Mechanism of toxicity of ionic copper and copper complexes to algae

The mechanism of toxicity of ionic copper and copper complexes to growth, photosynthesis, respiration, ATP levels and mitochondrial electron-transport chain-activity in two marine diatoms, Nitzschia closterium (Ehrenberg) W. Smith (Hasle, 1964) and Asterionella glacialis Castracane, and one freshwater green alga, Chlorella pyrenoidosa Chick was investigated. Copper ions depressed both cell division and photosynthesis in A. glacialis and C. pyrenoidosa, whereas ionic copper concentrations which were inhibitory to cell division in N. closterium had no effect on photosynthesis, respiration, ATP production, electron transport or membrane ultrastructure. This suggests that in N. closterium, copper does not act on the chloroplast, the mitochondrion, or the cell membrane, since if it did, the above parameters should be affected. Copper-ethylxanthogenate was exceptional amongst the copper complexes in that it stimulated respiration, mitochondrial electrontransport and ATP formation in N. closterium under conditions of strongly inhibited cell division and slightly stimulated photosynthesis. Ionic copper toxicity may result from an intracellular reaction between copper and reduced glutathione (GSH), leading to a lowering of the GSH:GSSG ratio and suppression of mitosis. In addition, copper inhibits the enzyme catalase and reduces cell defence mechanisms against H₂O₂ and oxygen-free radicals. Lipid-soluble copper complexes are more toxic than ionic copper because both the metal and the ligand are introduced into the cell. Toxicity of ionic copper is ameliorated by trivalent metal ions in the growth medium, including those of Mn, Co, Al, Fe and Cr which form a layer of metal (III) hydroxide around the algal cell, adsorb copper and reduce its penetration into the cell. The degree of insolubility of the metal (III) hydroxide is related to its ability to protect against copper toxicity. In addition, manganese and cobalt catalytically scavenge damaging H₂O₂ and superoxide radicals, respectively, produced by the cell.     

The effect of limiting nutrient on metal toxicity to selenastrum capricornutum

Metal toxicity on selenastrum capricornutum were examined by batch culture and chemostat culture system. EC₅₀ values under nutrient‐limited condition and saturated condition were compared. The two EC_5Os differed by a factor of 10.6 for Cd in batch test, and differed by a factor of 6.5 in continuous test. Both batch and continuous test results indicate that inhibition on algal growth is more severe under nutrient‐limited conditions compared to that under near saturation conditions.

Strong correlation, as characterized by the correlation coefficient ρ, were found between the activation level (nutrient requirement) and the tolerance of microorganisms to the toxicant. ρ is equal to 0.9 for the case of Cd in batch test and 0.6 in continuous test. The theory in this study provides generally good estimations to the dose‐response relationship at limiting nutrient conditions. The study indicate that nutrient conditions constitute an important factor in algal toxicity tests.     

Determination of types of interactions exhibited by binary mixtures of heavy metals tested against the hermit crab,Clibanarius africanus

Single and joint action toxicity of heavy metal compounds (ZnCO₃, CuSO₄ and CdSO₄) were carried out against the hermit crab, Clibanarius africanus in laboratory bioassays. On the basis of 96?h LC₅₀ values, CuSO₄ (96?h LC₅₀?=?0.03?mM) was found to be the most toxic followed by CdSO₄ and ZnCO₃ (96?h LC₅₀?=?0.24?mM) in decreasing order of toxicity. The evaluation of the metal compounds in mixtures of ZnCO₃–CuSO₄, ZnCO₃–CdSO₄ and CdSO₄–CuSO₄ in pre-defined molar ratios revealed that interactions between Zn–Cu and Zn–Cd test mixtures were in conformity with the models of antagonism (reduction in toxicity of constituent metals). The synergistic ratios (S.R.s) and relationship between observed/predicted 96?h LC₅₀ values (relative toxic unit, RTU) were consistently below 1, indicating antagonistic interactions. Analysis using isobolograms also indicated the interactions between the mixture components to be antagonistic. However, with regards to CdSO₄–CuSO₄ test mixtures, the interactions between these metals were mainly in conformity with the model of synergism. The S.R. and RTU values were largely above 1, indicating synergistic interactions. The importance of the joint action evaluations for the management and protection of aquatic ecosystems, particularly in the setting up of water quality criteria was discussed.     

