Multi-linear regression models predict the effects of water chemistry on acute lead toxicity to Ceriodaphnia dubia and Pimephales promelas

The current study examined the acute toxicity of lead (Pb) to Ceriodaphnia dubia and Pimephales promelas in a variety of natural waters. The natural waters were selected to range in pertinent water chemistry parameters such as calcium, pH, total CO(2) and dissolved organic carbon (DOC). Acute toxicity was determined for C. dubia and P. promelas using standard 48h and 96h protocols, respectively. For both organisms acute toxicity varied markedly according to water chemistry, with C. dubia LC50s ranging from 29 to 180μg/L and P. promelas LC50s ranging from 41 to 3598μg/L. Additionally, no Pb toxicity was observed for P. promelas in three alkaline natural waters. With respect to water chemistry parameters, DOC had the strongest protective impact for both organisms. A multi-linear regression (MLR) approach combining previous lab data and the current data was used to identify the relative importance of individual water chemistry components in predicting acute Pb toxicity for both species. As anticipated, the P. promelas best-fit MLR model combined DOC, calcium and pH. Unexpectedly, in the C. dubiaMLR model the importance of pH, TCO(2) and calcium was minimal while DOC and ionic strength were the controlling water quality variables. Adjusted R(2) values of 0.82 and 0.64 for the P. promelas and C. dubia models, respectively, are comparable to previously developed biotic ligand models for other metals.     

Influence of Ca, humic acid and pH on lead accumulation and toxicity in the fathead minnow during prolonged water-borne lead exposure

The present study examines the influence of Ca2+ as (CaSO4), dissolved organic carbon (DOC) and pH on chronic water-borne lead (Pb) toxicity to the larval fathead minnow (Pimephales promelas) under flow-through conditions. The 30 day LC50 for low hardness basic test water (19 mg CaCO3 L(-1)) was 39 (range: 27-51) microg dissolved Pb L(-1) and was greatly increased by increasing concentrations of CaSO4 and DOC to as much as 1903 (range: 1812-1992) mug dissolved Pb L(-1). Both reduced and increased pH (6.7 and 8.1, respectively) compared to control pH of 7.4 appeared to increase Pb toxicity substantially. Whole body Pb accumulation did not reflect water chemistry and thus exhibited no correlation with Pb induced mortality. One possible explanation for this lack of correlation is that mortality occurred predominantly during the first 4-6 days of exposure, whereas Pb accumulation was determined in surviving fish at the end of 30 days of exposure. Chronic Pb exposure resulted in a general iono-regulatory disturbance affecting K+, Na+ and Ca2+ homeostasis. However, recovery of Na+ and K+ levels and reversal of effects on Ca2+ homeostasis during continued exposure strongly suggest fathead minnow can acclimate to Pb. The gills accumulate the highest Pb concentrations during chronic exposure but the skeleton contains the largest mass of Pb by contributing up to approximately 80% of whole body Pb. In conclusion, water chemistry characteristics like Ca2+ and DOC should be considered for chronic water quality criteria.     

The toxicity and physiological effects of copper on the freshwater pulmonate snail, Lymnaea stagnalis

Several recent studies have demonstrated that the freshwater pulmonate snail Lymnaea stagnalis is extremely sensitive to metals (Co, Ni, Pb) in chronic exposures. The objective of the current study was to evaluate the acute and chronic sensitivity of L. stagnalis to Cu and investigate the underlying mechanism(s) of toxic action. A 96-h LC50 of 31μg L(-1) Cu was estimated indicating L. stagnalis was moderately acutely sensitive to Cu relative to other aquatic organisms. However, in a 30-day chronic exposure using juvenile snails an EC20 of 1.8μg L(-1) Cu was estimated for snail growth making L. stagnalis the most sensitive organism tested to date for Cu. Hardness-based and BLM-based water quality criteria for Cu at the water quality conditions used in this study were 7.8 and 1.5μg L(-1), respectively, indicating L. stagnalis is significantly under-protected by hardness-based WQC. Investigations into the mechanism(s) of toxic action for Cu were conducted on young adult snails necessitating higher Cu exposures. Exposure to Cu at 12μg L(-1) resulted in no detectable effects on hemolymph osmolality, net Ca(2+) uptake, titratable acid excretion, or ammonia excretion. Exposure to 48μg L(-1) Cu was shown to significantly reduce (91%) net Ca(2+) uptake which is strongly correlated with shell deposition and corresponding snail growth. Snails exposed to 48μg L(-1) Cu also exhibited reduced ammonia excretion, a marked hemolymph acidosis, and a compensatory increase in titratable acid excretion. The reduction in net Ca(2+) uptake was hypothesized to be a secondary effect of Cu-induced inhibition of carbonic anhydrase, but no reduction in carbonic anhydrase activity was detected. Overall, it remains unclear whether inhibition of Ca(2+) uptake is a direct result of Cu exposure or, along with the other observed physiological effects, is secondary to an unidentified primary mode of toxic action. Given the hypersensitivity of L. stagnalis to Cu, further study into the mechanisms of action and effects of varying water chemistry on Cu toxicity is clearly warranted.     

