Cholinesterase activity and effects of its inhibition by neurotoxic drugs in Dictyostelium discoideum

Recently, we found acetylcholinesterase (AChE) activity in the ciliate protozoan Paramecium primaurelia. As in the slime mould Dictyostelium discoideum the presence of a serine esterase was found with strong sequence identity to Torpedo AChE, we extended to D. discoideum the investigation on the characterization and possible functions of cholinesterases (ChEs). In amoeboid cells, histochemical, biochemical, and electrophoresis analyses evidenced both a ChE activity able to hydrolyze the substrate PrTChI, and AChE (E.C. 3.1.1.7.) activity similar to Electrophorus electricus AChE. Conversely, butyrylcholinesterase activity was nearly absent, according to our previous results on P. primaurelia. Moreover, the possibility to utilize D. discoideum in a bioassay for the pre-chemical screening both of moist environments and fresh waters, in relation to the occurrence of the neurotoxic organophosphate drugs, such as "basudin", inhibiting ChE activity, was investigated. Exposure to basudin inhibited propionylcholinesterase (PrChE) activity in a dose-dependent manner in the range 10(-1)-10(-7) M (60% at 10(-4) M), without any significant effect on AChE activity. PrChE activity was inhibited slightly by 10(-5) M eserine, and reduced significantly both by 10(-5) M iso-OMPA and BW284C51, classically used to discriminate the different ChE molecular forms. The effects on cell morphology, cell density, and differentiation were evaluated in cultures exposed to PrTChI 10(-5) M or basudin 10(-4) M for a three-day period. The PrTChI-exposed sample exhibited cell morphology, cell density, ability to aggregate, and to form fruiting bodies similar to the control; whereas, the basudin-exposed sample showed anomalies in cell morphology and lower cell density than the control, together with inability to aggregate.     

Effects of long-term contamination of DDT on soil microflora with special reference to soil algae and algal transformation of DDT

DDT (1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane) and its principle metabolites, DDE (1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene) and DDD (1,1-dichloro-2,2-bis(p-chlorophenyl)ethane) are widespread environmental contaminants but little information is available concerning their effects on non-target microflora (especially microalgae and cyanobacteria) and their activities in long-term contaminated soils. For this reason a long-term DDT-contaminated soil was screened for DDT residues and toxicity to microorganisms (bacteria, fungi, algae), microbial biomass and dehydrogenase activity. Also, five pure cultures isolated from various sites (two unicellular green algae and three dinitrogen-fixing cyanobacteria) were tested for their ability to metabolise DDT. Viable counts of bacteria and algae declined with increasing DDT contamination while fungal counts, microbial biomass and dehydrogenase activity increased in medium-level contaminated soil (27 mg DDT residues kg(-1) soil). All the tested parameters were greatly inhibited in high-level contaminated soil (34 mg DDT residues kg(-1) soil). Species composition of algae and cyanobacteria was altered in contaminated soils and sensitive species were eliminated in the medium and high contaminated soils suggesting that these organisms could be useful as bioindicators of pollution. Microbial biomass and dehydrogenase activity may not serve as good bioindicators of pollution since these parameters were potentially influenced by the increase in fungal (probably DDT resistant) counts. All the tested algal species metabolised DDT to DDE and DDD; however, transformation to DDD was more significant in the case of dinitrogen-fixing cyanobacteria.     

Anaerobic biodegradation of DDT residues (DDT, DDD, and DDE) in estuarine sediment.

The potential for anaerobic biodegradation of 1,1,1-trichloro-2,2-bischlorophenylethane (DDT), 1,1-dichloro-2,2,-bischlorophenylethane (DDD), and dichlorodiphenylchloroethylene (DDE) in anoxic sediment slurries collected from the Keelung River was investigated in this study. o,p'- and p,p'-DDT were dechlorinated to o,p'- and p,p'-DDD, respectively, and then transformed to other compound(s). 1-Chloro-2,2-bis (p-chlorophenyl) ethylene (DDMU) and trace amount of dichlorobenzophenone (DBP) were detected in sediment slurries amended with p,p'-DDT or p,p'-DDD. DDMU was also detected in sediment slurries amended with p,p'-DDE. The relative transformation rates for both o,p'- and p,p'-isomers of DDT, DDD, and DDE were DDT>DDD>DDE. Re-addition of DDT, DDD, or DDE to the sediment slurries after initial removal enhanced the respective dechlorination rates. The transformation rates of the p,p'-isomers of both DDT and DDD were faster than those of the respective o,p'-isomers. p,p'-DDT dechlorination in the p,p'-DDT-adapted sediment slurries were inhibited by the addition of molybdate, or molybdate plus sulfate, but not inhibited by the addition of sulfate. Addition of bromoethane-sulfonic acid (BESA) slightly inhibited p,p'-DDT dechlorination. Non-adapted sediment slurries lost the ability to dechlorinate pentachlorophenol during adaptation to p,p'-DDT. p,p'-DDD was the major transformation product of p, p'-DDT in 3,4,4',5-tetrachlorobiphenyl-adapted sediment slurries, which suggested that the microbial community in the 3,4,4',5-CB-adapted sediment was unable to remove chlorine from the aromatic rings of p,p'-DDT.     