Simultaneously recovering electricity and water from wastewater by osmotic microbial fuel cells: Performance and membrane fouling

Since the concept of the osmotic microbial fuel cell (OsMFC) was introduced in 2011, it has attracted growing interests for its potential applications in wastewater treatment and energy recovery. However, forward osmosis (FO) membrane fouling resulting in a severe water flux decline remains a main obstacle. Until now, the fouling mechanisms of FO membrane especially the development of biofouling layer in the OsMFC are not yet clear. Here, the fouling behavior of FO membrane in OsMFCs was systematically investigated. The results indicated that a thick fouling layer including biofouling and inorganic fouling was existed on the FO membrane surface. Compared to the inorganic fouling, the biofouling played a more important role in the development of the fouling layer. Further analyses by the confocal laser scanning microscopy (CLSM) implied that the growth of biofouling layer on the FO membrane surface in the OsMFC could be divided into three stages. Initially, microorganisms associated with ß-D-glucopyranose polysaccharides were deposited on the FO membrane surface. After that, the microorganisms grew into a biofilm caused a quick decrease of water flux. Subsequently, some of microorganisms were dead due to lack of nutrient source, in the meantime, polysaccharide and proteins in the biofouling layer were decomposed as nutrient source, thus leading to a slow development of the biofouling layer. Moreover, the microorganisms played a significant role in the formation and development of the biofouling layer, and further studies are needed to mitigate the deposition of microorganisms on FO membrane surfaces in OsMFCs.

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Evaluation of the toxicity of tetrabromobisphenol A and some of its oxidation products using a micro-scale algal growth inhibition test

A micro-scale algal growth inhibition (μ-AGI) test using a common micro-plate based fluorometric detection was used to demonstrate the effects of humic substances (HSs) on the toxicity of tetrabromobisphenol A (TBBPA) and its oxidative decomposition products 2,5-dibromo-1,4-benzoquinone (2,5-DBBQ), 2,5-dibromohydroquinone (2,5-DBHQ), 2,6-dibromobenzoquinone (2,6-DBBQ), and 2,6-dibromophenol (2,6-DBP) to Pseudokirchneriella subcapitata. The EC₅₀ values were: EC_50(TBBPA) = 7 mg L^?1, EC_50(2,5-DBHQ) = 7 mg L^?1, EC_50(2,5-DBBQ) = 19 mg L^?1, EC_50(2,6-DBP) = 49 mg L^?1, and EC_50(2,6-DBBQ) = 13 mg L^?1. The toxicity of the chemicals was slightly lower in the presence of HA. The toxicity of TBBPA decomposed by a biomimetic catalytic system consisting of iron (III) 5,10,15,20-tetrakis (p-sulfonatophenyl) porphyrin (Fe(III)-TPPS) and KHSO₅ was also evaluated using P. subcapitata and Chlamydomonas reinhardtii.     

The influence of bacterial quorum-sensing inhibitors against the formation of the diatom-biofilm

Quorum-sensing inhibitor (QSI) is one of the most promising and environmentally friendly agents for marine antifouling. In this study, the activities of three kinds of QSIs 3,4-dibromo-2(5)H-furanone, 4-nitropyridine-N-oxide and indole were evaluated on the growth of two marine diatoms Cylinthrotheca sp. and Nitzschia closterium. At the same time, the effects of QSIs on the formation of the diatom-biofilm were also discussed. All the results showed that QSIs significantly inhibited the growth, and the effects depended on the dose and diatom species. The extracellular polymeric substance contents in the diatom-biofilm were significantly reduced by QSIs. However, the contents of polysaccharide in culture mediums were increased, which might result in the destruction of diatom cells. Combined with the results of crystal violet staining-biofilm and images of scanning electron microscopy, it was further demonstrated that QSIs inhibited the biofilm formation of Cylindrotheca sp., and the inhibitory effect of 4-nitropyridine-N-oxide was superior to that of 3,4-dibromo-2(5)H-furanone and indole.     

