Pollution-induced community tolerance of soil microbes in response to a zinc gradient

The long-term accumulation of Zn in soils provides the microbial community time to adapt to this heavy metal. To assess the effects of long-term exposure to Zn on the metabolic diversity and tolerance to Zn of soil microbial community, the pollution-induced community tolerance (PICT) method, based on the Biolog microtitre plate system, was used. It especially is useful to study gradients of pollutants for detecting PICT. Such a Zn gradient was found by selecting soils at increasing distances from galvanized electricity pylons at two sites where Zn accumulation had occurred over a period of decades. Soil metabolic profiles showed a humpbacked response to increasing soil Zn concentrations, indicating that accumulation of Zn up to 2,000 mg/kg did not decrease the metabolic biodiversity in the culturable fraction of the microbial community. This fraction of the microbial community showed significant PICT, because the effective concentration that reduces the metabolic biodiversity by 50% (EC50), values for Zn added to the Biolog wells increased by up to two orders of magnitude with increasing soil-Zn concentration along the transects. Significant PICT was detectable at soil Zn concentrations above approximately 300 mg/kg. The EC50 values correlated more closely with soil total Zn than with soil pore water Zn. The results suggest that, although long-term exposure to Zn imposes stress on soil microbes, resulting in an increased tolerance, detectable PICT does not necessarily lead to a decrease in metabolic diversity.     

Location-specific ecotoxicological risk assessment of metal-polluted soils

When chemical analysis indicates metal pollution, a second-tier method is needed to evaluate whether toxic effects occur at the polluted sites. A method based on pollution-induced community tolerance (PICT) was developed using samples taken from locations polluted with sewage more than 20 years ago. Microorganisms extracted from soil samples were exposed to a concentration range of zinc, nickel, copper, chromium (III), or chromium (VI) salts in a buffer suspension. The remaining activity of the intoxicated microorganisms was determined by color formation with 31 different organic substrates in microtiter plates. Microorganisms from moderately Zn-polluted sites (>45 mg/kg) showed an increased tolerance for zinc. Nickel tolerance was observed at 51 mg Ni/kg soil, chromium (VI) tolerance at 923 mg Cr/kg. In most cases, tolerance also was observed at higher concentrations. High concentrations of 1,494 mg Cu/kg or 3,935 mg Cr/kg did not show PICT, indicating a limited bioavailability of Cu and Cr at these sites. The benefits of our method are its greater sensitivity compared to other tests used at these sites, and its specificity for those metals that exceed allowable levels.     

Use of pollution-induced community tolerance of the bacterial community to detect phenol toxicity in soil

Pollution-induced community tolerance (PICT) was used to study effects of phenol on soil bacteria. Phenol was added to an agricultural soil in a microcosm experiment. The effects were studied for up to four months. Bacterial growth rates were estimated with the leucine incorporation technique. This technique was also used as detection method for PICT. Changes in community structure were studied using the phospholipid fatty acid (PLFA) pattern. Increased phenol PICT of the bacterial community was found at phenol concentrations above 1 micromol/g wet weight soil. Direct inhibiting effect on bacterial growth rates 1 d after adding phenol was correlated to PICT. Phenol toxicity was reflected by changes in the structure of the bacterial community, although PICT appeared more sensitive than the PLFA method. In soil amended with 1 to 10 micromol phenol/g soil, bacterial growth recovered within one week. In the soil amended with the highest phenol concentration (30 micromol/g soil), bacterial growth rate recovered from total inhibition after 27 d, eventually reaching values six times higher than in the control. However, PICT did not change during the four months the experiment was performed. The specificity of PICT was also studied by examining cotolerance to 2-chlorophenol, 2,4-dichlorophenol, 2,3,6-trichlorophenol, Cu, and Zn. Adding phenol induced cotolerance of the bacterial community to the other phenols, although always at a lover level than to phenol. No cotolerance was found to metals in phenol-polluted soil. We conclude that the PICT concept is a valuable tool in determining phenol toxicity to bacterial communities, especially in situations where bacterial growth has recovered. Cotolerance between different phenols can, however, make interpretations of PICT more complicated.     

