The gut microbiota: a major player in the toxicity of environmental pollutants?

Exposure to environmental chemicals has been linked to various health disorders, including obesity, type 2 diabetes, cancer and dysregulation of the immune and reproductive systems, whereas the gastrointestinal microbiota critically contributes to a variety of host metabolic and immune functions. We aimed to evaluate the bidirectional relationship between gut bacteria and environmental pollutants and to assess the toxicological relevance of the bacteria–xenobiotic interplay for the host. We examined studies using isolated bacteria, faecal or caecal suspensions—germ-free or antibiotic-treated animals—as well as animals reassociated with a microbiota exposed to environmental chemicals. The literature indicates that gut microbes have an extensive capacity to metabolise environmental chemicals that can be classified in five core enzymatic families (azoreductases, nitroreductases, β-glucuronidases, sulfatases and β-lyases) unequivocally involved in the metabolism of >30 environmental contaminants. There is clear evidence that bacteria-dependent metabolism of pollutants modulates the toxicity for the host. Conversely, environmental contaminants from various chemical families have been shown to alter the composition and/or the metabolic activity of the gastrointestinal bacteria, which may be an important factor contributing to shape an individual’s microbiotype. The physiological consequences of these alterations have not been studied in details but pollutant-induced alterations of the gut bacteria are likely to contribute to their toxicity. In conclusion, there is a body of evidence suggesting that gut microbiota are a major, yet underestimated element that must be considered to fully evaluate the toxicity of environmental contaminants.

Single versus multiple occupancy - Effects on toxicity parameters measured on Eisenia fetida in lead nitrate-treated soil

The mortality (7 and 14 d), weight change (7 and 14 d), and metal uptake of Eisenia fetida (Savigny, 1826) kept in Pb(NO3)(2)-treated Kettering loam soil in single- and multiple-occupancy (10 earthworms) test containers were determined. The number of earthworms to dry mass (g) ratio of soil was 1:50 in both sets of test containers. Lead concentrations were in the nominal range of 0 to 10,000 mg Pb/kg soil (mg/kg hereafter). Levels of mortality at a given concentration were statistically identical between the single- and multiple-occupancy tests, except at 1,800 mg/kg, at which significantly (p less than or equal to 0.05) more mortality occurred in the multiple-occupancy tests. Death of individual earthworms in the multiple-occupancy tests did not trigger death of the other earthworms in that soil. The LC50 values (concentration statistically likely to kill 50% of the population) were identical between the multiple- and single-occupancy soils: 2,662 mg/kg (2,598-2,984, 7 d) and 2,589 mg/kg (2,251-3,013, 14 d) for the multiple-occupancy soils and 2,827 mg/kg (2,443-3,168, both 7 and 14 d) for the single-occupancy soils (values in brackets represent the 95% confidence intervals). Data were insufficient to calculate the concentration statistically likely to reduce individual earthworm mass by 50% (EC50), but after 14 d, the decrease in earthworm weight in the 1,800 and 3,000 mg/kg tests was significantly greater in the multiple- than in the single-occupancy soils. At 1,000, 1,800, and 3,000 mg/kg tests, earthworm Pb tissue concentration was significantly (p less than or equal to 0.05) greater in earthworms from the multiple-occupancy soils. The presence of earthworms increased the NH3 content of the soil; earthworm mortality increased NH3 concentrations further but not to toxic levels.

Is the OECD acute worm toxicity test environmentally relevant? The effect of mineral form on calculated lead toxicity

In a series of experiments the toxicity of lead to worms in soil was determined following the draft OECD earthworm reproduction toxicity protocol except that lead was added as solid lead nitrate, carbonate and sulphide rather than as lead nitrate solution as would normally be the case. The compounds were added to the test soil to give lead concentrations of 625-12500 pg Pb g-1 of soil. Calculated toxicities of the lead decreased in the order nitrate > carbonate > sulphide, the same order as the decrease in the solubility of the metal compounds used. The 7-day LC50 (lethal concentration when 50% of the population is killed) for the nitrate was 5321 +/- 275 mug Pb g(-1) of soil and this did not change with time. The LC50 values for carbonate and sulphide could not be determined at the concentration ranges used. The only parameter sensitive enough to distinguish the toxicities of the three compounds was cocoon (egg) production. The EC50S for cocoon production (the concentration to produce a 50% reduction in cocoon production) were 993, 8604 and 10 246 mug Pb g(-1) of soil for lead nitrate, carbonate and sulphide, respectively. Standard toxicity tests need to take into account the form in which the contaminant is present in the soil to be of environmental relevance. (C) 2002 Elsevier Science Ltd. All rights reserved.

