Do bioassays adequately predict ecological effects of pollutants?

With some notable exceptions, such as the echinoderm and oyster larvae tests, the species traditionally used in bioassays are not sufficiently sensitive to detect subtle ecological effects of pollutants. It is suggested that by using ecological criteria, species can be identified from any pollution gradient that are sensitive to subtle effects of pollution. Examples are given using gradients of oil, sewage and titanium dioxide pollution, showing how ecologically sensitive species for use in laboratory bioassays can be selected objectively.Many marine molluscs show microgrowth bands, which can be used as in situ field bioassays. An example is given using the bivalve Cerastoderma edule.     

Water sample treatments and their effects on bioassays using Stigeoclonium helveticum vischer

The effects of different pretreatments on the chemical properties of test waters and their effects on bioassays using Stigeoclonium helveticum Vischer were evaluated. All treatments removed or destroyed indigenous algae, and in addition each method altered the water chemistry. Filtration removed 78–98% of the iron, and up to 20% of the total phosphorus concentration; autoclaving caused precipitation of salts and removed up to 20% of total phosphorus. The effect was enhanced up to 60% when a combination of both methods was applied. The inorganic nitrogen concentration did not alter significantly. The various sample treatments changed the availability of the limiting nutrients. This was manifested in the bioassays using Stigeoclonium helveticum.     

Chemical and physical defenses of Singapore gorgonians (Octocorallia: Gorgonacea)

Gorgonians are abundant in tropical waters and their polyps are seldom predated on. This study investigates how gorgonians defend themselves chemically and physically against fish predation. Gorgonian extracts and sclerites were incorporated into fish feed and tested on reef fishes. Laboratory bioassays using Greyhead wrasses, Halichoeres purpurescens, as well as field bioassays showed five gorgonian species from the family Ellisellidae and three from the family Plexauridae collected from Singapore reefs to be deterrent towards fishes. Bioassays of fractions obtained from subsequent fractionation suggested synergistic or additive effects between compounds present in gorgonians. Sclerites incorporated into fish feed in their natural concentrations were also tested for fish deterrence and were positive for only two gorgonian species from the family Ellisellidae.     

Use of a Battery of Chemical and Ecotoxicological Methods for the Assessment of the Efficacy of Wastewater Treatment Processes to Remove Estrogenic Potency

Endocrine Disrupting Compounds pose a substantial risk to the aquatic environment. Ethinylestradiol (EE2) and estrone (E1) have recently been included in a watch list of environmental pollutants under the European Water Framework Directive. Municipal wastewater treatment plants are major contributors to the estrogenic potency of surface waters. Much of the estrogenic potency of wastewater treatment plant (WWTP) effluents can be attributed to the discharge of steroid estrogens including estradiol (E2), EE2 and E1 due to incomplete removal of these substances at the treatment plant. An evaluation of the efficacy of wastewater treatment processes requires the quantitative determination of individual substances most often undertaken using chemical analysis methods. Most frequently used methods include Gas Chromatography-Mass Spectrometry (GCMS/MS) or Liquid Chromatography-Mass Spectrometry (LCMS/MS) using multiple reaction monitoring (MRM). Although very useful for regulatory purposes, targeted chemical analysis can only provide data on the compounds (and specific metabolites) monitored. Ecotoxicology methods additionally ensure that any by-products produced or unknown estrogenic compounds present are also assessed via measurement of their biological activity. A number of in vitro bioassays including the Yeast Estrogen Screen (YES) are available to measure the estrogenic activity of wastewater samples. Chemical analysis in conjunction with in vivo and in vitro bioassays provides a useful toolbox for assessment of the efficacy and suitability of wastewater treatment processes with respect to estrogenic endocrine disrupting compounds. This paper utilizes a battery of chemical and ecotoxicology tests to assess conventional, advanced and emerging wastewater treatment processes in laboratory and field studies.     