Comparison of aerobic and anaerobic [3H]leucine incorporation assays for determining pollution-induced bacterial community tolerance in copper-polluted, irrigated soils

Pollution-induced community tolerance (PICT) constitutes a sensitive and ecologically relevant impact parameter in ecotoxicology. We report the development and application of a novel anaerobic [(3) H]leucine incorporation assay and its comparison with the conventional aerobic [(3) H]leucine incorporation assay for PICT detection in soil bacterial communities. Selection of bacterial communities was performed over 42 d in bulk soil microcosms (no plants) and in rice (Oryza sativa) rhizosphere soil mesocosms. The following experimental treatments were imposed using a full factorial design: two soil types, two soil water regimes, and four Cu application rates (0, 30, 120, or 280 μg g(-1)). Bacterial communities in bulk soil microcosms exhibited similar Cu tolerance patterns when assessed by aerobic and anaerobic PICT assays, whereas aerobic microorganisms tended to be more strongly selected for Cu tolerance than anaerobic microorganisms in rhizosphere soil. Despite similar levels of water-extractable Cu, bacterial Cu tolerance was significantly higher in acid sulfate soil than in alluvial soil. Copper amendment selected for significant PICT development in soils subjected to alternate wetting and drying, but not in continuously flooded soils. Our results demonstrate that soil bacterial communities subjected to alternate wetting and drying may be more affected by Cu than bacterial communities subjected to continuous flooding. We conclude that the parallel use of anaerobic and aerobic [(3) H]leucine PICT assays constitutes a valuable improvement over existing procedures for PICT detection in irrigated soils and other redox gradient environments such as sediments and wetlands.     

Response of Pistia stratiotes to Heavy Metals (Cr, Ni, and Zn) and Phosphorous

The effects of Cr, Ni, Zn, and P exposure on the root anatomic structure, growth, and chlorophyll a concentration of Pistia stratiotes L. were studied. Plastic aquaria containing 50 g of wet plants and 5 L of pond water added with the contaminant(s) were disposed. The treatments were: (1) Cr, (2) Ni, (3) Zn, (4) P, (5) Cr + Ni + Zn, (6) Cr + Ni + Zn + P, and (7) control. Contaminant additions were done seven times. In each addition, concentrations of 1 mg of metals or 5 mg of P per liter of water were added. Chlorophyll a was an indicator more sensitive to Zn and Cr toxicity than the relative growth rate. Ni and Cr + Ni + Zn treatments were the most toxic ones, in which biomass and the root anatomical parameters (root length, cross-sectional areas [CSAs] of root, stele, and metaxylem vessels) decreased significantly. The addition of P to the treatment with combined metals attenuated the decrease in plant growth and root length, and caused a significant increase in CSAs of total metaxylem vessels, suggesting that P increased the tolerance of P. stratiotes to metals. This fact has important implications for the use of this macrophyte in constructed wetlands for industrial wastewater treatment.     

