Ecotoxicological evaluation of sodium fluoroacetate on aquatic organisms and investigation of the effects on two fish cell lines

Sodium monofluoroacetate (compound 1080) is one of the most potent pesticides. It is also a metabolite of many other fluorinated compounds, including anticancer agents, narcotic analgesics, pesticides or industrial chemicals. Other sources of water contamination are the atmospheric degradation of hydrofluorocarbons and hydrochlorofluorocarbons. However, there is little information available about the adverse effects of sodium fluoroacetate in aquatic organisms. Firstly, the bacterium Vibrio fischeri (decomposer), the alga Chlorella vulgaris (1st producer) and the cladoceran Daphnia magna (1st consumer) were used for the ecotoxicological evaluation of SMFA. The most sensitive models were C. vulgaris and D. magna, with a NOAEL of 0.1 and an EC50 of 0.5 mM at 72 h, respectively. According to the results after the acute exposure and due to its high biodegradation rate and low bioaccumulation potential, sodium fluoroacetate is most unlikely to produce deleterious effects to aquatic organisms. Secondly, two fish cell lines were employed to investigate the effects and mechanisms of toxicity in tissues from 2nd consumers. The hepatoma fish cell line PLHC-1 was more sensitive to SMFA than the fibroblast-like fish cell line RTG-2, being the uptake of neutral red the most sensitive bioindicator. Lysosomal function, succinate dehydrogenase and acetylcholinesterase activities were inhibited, glucose-6-phosphate dehydrogenase activity was particularly stimulated, and metallothionein and ethoxyresorufin-O-deethylase levels were not modified. Intense hydropic degeneration, macrovesicular steatosis and death mainly by necrosis but also by apoptosis were observed. Moreover, sulphydryl groups and oxidative stress could be involved in PLHC-1 cell death induced by SMFA more than changes in calcium homeostasis.     

Online Water Monitoring Method as a Water Security Tool: A Feasibility View

Since the tragic event on September 11, 2001 (9/11), homeland security has been the center of major attention not only in the United States but also around the world. Among homeland security agenda, more concerns on drinking water system have been drawn into the forefront attention from the public and water industry. Governmental agencies have been called upon to strongly protect the water resources from becoming a possible terror target. The online monitoring of a water system offers the potential to reduce the possible danger from a terrorist contamination as well as from unintentional chemical spills. As potential terrorist contaminants, seven chemical compounds (aldicarb, cycloheximide, dicrotophos, nicotine, sodium arsenate, sodium cyanide, and sodium fluoroacetate) were studied at different doses, in order to determine their detectability when they are present in a water system, including intake, treatment, and distribution. These contaminants were monitored by measuring simple water quality parameters such as conductivity, pH, chlorine residual, turbidity, total organic carbon (TOC), and UV254. Results indicated that the contaminants used for the study were detected at certain toxicity concentrations through the online water quality monitoring method. This method provides a useful tool for watching water resources against possible terror attacks, and also keeping safe water quality.     

Stereoselective biosynthesis of chloroarylpropane diols by the basidiomycete Bjerkandera adusta

Previously we have shown that 1-arylpropane-1,2-diols are catabolic products of L-phenylalanine during idiophasic metabolism of B. adusta that are stereoselectively biosynthesized from a C(7)-unit (ring+benzylic carbon) and a C(2)-unit as predominantly erythro 1R, 2S enantiomers.In order to probe the mechanism of 1-arylpropane-1,2-diol formation, the products of the incubation of isotopically labelled aromatic aldehydes as substrates with Bjerkandera adusta (DAOM 215869) have been characterized. The aromatic aldehydes were benzaldehyde (ring D(5)) and 4-methoxy- and 4-hydroxybenzaldehydes (ring 13C(6)). These aldehydes were all stereoselectively incorporated into the corresponding 1-arylpropane-1,2-diols, including the chloro analogues, as well as into the corresponding alpha-ketols (phenyl acetyl carbinols (PAC's) and 2-hydroxy propiophenones (2-HPP's)) the presumed precursors of the diols. Benzoic acid (ring D(5)) was likewise incorporated into the diols, chlorodiols and alpha-ketols.These results lead us to conclude that the aromatic aldehydes benzaldehyde, 4-hydroxybenzaldehyde and 4-methoxybenzaldehyde are likely C(7)-unit precursors in the carboligation reaction(s) that leads to 1-arylpropane-1,2-diol biosynthesis. The metabolic role of the diols remains to be elucidated but they may be important intermediates in CAM (chlorinated anisyl metabolite) aldehyde-alcohol cycling and also act as substrates for the chlorination/hydroxylation enzymes yet to be identified in white rot fungi.     