Experimental Comparison of the Reproductive Outcomes and Early Development of the Offspring of Rats Given Five Common Types of Drinking Water

Tap water (unfiltered), filtered tap water and processed bottled water (purified water, artificial mineralized water, or natural water) are now the five most widely consumed types of drinking water in China. However, the constituents (organic chemicals and inorganic ingredients) of the five waters differ, which may cause them to have different long-term health effects on those who drink them, especially sensitive children. In order to determine which type of water among the five waters is the most beneficial regarding reproductive outcomes and the developmental behaviors of offspring, two generations of Sprague–Dawley rats were given these five waters separately, and their reproductive outcomes and the developmental behaviors of their offspring were observed and compared. The results showed that the unfiltered tap water group had the lowest values for the maternal gestation index (MGI) and offspring''s learning and memory abilities (OLMA); the lowest offspring survival rate was found in the purified water group; and the highest OLMA were found in the filtered tap water group. Thus, the best reproductive and offspring early developmental outcomes were found in the group that drank filtered tap water, which had the lowest levels of pollutants and the richest minerals. Therefore, thoroughly removing toxic contaminants and retaining the beneficial minerals in drinking water may be important for both pregnant women and children, and the best way to treat water may be with granular activated carbon and ion exchange by copper zinc alloy.     

Runoff from tomato cultivation in the estuarine environment: biological effects of farm management practices

The use of plastic row covers (plastic mulch) on vegetable farms increases runoff of pesticides after rainfall events and has been linked to toxic events in adjacent tidal waters. In coastal Virginia, USA, runoff from tomato fields with plastic mulch was suspected of causing mortality of commercial hard clam larvae at a hatchery located downstream of farming operations. Concern about the putative impacts of this practice on local waters resulted in a collection of studies to: (1) determine the sensitivity of early life stages of bivalves to copper, a commonly used fungicide; (2) examine acute and chronic biological effects of runoff on tidal creeks; and (3) examine the efficacy of management practices designed to reduce the delivery of pesticides to adjacent creeks. Laboratory bioassays revealed that 48-h LC₅₀ values for embryonic clams Mulinia lateralis and Mercenaria mercenaria were 38 and 20 μg/l, respectively. In situ bioassays with Palaemonetes pugio showed that pulsed toxic conditions sometimes occur downstream of some tomato farms in plastic mulch following rainfall events. Growth, mortality rates and bioaccumulation of copper and organic pesticides in oysters were not correlated with the use of plastic mulch in watersheds. Sediment bioassays indicated potential toxicity in sediment collected downstream of some tomato fields in plastic mulch, but the effects were not consistent between years. Closer examination of management practices on the farms suggests that controlling runoff can prevent toxic impacts. Elevated levels of crop protectants measured at the outflow of farm ponds suggested that they may do little to reduce loadings of some pesticides. However, forested buffer zones and ephemeral sedimentation basins appeared to be effective in reducing pesticide concentrations in runoff and pulsed toxicity in tidal creeks.     

Acute Toxicity Tests with Cadmium,Lead, Sodium Dodecyl Sulfate,and Bacillus thuringiensis on a Temporary Pond Ostracod

A complete assessment of effects of pollutants in the environment requires the use of a wide assemblage of test organisms. Crustaceans have been widely used in aquatic environments, but within this group, ostracods have been overlooked, even though they are one of the commonest groups of animals in temporary waters. Within this framework, static acute toxicity bioassays were conducted with heavy metals Cd and Pb, the surfactant SDS and spores of the bacteria Bacillus thuringiensis var. israeliensis using adult parthenogenetic females of the temporary pond ostracod Heterocypris bosniaca. Experiments were run at 20 °C and in a 12 by 12 h photoperiod. The toxicity tests results are expressed as LC₅₀ and EC₅₀ at 48 h and 96 h using probit analysis. The sensitivity of H. bosniaca at 48 h decreased in the following order Cd > Pb > SDS > Bti, while Pb and SDS exchanged their positions at 96 h. Comparison of the LC₅₀ values for this species with information available in the literature reveals that it is generally more resistant than other crustaceans. (© 2012 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Zooplankters as indicators of ecosystem health: past findings and future directions

