Nutrient-toxicant interactions in natural and constructed phytoplankton communities: results of experiments in semi-continuous and batch culture

Natural communities and clonal cultures of phytoplankton derived from large lakes in Yellowstone National Park (WY, USA) were employed to explore the effects of interactions between resource availability and toxic metals (divalent Cu and Cd) on pelagic plant communities. Results of semi-continuous competition experiments demonstrated strong direct and interactive effects of resource availability and additions of toxic metals (25-150 nmol l(-1)) in both natural and laboratory constructed four-species communities. Both endpoint community composition and population dynamics of individual species elicited responses to interaction between limiting resources and toxic metals. N limited growth in low-density batch cultures was suppressed by addition of Cu and these effects were specific to both algal species and level of available nitrate. Measures of semi-continuous culture effluent pH and pCu demonstrated that high levels of productivity can allow phytoplankton to selectively alter bulk water chemistry in the presence of elevated levels of metals to counteract their toxic effects. Concentrations of both limiting nutrients and toxicants employed were within the range of field-measured levels, suggesting that these types of interactions may be commonplace in natural environments. As such, environmental assessments considering potential impacts of toxic agents should take into account the nutrient status of the aquatic environment, and the interactions among stressors as demonstrated here.     

The combined influence of two agricultural contaminants on natural communities of phytoplankton and zooplankton

Concentrations of glyphosate observed in the environment are generally lower than those found to exert toxicity on aquatic organisms in the laboratory. Toxicity is often tested in the absence of other expected co-occurring contaminants. By examining changes in the phytoplankton and zooplankton communities of shallow, partitioned wetlands over a 5 month period, we assessed the potential for direct and indirect effects of the glyphosate-based herbicide, Roundup WeatherMax^© applied at the maximum label rate, both in isolation and in a mixture with nutrients (from fertilizers). The co-application of herbicide and nutrients resulted in an immediate but transient decline in dietary quality of phytoplankton (8.3 % decline in edible carbon content/L) and zooplankton community similarity (27 % decline in similarity and loss of three taxa), whereas these effects were not evident in wetlands treated only with the herbicide. Thus, even at a worst-case exposure, this herbicide in isolation, did not produce the acutely toxic effects on plankton communities suggested by laboratory or mesocosm studies. Indirect effects of the herbicide-nutrient mixture were evident in mid-summer, when glyphosate residues were no longer detectable in surface water. Zooplankton abundance tripled, and zooplankton taxa richness increased by an average of four taxa in the herbicide and nutrient treated wetlands. The lack of significant toxicity of Roundup WeatherMax alone, as well as the observation of delayed interactive or indirect effects of the mixture of herbicide and nutrients attest to the value of manipulative field experiments as part of a comprehensive, tiered approach to risk assessments in ecotoxicology.     

Chronic and pulse exposure effects of silver nanoparticles on natural lake phytoplankton and zooplankton

The increasing use of silver nanoparticles (AgNPs) in consumer products raises concerns regarding the environmental exposure and impact of AgNPs on natural aquatic environments. Here, we investigated the effects of environmentally relevant AgNP concentrations on the natural plankton communities using in situ enclosures. Using twelve lake enclosures, we tested the hypotheses that AgNP concentration, dosing regimen, and capping agent (poly-vinyl pyrrolidone (PVP) vs. citrate) exhibit differential effects on plankton communities. Each of the following six treatments was replicated twice: control (no AgNPs added), low, medium, and high chronic PVP treatments (PVP-capped AgNPs added continuously, with target nominal concentrations of 4, 16, and 64?μg/L, respectively), citrate treatment (citrate-capped AgNPs added continuously, target nominal concentrations of 64?μg/L), and pulse treatment (64?μg/L PVP-AgNPs added as a single dose). Although Ag accumulated in the phytoplankton, no statistically significant treatment effect was found on phytoplankton community structure or biomass. In contrast, as AgNP exposure rate increased, zooplankton abundance generally increased while biomass and species richness declined. We also observed a shift in the size structure of zooplankton communities in the chronic AgNP treatments. In the pulse treatments, zooplankton abundance and biomass were reduced suggesting short periods of high AgNP concentrations affect zooplankton communities differently than chronic exposures. We found no evidence that capping agent affected AgNP toxicity on either community. Overall, our study demonstrates variable AgNP toxicity between trophic levels with stronger AgNP effects on zooplankton. Such effects on zooplankton are troubling and indicate that AgNP contamination could affect aquatic food webs.     