Assessment of Sediment Toxicity Using Different Trophic Organisms

The main aim of the present project is to study the feasibility of using different trophic organisms for evaluating the toxicity of dredged sediments arising in Hong Kong. A total of eight sediment samples (duplicate samples collected from four selected sites: Kowloon Bay, Tsing Yi, Chek Lap Kok, and Double Haven) of Hong Kong coastal waters were analyzed for the total concentrations of As, Cd, Cr, Cu, Hg, Ni, Pb, and Zn, total organic carbon, acid volatile sulfides, simultaneously extracted metals, redox potential, and 12 organic micropollutants. The sediment elutriates were also analysed for the various metal concentrations, as well as contents of ammonia-N, nitrate, total sulfide, sulfate, and total organic carbon. Elutriate Sediment Toxicity Tests (ESTT) were also conducted, using two microalgae (Skeletonema costatum, a diatom and Dunaliella tertiolecta, a flagellate), juvenile shrimp (Metapenaeus ensis) and juvenile fish (Trachinotus obtaus). Two commercially available tests using bacteria (Microtox Test and Toxi-Chromotest) also were employed to test both the solid phase and elutriates of the sediments. The results of Microtox test on the solid phase, and bioassay tests using diatom on the sediment elutriate, especially the former, were correlated significantly (p < 0.05) with a number of physico-chemical properties of sediments and elutriates. It is recommended that a combination of a liquid-phase bioassay using diatom and a solid-phase bioassay using Microtox test should be used for screening a large number of sediment samples. However, the presence of ammonia in the sediments containing a high content of organic matter seemed to interfere the detection of contamination impacts. Received: 12 March 1996/Revised: 27 August 1996     

Variability in zinc tolerance,measured as incorporation of radio-labeled carbon dioxide and thymidine,in periphyton communities sampled from 15 European river stretches

Fifteen European rivers and streams belonging to watersheds in Sweden, the Netherlands, and Spain respectively, were sampled by allowing periphyton to colonize submerged glass substrata. Their zinc tolerances were quantified in short-term laboratory tests, where inhibition of photosynthesis in microalgae and thymidine incorporation in bacterial DNA was measured, and expressed as EC50 values. The variability in zinc tolerances was high reaching 1.5–2.5 orders of magnitude, ranging from 25–8145 μM for photosynthesis and 15–467 μM for thymidine assays. Based on the observed variability, uncertainty factors were estimated for the extrapolation of zinc toxicity data from river to river, both regionally and interregionally. Under the assumption to protect 95% of the observed communities the regional uncertainty factors were 1.7–4.3 and the interregional 2.4–8.6. The sampling sites were characterized in terms of biotope physiography, water chemistry, periphyton biomass, trace element content, and species composition. Multivariate analysis of the data using PLS (Projection to Latent Structure), was used to generate hypotheses about the relation between periphyton zinc tolerance and the 123 so-called predictor variables. Zinc contamination, phosphate, nitrogen nutrients, pH, calcium, bicarbonate, dissolved organic carbon, and various diatom species are important predictors for zinc tolerance in the entire data set representing all 15 river stretches. Regional models suggested that very different factors determined the zinc tolerance in the Swedish and Dutch periphyton. The results are interpreted in terms of Pollution-Induced Community Tolerance (PICT) and the bioavailability of zinc. Received: 16 December 2001/Accepted: 24 July 2002     

On the limits of toxicant-induced tolerance testing: cotolerance and response variation of antibiotic effects

Pollution-induced community tolerance (PICT) as an ecotoxicological test system has been claimed to detect pollutant effects highly specifically and sensitively. However, the specificity might be limited by the occurrence of cotolerance. Another limitation of the application of any ecotoxicological test system lies in variation of the measured responses. We tested the variation and the occurrence of cotolerance experimentally, using antibiotics as toxicants, soil microcosms as microbial communities, and tolerance determination in Biolog plates as PICT detection test. Bacteria have been discussed as being prone to multiple tolerances due to the possible accumulation of multiple resistance genes on mobile genetic elements. However, in our experiments, cotolerance occurred only between antibiotics of the same group (oxytetracycline and tetracycline), as expected from their identical mode of action. Cotolerance between oxytetracycline and tylosin in soil microcosms exposed to oxytetracycline was low, as was cotolerance to oxytetracycline in tylosin-exposed microcosms. We conclude that tolerance development to antibiotics in soils reflects the actual selection pressure rather than a general pattern of multiple resistances. Concerning variation, the PICT effect of tetracycline was well reproducible in two consecutive years. The response variation linked to PICT experiments in controlled microcosms was comparable to that of ecotoxicological test systems of equivalent complexity. In conclusion, our results support an application of the PICT methodology as an effective means to study the soil ecotoxicology of antibiotics.     