Influences of water chemistry on the acute toxicity of lead to Pimephales promelas and Ceriodaphnia dubia

The acute toxicity of lead (Pb) was examined for fathead minnows (Pimephales promelas; 96-h) and daphnids (Ceriodaphnia dubia; 48-h) in waters modified for hardness (as CaSO₄), dissolved organic carbon (DOC; as Aldrich humic acid) and alkalinity (as NaHCO₃) for parameterization of an acute freshwater biotic ligand model (BLM). Additionally, acute (96-h) and chronic (30-d) bioassays were performed for P. promelas to more clearly define the influence of pH (5.5–8.3) on Pb toxicity as modified by addition of HCl or NaOH using an automated titration system. Results indicate that Ca²⁺ is protective against acute Pb toxicity to P. promelas but not C. dubia. Strong protection was afforded by DOC and NaHCO₃ against acute Pb toxicity to P. promelas, whereas milder protection was observed for C. dubia with both parameters. Dissolved Pb LC50s from the P. promelas pH bioassays revealed a complex effect of pH on Pb toxicity, likely explained in part by Pb speciation and the competitive interaction of H⁺ with ionic Pb²⁺. Chronic pH bioassays also demonstrated that 30-d growth is not impaired in fathead minnows at relevant Pb concentrations. The findings reported herein suggest that development of separate BLMs for P. promelas and C. dubia should be considered.     

The influence of algal densities on the toxicity of chromium for Ceriodaphnia dubia Richard (Cladocera, Crustacea)

Food availability may affect metal toxicity for aquatic organisms. In the present study, the influence of high, medium and low densities of the algae Pseudokirchneriella subcapitata (10(6), 10(5) and 10(4) cells.mL(-1), respectively) on the chronic toxicity of chromium to the cladoceran Ceriodaphnia dubia was investigated. C. dubia was exposed to a range of chromium concentration from 2.71 to 34.04 microg.L(-1) and fed with algae at various densities. In another experiment, the green alga was exposed to chromium concentrations (94 to 774 microg.L(-1)) and supplied as food in different densities to zooplankton. The survival and reproduction of the cladoceran were measured in these toxicity tests. The IC50 for Cr to P. subcapitata and metal accumulated by algal cells were determined. The results of a bifactorial analysis (metal versus algal densities) showed that metal toxicity to zooplankton was dependent on algal densities. Significant toxic effects on the reproduction and survival of C. dubia were observed at 8.73, 18.22 and 34.04 microg.L(-1) Cr when the test organisms were fed with 10(6) cells.mL(-1) of P. subcapitata. Although the chlorophyta retain low chromium content, a decrease in the reproduction and survival of C. dubia occurred when they were fed with high algal density contaminated with 774 microg.L(-1) Cr. It was concluded that high algal density have an appreciable influence on chromium toxicity to daphnids.     