Both acute (ingestion, respiration) and chronic bioassays (reproduction, survival) have been used to identify sources of pollutants. A mass-balance analysis suggests that acute tests be paired, using important indicator species asDaphnia, Ceriodaphnia and potentially others, to estimate the impact of contaminants upon the zooplankton community. Eventually groups of community bioassays may be combined to approximate an ecosystem bioassay. Hormesis or the stimulation of a physiological process by a compound which is toxic at high concentrations is characteristic of several bioassays; in this paper the ecotoxicology community is challenged to keep detailed records of the species, toxic compound, and physiological response involving hormesis in order to understand it; and ultimately to use it to simplify interpretation of bioassays. Life history characteristics of the cladoceran zooplankton, including early reproduction, high net reproductive rates, and the potential for many parthenogenetic generations with constant genotypes and low mutation rates make good choices for environmental bioassays. In contrast, high mutation rates of rotifers make them questionable choices. Five innovations, one or more of which may improve our ability to detect and identify pollutants, are suggested for ecotoxicologists using zooplankton. These include (a) the use of strains of known genotype; (b) determination of the genetic adaptation of clones to common toxins; (c) the use of common behaviors, including responses to light in detection of non-lethal chemicals at ambient levels; (d) record keeping on occurrence of cladoceran tumors; and (e) the determination of precise toxins responsible for the inhibition of zooplankton function and behavior.     

Ecosystem health as measured from the molecular to the community level of organization,with reference to sediment bioassessment

The current recognition that chemical measurements are uncertain indicators of biological consequences of pollution has shifted the emphasis away from assessing environmental chemistry alone toward the inclusion of measurements of the health of organisms. Effects of pollutants begin with the individual, have subsequent repercussions on population level processes, and ramifications for community structure and functions. Pollutants act at a molecular level and the biochemical lesions is the first step in the manifestation of effects. Technologies that operate at the cellular level assist in elucidating toxicity. Higher levels of integration include an organism's capacity for growth. Laboratory bioassays andin situ research can monitor physiological incapacities and assist in predicting population level effects. A yet higher level of organization is that of the ecological community.     

Susceptibility of immature Ixodes scapularis (Acari: Ixodidae) to desiccants and an insecticidal soap

Efficacy of commercially available formulations of desiccants (Drione, Dri-dic, diatomaceous earth) and insecticidal soap (Safer's) comprised of active ingredients were evaluated against Ixodes scapularis Say immatures in petri dish and grass turf bioassays. In petri dish bioassays at label rate, all products, except diatomaceous earth, provided a high degree of control by 24 and 48 h (71–100% mortality). Mortality of larvae exceeded 94% by 4 h after treatment with Drione (10× label rate held at 85% RH), Dri-die (label rate held at both 85% and 98% RH) and Safer's insecticidal soap (10× label rate held at 98% RH). Nymphal mortality was highest by 4 h after treatment with Safer's insecticidal soap (up to 96%). Larvae were more susceptible than nymphs to desiccants, however, nymphs were more susceptible than larvae to Safer's insecticidal soap. Immature ticks treated with Safer's insecticidal soap exhibited sublethal effects with significantly decreased attachment to hosts and no engorgement. In grass turf bioassays, mortality of nymphs was equivalent (85–95%) between treatments of chlorpyrifos (0.6 kg [AI]/ha, Safer's insecticidal soap, and Drione. In the same bioassays, nymphal mortality was moderate after treatment with Dri-die (23–29%) and low following application with diatomaceous earth (16%) and for untreated controls (6%).     

Aquatic plant bioassays used in the assessment of water quality in German rivers

A novel aquatic bioassay with Lemna minor (duckweed), a representative of higher plants, was applied to pore waters (interstitial waters) from river sediments. The results have been used for the ecotoxicological assessment of sediments in several German rivers. The aim of the present study is to demonstrate the usefulness of a macrophyte bioassay, as a supplement to the standardised algae bioassays, with regard to the ecotoxicological assessment of aquatic systems. Results relating to the development of a sediment contact test (using the whole sediment) with Lemna minor are also presented.     

Effects of pesticides and related organic compounds in the sea

The majority of organic chemicals identified so far in the sea are pesticides and products of technical use; most contain chlorine. Only a limited amount of the actual pollutant load is detectable because few data for “unconventional ” pollutants are available. In view of the considerable structural variety of the large number of chemicals produced, there is a need for prediction measurements of bioconcentration and toxic effects. Physico-chemical data may be used for predicting bioconcentration and life-cycle toxicity tests for the estimation of safe levels. The degree of biomagnification via food chains increases with half lives of the pollutants. When comparing pollutant concentrations with toxicological data it becomes apparent that estuaries and coastal areas deserve special concern, whereas pollutant levels of open ocean waters are unlikely to endanger marine life at present.     

Détection à l'aide d'un bio-essai (Selenastrum capricornutum) des répercussions d'un rejet minier sur l'environnement aquatique: toxicité ou enrichissement en substance essentielle?