Effect of TiO2 nanoparticles and multiwall carbon nanotubes on the freshwater diatom Nitzschia frustulum: evaluation of growth,cellular components and morphology

Great attention should be paid to the possible impacts of various nanoparticles on aquatic organism. Freshwater diatoms are essential components of phytoplankton and play a critical role in bioassessment of nanoparticle exposure in the environment. In this study, cell growth was inhibited by TiO₂-NPs and MWCNTs, the 24?h EC₅₀ values of TiO₂-NPs and MWCNTs to Nitzschia frustulum were 20.75 and 24.64?mg?L^?1, respectively. Significant decreases of Chl a content after TiO₂-NPs exposures were detected and the Chl a content of N. frustulum was obviously increased by MWCNTs treatment at lower concentration. The ROS was detected in N. frustulum after TiO₂-NPs and MWCNTs exposures. The MDA content was significantly induced by TiO₂-NPs at lower concentrations of 24 and 48?h exposure; meanwhile, it increased at all tested concentrations at 24?h MWCNTs exposure. The SOD enzyme was induced by 72 and 96?h TiO₂-NPs exposure, and increased by MWCNTs treatment at 96?h in N. frustulum. Scanning electron microscopy results revealed that N. frustulum had obvious cell deformation after TiO₂-NPs treatment. The result showed that the physiological and biochemical response mechanisms after NPs exposure of diatom were species-specific, and in relation to the exposure concentration and time.     

脱氧雪腐镰刀菌烯醇、黄曲霉毒素B_1和玉米赤霉烯酮对秀丽隐杆线虫的联合毒性研究

脱氧雪腐镰刀菌烯醇(DON)、黄曲霉毒素B_1(AFB_1)和玉米赤霉烯酮(ZEN)是最为常见的粮食真菌毒素,易共存于谷物产品和动物饲料中,而目前对其联合毒性的研究较少,且研究结果并不完全一致。为探究DON、AFB_1和ZEN的联合毒性作用,本论文以秀丽隐杆线虫(C.elegans)为模型,分别评估了毒素混合物AFB_1+DON、AFB_1+ZEN、DON+ZEN和AFB_1+DON+ZEN对C.elegans的生长发育(体长)和生殖能力(产卵量)的毒性作用,并用Chou-Talalay模型来判定毒素混合物的相互作用类型。研究表明,AFB_1、DON和ZEN单独染毒C.elegans时,其毒作用强弱为AFB_1ZENDON。联合染毒时,AFB_1+DON对C.elegans产生协同作用,而DON+ZEN则产生拮抗作用;AFB_1+ZEN对体长(24 h)和产卵量的毒作用随着暴露浓度的增加,由弱拮抗变为协同作用,而在毒素暴露48 h后,对线虫的生长发育呈协同作用;AFB_1+DON+ZEN除在EC50-24 h和EC75-24 h时对体长产生明显的毒性增强作用外,其他普遍表现出拮抗作用。由此表明,DON、AFB_1和ZEN对C.elegans的联合毒性作用与剂量和时间相关。     

Effects of altered photic regimes on diel patterns of species-specific photosynthesis

Diel patterns of photosynthesis were measured for two polar diatoms (Coscinodiscus sp. and Porosira pseudodenticulata) collected in September 1985 from McMurdo Sound, Antarctica, and four temperate dinoflagellates (Gonyaulax hyalina, Gymnodinium splendens, Dinophysis caudata, and Glenodinium sp.) collected in July 1985 and January 1986 from the Southern California Bight, California, USA. For phytoplankton incubated under three combinations of photoperiod and irradiance, distinct diel patterns of light-saturated (P _max) and light-limited (P _L ) photosynthesis were found for (i) different species isolated from the same environment, and (ii) polar diatoms and temperate dinoflagellates. The time of day when the maximum rate of P _max occurred was influenced by both irradiance and daylength for the polar diatoms but not by daylength for three out of four temperate dinoflagellates. The range of values of the ratio of maximum to minimum rates of photosynthesis (P _max:P _min) was similar for polar diatoms and temperate dinoflagellates. The results of this study suggest that changes in irradiance or photoperiod could influence species-specific patterns of photosynthesis in nature. As a consequence, in light-limited environments differential reproductive success could result from these diel patterns, and ultimately be reflected in temporal and spatial differences in community structure.     