Extension of the biotic ligand model of acute toxicity to a physiologically-based model of the survival time of rainbow trout (Oncorhynchus mykiss) exposed to silver

Chemical speciation controls the bioavailability and toxicity of metals in aquatic systems and regulatory agencies are recognizing this as they develop updated water quality criteria (WQC) for metals. The factors that affect bioavailability may be quantitatively evaluated with the biotic ligand model (BLM). Within the context of the BLM framework, the 'biotic ligand' is the site where metal binding results in the manifestation of a toxic effect. While the BLM does account for the speciation and complexation of dissolved metal in solution, and competition among the free metal ion and other cations for binding sites at the biotic ligand, it does not explicitly consider either the physiological effects of metals on aquatic organisms, or the direct effect of water chemistry parameters such as pH, Ca(2+)and Na(+) on the physiological state of the organism. Here, a physiologically-based model of survival time is described. In addition to incorporating the effects of water chemistry on metal availability to the organism, via the BLM, it also considers the interaction of water chemistry on the physiological condition of the organism, independent of its effect on metal availability. At the same time it explicitly considers the degree of interaction of these factors with the organism and how this affects the rate at which cumulative damage occurs. An example application of the model to toxicity data for rainbow trout exposed to silver is presented to illustrate how this framework may be used to predict survival time for alternative exposure durations. The sodium balance model (SBM) that is described herein, a specific application of a more generic ion balance model (IBM) framework, adds a new physiological dimension to the previously developed BLM. As such it also necessarily adds another layer of complexity to this already useful predictive framework. While the demonstrated capability of the SBM to predict effects in relation to exposure duration is a useful feature of this mechanistically-based framework, it is envisioned that, with suitable refinements, it may also have utility in other areas of toxicological and regulatory interest. Such areas include the analysis of time variable exposure conditions, residual after-effects of exposure to metals, acclimation, chronic toxicity and species and genus sensitivity. Each of these is of potential utility to longer-term ongoing efforts to develop and refine WQC for metals.     

Derivation of a toxicity-based model to predict how water chemistry influences silver toxicity to invertebrates

The effect of altering water chemistry on acute silver toxicity to three invertebrate species, two Daphnids, Daphnia magna and Daphnia pulex, as well as an amphipod Gammarus pulex was assessed. In addition, the physiological basis of Ag(I) toxicity to G. pulex was examined. Daphnia magna and D. pulex were more sensitive than G. pulex and 48 h LC(50) values in synthetic ion poor water were 0.47, 0.65 and 2.1 microg Ag(I) l(-1), respectively. Increasing water [Cl(-)] reduced Ag(I) toxicity in all species, and increasing water [Ca(2+)] from 50 to 1,500 microM reduced Ag(I) toxicity in G. pulex. Whole body Na(+) content, but not K(+) or Ca(2+) was significantly reduced in G. pulex exposed to 6 microg Ag(I) l(-1) for 24 h, but there was no inhibition of whole body Na(+)/K(+)-ATPase activity. Both increasing water [Cl(-)] and [Ca(2+)] reduced this Ag(I)-induced Na(+) loss. For D. magna, the presence of 10 mg l(-1) humic acid or 0.5 microM 3-mercaptoproprionic acid (3-MPA) increased the 48 h LC(50) values by 5.9 and 58.5-fold, respectively, and for D. pulex the presence of 1 microM thiosulfate increased the 48 h LC(50) value by four-fold. The D. magna toxicity data generated from this study were used to derive a Daphnia biotic ligand model (BLM). Analysis of the measured LC(50) values vs. the predicted LC(50) values for toxicity data from the present and published results where water Cl(-), Ca(2+), Na(+) or humic acid were varied showed that 91% of the measured toxicity data fell within a factor of two of the predicted LC(50) values. However, the daphnid BLM could not accurately predict G. pulex toxicity. Additionally, the Daphnia BLM was under-protective in the presence of the organic thiols 3-MPA or thiosulphate and predicted an increase in the LC(50) value of 114- and 74-fold, respectively. The Daphnia toxicity based BLM derived from the present data set is successful in predicting Daphnia toxicity in laboratory data sets in the absence of sulfur containing compounds, but shows its limitations when applied to waters containing organic thiols or thiosulphate.     

The effects of the slug biological control agent, Phasmarhabditis hermaphrodita (Nematoda), on non-target aquatic molluscs

The nematode Phasmarhabditis hermaphrodita is used as a commercial biological control agent of slugs in the UK. Although it is known to affect other terrestrial mollusc species, its effects on freshwater molluscs are not known. The present study investigated the effects of P. hermaphrodita on the survival of juvenile Lymnaea stagnalis and Physa fontinalis, two common freshwater snails, at 'spray tank' concentration and a 50% diluted 'spray tank' concentration over a 14-day period. Survival of L. stagnalis was significantly reduced at both application levels but P. fontinalis suffered no mortalities over the experimental period. The possible differential mechanisms of pathology between the two host species are discussed.     