Assessing cortisol and dehydroepiandrosterone (DHEA) in saliva: effects of collection method

An increasing number of studies are utilizing saliva sampling as a method of assessing adrenal steroid secretion. Saliva samples have certain advantages over plasma, being non-invasive and easily collected. However, some methods of collection may compromise the accuracy of the assay, particularly those which employ aids to stimulate saliva production. We sought to compare the accuracy of cortisol and dehydroepiandrosterone (DHEA) measurement by examining the association between plasma levels, saliva and saliva collected using a citric acid-treated salivette device. Twenty six healthy male volunteers were recruited for the study. To increase the range of steroid levels in the samples collected, half the subjects were pre-treated with hydrocortisone (20mg, twice a day for 7 days) and half with placebo. Saliva samples were then collected from each subject using both a 'passive drool' method and a citric acid-treated salivette. A plasma sample was also collected. Cortisol and DHEA levels were measured by radioimmunoassay. For cortisol levels, both methods of saliva collection correlated highly with plasma levels and with each other (r 0.85; R(2) 0.72 for all). For DHEA levels, only saliva samples collected using the unstimulated collection method correlated with plasma levels. DHEA collected using the salivette device did not correlate significantly with either plasma or the unstimulated saliva (r 0.2;R(2) 0.04). It is crucial that future studies are aware of these issues and are cognizant of the effects of the method of collection when examining steroid levels in saliva.     

The ecological effects of naphthenic acids and salts on phytoplankton from the Athabasca oil sands region

To better elucidate the ecological effects of naphthenic acids and major ions liberated in oil sands development, the summer-time composition of phytoplankton communities in ten water bodies near Fort McMurray (northeastern Alberta) was studied in 1997. The water bodies varied in degree of process water influence, and in age, size and ancillary chemical characteristics. Community biomass of phytoplankton was not systematically related to naphthenic acid or major ion concentrations, even though the higher naphthenate concentrations exceeded published EC50's for acute effects on several different aquatic species. Chlorophyta were frequently dominant, particularly where naphthenate and major ion concentrations were highest. Canonical Correspondence Analysis (CCA) revealed gradients in taxonomic composition at a finer (genus and species) taxonomic level. Despite the simultaneous and uncontrolled variation of other environmental factors, naphthenate and major ion concentrations (as indexed by conductivity) explained a highly-significant 40% of the variation in taxonomic composition. Systems with naphthenates <6.5 mg l(-1) and conductivity <800 PhiS cm(-1) were clustered together near the origin of the CCA plots, suggesting little ecological effect at such concentrations. Taxa associated with elevated naphthenate and/or major ion concentrations were derived from six different algal divisions and included many that were identified as tolerant in previous bioassay experiments. Over the range of concentrations encountered (1.5-45 and 100-3000 mg l(-1) for naphthenates and ions, respectively), CCA indicated that the ecological effect of major ions appeared to be at least as great as that of naphthenates.     

Assessing the effects of acetochlor on soil fungal communities by DGGE and clone library analysis

We assessed the potential eco-toxicological risks of the herbicide acetochlor on fungal communities in the microcosm of black soil using 28S rRNA gene-PCR-DGGE and clone library analysis. The acetochlor was applied to black soil at four concentrations (0-control, 50, 150, and 250 mg/kg). The DGGE fingerprint patterns indicated that acetochlor stimulated fungal communities at day 7 after application, after which there was a suppression effect. The fungal communities in acetochlor-treated soil gradually became more like that of the control during the 60-day experimental period. Diversity indices in the 50 and 150 mg/kg acetochlor treatments changed more rapidly than in the 250 mg/kg acetochlor treatment. The cluster analysis indicated a significant change in fungal community structure after application of acetochlor. The impacts were markedly greater in the 150 and 250 mg/kg acetochlor treatments compared with the 50 mg/kg acetochlor treatment. Sequencing of clones showed that acetochlor application resulted in an increase in pathogenic and non-cultivatable fungal populations, which could increase the risk of plant disease outbreaks.     