The influence of time on lead toxicity and bioaccumulation determined by the OECD earthworm toxicity test

Internationally agreed standard protocols for assessing chemical toxicity of contaminants in soil to worms assume that the test soil does not need to equilibrate with the chemical to be tested prior to the addition of the test organisms and that the chemical will exert any toxic effect upon the test organism within 28 days. Three experiments were carried out to investigate these assumptions. The first experiment was a standard toxicity test where lead nitrate was added to a soil in solution to give a range of concentrations. The mortality of the worms and the concentration of lead in the survivors were determined. The LC(50)s for 14 and 28 days were 5311 and 5395 mug(Pb) g(soil)(-1) respectively. The second experiment was a timed lead accumulation study with worms cultivated in soil containing either 3000 or 5000 mug(Pb) g(soil)(-1). The concentration of lead in the worms was determined at various sampling times. Uptake at so' Sol both concentrations was linear with time. Worms in the 5000 mug g(-1) soil accumulated lead at a faster rate (3.16 mug Pb g(tissue)(-1) day(-1)) tiss than those in the 3000 mug g(-1) soil (2.21 mug Pb-tissue g(-1) day(-1)). The third experiment was a timed experiment with worms cultivated in tiss soil containing 7000 mugPb g(soil)(-1). Soil and lead nitrate solution were mixed and stored at 20 degreesC. Worms were added at various times over a 35-day period. The time to death increased from 23 h, when worms were added directly after the lead was added to the soil, to 67 It when worms were added after the soil had equilibrated with the lead for 35 days. In artificially Pb-amended soils the worms accumulate Pb over the duration of their exposure to the Pb. Thus time limited toxicity tests may be terminated before worm body load has reached a toxic level. This could result in under-estimates of the toxicity of Pb to worms. As the equilibration time of artificially amended Pb-bearing soils increases the bioavailability of Pb decreases. Thus addition of worms shortly after addition of Pb to soils may result in the over-estimate of Pb toxicity to worms. The current OECD acute worm toxicity test fails to take these two phenomena into account thereby reducing the environmental relevance of the contaminant toxicities it is used to calculate. (C) 2002 Elsevier Science Ltd. All rights reserved.

Measuring and modelling mixture toxicity of imidacloprid and thiacloprid on Caenorhabditis elegans and Eisenia fetida

While the standard models of concentration addition and independent action predict overall toxicity of multicomponent mixtures reasonably, interactions may limit the predictive capability when a few compounds dominate a mixture. This study was conducted to test if statistically significant systematic deviations from concentration addition (i.e. synergism/antagonism, dose ratio- or dose level-dependency) occur when two taxonomically unrelated species, the earthworm Eisenia fetida and the nematode Caenorhabditis elegans were exposed to a full range of mixtures of the similar acting neonicotinoid pesticides imidacloprid and thiacloprid. The effect of the mixtures on C. elegans was described significantly better (p<0.01) by a dose level-dependent deviation from the concentration addition model than by the reference model alone, while the reference model description of the effects on E. fetida could not be significantly improved. These results highlight that deviations from concentration addition are possible even with similar acting compounds, but that the nature of such deviations are species dependent. For improving ecological risk assessment of simple mixtures, this implies that the concentration addition model may need to be used in a probabilistic context, rather than in its traditional deterministic manner. Crown Copyright (C) 2008 Published by Elsevier Inc. All rights reserved.