The relation between extrapolated risk, expressed as potentially affected fraction, and community effects, expressed as pollution-induced community tolerance

The results of toxicity tests can be used to calculate the potentially affected fraction (PAF) of species in an ecosystem at a given pollutant concentration using statistical extrapolation methods. The PAF curve indicates the fraction of species from the original community that may become inhibited at each elevated pollutant concentration and is a measure of the ecotoxicological risk. Pollution-induced community tolerance (PICT) is a true community response that is measured under controlled conditions in the laboratory, using organisms from contaminated field sites. Microorganisms from experimental field plots with added Zn were exposed to various concentrations of Zn in the laboratory and the mineralization of 14C acetate was monitored. Microorganisms from plots with Zn concentrations above 124 mg/kg showed a significant increase in the effect concentration 10% (EC10) and, therefore, had a significant PICT. The pore-water concentrations of Zn in these field soils were in the same magnitude as the EC10 of the microorganisms from these soils. The PAF curve was calculated from previously reported toxicity tests with five different microbial species using the average and the standard deviation of the logarithmically transformed EC10 values. The average sensitivity of this PAF curve was similar to the EC50 of the acetate mineralization curve from the field plot without added Zn2+, but the PAF curve was less steep. Our experiments indicated that 27 to 84% of the original microbial species were inhibited at Zn concentrations from 334 to 1,858 mg/kg soil, respectively. Our results suggest that the PICT method can now also be used to quantify the fraction of the original species composition that is inhibited at a specific pollutant concentration.     

Impact of Heavy Metals (Copper,Zinc, and Lead) on the Chlorophyll Content of Some Mosses

The effects of the heavy metals copper (Cu), zinc (Zn), and lead (Pb) on the chlorophyll content of two mosses Thuidium delicatulum (L.) Mitt. and T. sparsifolium (Mitt.) Jaeg., as well as leafy liverwort Ptychanthus striatus (Lehm. & Linderb.) were examined to understand the impact of metal accumulation on the chlorophyll content of mosses and leafy liverwort, which are found only in uncontaminated sites of the Kathmandu valley. These plants were treated with different concentrations of CuCl_2, ZnCl₂, and Pb(NO₃)₂, ranging from 10⁻¹⁰ M to 10⁻² M, in isolation and in combination under experimental conditions. Metal accumulation in the plant bodies increased with metal concentrations. Cu accumulation showed a significant inhibitory effect on chlorophyll-a, chlorophyll-b, and total chlorophyll in the mosses and the leafy liverwort. An insignificant decrease in chlorophyll content in both Thuidium species, but a significant decrease in leafy liverwort, was observed after Zn and Pb accumulation. Chlorophyll-a decreased significantly in T. sparsifolium; chlorophyll-b and total chlorophyll decreased significantly in T. delicatulum; and all chlorophyll contents decreased insignificantly in P. striatus after accumulation of Cu+Zn+Pb ions together from mixed metal solution. The ratio of chlorophyll-a to -b decreased more rapidly in both Thuidium species, with higher concentrations occurring when Cu+Zn+Pb ions were together than when Cu, Zn, or Pb ions were alone. This indicated a more destructive effect of Cu metals on the chlorophyll contents of both Thuidium species. High concentrations of Cu are known to activate oxidative damage and alter cell-membrane properties by lipid peroxidation, thereby demonstrating the inhibitory effect on the enzymes involved in chlorophyll production. The greater loss of chlorophyll from heavy-metal accumulation in P. striatus than in T. delicatulum and T. sparsifolium may have been caused by relatively more K⁺ efflux in leafy liverwort than in the mosses, thus indicating their differences in membrane integrity.     