Physical and chemical studies and algal bioassays were conducted to assess the repercussions on waterways receiving the treated effluent wastes of an iron ore mining operation located in north-eastern Quebec, Canada.In the area directly below waste input, conductivity, turbidity, suspended solids, nitrite + nitrate nitrogen and phosphorus levels, as well as algal growth potential generally displayed higher values than elsewhere. With these parameters only, however, no perceptible mining impact on two major receiving waterways (aux Pékans and Moisie rivers) could be shown, except for nitrite + nitrate nitrogen values in the aux Pékans River.The presence of heavy metals did not result in any toxic effects as indicated by bioassays conducted with S. capricornutum. These tests when carried out on samples collected during low water levels, spiked with nitrogen + phosphorus, as well as with modified AAP medium, clearly indicated the contribution of the mining wastes to the enrichment of the receiving waters. This effect was linked to essential element other than nitrogen or phosphorus. For samples collected during high water levels, it is suspected that the increased presence of organic matter interferes by complexation of trace metals.Cautious interpretation of results is therefore recommended when using algal bioassays in order to distinguish between effects of growth-limiting essential element and the presence of toxic substances.

     

The Toxicology of PCB's—An Overview for Clinicians

Polychlorinated biphenyls (PCB''s) have low acute toxicity but are of public health concern because of their persistence in the environment, the bioaccumulation in human and animal tissues, and their potential for chronic or delayed toxicity.Although many questions remain unanswered, it is clear that occupational exposure, at a minimum, can produce dermatologic effects and liver dysfunction. The long half-life of PCB''s and their presence in various human tissues leaves open the possibility of substantial chronic and delayed effects analogous to those seen in animals. PCB''s are potent inhibitors of reproductive function in both rodents and nonhuman primates and produce liver tumors in animal cancer bioassays. As potent inducers of hepatic enzyme systems, PCB''s may have additional unpredictable long-term health effects.These effects have only recently begun to be studied in a rigorous manner, and although the epidemiological evidence is neither complete nor entirely consistent, there can be no question of the necessity to keep human exposures to the lowest feasible levels.     

菲、芘、1,2,4-三氯苯对土壤高等植物根伸长抑制的生态毒性效应

测定了草甸棕壤条件下,菲、芘、１,２,４—三氮苯对高等植物（小麦、白菜、西红柿）根伸长抑制串以及复合污染毒性效应。结果表明,菲、芘、１,２,４—三氮苯浓度与植物根伸长抑制串呈显著线性或对数相关（ｐ＝０．０５）。３种化学品对植物根伸长抑制的强弱顺序为１,２,４—三氮苯＞菲＞芘。这与３种化学品的水中溶解度大小显著相关。小麦是３种供试植物中对有机污染物最敏感植物。菲、芘、１,２,４—三氮苯复合污染主表现为协同作用。     

Genotoxicity of leachates from a landfill using three bioassays.

In the city of Queretaro, around 500 tons of solid wastes are produced everyday and are deposited in a landfill. This is the result of social and economic activities of human beings or from their normal physiological functions. As a result of rain, leachates are produced, which, if not handled and treated correctly, may pollute the underground water. Among the bioassays developed for the detection of mutagenicity in environmental pollutants, plant systems have been proven to be sensitive, cheap, and effective. The purpose of this study was to determine the presence of genotoxic agents in the leachates of the landfill of the city using three bioassays: Tradescantia-micronucleus (Trad-MCN), Tradescantia stamen hair mutations (Trad-SHM) and Allium root anaphase aberrations (AL-RAA) and make a comparison of the results in the three assays. Leachates were sampled during both the dry and rainy seasons. Plant cuttings of Tradescantia or the roots of Allium were treated by submerging them in the leachates. Three replicates of each sample were analyzed in each of the three bioassays. As expected the samples of leachates collected during the dry season showed a higher genotoxicity than those collected during the rainy season. In conclusion, there are substances present in the leachates capable of inducing genotoxicity in the plant assays. On the other hand, the plant assays showed different degrees of sensitivity: the more sensitive was the Trad-MCN bioassay and the less sensitive the Trad-SHM assay. Therefore, when analyzing environmental pollutants it is recommended to use a battery of bioassays.     