Survival, Pb-uptake and behaviour of three species of earthworm in Pb treated soils determined using an OECD-style toxicity test and a soil avoidance test

Mature (clitellate) Eisenia andrei Bouche (ultra epigeic), Lumbricus rubellus Hoffmeister (epigeic), and Aporrectodea caliginosa (Savigny) (endogeic) earthworms were placed in soils treated with Pb(NO3)(2) to have concentrations in the range 1000 to 10 000 mg Pb kg(-1). After 28 days LC50(-95%confidence limit) (+95%confidence limit) values were E. andrei 5824(-361)(+898) mg Pb kg(-1), L. rubellus 2867(-193)(+145) mg Pb kg(-1) and A. caliginosa 2747(-304)(+239) mg Pb kg(-1) and EC50s for weight change were E. andrei 2841(-68)(+150) Pb kg(-1), L. rubellus 1303(-201)(+204) mg Pb kg(-1) and A. caliginosa 1208(-206)(+212) Mg Pb kg(-1). At any given soil Pb concentration, Pb tissue concentrations after 28 days were the same for all three earthworm species. In a soil avoidance test there was no difference between the behaviour of the different species. The lower sensitivity to Pb exhibited by E. andrei is most likely due to physiological adaptations associated with the modes of life of the earthworms, and could have serious implications for the use of this earthworm as the species of choice in standard toxicological testing. (c) 2005 Elsevier Ltd. All rights reserved.

Inherited resistance to arsenate toxicity in two populations of Lumbricus rubellus

No unequivocal evidence exists of genetically inherited resistance to metals/metalloids in field populations of earthworms. We studied cocoon production in adult Lumbricus rubellus Hoffmeister collected from an abandoned arsenic and copper mine (Devon Great Consols, Devon, UK), and abandoned tungsten mine (Carrock Fell, Cumbria, UK) and an uncontaminated cultured population. The earthworms were kept in uncontaminated soil for nine weeks. From a total of 42 L. rubellus from each site, Devon Great Consols adults produced 301 cocoons, of which 42 were viable; Carrock Fell 60 cocoons, of which 11 were viable; and the reference population 101 cocoons, of which 62 were viable. The hatchlings were collected and stored at 4degreesC at weekly intervals. After 12 weeks, all hatchlings were transferred to clean soil and maintained at 15degreesC for 20 weeks until they showed evidence of a clitellum. In toxicity trials, F1 generation L. rubellus were exposed to 2,000 mg As/kg as sodium arsenate or 300 mg Cu/kg as copper chloride for 28 d. The F1 generation L. rubellus from Devon Great Consols mine demonstrated resistance to arsenate but not copper. All L. rubellus from Devon Great Consols kept in soil treated with sodium arsenate remained in good condition over the 28-d period but lost condition rapidly and suffered high mortality in soil treated with copper chloride. The control population suffered high mortality in soil treated with sodium arsenate and copper chloride. Previous work has shown that field-collected adults demonstrate resistance to both arsenate and Cu toxicity under these conditions. Thus, while arsenate resistance may be demonstrated in F1 generation L. rubellus from one of the contaminated sites, Cu resistance is not. The F1 adults and F2 cocoons did not have significantly higher levels of As than the control population, with no residual As tissue burden, suggesting that resistance to As in these populations may be inherited.

MEASUREMENT AND MODELING OF THE TOXICITY OF BINARY MIXTURES IN THE NEMATODE CAENORHABDITIS ELEGANS-A TEST OF INDEPENDENT ACTION

Ecological risk assessments must increasingly consider the effects of chemical mixtures on the environment as anthropogenic pollution continues to grow in complexity. Yet testing every possible mixture combination is impractical and unfeasible; thus, there is an urgent need for models that can accurately predict mixture toxicity from single-compound data. Currently, two models are frequently used to predict mixture toxicity from single-compound data: Concentration addition and independent action (IA). The accuracy of the predictions generated by these models is currently debated and needs to be resolved before their use in risk assessments can be fully justified. The present study addresses this issue by determining whether the IA model adequately described the toxicity of binary mixtures of five pesticides and other environmental contaminants (cadmium, chlorpyrifos, diuron, nickel, and prochloraz) each with dissimilar modes of action on the reproduction of the nematode Caenorhabditis elegans. In three out of 10 cases, the IA model failed to describe mixture toxicity adequately with significant or antagonism being observed. In a further three cases, there was an indication of synergy, antagonism, and effect-level-dependent deviations, respectively, but these were not statistically significant. The extent of the significant deviations that were found varied, but all were such that the predicted percentage effect seen on reproductive output would have been wrong by 18 to 35% (i.e., the effect concentration expected to cause a 50% effect led to an 85% effect). The presence of such a high number and variety of deviations has important implications for the use of existing mixture toxicity models for risk assessments, especially where all or part of the deviation is synergistic.

