Hydrolysis of di-n-butyl phthalate, butylbenzyl phthalate and di(2-ethylhexyl) phthalate in human liver microsomes

Diester phthalates are industrial chemicals used primarily as plasticizers to import flexibility to polyvinylchloride plastics. In this study, we examined the hydrolysis of di-n-butyl phthalate (DBP), butylbenzyl phthalate (BBzP) and di(2-ethylhexyl) phthalate (DEHP) in human liver microsomes. These diester phthalates were hydrolyzed to monoester phthalates (mono-n-butyl phthalate (MBP) from DBP, mono-n-butyl phthalate (MBP) and monobenzyl phthalate (MBzP) from BBzP, and mono(2-ethylhexyl) phthalate (MEHP)) by human liver microsomes. DBP, BBzP and DEHP hydrolysis showed sigmoidal kinetics in V-[S] plots, and the Hill coefficient (n) ranged 1.37-1.96. The S₅₀, V_max and CL_max values for DBP hydrolysis to MBP were 99.7 μM, 17.2 nmol min⁻¹ mg⁻¹ protein and 85.6 μL min⁻¹ mg⁻¹ protein, respectively. In BBzP hydrolysis, the values of S₅₀ (71.7 μM), V_max (13.0 nmol min⁻¹ mg⁻¹ protein) and CL_max (91.3 μL min⁻¹ mg⁻¹ protein) for MBzP formation were comparable to those of DBP hydrolysis. Although the S₅₀ value for MBP formation was comparable to that of MBzP formation, the V_max and CL_max values were markedly lower (<3%) than those for MBzP formation. The S₅₀, V_max and CL_max values for DEHP hydrolysis were 8.40 μM, 0.43 nmol min⁻¹ mg⁻¹ protein and 27.5 μL min⁻¹ mg⁻¹ protein, respectively. The S₅₀ value was about 10% of DBP and BBzP hydrolysis, and the V_max value was also markedly lower (<3%) than those for DBP hydrolysis and MBzP formation for BBzP hydrolysis. The ranking order of CL_max values for monoester phthalate formation in DBP, BBzP and DEHP hydrolysis was BBzP to MBzP ? DBP to MBP > DEHP to MEHP > BBzP to MBP. These findings suggest that the hydrolysis activities of diester phthalates by human liver microsomes depend on the chemical structure, and that the metabolism profile may relate to diester phthalate toxicities, such as hormone disruption and reproductive effects.     

Anaerobic degradation of diethyl phthalate, di-n-butyl phthalate, and di-(2-ethylhexyl) phthalate from river sediment in Taiwan

We investigated anaerobic degradation rates for three phthalate esters (PAEs), diethyl phthalate (DEP), di-n-butyl phthalate (DBP), and di-(2-ethylhexyl) phthalate (DEHP), from river sediment in Taiwan. The respective anaerobic degradation rate constants for DEP, DBP, and DEHP were observed as 0.045, 0.074, and 0.027 1/day, with respective half-lives of 15.4, 9.4, and 25.7 days under optimal conditions of 30 °C and pH 7.0. Anaerobic degradation rates were enhanced by the addition of the surfactants brij 35 and triton N101 at a concentration of 1 critical micelle concentration (CMC), and by the addition of yeast extract. Degradation rates were inhibited by the addition of acetate, pyruvate, lactate, FeCl₃, MnO₂, NaCl, heavy metals, and nonylphenol. Our results indicate that methanogen, sulfate-reducing bacteria, and eubacteria are involved in the degradation of PAEs.     

Di(2-ethylhexyl)phthalate and mono(2-ethylhexyl)phthalate in media for in vitro fertilization

In vitro fertilization (IVF) is one of the most important treatments of infertility to provide a chance of conceiving. In IVF treatment, sperm are washed and motile sperm are isolated with sperm washing media (SWM) for the purpose of fertilization; fertilized ova are then incubated for a maximum of 5 or 6 d in media for IVF (IVFM). The exposure of fertilized ova to chemicals via such media has not been studied. We determined the concentrations of two contaminants; di(2-ethylhexyl)phthalate (DEHP) and its hydrolyzed product mono(2-ethylhexyl)phthalate (MEHP) in IVFM, SWM, and protein sources (PS: human serum albumin or serum substitute) for IVFM and SWM. The DEHP and MEHP in these media were extracted by a liquid-liquid extraction method and their concentrations determined by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Fifteen IVFM, nine SWM, and six PS obtained in Japan were examined. The concentrations of DEHP and MEHP in IVFM and SWM were <10-114 and <2.0-263 ng mL⁻¹, respectively. The concentrations of both DEHP and MEHP were higher in the media containing PS than in those without PS. Either MEHP alone or both DEHP and MEHP were detected in PS. The concentrations of DEHP and MEHP in PS were <10-982 and 47.0-1840 ng mL⁻¹, respectively. The DEHP and MEHP detected in these media were derived from PS. This is the first study on the chemical contamination of IVFM, SWM, and PS.     