Toxigenic effects of diatoms on grazers, phytoplankton and other microbes: a review

Traditionally, diatoms have been regarded as providing the bulk of the food that sustains the marine food chain and important fisheries. However, this view was challenged almost two decades ago on the basis of laboratory and field studies showing that when copepods, the principal predators of diatoms, feed on certain diatom diets, they produce abnormal eggs that either fail to develop to hatching or hatch into malformed (i.e. teratogenic) nauplii that die soon afterwards. Over the years, many explanations have been advanced to explain the causes for reproductive failure in copepods and other marine and freshwater invertebrates including diatom toxicity, or nutritional deficiency and poor assimilation of essential compounds in the animal gut. Here we review the literature concerning the first possibility, that diatoms produce cytotoxic compounds responsible for growth inhibition and teratogenic activity, potentially sabotaging future generations of grazers by inducing poor recruitment. The cytotoxic compounds responsible for these effects are short chain polyunsaturated aldehydes (PUAs) and other oxygenated fatty acid degradation products such as hydroxides, oxo-acids, and epoxyalcohols (collectively termed oxylipins) that are cleaved from fatty acid precursors by enzymes activated within seconds after crushing of cells. Such toxins are suggested to have multiple simultaneous functions in that they not only deter herbivore feeding but some also act as allelopathic agents against other phytoplankton cells, thereby affecting the growth of competitors, and also signalling population-level cell death and termination of blooms, with possible consequences for food web structure and community composition. Some oxylipins also play a role in driving marine bacterial community diversity, with neutral, positive or negative interactions depending on the species, thereby shaping the structure of bacterial communities during diatom blooms. Several reviews have already been published on diatom-grazer interactions so this paper does not attempt to provide a comprehensive overview, but rather to consider some of the more recent findings in this field. We also consider the role of diatom oxylipins in mediating physiological and ecological processes in the plankton and the multiple simultaneous functions of these secondary metabolites.     

浮游植物叶绿素a4种提取方法的比较

目的优选地表水中浮游植物叶绿素a的提取方法。方法实验室制备发生水华的水样,分别选用丙酮研磨法、冻融法、丙酮加热法、混合溶剂法4种方法提取叶绿素a,进行叶绿素a含量的测定。结果冻融法、丙酮加热法、混合溶剂法对样品中叶绿素a的提取效果及方法精密度优于丙酮研磨法（P〈0.05）,差异有统计学意义;丙酮加热法和混和溶剂法耗时较短,叶绿素a在丙酮和混合溶剂中稳定性差异无统计学意（P〉0.05）。结论丙酮加热法和混和溶剂法可为常用叶绿素a测量方法,尤其适用于大批量水环境样品的测定。     

Assessing learning effects and nonrandom dropout in a contraceptive device trial

When there is a new device involved in a randomized trial, participants in the new device group might need more time to become skilled at using the new device than those who are in the control group. In addition, participants who are not used to the new device might drop out from the study at a different rate than participants in the control group. Such learning effects and possible nonrandom dropout can substantially influence the evaluation of treatment effects. Without considering these factors, the conclusion of the main study result is vulnerable to potential bias. In this paper we propose a transition model under a Markov process to assess the effects of learning and nonrandom dropout in a randomized control device trial. The transition probabilities of the proposed model can be obtained by maximum likelihood approaches, and then fit by weighted local regressions to describe patterns of learning and participant dropout. A generalized estimating equation approach is applied to evaluate the impact of prognostic factors on those patterns. We use data from a randomized contraceptive device trial to illustrate the usefulness of the proposed method. The potential impact of learning effects and nonrandom dropout on the primary study objective is discussed.     

An improved method for detecting drug effects in the open field

A double test cross-over design was applied to the testing of rats in the open field. When used to examine the effects of atropine, chlorpromazine and lysergic acid diethylamide (LSD) on open field behaviour, this design proved 4–40 times more sensitive than the previously popular single test design. In no case was the double test design less sensitive. Results are discussed in relation to screening of medically useful compounds.     

Seasonal variations in the effect of zinc pyrithione and copper pyrithione on pelagic phytoplankton communities

The relationship between environmental factors, community composition and the sensitivity of pelagic phytoplankton to the antifouling agents zinc pyrithione (ZPT) and copper pyrithione (CPT) was studied using phytoplankton communities collected from March until August 2001 in Roskilde fjord, Denmark. Sensitivity to ZPT and CPT was measured as EC50 values obtained from dose-response curves of photosynthesis to ZPT and CPT. EC50 for ZPT and CPT varied between 2 and 60 nM and 4 and 25 nM, respectively. Changes in sensitivity throughout the season were related to changes in phytoplankton community composition and density, and to nutrient levels. It was found that the variation in sensitivity of ZPT and CPT was related to the abundance of the groups Cryptophyceae, Bacillariophycaea and Dinophyceae when they were dominating the community. Furthermore, the sensitivity to ZPT was increased at low concentrations of phosphate per cell (<0.2 nmol/cell). For CPT there was a negative correlation between toxicity and phosphate concentration in the water. Consequently, in aquatic environments where phytoplankton is phosphate limited the effect of ZPT and CPT may be enhanced.     