Nutrient addition effects on chlorophyll a,phytoplankton biomass,and heterocyte formation in Lake Erie’s central basin during 2014–2017: Insights into diazotrophic blooms in high nitrogen water

1. Phosphorus (P) usually is the primary limiting nutrient of phytoplankton biomass, but attention towards nitrogen (N) and trace nutrients, such as iron (Fe), has surfaced. Additionally, N-fixing cyanobacterial blooms have been documented to occur in N-rich, P-poor waters, which is counterintuitive from the paradigm that low N and high P promotes blooms. For example, Lake Erie's central basin has Dolichospermum blooms when nitrate concentrations are high, which raises questions about which nutrient(s) are selecting for Dolichospermum over other phytoplankton and why an N-fixer is present in high N waters?
2. We conducted a 4-year (2014–2017) study in Lake Erie's central basin to determine which nutrient (P, N, or trace nutrients such as Fe, molybdenum [Mo], and boron [B]) constrained chlorophyll concentration, phytoplankton biovolume, and nitrate assimilation using nutrient enrichment bioassays. The enriched lake water was incubated in 1-L bottles in a growth chamber programmed at light and temperatures of in situ conditions for 4–7 days. We also quantified heterocytes when N-fixing cyanobacteria were present.
3. Compared to the non-enriched control, the P-enriched (+P) treatment had significantly higher chlorophyll and phytoplankton biovolume in c. 75% of experiments. Combination enrichments of P with ammonium-N, nitrate-N, Fe, Mo, and B were compared to the +P treatment to determine secondary limitations. +P and ammonium-N and +P nitrate-N resulted in higher chlorophyll in 50% of experiments but higher phytoplankton biovolume in only 25% of experiments. These results show that P was the primary limiting nutrient, but there were times when N was secondarily limiting.
4. Chlorophyll concentration indicated N secondary limitation in half of the experiments, but biovolume indicated only N secondary limitation in 25% of the experiments. To make robust conclusions from nutrient enrichment bioassays, both chlorophyll and phytoplankton biovolume should be measured.
5. The secondary effects of Fe, Mo, and B on chlorophyll were low (<26% of experiments), and no secondary effects were observed on phytoplankton biovolume and nitrate assimilation. However, +P and Fe resulted in more chlorophyll than +P in experiments conducted during Dolichospermum blooms, and +P and B significantly increased the number of heterocytes in Dolichospermum. These results indicate that low Fe availability might select for Dolichospermum, and low B constrains heterocyte formation in the central basin of Lake Erie. Furthermore, these results could apply to other lakes with high N and low P where diazotrophic cyanobacterial blooms occur.

     

Genotoxicity of soil from farmland irrigated with wastewater using three plant bioassays.

Three well known plant bioassays, the Allium root chromosome aberration (AL-RAA) assay, the Tradescantia micronucleus (Trad-MCN) assay, and the Tradescantia stamen hair (Trad-SHM) mutation assay were validated in 1991 by the International Programme on Chemical Safety (IPCS) under the auspices of the World Health Organization, and the United Nations Environment Programme (UNEP). These plant bioassays have proven to be efficient tests for chemical screening and especially for in situ monitoring for genotoxicity of environmental pollutants. As a result of this validation study, standard protocols of these three plant bioassays were used by some of the 11 participating countries in the IPCS to carry on genotoxicity tests on air, water and soil as a follow up activity. In the city of Queretaro, Mexico, wastewater coming from both industrial and domestic sources and without any treatment is used to irrigate the farm crops, polluting the soil. Potentially the pollutants could reach the food chain. For the above reason, soil irrigated with wastewater was sampled and monitored for the presence of genotoxic agents using the above three bioassays. Extracts from soil samples were made using distilled water and organic solvents by shaking the sample for about 12 h under a relatively low temperature (15-20 degrees C). Plant cuttings of Tradescantia or the roots of Allium were treated by submerging them in the extracts. Three replicates of each sample were analyzed in each of the three bioassays. Extracts using DMSO, ethanol and distilled water tested positive in the three bioassays and there were no differences for the genotoxicity of the extracts with the different solvents.     

The role of aquatic toxicity tests in predicting and monitoring pollution effects

The complementary role of aquatic toxicity tests in relation to other forms of pollution assessment is discussed. These tests may be predictive and designed to estimate hazard, or used for monitoring waters to assess compliance with standards. The effects of pollutants are significantly modified by certain water quality characteristics and by a variety of biological factors. The latter, particularly those causing physiological stress, are discussed.