Radiotracer Studies on Waterborne Copper Uptake,Distribution, and Toxicity in Rainbow Trout and Yellow Perch: A Comparative Analysis

Rainbow trout (Oncorhynchus mykiss) are often used to estimate important biotic ligand model (BLM) parameters, such as metal-binding affinity (log K) and capacity (B_max). However, rainbow trout do not typically occupy metal-contaminated environments, whereas yellow perch (Perca flavescens) are ubiquitous throughout most of North America. This study demonstrates that dynamic processes that regulate Cu uptake at the gill differ between rainbow trout and yellow perch. Rainbow trout were more sensitive to acute aqueous Cu than yellow perch, and toxicity was exacerbated in soft water relative to similar exposures in hard water. Whole body Na loss rate could account for acute Cu toxicity in both species, as opposed to new Cu uptake rate that was not as predictive. Time course experiments using radiolabelled Cu (⁶⁴Cu) revealed that branchial Cu uptake was rather variable within the first 12 h of exposure, and appeared to be a function of Cu concentration, water hardness, and fish species. After 12 h, new branchial Cu concentrations stabilized in both species, suggesting that metal exposures used to estimate BLM parameters should be increased in duration from 3 h to 12+ h. In rainbow trout, 71% of the new Cu bound to the gill was exchangeable (i.e., able to either enter the fish or be released back to the water), as opposed to only 48% in yellow perch. This suggests that at equal exposure concentrations, proportionally more branchial Cu can be taken up by rainbow trout than yellow perch, which can then go on to confer toxicity. These qualitative differences in branchial Cu handling between the two species emphasize the need to develop BLM parameters for each species of interest, rather than the current practice of extrapolating BLM results derived from rainbow trout (or other laboratory-reared species) to other species. Data reported here indicate that a one-size-fits-all approach to predictive modeling, mostly based on rainbow trout studies, may not suffice for making predictions about metal toxicity to yellow perch—that is, a species that inhabits metal-contaminated lakes around northern Canadian industrial operations.     

Toxicity of dissolved Cu, Zn, Ni and Cd to developing embryos of the blue mussel (Mytilus trossolus) and the protective effect of dissolved organic carbon

Marine water quality criteria for metals are largely driven by the extremely sensitive embryo-larval toxicity of Mytilus sp. Here we assess the toxicity of four dissolved metals (Cu, Zn, Ni, Cd) in the mussel Mytilus trossolus, at various salinity levels while also examining the modifying effects of dissolved organic carbon (DOC) on metal toxicity. In 48 h embryo development tests in natural seawater, measured EC50 values were 6.9-9.6 microg L(-1) (95% C.I.=5.5-10.8 microg L(-1)) for Cu, 99 microg L(-1) (86-101) for Zn, 150 microg L(-1) (73-156) for Ni, and 502 microg L(-1) (364-847) for Cd. A salinity threshold of >20 ppt (approximately 60% full strength seawater) was required for normal control development. Salinity in the 60-100% range did not alter Cu toxicity. Experimental addition of dissolved organic carbon (DOC) from three sources reduced Cu toxicity; for example the EC50 of embryos developing in seawater with 20 mg C L(-1) was 39 microg Cu L(-1) (35.2-47.2) a 4-fold increase in Cu EC50. The protective effects of DOC were influenced by their distinct physicochemical properties. Protection appears to be related to higher fulvic acid and lower humic acid content as operationally defined by fluorescence spectroscopy. The fact that DOC from freshwater sources provides protection against Cu toxicity in seawater suggests that extrapolation from freshwater toxicity testing may be possible for saltwater criteria development, including development of a saltwater Biotic Ligand Model for prediction of Cu toxicity.     