Evaluating pesticide effects on freshwater invertebrate communities in alpine environment: a model ecosystem experiment

Pesticide loads in streams are potentially one of the most relevant stressors for macroinvertebrate communities. Nevertheless, real effects provoked at the community level are still largely unknown. Model ecosystems are frequently used as tools for the risk assessment of pesticides, especially for their regulation, however, they can be also applied to site-specific risk assessment in order to gain better understanding of the responses of aquatic ecosystems to chemical stress. In the present work, an experimental system was composed of 5 artificial streams that reproduced a mountain lotic environment under controlled conditions. This study was aimed to better understand, whether (and how) the biological community was influenced by pesticides pulse exposures. 5 mixture load events were simulated over the productive season (March-July 2010): biological community was regularly sampled and nominal concentrations of water were tested. The results were interpreted comparing the output of different metrics and statistical methodologies. The sensitivity of different metrics was analyzed considering single exposure events (maximum Toxic Units) as well as overall temporal trends. Results showed how some common taxonomic metrics (e.g. taxa richness, Shannon's index, total abundance of organisms, and the Extended Biotic Index) were not suitable to identify the effects of pesticides at community level. On the contrary EPT%, SPEAR(pesticide) and the Principal Response Curve methodology proved to be sensitive to this kind of stress, providing comparable results. Temporal trends of these metrics proved to be related to the concentration of chemicals. Remarkably, the first Principal Response Curve illustrates the trend followed by the most vulnerable species, while the second is more related to the trend of opportunistic species. A high potential risk for the invertebrate community was highlighted by a statistically significant decline of 40 points (comparison with the control) in both SPEAR(pesticide) and EPT%.     

Assessing a vigilance decrement in aged rats: effects of pre-feeding,task manipulation,and psychostimulants

Abstract Rationale. In studies assessing sustained attention in humans, performance is often characterised by a decline in function over time. This response pattern, termed the vigilance decrement, is sensitive to manipulations affecting task difficulty, and to reversal with psychostimulant drugs. A valid test of attention in non-human species requires both comparable characteristics of performance, and sensitivity to similar psychoactive drugs. The five-choice serial reaction time task (5-CSRTT) has been described as a test of sustained attention in the rat, however, studies describing vigilance decrements and performance effects of psychostimulants in this task are scarce. Objectives. We manipulated the standard 5-CSRTT to determine under which conditions a replicable vigilance decrement could be observed, and sought to determine whether this pattern of changes was sensitive to psychostimulant administration. Methods. One and two-year-old rats performed in five-choice sessions extended to 250 trials. Task difficulty was manipulated by either increasing or decreasing the duration of stimulus presentation, and pre-feeding studies were performed to control for effects of the additional food earned. In the two-year-old group dose-responses were then derived for nicotine (0.1–0.4 mg/kg), amphetamine (0.05–0.4 mg/kg) and caffeine (3–10 mg/kg). Results. Extending five-choice sessions revealed a decline in the performance of two-year-old rats as a function of trial number. Increasing task difficulty induced a response-decrement in one-year old rats; whilst reducing it enhanced the performance of two-year-old rats to that observed in younger subjects. Pre-feeding did not alter the response patterns observed in either group. Nicotine, amphetamine, and caffeine all reversed the performance decrement observed. Conclusions. These results demonstrate that similar performance characteristics can be observed in both human and rat, serving to validate further the 5-CSRTT as a measure of sustained attention. Electronic Publication     

Depleted and natural uranium: chemistry and toxicological effects

Depleted uranium (DU) is a by-product from the chemical enrichment of naturally occurring uranium. Natural uranium is comprised of three radioactive isotopes: (238)U, (235)U, and (234)U. This enrichment process reduces the radioactivity of DU to roughly 30% of that of natural uranium. Nonmilitary uses of DU include counterweights in airplanes, shields against radiation in medical radiotherapy units and transport of radioactive isotopes. DU has also been used during wartime in heavy tank armor, armor-piercing bullets, and missiles, due to its desirable chemical properties coupled with its decreased radioactivity. DU weapons are used unreservedly by the armed forces. Chemically and toxicologically, DU behaves similarly to natural uranium metal. Although the effects of DU on human health are not easily discerned, they may be produced by both its chemical and radiological properties. DU can be toxic to many bodily systems, as presented in this review. Most importantly, normal functioning of the kidney, brain, liver, and heart can be affected by DU exposure. Numerous other systems can also be affected by DU exposure, and these are also reviewed. Despite the prevalence of DU usage in many applications, limited data exist regarding the toxicological consequences on human health. This review focuses on the chemistry, pharmacokinetics, and toxicological effects of depleted and natural uranium on several systems in the mammalian body. A section on risk assessment concludes the review.     