Biological dehalogenation and halogenation reactions

A large number of halogenated compounds is produced by chemical synthesis. Some of these compounds are very toxic and cause enormous problems to human health and to the environment. Investigations on the degradation of halocompounds by microorganisms have led to the detection of various dehalogenating enzymes catalyzing the removal of halogen atoms under aerobic and anaerobic conditions involving different mechanisms. On the other hand, more than 3500 halocompounds are known to be produced biologically, some of them in great amounts. Until 1997, only haloperoxidases were thought to be responsible for incorporation of halogen atoms into organic compounds. However, recent investigations into the biosynthesis of halogenated metabolites by bacteria have shown that a novel type of halogenating enzymes, FADH(2)-dependent halogenases, are involved in biosyntheses of halogenated metabolites. In every gene cluster coding for the biosynthesis of a halogenated metabolite, isolated so far, one or several genes for FADH(2)-dependent halogenases have been identified.     

Isolation of mesotrione-degrading bacteria from aquatic environments in Brazil

Mesotrione is a benzoylcyclohexane-1,3-dione herbicide that inhibits 4-hydroxyphenyl pyruvate dioxygenase in target plants. Although it has been used since 2000, only a limited number of degrading microorganisms have been reported. Mesotrione-degrading bacteria were selected among strains isolated from Brazilian aquatic environments, located near corn fields treated with this herbicide. Pantoea ananatis was found to rapidly and completely degrade mesotrione. Mesotrione did not serve as a sole C, N, or S source for growth of P. ananatis, and mesotrione catabolism required glucose supplementation to minimal media. LC-MS/MS analyses indicated that mesotrione degradation produced intermediates other than 2-amino-4-methylsulfonyl benzoic acid or 4-methylsulfonyl-2-nitrobenzoic acid, two metabolites previously identified in a mesotrione-degrading Bacillus strain. Since P. ananatis rapidly degraded mesotrione, this strain might be useful for bioremediation purposes.     

雪貂和旱獭作为胃癌病因研究的实验模型

本项研究是以雪貂（Ｆｅｒｒｅｔ）和旱獭（Ｗｏｏｄｃｈｕｃｋ）作为实验动物模型，由于这两种动物的某些器官特征在解剖学上和生理学上与人类有其相似之处，试图通过雪貂和旱獭体内生物合成亚硝胺，说明人类暴露于化学致场物亚硝胺的另外一个来源是生物体内合成。同时，用烷基化学致癌物二甲基亚硝胺对旱獭进行ＤＮＡ烷基化的研究，发现由于被旱獭肝炎病毒（ＷＨＶ，ＷｏｏｄｃｈｕｃｋＨｅｐａｔｉｔｉｓＶｉｒｕｓ）感染的旱獭，不仅体内易于合成亚硝胺，而且其肝脏组织ＤＮＡ烷基化的水平高于对照组。以ＤＮＡ烷基化反应产生的烷化核酸碱基作为生物标志（Ｒｉｏｍａｒｋｅｒｓ），对于胃癌高发区人群尿中的烷化核酸碱基进行定量测定，并与低发区相对照，显示两者之间有差异性。本项研究为从动物实验外推到人群的设想提供了启示，还提出了以生物标志作为建立分子流行病学的依据。     

Enzymatic and microbial transformation assays for the evaluation of the environmental fate of diclofenac and its metabolites