Pollution-induced community tolerance and functional redundancy in a decomposer food web in metal-stressed soil

Pollution may lead to the development of pollution-induced community tolerance (PICT) in a stressed community. We studied the presence of PICT in soil food webs using soil microcosms. Soil microcosms containing soil invertebrates and microbes were collected from polluted and unpolluted areas and exposed to a range of soil zinc concentrations. A pine seedling was planted in each microcosm to measure the effects of the origin of the community and Zn pollution on above-ground plant production. The effects of the treatments on nutrient content in the soil were also measured. The diversity of soil microarthropods and the soil's mineral nutrient content were low at the Zn-polluted site. We did not observe an increasing Zn tolerance among the soil organisms in the polluted soil. However, low population growth rates of soil invertebrates from the polluted site may indicate the deleterious effects on fitness of long-lasting pollution. In the soil from the nonpolluted site, Zn additions caused changes in the invertebrate food web structure. These changes were explained by the good physiological condition of the animals and their insensitivity to Zn. The fact that the food web structure in soil from the polluted site did not change can be used as a rough indicator of PICT. Structural stability is presumed by the lack of Zn-sensitive species at this site and the inability of populations to acclimate by altering their growth or reproduction patterns in response to changing soil conditions. Although microbial-based soil decomposer systems may have a high functional redundancy, our results indicate that metal stress at the polluted site exceeds the tolerance limits of the system. As a consequence, ecosystem function at this site is endangered. This study also shows that the evolution of metal tolerance by soil decomposer organisms may not be a common reaction to soil pollution, although changes of population and community structure indicated severe metal stress on organisms.     

Responses of Antioxidant Systems After Exposition to Rare Earths and Their Role in Chilling Stress in Common Duckweed (Lemna minor L.): A Defensive Weapon or a Boomerang?

Extensive agriculture application of rare earth elements (REEs) in Far East countries might cause spreading of these metals in aquatic and terrestrial ecosystems, thus inducing a growing concern about their environmental impact. In this work the effects of a mix of different REE nitrate (RE) and of lanthanum nitrate (LA) on catalase and antioxidant systems involved in the ascorbate–glutathione cycle were investigated in common duckweed Lemna minor L. The results indicated that L. minor shows an overall good tolerance to the presence of REEs in the media. Treatments at concentrations up to 5 mM RE and 5 mM LA did not cause either visible symptoms on plants or significant effects on reactive oxygen species (ROS) production, chlorophyll content, and lipid peroxidation. Toxic effects were observed after 5 days of exposition to 10 mM RE and 10 mM LA. A remarkable increase in glutathione content as well as in enzymatic antioxidants was observed before the appearance of the stress symptoms in treated plants. Duckweed plants pretreated with RE and LA were also exposed to chilling stress to verify whether antioxidants variations induced by RE and LA improve plant resistance to the chilling stress. In pretreated plants, a decrease in ascorbate and glutathione redox state and in chlorophyll content and an increase in lipid peroxidation and ROS production levels were observed. The use of antioxidant levels as a stress marker for monitoring REE toxicity in aquatic ecosystems by means of common duckweed is discussed.     

Effects of Copper on Wild and Tolerant Strains of the Lichen Photobiont Trebouxia erici (Chlorophyta) and Possible Tolerance Mechanisms

The influence of copper was assessed on wild and tolerant strains of the lichen photobiont Trebouxia erici and shown to have multiple toxic effects. Addition of 4 mM copper chloride into Trebouxia media reduced growth rates of the wild type to less than 1% of control levels. It also injured cell membranes, inhibited dehydrogenase activity, altered pigment composition, and reduced activity of photosystem II. Membrane damage was assessed through measuring electrical conductivity and potassium content, dehydrogenase activity by degree of conversion of 2,3,5-triphenyl tetrazolium chloride to red triphenylformazan, and functioning of PS II by chlorophyll a fluorescence. In respect to most parameters the tolerant strain was usually less affected by copper than the wild strain and, in some cases, not at all. We demonstrated by inductively coupled plasma atomic emission spectrometry and energy-dispersive X-ray microanalysis that enriched copper in the medium resulted in increased uptake by both wild and tolerant photobiont cells, but extracellular concentrations were significantly higher in the tolerant strain. This suggested that, at least in vitro, extracellular deposition was a mechanism of copper tolerance.     