利用霉菌净化糖厂滤泥方法的优化

为净化糖厂滤泥,使其得到更好的资源化利用,研究pH、时间、温度、转速和接种量对霉菌净化糖厂滤泥效果的影响。在单因素基础之上,选取温度、pH和接种量为自变量,耗糖量为响应值,采用响应面优化法优化霉菌净化糖厂滤泥的条件。利用Box-Behnken设计和方差分析,研究各自变量及其交互作用对耗糖量的影响,模拟得到二次多项式回归方程的预测模型。结果表明,温度30℃,pH为8,接种量为6%为最佳优化条件,在此条件下,耗糖量的实际值为1 040 mg/20g滤泥,理论值为1 061.39 mg/20 g滤泥,两者的相对误差为1.97%。二次响应曲面方差分析结果表明,回归模型达到显著性水平,在研究区域内拟合较好,模型的精密度、可信度和精确度均在可行范围内。说明响应面法优化霉菌净化糖厂滤泥的条件切实可行。     

Growth and development of smooth bromegrass and tall fescue in TNT-contaminated soil

Plants can be used for effective and economical remediation of soil provided they are tolerant or resistant to contaminants. This study was conducted to determine effects of 2,4,6-trinitrotoluene (TNT) on growth and development of smooth bromegrass and tall fescue. Seeds of both species were grown in contaminated and non-contaminated soil mixed at ratios to obtain a range of concentrations and also in non-contaminated soil underlain by contaminated and non-contaminated soil mix. Germination, shoot and root dry weight, root length and area were measured. Germination and height of both species decreased with increasing TNT concentration. Shoot dry weight from tall fescue was 50% greater than smooth bromegrass at a given TNT concentration. Root length, area and dry weight of both species decreased with increasing TNT concentration. Root area and dry weight were greater for smooth bromegrass compared to tall fescue. This research indicates tall fescue and smooth bromegrass can germinate and grow in soils with concentrations less than 31 and 24 mg TNT l(-1), respectively.     

土壤中六六六和滴滴涕的堆肥生物修复研究

采用堆肥生物修复法对土壤中六六六(HCH)和滴滴涕(DDT)的降解效果进行了研究.试验结果表明,六六六和滴滴涕的降解先后经历了快速降解阶段和缓慢降解阶段;堆肥第21天后,六六六和滴滴涕降解率均达60%以上;γ-HCH降解效果最好,其次为β-HCH,α-HCH和δ-HCH的降解效果差别较大;p,p'-DDE的降解效果要好于p,p'-DDD,初始质量浓度为0.03 mg/kg的p,p'-DDT未得到降解;微生物对六六六和滴滴涕的降解作用均较明显.     

Lung subcellular fractions and surfactant lipid metabolism of rats exposed with DDT or endosulfan intratracheally

The effect of intratracheally administered DDT (5 mg/100 g body weight) or endosulfan (1 mg/100 g body weight) for three consecutive days has been studied on lipid metabolism of rat lung subcellular fractions. Both the insecticides did not affect the lung weight and the protein contents of microsomes, lamellar bodies and surfactant but significantly increased the phospholipid contents of microsomal and surfactant system. Most of the neutral lipid components of lung subcellular fractions were also increased by DDT or endosulfan treatments, except that of surfactant triglycerides which were decreased by DDT treatment. DDT or endosulfan both increased the incorporation of radioactive [methyl-3H]choline into microsomal phosphatidylcholine (PC) and surfactant dipalmitoylphosphatidylcholine (DPPC) without affecting the incorporation of radioactive [methyl-14C]methionine, showing the increased synthesis of PC via CDPcholine pathway. The results presented in this communication showed that DDT and endosulfan, the two different chloroinsecticides have similar effects on microsomal lipid metabolism but produce different biochemical manifestations on the secretion of surfactant phospholipids.     