Environmental impact of genetically modified cotton in South Africa

This paper presents the results of a large-scale study designed to monitor the impact arising from the introduction of insect-resistant Bt cotton in the Makhathini Flats, Republic of South Africa. Bt cotton provides a degree of resistance to cotton bollworm complex (Lepidoptera). Data were collected on the use of insecticides (type and quantity) as well as the farm-level economics of production from over 2200 farmers in three growing seasons (1998/1999, 1999/2000 and 2000/2001). and the results are discussed within the context of environmental impact brought about by insecticide. Over the three seasons of the study it was clear that Bt cotton provided benefits in terms of higher yield and gross margin relative to farmers growing conventional (non-Bt) cotton, and the benefits were particularly apparent for the smallest producers. Bt growers also used significantly less insecticide than growers of non-Bt cotton. Once quantities of insecticide applied to Bt and non-Bt cotton were converted into a Biocide Index and an Environmental Impact Quotient (EIQ) in order to allow for differences in terms of toxicity and persistence in the environment, it was apparent that the growing of Bt had a less negative impact on the environment. While this points to beneficial impacts on agricultural sustainability there are wider concerns regarding the vulnerability of resource-poor farmers in an area with limited (as yet) marketing options for their product and options for livelihood diversification both within and outside agriculture. Cotton producers in Makhathini are vulnerable as they rely on just One company for inputs (including, credit) and for their market. While Bt cotton provides benefits it does not in itself address some of the structural limitations that farmers face. (c) 2006 Elsevier B.V. All rights reserved.

The use of life cycle assessment to compare the environmental impact of production and feeding of conventional and genetically modified maize for broiler production in Argentina

The paper presents the methods and results of a life-cycle assessment (LCA) applied to the production of maize grain from a conventional variety compared with maize grain from a variety genetically modified to be herbicide tolerant and insect protected and to contain an enhanced oil and lysine content, and its impact when fed to broiler chickens. The findings show that there are both environmental and human health benefits of growing GM maize including lower impacts on global warming, ozone depletion, freshwater ecotoxicity and human toxicity. However, when considered in terms of the use of maize as a feed input to broiler chicken production, the benefits of the GM alternative become negligible compared to the use of conventional maize.

Environmental impact of genetically modified cotton in South Africa

This paper presents the results of a large-scale study designed to monitor the impact arising from the introduction of insect-resistant Bt cotton in the Makhathini Flats, Republic of South Africa. Bt cotton provides a degree of resistance to cotton bollworm complex (Lepidoptera). Data were collected on the use of insecticides (type and quantity) as well as the farm-level economics of production from over 2200 farmers in three growing seasons (1998/1999, 1999/2000 and 2000/2001). and the results are discussed within the context of environmental impact brought about by insecticide. Over the three seasons of the study it was clear that Bt cotton provided benefits in terms of higher yield and gross margin relative to farmers growing conventional (non-Bt) cotton, and the benefits were particularly apparent for the smallest producers. Bt growers also used significantly less insecticide than growers of non-Bt cotton. Once quantities of insecticide applied to Bt and non-Bt cotton were converted into a Biocide Index and an Environmental Impact Quotient (EIQ) in order to allow for differences in terms of toxicity and persistence in the environment, it was apparent that the growing of Bt had a less negative impact on the environment. While this points to beneficial impacts on agricultural sustainability there are wider concerns regarding the vulnerability of resource-poor farmers in an area with limited (as yet) marketing options for their product and options for livelihood diversification both within and outside agriculture. Cotton producers in Makhathini are vulnerable as they rely on just One company for inputs (including, credit) and for their market. While Bt cotton provides benefits it does not in itself address some of the structural limitations that farmers face. (c) 2006 Elsevier B.V. All rights reserved.

A comparative study on the relationship between acetyl cholinesterase activity and acute toxicity in Daphnia magna exposed to anticholinesterase insecticides