An Evaluation of the Use of Colonized Periphyton as an Indicator of Wastewater Impact in Near-Coastal Areas of the Gulf of Mexico

Receiving water impacts of point source discharges to the Gulf of Mexico are seldom reported on indigenous flora. The objective of this research was to evaluate the ability of colonized periphyton to provide this information. Water quality and biomass and pigment concentrations of the periphyton were determined at 27 stations located above and below 8 wastewater discharges. Most physicochemical parameters and concentrations of pesticides and PCBs were either unchanged or below detection in the receiving waters, which contrasted occasional increases in concentrations of several trace metals and nutrients. The response of the periphyton was specific to the wastewater, colonization station, response parameter, and colonization period. Statistically significant differences in biomass and pigment content occurred for at least one colonization station located below each of the eight outfalls. This represented a total of 18 of the 21 stations located in wastewater-impacted areas. Phytostimulation was more common than inhibition. Ash-free dry weight increased, on average, by 181% (± 1 SD = 123%) and chlorophyll a increased by 356% (± 593%) in wastewater-impacted areas. The in situ phytostimulation paralleled the stimulatory trend observed in standardized NPDES whole effluent tests conducted with cultured microalgae for four of eight wastewaters. The use of colonized periphyton as an indicator of wastewater impact was not simple. Spatial variation in response needs consideration to ensure relevancy of the results if this assessment methodology is used for near-coastal wastewater hazard evaluations. Received: 29 September 2001/Accepted: 7 January 2002     

Effects of organic solvents and solvent-atrazine interactions on two algae, Chlorella vulgaris and Selenastrum capricornutum

When assessing pesticides toxicity on test organisms, the use of organic solvents is frequently required to formulate solutions of pesticides having low or moderate water solubility. These solvents may influence the results of toxicity tests. This study examined the effects of ethanol and dimethyl sulfoxide (DMSO) on two species of algae, Chlorella vulgaris and Selenastrum capricornutum. The interactives effects of these organic solvents with various concentrations of atrazine were tested to analyse how these solvents can affect the toxicity of atrazine. Toxicity was measured as the change in chlorophyll (a) content (estimated via fluorescence) in cultures of the test organism that were incubated over a 96-h period at standard conditions of temperature and light. In the absence of atrazine, ethanol was toxic to both algae, yielding significant inhibition of chlorophyll (a) content at concentrations as low as 0.05%. Morever, S. capricornutum was less sensitive to ethanol than was C. vulgaris. DMSO did not show any toxic effects on either chlorophycea. At concentrations up to 0.5%, DMSO interacted additively with atrazine for both chlorophycea. An additive response was also observed with ethanol towards S. capricornutum. For C. vulgaris, ethanol interacted antagonistically at most of atrazine concentrations and gave few additive and synergistic interactions. An additive response of an atrazine-solvent mixture indicate the inherent toxicity of the atrazine. Since DMSO generally gave additive responses for both algae, it can be considered as an adequate organic solvent to use in bioassays.     

Effects of Exposing Two Non-Target Crustacean Species, Asellus aquaticus L., and Gammarus fossarum Koch., to Atrazine and Imidacloprid

The physiological responses of two freshwater crustaceans, Asellus aquaticus L. and Gammarus fossarum Koch., following in vitro exposure to two pesticides were measured. Both species responded to short-term exposure with elevated levels of Respiration (R) and/or lower levels of Electron Transport System (ETS) activity. 1 h exposure to concentrations of up to 10 mg L⁻¹ showed an effect in both test species. Laboratory tests confirmed that G. fossarum is more sensitive to short-term pesticide exposure than A. aquaticus. ETS/R ratio may be used as a quick predictor of effects on organisms exposed to pesticides.     