Lung subcellular fractions and surfactant lipid metabolism of rats exposed with DDT or endosulfan intratracheally

Abstract

The effect of intracheally administered DDT (5 mg/100 g body weight) or endosulfan (1 mg/100 g body weight) for three cosecutive days have been studied on lipid metabolism of rat lung subcellular fractions. Both the insecticides did not affect the lung weight and the protein contents of microsomes, lamellar bodies and surfactant but significantly increased the phospholipid contents of microsomal and surfactant system. Most of the neutral lipid components of lung subcellular fractions were also increased by DDT or endosulfan treatments, except that of surfactant triglycerides which were decreased by DDT treatment. DDT or endosulfan both increased the incorporation of radioactive [methyl‐³H]choline into microsomal phosphatidylcholine (PC) and surfactant dipalmitoylphosphatidylcholine (DPPC) without affecting the incorporation of radioactive [methyl‐¹⁴C]methionine, showing the increased synthesis of PC via CDPcholine pathway. The results presented in this communication showed that DDT and endosulfan, the two different chloroinsecticides have similar effects on microsomal lipid metabolism but produce different biochemical manifestations on the secretion of surfactant phospholipids.     

Behaviour of DDT under laboratory and outdoor conditions in Germany

Abstract

Experiments were conducted on adsorption, volatilization and UV‐degradation of p,p'‐DDT on soil surface, and leaching and degradation in sand columns. p,p'‐DDT was shown to adsorb stronger to soils with higher organic content. UV irradiation at 290 nm for 10 hours mineralized less than 0.1% of DDT in soil.

Results show that only 0.1% of DDT volatilized in a sun‐exposed semi‐closed quartz system. Polar compounds accounted from 1.4% after 55 days. The rate of volatilization and degradation in an open system was much higher; only 15% DDT and 7% DDE were recovered after 6 weeks in the organic extract. p,p'‐DDT was adsorbed to a great extent on the top layers of sand columns; 86% in the top 8 cm.     

Influence of arsenic co-contamination on DDT breakdown and microbial activity

The impacts of arsenic co-contamination on the natural breakdown of 1,1,l1-trichloro-2,2-bis(4-chlorophenyl)ethane (DDT) in soil are investigated in a study of 12 former cattle dip sites located in northeastern NSW, Australia. This study examines the relationship between the intrinsic breakdown of DDT to 1,1 -dichloro-2,2-bis(4-chlorophenyl)ethane (DDD) and 1,l-dichloro-2,2-bis(4-chlorophenyl)ethylene (DDE), and the impacts of arsenic co-contamination on this breakdown. Between-site analysis demonstrated that arsenic at 2000 mg/kg gave a 50% reduction in the concentration of DDD compared to background arsenic of 5 mg/kg.Within-site analysis also showed the ratio of DDT:DDD increased in soils as arsenic concentrations increased. This within-site trend was also apparent with the DDT:DDE ratio, suggesting inhibition of DDT breakdown by arsenic co-contamination. Microbial activity was inhibited as residues of total DDTs and arsenic increased. Hence arsenic co-contamination and high concentrations of DDT in soil may result in an increased persistence of DDT in the environment studied.     

Emission characteristics of granulated fuel produced from sewage sludge and coal slime

The neutralization of wastewater treatment residues is an issue for many countries. The European Union (EU) legal regulations have limited the use of the residues in agriculture and implemented a ban for their disposal. Therefore, urgent action should be taken to find solutions for the safe disposal of sewage sludge. The problem refers in particular to the new EU member countries, including Poland, where one can now observe an intensive development of sewage system networks and new sewage treatment plants. At the same time, these countries have few installations for thermal sewage sludge utilization (e.g., there is only one installation of that type in Poland). Simultaneously, there are many coal-fired mechanical stoker-fired boilers in some of these countries. This paper presents suggestions for the production of granulated fuel from sewage sludge and coal slime. Additionally, among others, lime was added to the fuel to decrease the sulfur compounds emission. Results are presented of research on fuel with two average grain diameters (approximately 15 and 35 mm). The fuel with such diameters is adapted to the requirements of the combustion process taking place in a stoker-fired boiler. The research was aimed at identifying the behavior of the burning fuel, with special attention paid to its emission properties (e.g., to the emissions of oxides of nitrogen [NO(x)], sulfur dioxide [SO2], and carbon monoxide [CO], among others). The concentration and emission values were compared with similar results obtained while burning hard coal. The combustion process was carried out in a laboratory stand where realization of the large-scale tests is possible. The laboratory stand used made simulation possible for a wide range of burning processes in mechanical stoker-fired boilers.     