Environmental effects of nanosilver: impact on castor seed germination, seedling growth, and plant physiology

Increasing use of nanoparticles in daily products is of great concern today, especially when their positive and negative impact on environment is not known. Hence, in current research, we have studied the impact of silver nanoparticle (AgNPs) and silver nitrate (AgNO₃) application on seed germination, root, and shoot length of castor bean, Ricinus communis L. plant. Silver nanoparticles had no significant effects on seedling growth even at higher concentration of 4,000 mg L^?1, while the silver in bulk form as AgNO₃ applied on the castor bean seeds inhibited the seed germination. Silver uptake in seedlings of the castor seeds on treatment with both the forms of silver was confirmed through atomic absorption spectroscopy studies. The silver nanoparticle and silver nitrate application to castor seeds also caused an enhanced enzymatic activity of ROS enzymes and phenolic content in castor seedlings. High-performance liquid chromatography analysis of individual phenols indicated enhanced content of parahydroxy benzoic acid. These kinds of studies are of great interest in order to unveil the movement and accumulation of nanoparticles in plant tissues for assessing future applications in the field or laboratory.     

土壤石油污染对植物种子萌发和幼苗生长的响应

研究了不同质量分数石油污染土壤对4种植物种子萌发和幼苗生长的影响,并对幼苗叶片的丙二醛(MDA)含量对污染胁迫的响应进行了研究,以期筛选适合石油污染土壤的潜在修复植物。设置了4组不同质量分数的石油污染土壤处理,对玉米(Zea mays L.)、高粱(Sorghum bicolor(L.)Moench)、披碱草(Elymus dahuricus Turcz.)和翅碱蓬(Suaeda hetroptera Kitag)4种供试植物进行了温室盆栽试验观测。结果表明,试验土属滨海盐渍土,在盐碱化和石油污染的双重胁迫下,不同的植物种子对石油污染表现出不同的耐受性;受试植物在10%(质量分数)石油污染处理下的发芽率均达80%以上,且石油的暴露水平越高,高粱、披碱草和翅碱蓬的种子发芽率越大;玉米和高粱的株高和根长均与土壤中石油浓度呈极显著正相关,而披碱草的根长则与之呈显著负相关;玉米的根冠比与土壤中石油浓度呈显著负相关,高粱和披碱草则与之相关性不明显;在石油污染土壤中生长的玉米和高粱,其叶片中MDA含量与空白对照组存在极显著差异,且随着污染土壤石油质量分数的升高呈现单峰响应特征;披碱草2个生长期(45、60d)叶片内的MDA含量均随土壤中石油浓度的升高而增加,土壤石油污染会对其造成毒害作用。总的来看,相比其他2种植物,高粱和披碱草具有较大的种子发芽率和石油污染耐受性,表明它们具有修复石油污染土壤的应用潜力。     

Effect of ultrasonication and Fenton oxidation on biodegradation of bis(2-ethylhexyl) phthalate (DEHP) in wastewater sludge

The presence of bis(2-ethylhexyl) phthalate (DEHP) and its metabolites, i.e. 2-ethylhexanol, 2-ethylhexanal, and 2-ethylhexanoic acid in wastewater sludge (WWS) were investigated during aerobic digestion and Bacillus thuringiensis (Bt)-based fermentation of WWS. Ultrasonication and Fenton oxidation pre-treatment was applied to improve biodegradability of WWS and bioavailability of the target compounds for digestion and fermentation. DEHP and 2-ethylhexanoic acid were observed at higher concentration, meanwhile 2-ethylhexanol and 2-ethylhexanal were observed at lower concentration in WWS. After 20-day aerobic digestion, DEHP removal was 72%, 89%, and 85%, and 2-ethylhexanoic acid removal was 71%, 84%, 79%, respectively for raw, ultrasonicated, and Fenton-oxidized sludges. Bt was found to degrade DEHP, leading to DEHP removal of 21%, 40%, and 30%, respectively for raw, ultrasonicated, and Fenton-oxidized sludges in the fermentation. The results suggested that aerobic stabilization and Bt-based fermentation can remove the phthalates, and pre-treatment of WWS was also effective in improvement of DEHP biodegradation. Hence, Bt-based biopesticide production from WWS can be applied safely when taking into consideration the phthalate contaminants.     