Influence of Selected Water Quality Characteristics on the Toxicity of λ-Cyhalothrin and γ-Cyhalothrin to <Emphasis Type="Italic">Hyalella azteca</Emphasis>

This study was conducted to assess the influence of suspended solids, dissolved organic carbon, and phytoplankton (as chlorophyll a) water quality characteristics on λ-cyhalothrin and γ-cyhalothrin aqueous toxicity to Hyalella azteca using natural water from 12 ponds and lakes in Mississippi, USA with varying water quality characteristics. H. azteca 48-h immobilization EC50 values ranged from 1.4 to 15.7 ng/L and 0.6 to 13.4 ng/L for λ-cyhalothrin and γ-cyhalothrin, respectively. For both pyrethroids, EC50 values linearly increased as turbidity, suspended solids, dissolved organic carbon and chlorophyll a concentrations increased.     

Effects of atrazine on freshwater microbial communities

A multispecies toxicity test system using naturally derived microbial communities on polyurethane foam substrates was used to evaluate the toxic effects of the herbicide atrazine. Both structural (e.g., protozoan species number, biomass) and functional (e.g., colonization rate, oxygen production) community responses were measured. Oxygen production and the ability of communities to sequester magnesium and calcium were the most sensitive measures of toxic stress due to atrazine (maximum allowable toxicant concentrations [MATCs]=17.9 g/L). Dissolved oxygen was 33% lower, and there was 15% less calcium and magnesium in communities at and above 32.0 g/L atrazine compared to controls. Species richness and estimates of biomass (total protein and chlorophyll a) were less sensitive (MATCs=193) to atrazine. At the highest atrazine concentration (337 g/L), species numbers were 30% lower than controls, and protein and chlorophyll a content of communities were reduced by 38 and 91%, respectively. Low levels of atrazine (3.2–32.0 g/L) resulted in a 46% increase in species numbers and a greater concentration of total protein and chlorophyll a (41 and 57%, respectively). Results compared well with other estimates of chronic toxicity for effects of atrazine on aquatic communities. Reported MATCs ranged from 70.7 to 3,400 g/L. The results from this test emphasize the importance of monitoring both structural and functional measures of community integrity in toxicity testing with multispecies.     

Construction of River Model Biofilm for Assessing Pesticide Effects

Due to the high importance of biofilms on river ecosystems, assessment of pesticides’ adverse effects is necessary but is impaired by high variability and poor reproducibility of both natural biofilms and those developed in the laboratory. We constructed a model biofilm to evaluate the effects of pesticides, consisting in cultured microbial strains, Pedobacter sp. 7-11, Aquaspirillum sp. T-5, Stenotrophomonas sp. 3-7, Achnanthes minutissima N71, Nitzschia palea N489, and/or Cyclotella meneghiniana N803. Microbial cell numbers, esterase activity, chlorophyll-a content, and the community structure of the model biofilm were examined and found to be useful as biological factors for evaluating the pesticide effects. The model biofilm was formed through the cooperative interaction of bacteria and diatoms, and a preliminary experiment using the herbicide atrazine, which inhibits diatom growth, indicated that the adverse effect on diatoms inhibited indirectly the bacterial growth and activity and, thus, the formation of the model biofilm. Toxicological tests using model biofilms could be useful for evaluating the pesticide effects and complementary to studies on actual river biofilms.     

In VivoChlorophyll A Fluorescence of Selenastrum capricornutum as a Screening Bioassay in Toxicity Studies

A method for the estimation of the effect of specific toxins on phytoplankton photosynthesis (index of toxicity) was investigated using in vivo chlorophyll a fluorescence. No meaningful results were obtained with the following substances; copper, cadmium, mercury, and gusathion, within a 4-h exposure period. It can, therefore, be concluded that in vivo chlorophyll a fluorescence are not a simple bioassay tool, due to the complexity of its origins, but a complex research tool. Received: 7 August 1997/Accepted: 8 February 1998