Analysis of the toxicity of glyphosate and Faena® using the freshwater invertebrates Daphnia magna and Lecane quadridentata

With the present contribution an evaluation of the toxicity of the pure herbicide glyphosate and its commercial formulation Faena® is reported using the cladoceran Daphnia magna Strauss and the rotifer Lecane quadridentata Ehrenberg. LC₅₀, EC₅₀, NOEC, and LOEC values for each toxicant and for both test organisms are obtained. Regarding acute toxicity, Faena was 11-fold more toxic to L. quadridentata than pure glyphosate and slightly more toxic to D. magna (1.7-fold). Inhibition of esterase activity in L. quadridentata by glyphosate was the most sensitive end-point; the EC₅₀ was 1500-fold smaller than the LC₅₀. The implications of these results and their comparison with established international and national limit values for glyphosate are discussed.     

δ<Superscript>15</Superscript>N as a natural tracer of particulate nitrogen effluents released from marine aquaculture

The flow of particulate nitrogen from marine net pen fish farm effluents to the surrounding biofouling community was quantified by means of stable isotopes of nitrogen. Plastic mesh substrates were deployed at 8 m depth near a sea bream fish farm and at a nearby reference site in the northern Gulf of Aqaba (Red Sea) to assess whether natural fouling organisms could sequester substantial quantities of farm-derived particulate nitrogen waste. A mixing equation, incorporating differences in nitrogen stable isotope composition, δ¹⁵N, between particulate organic matter (“source”) and fouling organisms (“sink”) at the fish farm and reference site, was used to estimate the amount of farm-derived nitrogen that was incorporated by the fouling community. Among the conspicuous fouling organisms examined, sponges, tunicates and polychaetes showed greatest uptake of fish farm N, where the mean fractions of farm-derived N estimated over the 2-year period of observation were 19±7, 22±6 and 31±8% of total organisms’ N content, respectively, with maximal recorded seasonal values of 68, 85 and 57%, respectively. Mean N uptake by mixed fouling communities (conspicuous + cryptic organisms) was as much as fivefold higher than that calculated for the sum of conspicuous taxa, suggesting that the retention efficiency is greater in mixed than in mono-specific biofouling communities.     

Monitoring toxicity of an azo dye methyl red and a heavy metal Cu,using plant and animal bioassays

Toxicities of an azo dye methyl red and a heavy metal copper (Cu) were quantified, using growth and mortality as end points, in four plant species and three animal species by subjecting them to short-term (4 days for animals, 10 days for plants) static bioassays. Lemna aequinoctialis Welwitch (EC₅₀: 7–16 ppm) was found to be the most sensitive species for methyl red, Ceratophyllum demersum L. (EC₅₀: 25 ppm) and Lactuca sativa L. (EC₅₀: 56 ppm) were intermediate, while Oryza sativa L. shows reduction in seedling vigor (9–27%) of <50%, being the least sensitive amongst the tested plant species. Methyl red toxicity is almost 3–5-fold higher in growing medium (pH = 5.8–6.0), even at high nutrient levels, while Cu toxicity is higher in nutrient-poor alkaline medium at alkaline pH (8.3–8.7; EC₅₀: Ceratophyllum = 104–200 ppb; Lemna = 100–170 ppb) compared to nutrient-rich acidic medium (pH = 5.4–5.7; EC₅₀: Ceratophyllum = 2600–3175 ppb; Lemna = 4350–4715 ppb). Rice tolerance (EC₅₀: 6500 ppb) was found to be higher than hydrophytes while lettuce was most tolerant to Cu. Fish sensitivity toward the test chemicals was almost parallel to Ceratophyllum and Lemna [Gambusia affinis Baird and Gerard (LC₅₀: 250 ppb for Cu) and Poecilia reticulata Peters (LC₅₀: 24 ppm for methyl red)]. Similar to the plants, dye toxicity increased markedly (LC₅₀: 7 ppm) in the acidic medium (pH = 6.0). Amongst the tested organisms, Daphnia was found to be most sensitive to methyl red (EC₅₀: 6 ppm) while its sensitivity to Cu (EC₅₀: 230 ppb) was similar to hydrophytes and fish. Initially, the combination of dye and Cu (at their sublethal concentrations) had additive effects in duckweed, while dye concentration ruled afterward. These results indicate that hydrophytes and animals are equally sensitive toward the test chemicals. Dye toxicity in hydrophytes and fish was pH dependent, while in the case of Cu, it is related to the nutrient status of the growth medium of plants.     

