20 days in clean water. Aluminium was measured using atomic absorption spectroscopy. Significant accumulation of Al occurred in the whole soft tissues, gut, digestive gland and kidney at the latest by day 10. High concentration factors were observed, ranging from 4.5 x 10(3) in the whole soft tissues to 6.3 x 10(4) in the kidney, corresponding to actual concentrations of 800 to 7500 microg g(-1), respectively. Proportionality between environmental (water) and tissue concentrations of Al was observed in the gut but not in the other tissues. Following transfer to clean water, rapid loss of Al from the whole soft tissues and gut was seen over the first 10 days. Loss of Al from the digestive gland was much less as a proportion of the total, with approximately 90% of the Al remaining in the tissue. In contrast, significant loss of Al from the kidney occurred between days 20 and 30, even in the continued presence of Al; little further loss occurred following transfer to clean water. Aluminium is clearly available to the snail at neutral pH, the most likely route of entry being the gut. This could facilitate entry of the metal into the food chain. The possible roles of the digestive gland and kidney in the handling of Al are discussed.     

Exposure of caged mussels to metals in a primary-treated municipal wastewater plume

The biological availability of metals in municipal wastewater effluents is strongly influenced by the physical and chemical conditions of both the effluent and the receiving water. Aquatic organisms are exposed to both dissolved and particulate (food ingestion) forms of these metals. In the present study, the distribution of metals in specific tissues was used to distinguish between exposure routes (i.e. dissolved vs. particulate phase) and to examine metal bioavailability in mussels exposed to municipal effluents. Caged Elliptio complanata mussels were deployed at sites located between 1.5 km upstream and 12 km downstream of a major effluent outfall in the St. Lawrence River. Metals in surface water samples were fractionated by filtration techniques to determine their dissolved, truly-dissolved (<10 kDa), total-particulate and acid-reactive-particulate forms. At the end of the exposure period (90 days), pooled mussel soft tissues (digestive gland, gills, gonad, foot and mantle) were analyzed for several metals. The results showed that gills and digestive gland were generally the most important target tissues for metal bioaccumulation, while gill/digestive gland metal ratios suggest that both exposure routes should be considered for mussels exposed to municipal effluents. We also found that Ag and Cd in the dispersion plume nearest the outfall, in contrast to other metals such as Cu and Zn, are more closely associated with colloids and were generally less bioavailable than at the reference site in the St. Lawrence River.     

Bioaccumulation of PCBs in the cuttlefish Sepia officinalis from seawater, sediment and food pathways

The cuttlefish Sepia officinalis was selected as a model cephalopod to study PCB bioaccumulation via seawater, sediments and food. Newly hatched, juvenile cuttlefish were exposed for 17 days to environmentally realistic concentrations of (14)C-labeled 2,2',4,4',5,5'-hexachlorobiphenyl (PCB#153) (18 ng PCB l(-1) seawater; 30 ng PCB g(-1) dry wt sediments; Artemia salina exposed to 18 ng PCB l(-1) seawater). Accumulation of PCB#153 was followed in three body compartments: digestive gland, cuttlebone and the combined remaining tissues. Results showed that (1) uptake kinetics were source- and body compartment-dependent, (2) for each body compartment, the accumulation was far greater when S. officinalis was exposed via seawater, (3) the cuttlebone accumulated little of the contaminant regardless of the source, and (4) the PCB congener showed a similar distribution pattern among the different body compartments following exposure to contaminated seawater, sediment or food with the lowest concentrations in the cuttlebone and the highest in the remaining tissues. The use of radiotracer techniques allowed delineating PCB kinetics in small whole organisms as well as in their separate tissues. The results underscore the enhanced ability of cephalopods to concentrate organic pollutants such as PCBs, and raise the question of potential risk to their predators in contaminated areas.     