Diclofenac has been of environmental concern due to the potential harmful effects on non-target organisms at environmentally relevant concentrations. In this study, we evaluated the transformation kinetics of diclofenac and its two major metabolites in two laboratory-scale experiments: the transformation of diclofenac in the presence of rat liver S9 fraction with co-factors, and the transformation of diclofenac, 4′-hydroxy-diclofenac and diclofenac β-O-acyl glucuronide in the inoculum used for the OECD 301C ready-biodegradability test. 4′-Hydroxy-diclofenac was identified as the major phase I metabolite and diclofenac β-O-acyl glucuronide was identified as the major phase II metabolite in the S9 assay. Transformation of diclofenac in the microbial degradation test did not occur significantly for 28 d, whereas 4′-hydroxy-diclofenac degraded slowly, indicating that the biological removal of diclofenac is not likely to occur in conventional STPs unless sorptive removal is significant. However, diclofenac β-O-acyl glucuronide deconjugated to form equimolar diclofenac within 7 d, in the microbial degradation test. The mixture of diclofenac and its two metabolites, formed after incubating diclofenac in S9 medium for 2 h, was spiked in the inoculum to link both assays. The concentrations of diclofenac and its metabolites, measured over time, agreed well with predicted values, using rate parameters obtained from independent experiments. The results show that phase II metabolites generated in mammals may deconjugate easily in conventional STPs to form a parent compound and that these processes should be considered during the environmental monitoring and risk assessment of diclofenac.     

Transformation of 4-nonylphenol isomers during biosolids composting

4-Nonylphenol, a degradation intermediate of commercial surfactant and known endocrine disruptor, has been frequently detected at levels up to several thousand microgl(-1) in surface waters and up to several hundred mgkg(-1) (dry weight) in soil and sediment samples. Large quantities of 4-NP can be quickly sorbed by the organic rich solid phase during wastewater treatment and are concentrated in biosolids, a possible major source for 4-NP in the environment. Microbial transformation in culture studies followed different mechanisms for different 4-NP isomers, which have different estrogenic activity. Composting is a process of solid matrix transformation where biological activity is enhanced by process control. This approach has been used successfully in remediation of contaminated soils and sludges. In this study, the transformation kinetics of 4-NP and its isomers were characterized during biosolids composting. Five distinctive 4-NP isomer groups with structures relative to alpha- and beta-carbons of the alkyl chain were identified in biosolids. Composting biosolids mixed with wood shaving at a dry weight percentage ratio of 43:57 (C:N ratio of 65:1) removed 80% of the total 4-NP within two weeks. At this biosolids/wood shaving ratio (B:WS), the transformation of total 4-NP and its isomers followed second-order kinetic. Higher B:WS ratios yielded significantly slower 4-NP transformation which followed first-order kinetic. Isomers with alpha-methyl-alpha-propyl structure transformed significantly slower than those with less branched tertiary alpha-carbon and those with secondary alpha-carbon, suggesting isomer-specific degradation of 4-NP during biosolids composting.     

"Ni-struvite" - a new biomineral formed by a nickel resistant Streptomyces acidiscabies

Biomineralization dependent on bacterial activity has been described for struvite which is formed in soils, guano, putrescent matter and sediments. A new biomineral containing nickel instead of magnesium, Ni(NH4)(PO4) . 6H2O, has been identified. It was formed by nickel resistant Streptomyces acidiscabies E13, and putatively named nickel struvite. The mineral formation is dependent on biological activity since non-viable bacterial cells are not capable to induce formation of Ni-struvite under identical conditions. Formation of Ni-struvite was observed on colony surfaces upon prolonged incubation of solid minimal or complex media containing elevated concentrations of 8-15mM NiCl2. The formation of magnesium containing crystals was not observed although Mg2+ is present in the medium. However, the process was not depending on desiccation since small crystals attached to the mycelial biomass of the bacteria were observed microscopically also in liquid cultures of nickel supplemented minimal and complex media after two weeks of incubation. The capacity to induce biomineralization of a nickel containing mineral is postulated to constitute a resistance factor, allowing the soil bacterium to withstand high nickel concentrations. The strain shows nickel resistance as an adaption to its habitat, since this bacterium was isolated from a former uranium mining site in Eastern Thuringia, Germany, where nickel concentrations of up to 2000ppm (translating to appr. 30mM) occur as a result of former mining activities.     