Toxic effects of cadmium and zinc on the transmission of Echinoparyphium recurvatum cercariae

The toxicity of cadmium, zinc and Cd/Zn mixtures to the transmission of Echinoparyphium recurvatum (Digenea: Echinostomatidae) cercariae into the snail second intermediate hosts was investigated at concentrations ranging from 100 microg l-1 to 10 000 microg l-1 in both soft and hard water. A differential response in the infectivity of metal-exposed cercariae into Lymnaea peregra and Physa fontinalis was demonstrated which was dependent on the snail species being infected. Exposure of L. peregra, P. fontinalis, and L. stagnalis to heavy metals caused a differing susceptibility to E. recurvatum cercariae depending on the snail species being exposed. The mechanism and effects of metal toxicity, together with the importance of the parasite/host strain on cercarial transmission are discussed.     

Effects of water chemistry variables on gill binding and acute toxicity of cadmium in rainbow trout (Oncorhynchus mykiss): A biotic ligand model (BLM) approach

This study investigated the short-term (3 h) cadmium binding characteristics of the gills, as well as the influence of various water chemistry variables [calcium, magnesium, sodium, pH, alkalinity and dissolved organic carbon (DOC)] on short-term gill accumulation and acute toxicity of cadmium in juvenile freshwater rainbow trout. The cadmium binding pattern revealed two types of cadmium binding sites in the gill: (i) saturable high affinity sites operating at a low range of waterborne cadmium concentration, and (ii) non-saturable low affinity sites operating at a higher range of cadmium concentration. Among the water chemistry variables tested, only calcium and DOC significantly reduced both gill accumulation and toxicity of cadmium. Interestingly, alkalinity (15-90 mg L(-1) as CaCO(3)) did not influence the gill cadmium accumulation but a significant increase in toxicity was recorded at a higher alkalinity level (90 mg L(-1)). Affinity constants (log K) for binding of competing cations (Cd(2+) and Ca(2+)) to the biotic ligand and for binding of Cd(2+) to DOC were derived separately from the 3 h gill binding tests and the 96 h toxicity tests. In general, the values agreed well, indicating that both tests targeted the same population of high affinity binding sites, which are likely Ca(2+) uptake sites on the gills. These parameters were then incorporated into a geochemical speciation model (MINEQL+) to develop a biotic ligand model for predicting acute toxicity of cadmium in trout. The model predictions exhibited a good fit with the measured toxicity data except for high alkalinity and pH.     

Variations in the retinal designs of pulmonate snails (Mollusca, Gastropoda): squaring phylogenetic background and ecophysiological needs (I)

Abstract. The eyes of aquatic pulmonates differ from those of terrestrial pulmonates; the latter, in species such as Cepaea nemoralis and Trichia hispida , possess conventional, cup-shaped retinas, but the aquatic species Lymnaea stagnalis, Radix peregra, Physa fontinalis , and Planorbarius corneus have retinas that are partitioned into dorsal and ventral depressions ("pits"). The pits are separated by an internal ridge, called the "crest", and on account of their pigmentation can be seen in vivo . The dominant cellular components of the retinae of terrestrial as well as aquatic snails are pigmented cells and microvillar photoreceptors, the latter occurring in two morphologically distinct types (I and II). Aquatic snails with preferences for shallow water possess eyes with both type I and type II photoreceptive cells, but Pl. corneus , an inhabitant of deeper water, only has type-I receptors, supporting an earlier finding that type I cells represent dim- and type II cells bright-light receptors. On the basis of histological and optical comparisons, we conclude that the eyes of L. stagnalis and R. peregra , species that are known to escape and seek temporary refuge above the water surface, are well adapted to function in water as well as air, but that the eyes of P. fontinalis and Pl. corneus are less modified from those of their terrestrial ancestors.     

Models of Cadmium Accumulation and Toxicity to Hyalella azteca during 7- and 28-Day Exposures

The inhibition of Cd accumulation by Ca in the amphipod Hyalella azteca in short-term (7-d) exposures appears to follow anti-competitive, rather than competitive, inhibition. Increasing Ca reduces Cd accumulation more at high than at low Cd concentrations. Cadmium accumulation and toxicity in chronic exposures can be predicted using the 7-d model to which the effects of acclimation, Cd inhibition of acclimation, and growth dilution are added. The resultant model is complex and species-specific, making it unwieldy for direct application in water quality guideline or criteria development. However, it does demonstrate that a mechanistic explanation of the relationship between short- and long-term accumulation and toxicity is possible, as well as suggest why the acute-to-chronic ratio changes with water chemistry. It is not, therefore, appropriate to estimate chronic Cd toxicity to H. azteca from acute toxicity assuming a constant acute-to-chronic ratio. The standard Biotic Ligand Model (BLM) can also be fit to the chronic bioaccumulation and toxicity data. This may be a more practical approach to guideline or criteria development, provided it is understood that this is an empirical fit of the model and that the underlying mechanisms are far more complex than those invoked in the standard BLM.     