Ecotoxicological effects of typical personal care products on seed germination and seedling development of wheat (Triticum aestivum L.)

Biochemical responses of wheat (Triticum aestivum L.) seedlings stressed by two typical personal care products (PCPs) – triclosan (TCS) and galaxolide (HHCB) were experimentally investigated to assess their ecological risks. The results showed that wheat shoot and root elongation was significantly inhibited by 50–250 mg L⁻¹ TCS and HHCB. Wheat roots were sensitive to TCS, while shoots were sensitive to HHCB. The median effect concentration (EC₅₀) of TCS and HHCB based on the inhibition of their sensitive sites were 147.8 and 143.4 mg L⁻¹, respectively. Moreover, the damage of wheat seedlings treated by low concentration of TCS and HHCB during a long period cannot be neglected. After a 21-d exposure, 0.2–3.0 mg L⁻¹ TCS and HHCB treatment caused the damage to the accumulation of chlorophyll (CHL), the synthesis of soluble protein (SP), and the activity of peroxidase (POD) and superoxide dismutases (SOD) in different degree. However, different changing trends of these physiological indexes treated by different PCPs were observed after 7-d to 14-d exposures, especially the activity of POD and SOD. The activity of POD and SOD in wheat leaves and roots decreased with an increase in the concentration of TCS and the exposure time. However, the enzyme activities in wheat leaves treated by 0.2–3.0 mg L⁻¹ HHCB increased after a 14-d exposure, and with the prolongation of exposure time, the enzyme activities significantly decreased. The variations in these physiological indexes of wheat could be considered as good biomarkers of serious stress by TCS and HHCB in the environment.     

Effects of mono-(2-ethylhexyl) phthalate (MEHP) on chicken germ cells cultured in vitro

In recent decades, many toxicological tests based on in vivo or in vitro models, mainly from mammalian (rat–mouse) and fish species, were used to assess the risks raised by contact or ingestion of molecules of pharmaceutical, agricultural, or natural origin. But no, or few, in vitro tests using other non-mammalian models such as bird have been explored despite their advantages: the embryonic gonads of birds have a high plasticity of development sensitive to estrogen, and sperm production is nearly two times faster than in rodents. Hence, we have established an in vitro culture of germ cells and somatic cells from chicken post-natal testis, and we have evaluated the sensitivity against the endocrine disruptor compound mono-(2-ethylhexyl) phthalate (MEHP) in comparison to previous studies using rodent and human models. After 96 h of exposure in presence of 10 μM MEHP, chicken seminiferous tubules cultures present a structural alteration, a reduction in cell proliferation and in germ cells population. Apoptosis of germ and somatic cells increases in presence of 1 μM MEHP. Furthermore, MEHP does not affect inhibin B and lactate production by Sertoli cells. These results are in accordance with previous studies using rat, mice, or human culture of testicular cells and in similar range of exposures or even better sensitivity for some “end-points” (biological parameters). In conclusion, the establishment of this postnatal testicular cells culture could be considered as an alternative method to in vivo experiments frequently used for evaluating the impact on the terrestrial wildlife species. This method could be also complementary to mammal model due to the limiting number of animals used and its elevated sensitivity.     

分子印迹型TiO2的制备及其对邻苯二甲酸二(2-乙基)己酯的光催化性能

以邻苯二甲酸二（2-乙基）己酯为模板分子、甲基丙烯酸为功能单体和乙二醇二甲基丙烯酸酯为交联剂,在TiO2表面合成DEHP分子印迹聚合物,制备了分子印迹型TiO2（MIP-TiO2）,优化了制备工艺条件。利用傅里叶红外光谱（FI-IR）和X-射线衍射（XRD）对MIP-TiO2进行表征,并考察了MIP-TiO2对DEHP的光催化降解性能。结果表明,MIP-TiO2的最佳制备工艺条件为模板分子、单体、交联剂和TiO2的摩尔比为1：4：10：1.875,反应温度为60 ℃;聚合时间为18 h。MIP-TiO2仍保持了和TiO2一样的锐钛矿型结构,对DEHP 的光催化效果明显高于TiO2,对浓度为5 mg·L-1的DEHP的光催化降解效率可达88.15%。     