Inhibition of animal acetyl-coenzyme a carboxylase by 2-(p-chlorophenoxy)-2-methylpropionic acid and 2-ethylhexanoic acid

Peroxisome proliferators are generally known as activators of fatty acid β-oxidation which is one of degradation pathways. However, since it is unknown whether peroxisome proliferators have an effect on biosynthesis of fatty acid or not, we commenced to study if acetyl-CoA carboxylase, the rate-limiting enzyme of fatty acid biosynthesis, is inhibited by 2-(p-chlorophenoxy)-2-methylpropionic acid or 2-ethylhexanoic acid which is a typical peroxisome proliferator. Acetyl-CoA carboxylase(s) from rat, mouse, hamster, rabbit, dog and monkey were completely inhibited by 2-(p-chlorophenoxy)-2-methylpropionic acid or 2-ethylhexanoic acid at the concentration of 10⁻³M. For both compounds, there was not a large species difference in the inhibition. However, there was only several times difference between I₅₀ values for the two compounds. The findings imply that inhibition of acetyl-CoA carboxylase by the chemicals leads to reduction of fatty acid biosynthesis when the chemicals are administered to the animals at high dose levels.     

Lead phosphate formation in soils

Pyromorphite (Pb5(PO4)3Cl) is one of the most thermodynamically stable lead minerals under the geochemical conditions prevailing in the surface environment. It is therefore expected to form in soils contaminated with Pb if sufficient phosphorus is available. Pyromorphite has previously been identified in mine-waste and industrially contaminated soils but has not previously been identified in urban soils. This paper reports on the presence of a Pb phosphate in urban and roadside soils. This phase has formed in the soil as a weathering product of Pb-bearing grains. Quantitative EDX analyses indicated that the Pb phosphate phase is pyromorphite with Ca frequently substituting for Pb between 21-31 atomic percent. However, positive identification of this phase by XRD was hindered by the deviation from pure end-member and possibly also by the poorly crystalline nature of the phase. Pyromorphite accounted for less than 2% of the total Pb in these soils. However, phosphate amendments to the soil could induce further formation of pyromorphite. As pyromorphite is a highly insoluble mineral, this may be effective in reducing the bioavailability of Pb in urban soils.     

Degradation of macrolide antibiotics by ozone: a mechanistic case study with clarithromycin

Macrolide antibiotics are widely used (in the order of 1g per person per year). They pass the body largely unchanged and are also not degraded in wastewater treatment plants. With not too much effort, they may be eliminated from their effluents by ozonation. The macrolide antibiotics have all a dimethylamino group at one of the carbohydrate residues in common. This functional group is the target of the ozone reaction, and clarithromycin has been selected here for a more detailed study. Since only the free amine reacts with ozone, the rate of reaction is pH dependent (at pH 7: k = 4 x 10(4) M(-1) s(-1)). In analogy to the ozonolysis of trimethylamine, the main reaction is a transfer of an O-atom yielding the N-oxide (identified by HPLC/MS-MS). A minor product (10%, based on formaldehyde yields) is demethylated clarithromycin (identified by HPLC/MS-MS). The dimethylamino group is thought to be essential for the binding of the macrolide antibiotics to their target. As a consequence, chemical changes of this functional group, notably the formation of the N-oxide that is no longer a proton acceptor, inactivates these drugs as assayed by the suppression of the growth of Pseudomonas putida. This is most important for wastewater treatment, as mineralization of clarithromycin by ozone would require 100 times as much ozone.     

Effects of sensitizers on the photodegradation of the systemic fungicide triadimenol

The photochemical behaviour of triadimenol (1) under various conditions has been examined. Significant degradation is obtained only in the presence of electron-acceptor sensitizers as 9,10-dicyanoanthracene or 2,4,6-triphenylpyrylium tetrafluoroborate, and long irradiation times are required. 1H-1,2,4-Triazole (2), 4-chlorophenyl formate (3), 4-chlorophenol (4), 1-(4-chlorophenoxy)-3,3-dimethyl-1-(1H-1,2,4-triazol-1-yl)butan-2-one (5), 4-chlorophenyl 2,2-dimethylpropanoate (6) and 4-chlorobenzoic acid (7) were identified as photoproducts by NMR and GC-MS.     