Application of the biotic ligand model to predicting zinc toxicity to rainbow trout,fathead minnow,and Daphnia magna

The Biotic Ligand Model has been previously developed to explain and predict the effects of water chemistry on the toxicity of copper, silver, and cadmium. In this paper, we describe the development and application of a biotic ligand model for zinc (Zn BLM). The data used in the development of the Zn BLM includes acute zinc LC50 data for several aquatic organisms including rainbow trout, fathead minnow, and Daphnia magna. Important chemical effects were observed that influenced the measured zinc toxicity for these organisms including the effects of hardness and pH. A significant amount of the historical toxicity data for zinc includes concentrations that exceeded zinc solubility. These data exhibited very different responses to chemical adjustment than data that were within solubility limits. Toxicity data that were within solubility limits showed evidence of both zinc complexation, and zinc-proton competition and could be well described by a chemical equilibrium approach such as that used by the Zn BLM.     

Silver speciation during chronic toxicity tests with the mysid,Americamysis bahia

A 28-day chronic toxicity test and two 7-day chronic estimation toxicity tests were conducted with silver nitrate (AgNO(3)) and the marine invertebrate, Americamysis bahia, in 20 per thousand (parts-per thousand) salinity seawater. One 7-day test was initiated with 7-day-old mysids and the second was initiated with <24-h-old mysids. There was very good agreement between the three toxicity tests. The no-observed-effect concentration (NOEC) values from the 28-day test, the 7-day test initiated with 7-day-old mysids, and the 7-day test initiated with <24-h-old mysids were 34, 65 and 38 microg/l silver, respectively. The 96-h LC50 values from the 28-day toxicity test and the 7-day toxicity test initiated with 7-day old mysids were 260 microg/l, and the 96-h LC50 value from the 7-day toxicity test initiated with <24-h-old mysids was 280 microg/l. Free ionic silver, Ag(+), concentrations measured with a silver electrode were in good agreement with concentrations calculated using total dissolved silver and chloride concentrations. Mean measured concentrations of Ag(+) in the test solutions ranged from 0.99 to 25 ng/l for the dissolved silver concentrations that ranged from 34 to 410 microg/l silver, indicating that free ionic silver varied from 0.003 to 0.006% of the silver dissolved in the 20 per thousand salinity seawater. Understanding the relationship of salinity and silver speciation, and the effect of this relationship on chronic invertebrate toxicity, will be useful for development of a marine biotic ligand model (BLM) and a water quality criterion for silver. This model could provide an important tool for improving the relationship of laboratory toxicity test results and predicted effects in natural environments, where variations in salinity may act to modify the toxicity of silver and other metals.     

Limnological and ecotoxicological studies in the cascade of reservoirs in the Tietê River (S?o Paulo, Brazil).

An evaluation was made of the quality of samples of water and sediment collected from a series of reservoirs in the Tietê River (SP), based on limnological and ecotoxicological analyses. The samples were collected during two periods (Feb and Jul 2000) from 15 sampling stations. Acute toxicity bioassays were performed using the test organism Daphnia similis, while chronic bioassays were carried out with Ceriodaphnia dubia and Danio rerio larvae. The water samples were analyzed for total nutrients, total suspended matter and total cadmium, chromium, copper and zinc concentrations, while the sediment samples were examined for organic matter, granulometry and potentially bioavailable metals (cadmium, chromium, copper and zinc). The results obtained for the limnological variable, revealed differences in the water quality, with high contribution of nutrients and metals for Tietê and Piracicaba rivers, besides the incorporation and sedimentation, consequently causing a reduction of materials in Barra Bonita reservoir, thus promoting the improvement of the water quality in the other reservoirs. The toxicity bioassays revealed acute toxicity for Daphnia similis only in the reservoirs located below Barra Bonita dam. On the other hand, chronic toxicity for Ceriodaphnia dubia and acute for Danio rerio showed a different pattern, decreasing in magnitude from Barra Bonita to Três Irm?os, demonstrating an environmental degradation gradient in the reservoirs.     