Application of microcosm and species sensitivity distribution approaches in the ecological hazard assessment of 4-tert-butylphenol

In order to evaluate the ecological hazard of 4-tert-butylphenol (4-TBP), species abundances and physicochemical properties were monitored for 63 days in a microcosm system containing planktonic algae, rotifers, shrimps and Daphnia. The 63 days-NOEC (no observed effect concentration) of 4-TBP for this system was calculated. At the same time, species sensitivity distribution curves were constructed based on the toxicity data of EC₁₀, EC₅₀ and NOEC derived from laboratory single-species toxicity tests, which further gives no obvious hazard concentrations for 95% species in the ecosystem (HC₅). In both the microcosm and the single-species tests, the shrimp (Neocaridina sp.) was the most sensitive species to 4-TBP exposure, while most algae species were relatively insensitive compared to the zooplankton. For the microcosm exposed to 4-TBP, significant changes occurred to the community structure instead of the community function, resulting in a NOEC_microcosm of 265.38?μg?L^–1 which approached the EC₁₀-based HC₅ (0.376?mg?L^–1). Nevertheless, EC₅₀-based HC₅ was higher than the NOEC_microcosm, and the NOEC-based HC₅ was lower. The data showed that the EC₁₀-based HC₅ was protective at the similar level with the result of microcosm for 4-TBP, providing supportive data for chemical risk assessment.     

Inhibition of the growth of micro-algae and bacteria by extracts of eelgrass (Zostera marina) leaves

The effects of the water-soluble fraction of dead leaves of the eelgrass Zostera marina L. on the growth of 8 species of micro-algae (pennate and centric diatoms, dinoflagellates, and a green flagellate) and a bacterium were studied on agar plates and in liquid culture. The extracts of leaves which had been dead from a few days to 2 wk inhibited growth and often killed cells in all test organisms. Extracts were lethal even at concentrations equivalent to as little as 0.25 mg dry leaf ml^-1, but inhibition decreased when extracts were prepared from leaves aged in the laboratory for 35 d (loss of anti-bacterial activity) or 90 d (loss of anti-algal activity). Extracts of leaves which had aged and dried several months in the field had no effect, except at very high concentrations (13 mg dry leaf ml^-1) when the lag phase of growth was prolonged several days in a culture of the chlorophyte Platymonas sp. The active fraction in eelgrass leaves may be important in controlling initial growth of micro-organisms on eelgrass detritus, and it could determine the composition and activity of the epiphytic community on living leaves.     

In vitro antioxidant and antibacterial activities of certain 3-{4-(5-mercapto-1,3,4-oxadiazole-2-yl)phenylimino}-indolin-2-one derivatives

A new series of 5- or 7-substituted 3-{4-(5-mercapto-1,3,4-oxadiazol-2-yl)phenylimino}-indolin-2-one derivatives were screened for in vitro antioxidant and antimicrobial activities. The results of this study revealed that all synthesized compounds were significantly scavenged DPPH free radicals in a concentration-dependant manner. The IC₅₀ values of all test compounds were found to be between 18.34 and 50.54?µM. All compounds significantly inhibited growth of bacteria and most of the minimum inhibitory concentration values were between 20 and 100?mg?mL^?1 indicating potent to moderate antibacterial activity. In conclusion, 3-{4-(5-mercapto-1,3,4-oxadiazole-2-yl)phenylimino)-5 or 7-substituted indolin-2-ones derivatives exhibited moderate antioxidant and antimicrobial activities.     

6种苯系物对球等鞭金藻和新月菱形藻的生长抑制

海洋环境中苯系物污染主要来源于海洋溢油事故以及沿海石油化工企业的废水排放。为探究苯系物对海洋微藻的毒性作用,选择球等鞭金藻和新月菱形藻作为受试生物,分别考察了苯、甲苯、乙基苯、邻-二甲苯、间-二甲苯和对-二甲苯6种苯系物对2种海洋微藻生长的影响。结果表明,在0.25~64.0 mg·L-1暴露浓度下,6种苯系物对2种微藻生长具有显著的抑制作用,随着暴露浓度的升高,抑制作用明显增强。苯、甲苯、乙基苯、邻-二甲苯、间-二甲苯、对-二甲苯对球等鞭金藻的24 h的半数效应浓度(24 h-EC50)分别为:17.07、12.88、7.58、0.55、0.36、0.27 mg·L-1;对新月菱形藻的24 h-EC50值分别为:1.03、0.68、0.46、0.40、0.42、0.38 mg·L-1。上述研究结果为确定苯系物海洋环境质量标准、保护海洋生态环境提供了基础数据。     