Contents and sources of DDT impurities in dicofol formulations in Turkey

Background, aim, and scope  Dicofol is widely used as a pesticide in agriculture applications. Since dicofol is mainly synthesized from dichlorodiphenyltrichlorethane (DDT), it contains DDT as an impurity. The European Community has forced Prohibition Directive 79/117/EEC to reduce DDT in dicofol formulations. Specifically, DDT content in a dicofol formulation cannot exceed 0.1%. The goal of this project was to determine the DDT content in dicofol formulations used in Turkey. Materials and methods  Samples of all the dicofol formulations in Turkey were collected to quantify DDT and DDT-related compounds. Four replicates were used for each sample. GC/MS/MS was used to analyze p,p′ and o,p′ isomers of DDT, DDD, and DDE. A HPLC was used to determine p,p′-Cl-DDT concentrations. Results  The total DDT content of the formulated dicofol was found between 0.3% and 14.3%. The concentration of p,p′-DDE ranged from 167 to 1,042 mg kg⁻¹ in dicofol samples. p,p′-DDT concentrations were found to be 32 to 183 mg kg⁻¹. The o,p’-DDT level ranged from 2 to 34 mg kg⁻¹ in the dicofol formulations analyzed. Discussion  It was estimated that 617.8 kg of DDT was released from dicofol. The main impurity was identified as p,p-Cl-DDT. Based on these results, dicofol serves as a continuing source of DDT contamination. Conclusions  All DDT concentrations in dicofol samples analyzed were higher than the permitted 0.1% level of Prohibition Directive 79/117/EEC. The reduction of dicofol is critical since it serves as a continual source of DDT contamination. Recommendations and perspectives  DDT has been found in soil, water, and air samples. Dicofol has been identified as a contributor to continued DDT contamination in soil and water. More studies are needed to ascertain the source of DDT in the air.     

高效脱水霉菌的筛选及其脱水机理分析

用常规分离纯化方法从某池塘底泥筛选分离霉菌,根据菌落颜色和形态特征的不同共分离得到49株菌株。通过剩余污泥脱水实验获得高效脱水霉菌菌株D41,在24 h内使污泥比阻降低49.2%,含水率由最初的82.9%降至78.3%。通过机理研究发现,霉菌D41的投入使剩余污泥中多糖含量降低36.9%,从而改善剩余污泥脱水性能。     

Contamination of bovine milk with DDT and HCH residues in relation to their usage in malaria control programme

The contribution of spraying DDT and HCH for malaria control towards the contamination of bovine milk was investigated by analysing milk samples collected from preselected localities sprayed with either DDT or HCH in the Punjab. A direct correlation was found between the amounts of residues of these insecticides in milk and their typical usage pattern for the mosquito control programme. It is considered that these results may have significant bearing on the regulatory control of DDT and HCH residues in milk.     

The degradation of parathion and DDT in aqueous systems containing organic additives

Abstract

Laboratory studies were conducted to investigate some of the factors influencing pesticide degradation in‐ aqueous systems. Parathion added to natural water in ethanol, acetone or without organic solvent, was completely degraded within 2 wk. While most of the parathion was reduced to amino‐parathion when added in ethanol, no amino‐parathion was detected in the presence of acetone or when no solvent was added, suggesting that in the latter two cases the insecticide was aerobically degraded to other metabolites. No paraoxon was detected. When ethanol concentration was increased from 1% to 2 and 4%, the rate of parathion degradation was inversely related to the ethanol concentration. In the presence of glucose as a carbon source, approximately 50% of the parathion was reduced to aminoparathion. DDT degradation in natural water was more rapid when it was added in ethanol than when added in acetone. The only DDT metabolite detected was TDE, with about 36% conversion in presence of ethanol, and 20% when the DDT was added in acetone.     

Effects of different treatments on soil-borne DDT and HCH dynamics and plant uptake