Acetylcholinesterase (AChE) activity was measured in Daphnia magna that had been exposed to four organophosphates (OPs; parathion, chlorpyrifos, malathion, and acephate) and one carbamate (propoxur) for 48 h. These results were related to acute toxicity (median effective concentration [EC50] for immobility). For the four OPs, the EC50s were 7.03 pM, 3.17 pM, 10.56 pM, and 309.82 muM, respectively. The EC50 for propoxur was 449.90 pM. Reduction in AChE activity was directly related to an increase in immobility in all chemicals tested. However, the ratio between the EC50 and the AChE median inhibiting concentration ranged from 0.31 to 0.90. A 50% reduction in AChE activity generally was associated with detrimental effects on mobility. However, for acephate, high levels of AChE inhibition (70%) were observed in very low concentrations and were not associated with immobility. In addition, increasing the concentration of acephate further had a slight negative effect oil AChE activity but a Strong detrimental effect on mobility. Binding sites other than AChE possibly are involved in acephate toxicity to D. magna. Our findings demonstrate different associations between AChE inhibition and toxicity when different chemicals are compared. Therefore, the value of using AChE activity as a biomarker in D. magna will be dependent on the chemical tested.

Multivariate relationships between groundwater chemistry and toxicity in an urban aquifer

Multivariate statistical methods were used to investigate file Causes of toxicity and controls on groundwater chemistry from 274 boreholes in an Urban area (London) of the United Kingdom. The groundwater was alkaline to neutral, and chemistry was dominated by calcium, sodium, and Sulfate. Contaminants included fuels, solvents, and organic compounds derived from landfill material. The presence of organic material in the aquifer caused decreases in dissolved oxygen, sulfate and nitrate concentrations. and increases in ferrous iron and ammoniacal nitrogen concentrations. Pearson correlations between toxicity results and the concentration of individual analytes indicated that concentrations of ammoinacal nitrogen, dissolved oxygen, ferrous iron, and hydrocarbons were important where present. However, principal component and regression analysis suggested no significant correlation between toxicity and chemistry over the whole area. Multidimensional Scaling was used to investigate differences in sites caused by historical use, landfill gas status, or position within the sample area. Significant differences were observed between sites with different historical land use and those with different gas status. Examination of the principal component matrix revealed that these differences are related to changes in the importance of reduced chemical species.

Is there a relationship between soil and groundwater toxicity?

Part IIA of the Environmental Protection Act 1990 requires environmental regulators to assess the risk of contaminants leaching from soils into groundwater (DETR, 1999). This newly introduced legislation assumes a link between soil and groundwater chemistry, in which rainwater leaches contaminants from soil into the saturated zone. As the toxicity of both groundwater and overlying soils is dependent upon the chemicals present, their partitioning and their bioavailability, similar patterns of soil, leachates and groundwater toxicity should be observed at contaminated sites. Soil and groundwater samples were collected from different contaminated land sites in an urban area, and used to determine relationships between soil chemistry and toxicity, mobility of contaminants, and groundwater chemistry and toxicity. Soils were leached using water to mimic rainfall, and both the soils and leachates tested using bioassays. Soil bioassays were carried out using Eisenia fetida, whilst groundwater and leachates were tested using the Microtox(TM) test system and Daphnia magna 48 h acute tests. Analysis of the bioassay responses demonstrated that a number of the samples were toxic to test organisms, however, there were no significant statistical relationships between soil, groundwater and leachate toxicity. Nor were there significant correlations between soil, leachates and groundwater chemistry.

A comparative study on the relationship between acetylcholinesterase activity and acute toxicity in Daphnia magna exposed to anticholinesterase insecticides

Acetylcholinesterase (AChE) activity was measured in Daphnia magna that had been exposed to four organophosphates (OPs; parathion, chlorpyrifos, malathion, and acephate) and one carbamate (propoxur) for 48 h. These results were related to acute toxicity (median effective concentration [EC50] for immobility). For the four OPs, the EC50s were 7.03 pM, 3.17 pM, 10.56 pM, and 309.82 microM, respectively. The EC50 for propoxur was 449.90 pM. Reduction in AChE activity was directly related to an increase in immobility in all chemicals tested. However, the ratio between the EC50 and the AChE median inhibiting concentration ranged from 0.31 to 0.90. A 50% reduction in AChE activity generally was associated with detrimental effects on mobility. However, for acephate, high levels of AChE inhibition (70%) were observed in very low concentrations and were not associated with immobility. In addition, increasing the concentration of acephate further had a slight negative effect on AChE activity but a strong detrimental effect on mobility. Binding sites other than AChE possibly are involved in acephate toxicity to D. magna. Our findings demonstrate different associations between AChE inhibition and toxicity when different chemicals are compared. Therefore, the value of using AChE activity as a biomarker in D. magna will be dependent on the chemical tested.