Changes in the cell size of the diatom Cylindrotheca closterium in a hyperhaline pond

Cell length of the diatom Cylindrotheca closterium (Ehr.) Reimann & Lewin was investigated on a fortnightly basis from January to April 2000 in a hyperhaline pond (Sardinia, Mediterranean Sea) and relationships, with changes in temperature, salinity, and nitrite concentrations were assessed using single and multiple regression analyses. Results of our study indicate that C. closterium cell size was inversely related to temperature changes, but that salinity and nitrite concentrations may have a major idiosyncratic effect on cell size variability.     

Larval settlement and metamorphosis of the mussel <Emphasis Type="Italic">Mytilus galloprovincialis</Emphasis> on different macroalgae

Settlement of mussels is commonly associated with macroalgae. The effects of 19 macroalgal species on the settlement and metamorphosis of pediveliger larvae of the mussel Mytilus galloprovincialis were investigated in the laboratory. Settlement and metamorphosis inducing activities of macroalgae Chlorodesmis fastigiata and Ceramium tenerrimum collected each month during the period between January 2005 and April 2006 were also investigated. Furthermore, C. fastigiata and C. tenerrimum were subjected to various treatments to investigate the roles of bacteria and diatoms on the algal surface in the induction of larval settlement and metamorphosis of M. galloprovincialis and the characteristics of the cues in these two macroalgae. Pediveliger larvae of M. galloprovincialis settled and metamorphosed in high percentages on Cladophora sp., Chlorodesmis fastigiata, Centroceras clavulatum, and Ceramium tenerrimum, all of which were filamentous in morphology. Macroalgae that were cylindrical, phylloid, flabellate, palmate and pinnate all showed low (<8%) percentages of post-larvae but four other filamentous macroalgae also had low mussel larval settlement, suggesting that chemical factors may also be involved. Seasonal variation had no effect on inductive activities of C. fastigiata and C. tenerrimum. Treatment of C. fastigiata and C. tenerrimum with formalin, ethanol and heat resulted in the significant decrease or loss of their inductive activities. Survival of algal cells within treated macroalgae also decreased significantly. Treatment of the two macroalgae with antibiotics and GeO₂ reduced the numbers of bacteria and diatoms on their surface but did not affect their inductive activities, indicating that the cue was produced by macroalgae and not by coexisting bacteria and diatoms. However, conditioned water and crude extracts of these two macroalgae did not induce larval settlement and metamorphosis. Thus, larvae of M. galloprovincialis settled and metamorphosed on specific filamentous macroalgae. The chemical cues produced by C. fastigiata and C. tenerrimum were susceptible to ethanol and heat treatments and were not recovered in the conditioned water nor in the extracts. The finding that inactive C. tenerrimum can be produced from culturing its apical segments provides a new tool to elucidate the chemical cue(s) from macroalgae through manipulation of their culture conditions.     

Assessing the toxicity of chemical compounds associated with marine land-based fish farms: The use of mini-scale microalgal toxicity tests

Many chemicals that are currently used in aquaculture have not been evaluated with regard to their specific effects on the aquatic environment. In the present study, the toxic effects of several chemicals associated with land-based marine fish farming activities were assessed using two species of marine microalgae (Phaeodactylum tricornutum and Isochrysis galbana). Mini-scale toxicity tests were performed with six antibiotics (amoxicillin, ampicillin, flumequine, oxytetracycline, streptomycin and sulfadiazine) and two disinfectants (formaldehyde and hypochlorite). Amoxicillin and streptomycin did not exert toxic effects. Sulfadiazine was the most toxic chemical; the EC₅₀ values were 0.11 mg/L and 1.44 mg/L for P. tricornutum and I. galbana respectively. As expected, the disinfectants displayed high toxicity, and P. tricornutum was particularly sensitive to these compounds. Although the differences in microalgal sensitivity depended on the chemical considered, both species were highly sensitive to most of the compounds tested. We recommend the inclusion of mini-scale microalgal toxicity tests in environmental risk assessment (ERA) and environmental monitoring plans because they are cost-effective and rapid.