邻苯二甲酸二(2-乙基己基)酯对硝化型曝气生物滤池的影响及其去除效能

为探究邻苯二甲酸二(2-乙基己基)酯(DEHP)对硝化反应的影响及其去除情况,采用2座下向流硝化型曝气生物滤池进行对比实验.其中一座滤池进水DEHP浓度为100 μg/L,另一座滤池进水中不添加DEHP作为空白对照.对比实验期间氨氮负荷先后调整为0.6 kg NH4+-N/(m3·d)和1.5 kg NH4+-N/(m3·d)2个工况.通过60 d的对比实验,结果发现,2种工况下进水加DEHP的滤池,出水亚氮积累率分别为83.92%和80.71%,氨氮去除率分别为97.52%和49.39%;进水不加DEHP的滤池,出水亚氮积累率分别为64.15%和49.37%,氨氮去除率分别为98.77%和49.34%.DEHP的加入均促进了滤池出水中亚氮的积累,但对氨氮的去除不产生影响.进水加DEHP的滤池在0.6 kg NH4+-N/(m3·d)氨氮负荷下运行时,氨氮去除率沿水流方向逐渐提高,在滤床深度为1 400 mm处达到98.40%.亚氮积累率沿水流方向呈现先升高后降低的趋势,在滤床800 mm处达到最高值94.21%.在2个工况条件下,下向流硝化型曝气生物滤池对DEHP的平均去除率分别为91.71%和89.38%.大部分DEHP是在滤池进水端200 mm之内去除的.     

Influence of lead acetate on soil microbial biomass and community structure in two different soils with the growth of Chinese cabbage (Brassica chinensis)

A greenhouse pot experiment was conducted to evaluate the impact of different concentrations of lead acetate on soil microbial biomass and community structure during growth of Chinese cabbage (Brassica chinensis) in two different soils. The field soils were used for a small pot, short-term 60-day growth chamber study. The soils were amended with different Pb concentrations, ranging from 0 to 900mgkg(-1) soil. The experimental design was a 2 soilx2 vegetation/non-vegetationx6 treatments (Pb)x3 replicate factorial experiment. At 60 days the study was terminated and soils were analyzed for microbial parameters, namely, microbial biomass, basal respiration and PLFAs. The results indicated that the application of Pb at lower concentrations (100 and 300mgkg(-1)) as lead acetate resulted in a slight increase in soil microbial biomass, whereas Pb concentrations >500mgkg(-1) caused an immediate gradual significant decline in biomass. However, the degree of impact on soil microbial biomass and basal respiration by Pb was related to management (plant vegetation) or the contents of clay and organic matter in soils. The profiles of 21 phospholipid fatty acids (PLFAs) were used to assess whether observed changes in functional microbial parameters were accompanied by changes in the composition of the microbial communities after Pb application at 0, 300 and 900mg Pbkg(-1) soil. The results of principal component analyses (PCA) indicated that there were significant increases in fungi biomarkers of 18:3omega6c, 18:1omega9c and a decrease in cy17:0, which is an indicator of gram-negative bacteria for the high levels of Pb treatments In a word, soil microbial biomass and community structure, therefore, may be sensitive indicators reflecting environmental stress in soil-Pb-plant system. However, further studies will be needed to better understand how these changes in microbial community structure might actually impact soil microbial community function.     

Effect of introduced phthalate-degrading bacteria on the diversity of indigenous bacterial communities during di-(2-ethylhexyl) phthalate (DEHP) degradation in a soil microcosm

Four previously isolated di-butyl-phthalate (DBP) degraders were tested for their abilities to degrade di-(2-ethylhexyl) phthalate (DEHP). In aqueous medium supplemented with 100mg/l of DEHP, both isolate G1 and Rhodococcus rhodochrous G2 showed excellent degradative activity; in three days they were able to degrade more than 97% of the added DEHP. Rhodococcus rhodochrous G7 degraded 32.5% of the added DEHP and Corynebacterium nitrilophilus G11 showed the least amount of DEHP degradation. The addition of surfactant Brij 30 at 0.1x critical micelle concentration (2mg/l) significantly improved DEHP degradation by Rhodococcus rhodochrous G2 (more than 90% of the added DEHP was degraded within 24 hours), but slightly inhibited the degradation of DEHP by the isolate G1 and Rhodococcus rhodochrous G7. Based on the 16S rDNA sequence data, isolate G1 was identified as Gordonia polyisoprenivorans. Soil inhibited DEHP degradation by G. polyisoprenivorans G1; fourteen days after a second addition of DEHP, 11.5% of the total added DEHP (i.e., 243.4 microg/g soil) remained detectable. Changes in the bacterial community were monitored using denaturing gradient gel electrophoresis (DGGE) and respective dendrogram analysis. It is clear that DEHP and DEHP plus G. polyisoprenivorans G1 substantially affected the bacterial community structure in the soils. However, as the population of indigenous DEHP degraders increased in the DEPH-treated soil, its bacterial communities resembled those in the DEHP plus G. polyisoprenivorans G1-inoculated soil by Day 17.     