Isolation and characterization of <Emphasis Type="Italic">Bradyrhizobium</Emphasis> sp. SR1 degrading two β-triketone herbicides

In this study, a bacterial strain able to use sulcotrione, a β-triketone herbicide, as sole source of carbon and energy was isolated from soil samples previously treated with this herbicide. Phylogenetic study based on16S rRNA gene sequence showed that the isolate has 100 % of similarity with several Bradyrhizobium and was accordingly designated as Bradyrhizobium sp. SR1. Plasmid profiling revealed the presence of a large plasmid (>50 kb) in SR1 not cured under nonselective conditions. Its transfer to Escherichia coli by electroporation failed to induce β-triketone degrading capacity, suggesting that degrading genes possibly located on this plasmid cannot be expressed in E. coli or that they are not plasmid borne. The evaluation of the SR1 ability to degrade various synthetic (mesotrione and tembotrione) and natural (leptospermone) triketones showed that this strain was also able to degrade mesotrione. Although SR1 was able to entirely dissipate both herbicides, degradation rate of sulcotrione was ten times higher than that of mesotrione, showing a greater affinity of degrading-enzyme system to sulcotrione. Degradation pathway of sulcotrione involved the formation of 2-chloro-4-mesylbenzoic acid (CMBA), previously identified in sulcotrione degradation, and of a new metabolite identified as hydroxy-sulcotrione. Mesotrione degradation pathway leads to the accumulation of 4-methylsulfonyl-2-nitrobenzoic acid (MNBA) and 2-amino-4 methylsulfonylbenzoic acid (AMBA), two well-known metabolites of this herbicide. Along with the dissipation of β-triketones, one could observe the decrease in 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibition, indicating that toxicity was due to parent molecules, and not to the formed metabolites. This is the first report of the isolation of bacterial strain able to transform two β-triketones.     

Trends in the management of waste tyres and recent experimental approaches in the analysis of polycyclic aromatic hydrocarbons (PAHs) from rubber crumbs

The health and environmental consciousness of waste tires has increased tremendously over the years. This has motivated efforts to develop secondary applications that will utilize tire when they reach the end of their life cycle and limit their disposal in landfills. Among the applications of waste tires which are discussed in this review, the use of rubber crumbs in artificial turf fields has gained worldwide attention and is increasing annually. However, there are serious concerns regarding chemicals that are used in the manufacturing process of tires, which ultimately end up in rubber crumbs. Chemicals such as polycyclic aromatic hydrocarbons (PAH) and heavy metals which are found in rubber crumbs have been identified as harmful to human health and the environment. This review paper is intended to highlight some of the methods which have been used to manage waste tire; it also looks at chemicals/materials used in tire compounding which are identified as possible carcinogenic.

     

Characterisation of the dilute HCl extraction method for the identification of metal contamination in Antarctic marine sediments

A regional survey of potential contaminants in marine or estuarine sediments is often one of the first steps in a post-disturbance environmental impact assessment. Of the many different chemical extraction or digestion procedures that have been proposed to quantify metal contamination, partial acid extractions are probably the best overall compromise between selectivity, sensitivity, precision, cost and expediency. The extent to which measured metal concentrations relate to the anthropogenic fraction that is bioavailable is contentious, but is one of the desired outcomes of an assessment or prediction of biological impact. As part of a regional survey of metal contamination associated with Australia's past waste management activities in Antarctica, we wanted to identify an acid type and extraction protocol that would allow a reasonable definition of the anthropogenic bioavailable fraction for a large number of samples. From a kinetic study of the 1 M HCl extraction of two Certified Reference Materials (MESS-2 and PACS-2) and two Antarctic marine sediments, we concluded that a 4 h extraction time allows the equilibrium dissolution of relatively labile metal contaminants, but does not favour the extraction of natural geogenic metals. In a regional survey of 88 marine samples from the Casey Station area of East Antarctica, the 4 h extraction procedure correlated best with biological data, and most clearly identified those sediments thought to be contaminated by runoff from abandoned waste disposal sites. Most importantly the 4 h extraction provided better definition of the low to moderately contaminated locations by picking up small differences in anthropogenic metal concentrations. For the purposes of inter-regional comparison, we recommend a 4 h 1 M HCl acid extraction as a standard method for assessing metal contamination in Antarctica.     

Degradation of the antifouling compound Irgarol 1051 by manganese peroxidase from the white rot fungus Phanerochaete chrysosporium

Irgarol 1051 (2-methylthio-4-tert-butylamino-6-cyclopropylamino-s-triazine), a derivative of s-triazine herbicide, is an antifouling compound used as an alternative to organotins. The compound is highly persistent and is known to be biodegraded only by the white rot fungus, Phanerochaete chrysosporium. We used partially purified manganese peroxidase (MnP) prepared from P. chrysosporium to evaluate its capacity to degrade Irgarol 1051. MnP degraded Irgarol 1051 to two major products, one identified as M1 (identical to GS26575, 2-methylthio-4-tert-butylamino-6-amino-s-triazine) and the other not identified but with same mass spectrum as M1 and a different ultraviolet spectrum. This report clearly demonstrates that this ligninolytic enzyme is involved in the degradation of Irgarol 1051.     