On the way to cyanobacterial blooms: impact of the herbicide metribuzin on the competition between a green alga (Scenedesmus) and a cyanobacterium (Microcystis)

The hypothesis that exposure to a common and widely applied photosynthesis-inhibiting herbicide, metribuzin, would alter the outcome of the competitive battle between susceptible green algae (Scenedesmus obliquus) and tolerant cyanobacteria (Microcystis aeruginosa) was tested. In a long-term (17 d) experiment, Scenedesmus and Microcystis populations as well as mixtures that started with different inoculum composition (i.e. 3:1, 1:1 and 1:3 Scenedesmus:Microcystis) were grown in the absence or presence of metribuzin (100 microg l-1). In the absence of metribuzin, Scenedesmus was competitively superior and out-competed Microcystis regardless the initial composition of the mixed communities. However, this competitive outcome was reversed completely in the presence of metribuzin, where despite growth inhibition Microcystis became dominant. Hence, photosynthesis-inhibiting herbicides may not only affect algal community structure, but also provide cyanobacteria founder populations a window for dominance and thus play an important role in promoting cyanobacteria blooms.     

Accumulation, transformation, and release of inorganic arsenic by the freshwater cyanobacterium Microcystis aeruginosa

Arsenic (As) as a major hazardous metalloid was affected by phytoplankton in many aquatic environments. The toxic dominant algae Microcystis aeruginosa was exposed to different concentrations of inorganic arsenic (arsenate or arsenite) for 15 days in BG11 culture media. Arsenic accumulation, toxicity, and speciation in M. aeruginos as well as the changes of As species in media were examined. M. aeruginosa has a general well tolerance to arsenate and a definite sensitivity to arsenite. Additionally, arsenate actively elevated As methylation by the algae but arsenite definitely inhibited it. Interestingly, the uptake of arsenite was more pronounced than that of arsenate, and it was correlated to the toxicity. Arsenate was the predominant species in both cells and their growth media after 15 days of exposure to arsenate or arsenite. However, the amount of the methylated As species in cells was limited and insignificantly affected by the external As concentrations. Upon uptake of the inorganic arsenic, significant quantities of arsenate as well as small amounts of arsenite, DMA, and MMA were produced by the algae and, in turn, released back into the growth media. Bio-oxidation was the first and primary process and methylation was the minor process for arsenite exposures, while bioreduction and the subsequent methylation were the primary metabolisms for arsenate exposures. Arsenic bioaccumulation and transformation by M. aeruginosa in aquatic environment should be paid more attention during a period of eutrophication.     

Tissue distributions and seasonal dynamics of the hepatotoxic microcystins-LR and -RR in a freshwater snail (Bellamya aeruginosa) from a large shallow, eutrophic lake of the subtropical China

Tissue distributions and seasonal dynamics of the hepatotoxic microcystins-LR and -RR in a freshwater snail (Bellamya aeruginosa) were studied monthly in a large shallow, eutrophic lake of the subtropical China during June-November, 2003. Microcystins (MCs) were quantitatively determined by High-Performance Liquid Chromatography (HPLC) with a qualitative analysis by a Finnigan LC-MS system. On the average of the study period, hepatopancreas was the highest in MC contents (mean 4.14 and range 1.06-7.42 microg g(-1)DW), followed by digestive tracts (mean 1.69 and range 0.8-4.54 microg g(-1)DW) and gonad (mean 0.715 and range 0-2.62 microg g(-1)DW), whereas foot was the least (mean 0.01 and range 0-0.06 microg g(-1)DW). There was a positive correlation in MC contents between digestive tracts and hepatopancreas. A constantly higher MC content in hepatopancreas than in digestive tracts indicates a substantial bioaccumulation of MCs in the hepatopancreas of the snail. The average ratio of MC-LR/MC-RR showed a steady increase from digestive tracts (0.44) to hepatopancreas (0.63) and to gonad (0.96), suggesting that MC-LR was more resistant to degradation in the snail. Since most MCs were present in the hepatopancreas, digestive tracts and gonad with only a very small amount in the edible foot, the risk to human health may not be significant if these toxic parts are removed prior to snail consumption. However, the possible transference of toxins along food chains should not be a negligible concern.