Fixed focal-length optics in pulmonate snails (Mollusca, Gastropoda): squaring phylogenetic background and ecophysiological needs (II)

Abstract. Our results suggest that freshwater pulmonates like Lymnaea stagnalis, Radix peregra, Physa fontinalis , and Planorbarius corneus have inherited from their terrestrial ancestors eyes with a spherical, immobile lens with fixed focal-length optics. Unable to change the dioptric apparatus to form an image under water, modifications to the retina had to occur if sharp vision was required. Computer-assisted calculations and 3-D eye reconstructions demonstrate that the photoreceptors in the deeper, ventral pit are in a position to perceive focused images under water. Vision in air, however, would favour photoreceptive cells located in the shallower, dorsal pit. On the basis of histological, ethological, and optical comparisons, we conclude that the eyes in L. stagnalis and R. peregra , species that are known to escape and seek temporary refuge above the water surface, are well adapted to function in water as well as air, but that the eyes in P. fontinalis and Pl. corneus are less modified from those of their terrestrial ancestors. We also conclude that good resolving power may be of greater importance in the aquatic pulmonates than the terrestrial species, since the former have to locate thin, vertical stems of reeds and sedges to ascend in order to reach the surface to breathe.     

川西亚高山森林凋落物不同分解阶段碳氮磷化学计量特征及种间差异

为了了解亚高山森林凋落物在不同分解阶段的化学计量特征,采用空间代替时间的方法,以自然状态下凋落物的3个层次--新鲜凋落物层（L）、发酵层（F）、腐殖质层（H））模拟凋落物分解的不同阶段,对川西亚高山不同林分类型（岷江冷杉天然林、粗枝云杉人工林、白桦天然林、杜鹃矮曲林）凋落物的碳氮磷（C、N、P）及可溶性碳氮磷（DOC、SN、SP）含量进行研究。结果表明：林分类型及分解阶段将显著影响凋落物分解过程中的碳氮磷含量及其化学计量比。亚高山森林凋落物可溶性有机碳、水溶性磷含量均随着分解过程的不断进行而降低,分解初期快速淋溶,而分解中后期释放变缓。4种林分比较而言,水溶性碳氮磷含量表现为：白桦 > 杜鹃 > 冷杉 > 云杉,阔叶树种凋落物的可溶性碳氮磷普遍高于针叶树种,尤其在分解初期。针叶树种凋落物SN在分解过程中呈现释放模式,而阔叶树种SN呈现先富集后释放模式。凋落物C含量随着分解的不断进行而降低,冷杉、白桦及杜鹃N含量呈现先富集后释放的趋势,分解阶段对云杉与白桦各层P含量影响不显著,但冷杉却呈现先降低后升高的现象,而杜鹃则是在分解后期P含量显著降低。从总体来看,亚高山森林凋落物C/P和N/P均显著小于全球平均水平,凋落物C/N、C/P、N/P、DOC/C、SN/N、SP/P均随着分解的不断进行呈现降低的趋势。分解初期白桦和杜鹃DOC/C显著降低,而冷杉则在分解后期显著降低。冷杉N/P先升高后降低,杜鹃N/P随着分解的不断进行呈现升高趋势。这些研究结果为深入理解亚高山森林生态系统的凋落物分解进程和养分循环提供了依据。     

Glucans of lichenized fungi: significance for taxonomy of the genera Parmotrema and Rimelia

The glucans of lichenized fungi are an important class of polysaccharides with structural and chemotaxonomic roles. The water-insoluble glucans of the genus Parmotrema (P. austrosinense, P. delicatulum, P. mantiqueirense, P. schindleri, and P. tinctorum) and those of Rimelia (R. cetrata and R. reticulata), were investigated in order to evaluate the significance in chemotyping, with nigeran [(1-->3),(1-->4)-alpha-glucan] and lichenan [(1-->3),(1-->4)-beta-glucan] characterized using (1)H and (13)C NMR, methylation analysis, and controlled Smith degradations. Results from all species were similar, suggesting that glucan chemistry does not support separation of Rimelia from Parmotrema.