Kinetics of particulate organic matter removal as a response to bioflocculation in aerobic biofilm reactors.

Recent research has identified that the major fraction of chemical oxygen demand in domestic wastewaters is in particulate form. The research presented herein develops the kinetics of particle removal as a response to bioflocculation at the surface of aerobic biofilms. This study focuses on the removal of particles that are maintained in aqueous suspension after 30 minutes of gravity settling. It is helpful to consider the particulate organics removal process in biofilms as the sum of four steps, namely (1) external transport of the particles to the biofilm surface, (2) bioflocculation, (3) organic particulate hydrolysis, and (4) diffusion and reaction of the solubilized organics by the bacterial cells comprising the biofilm. Organic (native corn starch) and inorganic particle (Min-U-Sil 10 [U.S. Silica Company, Berkeley Springs, West Virginia]) suspensions, with micronutrients, were continuously fed to a rotating disc biofilm reactor to verify a first-order kinetic expression that has been used to describe bioflocculation and to demonstrate that bioflocculation is the primary particle removal mechanism. Extracellular polymeric substances were extracted and quantified to describe the role they play in the bioflocculation process.     

PAHs in the bulk atmospheric deposition of the Seine river basin: source identification and apportionment by ratios, multivariate statistical techniques and scanning electron microscopy

The origin of polycyclic aromatic hydrocarbons (PAH) contamination in bulk atmospheric deposition at two sites of the Seine estuary, one urban and one industrial, has been investigated. The PAH profiles indicate that PAHs mainly have a pyrolytic origin, both in urban and industrial areas. PAH sources vary during the year with an increase of high molecular weight PAH proportions (especially for carcinogenic PAHs) in winter, that means an increase of combustion processes such as domestic heating. Ratios of indicator PAHs (FTH/FTH+PYR and IcdP/IcdP+BghiP) confirm the pyrolytic origin of PAHs. In summer, ratios show the presence of industrial sources. In addition to these two methods, a factor analysis/multiple linear regression model was applied and gave an approximation of PAH source apportionment. PAH were found to be associated predominantly with emissions from road traffic (gasoline and diesel), that accounts for 17-34%. Domestic heating is a very important PAH source in urban areas and accounts for up to 85% of PAHs in winter. Industrial emissions (refineries...) account for 25% in the industrial area in summer. Each is an identified source category for the region and these results are consistent with fly-ashes identified by scanning electron microscopy. This study demonstrates that a combination of source identification methods is a far more efficient than one method alone.     

Risk assessment for hazardous lubricants in machining industry

Knowing that over two-thirds of lubricant disposals return to the environment with no purification process, adequate strategies are demanded to reduce their risks. For this reason, the main focus of the present study is to describe an environment-friendly approach. In the first part, two widely used lubricants (mineral-based and vegetable-based) were introduced, and the reasons for their hazards were investigated. The composition of mineral-based lubricant was characterized by x-ray fluorescence elemental analysis. The result showed the presence of phosphorus, chlorine, and zinc dialkyl dithiophosphate in its composition that many scholars considered them to be the leading risk factor in the chemical composition of mineral-based lubricants. It has been focused on the potential risks of vegetable-based lubricants, which many researchers have identified as a safe lubricant. The Pseudomonas microbe was cultivated in the vegetable-based sample, and the result showed that although vegetable-based lubricants are compatible with humans, bacteria colony can quickly grow there without making any apparent changes that lead to harm to operators in a mysterious way. In the present work, the hypothesis of the safety of unreinforced vegetable-based lubricants has been rejected, and a new window on the environmental issues of vegetable-based lubricants has been presented. In the final, to eliminate environment-human risks and to reduce consumption of lubricant and natural resources to green manufacturing, a comprehensive study on the possibility of completely removing lubricant was performed. The results showed that the machining without lubricant could be replaced with the traditional method.