Pot experiments were conducted to examine the effects of various fertilizers, as well as soil dilution treatments on the dynamics of soil-borne DDTs [sum of dichlorodiphenyltrichloroethane (DDT), chlorodiphenyldichloroethylene (DDE) and di- chlorodiphenyldichloroethane (DDD)] and hexachlorocyclohexanes (HCHs, sum of α-HCH, β-HCH, γ-HCH and δ-HCH) and their subsequent impacts on the uptake of DDTs and HCHs by a test plant. The results show that the soil residual DDTs and HCHs concentrations in the iron-rich fertilizer-treated soil were significantly lower than those in other fertilizer-treated soils. There was a close relationship between the soil residual DDTs and the plant tissue DDTs. This suggests that the uptake rate of DDTs by the plant was dependent on the concentration of soil-borne DDTs. A less close relationship between soil residual HCHs and plant tissue HCHs was also observed. Dilution of pesticide-contaminated soil with the non-contaminated soil not only physically reduced the concentration of pesticides in the soil but also enhanced the loss of soil-borne pesticides, possibly through the improvement of soil conditions for microbial degradation. Soil dilution had a better effect on promoting the loss of soil-borne HCHs, relative to soil-borne-DDTs. The research findings obtained from this study have implications for management of heavily contaminated soils with DDTs and HCHs. Remediation of DDTs and HCHs-contaminated soils in a cost-effective way can be achieved by incorporating treatment techniques into conventional agricultural practices. Applications of iron-rich fertilizer and soil dilution treatments could cost-effectively reduce soil-borne DDTs and HCHs, and subsequently the uptake of these organochlorine pesticides by vegetables.     

Effects of different treatments on soil-borne DDT and HCH dynamics and plant uptake

Pot experiments were conducted to examine the effects of various fertilizers, as well as soil dilution treatments on the dynamics of soil-borne DDTs [sum of dichlorodiphenyltrichloroethane (DDT), chlorodiphenyldichloroethylene (DDE) and di- chlorodiphenyldichloroethane (DDD)] and hexachlorocyclohexanes (HCHs, sum of α-HCH, β-HCH, γ-HCH and δ-HCH) and their subsequent impacts on the uptake of DDTs and HCHs by a test plant. The results show that the soil residual DDTs and HCHs concentrations in the iron-rich fertilizer-treated soil were significantly lower than those in other fertilizer-treated soils. There was a close relationship between the soil residual DDTs and the plant tissue DDTs. This suggests that the uptake rate of DDTs by the plant was dependent on the concentration of soil-borne DDTs. A less close relationship between soil residual HCHs and plant tissue HCHs was also observed. Dilution of pesticide-contaminated soil with the non-contaminated soil not only physically reduced the concentration of pesticides in the soil but also enhanced the loss of soil-borne pesticides, possibly through the improvement of soil conditions for microbial degradation. Soil dilution had a better effect on promoting the loss of soil-borne HCHs, relative to soil-borne-DDTs. The research findings obtained from this study have implications for management of heavily contaminated soils with DDTs and HCHs. Remediation of DDTs and HCHs-contaminated soils in a cost-effective way can be achieved by incorporating treatment techniques into conventional agricultural practices. Applications of iron-rich fertilizer and soil dilution treatments could cost-effectively reduce soil-borne DDTs and HCHs, and subsequently the uptake of these organochlorine pesticides by vegetables.     

Levels and distribution of DDT in the Cinca River (Spain).

The evolution over time of the levels and distribution of dichlorodiphenyltrichloroethane (DDT) in water, surface sediments, and fish from the River Cinca (Spain), a tributary of the River Ebro, during the period 1999 to 2004, was investigated by means of gas chromatography coupled with mass spectrometry. The sampling site corresponded to a point downstream from Monzón, a heavily industrialized town with drainage into the river. This river has historically been a source of emissions of DDT and its metabolites. The highest levels were found in 1999 and 2000, although the concentrations of organic compounds in sediments and fish have decreased since then. The levels of DDT in water were below the quantification limit during the period of study. The average composition of DDT isomers measured in sediments and fish showed the prevalence of p,p'-DDE, the product of aerobic degradation of p,p'-DDT. Concentrations in fish were compared with sediment samples, and high quotients indicate that they are highly bioavailable.     

Level and distribution of DDT in surface soils from Tianjin, China

One hundred and eighty eight surface soil samples were collected from the Tianjin area to study the contamination of DDT and its metabolites. Measurements were taken for p,p'-DDE, p,p'-DDD, p,p'-DDT, o,p'-DDE, o,p'-DDD and o,p'-DDT for all samples. The results indicated that p,p'-DDT and p,p'-DDE were the predominant contaminant compounds in the surface soil samples, with mean concentrations of 27.5 and 18.8 ng g(-1) respectively. No significant differences in DDT concentrations were found between the soils from wastewater treated irrigated areas and other areas, suggesting that wastewater irrigation is not an important source of DDT in the area. However, the spatial distribution of soil DDTs levels in the area did correlate well with early direct application rates of pesticides. In addition, both pH level and organic carbon content are also known factors affecting the level of DDT and its metabolites. Although it was assumed that the use of these chemicals was banned in the early 1980s, the current concentration levels appear to be too high to be mere residuals after 20 years degradation.