Toxicological effects of cypermethrin to marine phytoplankton in a co-culture system under laboratory conditions

The growth of three marine phytoplankton species Skeletonema costatum, Scrippsiella trochoidea and Chattonella marina and the response of the antioxidant defense system have been investigated on exposure to commercial cypermethrin for 96 h and 32 days in a co-culture system. Growth of the three species was generally comparable over 96 h with an inoculation of 1:3:6.5 (C. marina:S. trochoidea:S. costatum), with stimulation at 5 μg l⁻¹ and inhibition under higher concentrations (50, 100 μg l⁻¹). However, when inoculating at ratios of 1:1:1 during a 32 day test, S. costatum became the most sensitive species and was significantly inhibited in all test groups under the dual stresses of cypermethrin and interspecies competition. The growth of C. marina was significantly inhibited at the concentrations higher than 5 μg l⁻¹, while the growth of S. trochoidea was significantly promoted at low concentrations. Superoxide dismutase (SOD) activities significantly increased during 6–12 h exposure periods in test treatments at low concentrations, and enhanced in the control as well due to interspecies competition. The lipid peroxidation product malondialdehyde was enhanced at high concentrations, but did not increase in control and low concentration cultures with high SOD activities, indicating that algal cells activated the antioxidant enzymes promptly to protect the cells from lipid membrane damage. Results from this study suggested that cypermethrin pollution in maricultural sea waters might lead to a shift in phytoplankton community structure from diatom to harmful dinoflagellate species, and thus potentially stimulatory for harmful algal blooms.     

Development of a field test kit for coliphage detection in natural waters

Coliphages may be suitable alternative indicators of fecal pollution in natural waters. The relationship between coliphage and fecal coliform numbers may be expressed by log₁₀ transformed linear regression equations in surface waters (correlation coefficient, r² = 0.5872; p < 0.001) and in well waters (r² = 0.4767; p < 0.001). Using the American Public Health Association 919C single agar layer method, coliphages can be enumerated easily and at less cost and more rapidly than coliform enumeration methods. In adapting this test for use in a field test kit, however, further modifications and simplifications are required. The use of 2,3,5-triphenyl tetrazolium chloride (TPTZ) to enhance plaque visibility while helpful, was not essential. Matched-pair t-test analysis showed no significant difference between the plaque numbers enumerated with or without TPTZ to help plaque visibility. The Escherichia coli C host bacteria can be maintained either as a frozen preparation or dried on to paper disks with protective milk colloids. In the latter procedure, a resuscitation period of 18–24 h in broth at 30°C was found to be satisfactory in giving plaque numbers in the test that were not significantly different than when frozen host was used. A portable field kit has been designed to carry out a total of eight tests. The kit includes the media, the bacterial host, petri dishes, syringes, receptacles, and a camping gas cylinder. It measures 38 cm by 25.5 cm by 33.5 cm and weighs about 6.4 kg. Using the kit, the test procedure for a water sample can be carried out in under 15 min.     

Some ecological implications of a neem (azadirachtin) insecticide disturbance to zooplankton communities in forest pond enclosures

A neem-based insecticide, Neemix 4.5, was applied to forest pond enclosures at concentrations of 10, 17, and 28 microg l(-1) azadirachtin (the active ingredient). At these test concentrations, significant, concentration-dependent reductions in numbers of adult copepods were observed, but immature copepod and cladoceran populations were unaffected. There was no evidence of recovery of adult copepods within the sampling season (May to October). The ecological significance of this disturbance to the zooplankton community was examined by determining biomass as a measure of food availability for higher predators, plankton community respiration, dissolved oxygen (DO) concentrations, and conductivity as functional indicators of ecosystem stress, and zooplankton food web stability as a measure of effects on trophic structure. The selective removal or reduction of adult copepods was sufficient to measurably reduce total zooplankton biomass for several weeks mid-season. During the period of maximal impact (about 4-9 weeks after the applications), total plankton community respiration was significantly reduced, and this appeared to contribute to significant, concentration-dependent increases in dissolved oxygen and decreases in conductivity among treated enclosures. The reductions in adult copepods resulted in negative effects on zooplankton food web stability through eliminations of a trophic link and reduced interactions and connectance. Comparing the results here to those from a previous study with tebufenozide, which was selectively toxic to cladocerans and had little effect on food web stability, indicates that differential sensitivity among taxa can influence the ecological significance of pesticide effects on zooplankton communities.