Phthalates and the aquatic environment: Part II The bioconcentration and depuration of di-2-ethylhexyl phthalate (DEHP) and di-isodecyl phthalate (DIDP) in mussels (Mytilus edulis)

Mussels ($\text{Mytilus edulis}$) were exposed to di-2-ethylhexyl phthalate (DEHP) and to di-isodecyl phthalate (DIDP) over a period of 28 days. The bioconcentration factor (BCF) as measured by ¹⁴C analysis, reached estimated plateau levels corresponding to mean BCF values of approximately 2500 and 3500 for the DEHP and DIDP respectively. The mussels were then held in clean seawater for a further 14 days and ¹⁴C analysis showed a depuration half-life of approximately 3.5 days for both phthalates. During the whole 42 days of the experiment general observations on the health of the animals showed no evidence of any adverse effects.     

Bio-herbicidal effects of five essential oils on germination and early seedling growth of velvetleaf (Abutilon theophrasti Medik.)

The evaluation of the inhibition effect exerted by the essential oils of basil (Ocimum basilicum), sage (Salvia officinalis), thyme (Thymus vulgaris), lemon balm (Melissa officinalis) and goldenrod (Solidago virgaurea) on seeds germination and early seedling growth of velvetleaf (Abutilon theophrasti Medik.) weed was examined in a laboratory bioassay. The essential oils were obtained by hydrodistillation and characterized chemically by gas chromatography coupled with both mass spectrometry and flame ionization detector. The working solutions of essential oil emulsified with Tween 20 and dissolved in distilled water were prepared at three concentration levels (0.01%, 0.1% and 1%, vol/vol). The results obtained showed that increase of essential oil concentration leads to decrease of seed germination, shoot and radical length of velvetleaf. The obtained data revealed a highly significant effect (P?<?0.05) between control and 1% and 0.1% oil concentrations in all treatments. The essential oils of basil, thyme and lemon balm exhibited more powerful bio-herbicidal effect compared to sage and goldenrod essential oils on the germination and early seedling growth of velvetleaf weed.     

Influence of seedling roots, environmental factors and soil characteristics on soil CO2 efflux rates in a 2-year-old loblolly pine (Pinus taeda L.) plantation in the Virginia Piedmont

To understand the role of managed forests in carbon sequestration an understanding of factors controlling soil CO2 efflux will be necessary. This study examined the influence of seedling roots, environmental factors, nutrient availability, and soil characteristics on soil CO2 efflux patterns in a 2-year-old pine plantation in the Virginia Piedmont. Efflux rates were measured both near the base of seedlings and midway between rows in plots that had received fertilization and mulch treatments in a factorial combination. Soil CO2 efflux rates were consistently higher near the base of seedlings, fertilization increased seedling growth with no significant effect on rates. and mulching increased winter efflux rates. In a regression analysis of seasonal soil CO2 efflux, soil temperature explained 42.2% of the variance followed by the interaction of soil temperature and moisture and of soil temperature and plot position, which together explained an additional 9.8% of the observed variance in seasonal rates. During March 2000 measurements, the spatial pattern of soil CO2 efflux between plots was most influenced by differences in soil nitrogen and pine root biomass. Furthermore, spatial differences observed in mean annual efflux rates were found to be highly influenced by the amount of soil coarse fragments in the upper soil profile.     

曝气生物滤池去除污水中邻苯二甲酸二(2-乙基己基)酯的效能

当前国内很多地方都将曝气生物滤池用于污水深度处理和回用.从保障再生水安全的角度出发,采用小试装置,对曝气生物滤池去除典型环境激素--邻苯二甲酸二(2-乙基己基)酯的效能进行了研究.结果发现,采用厚度1 000 mm的石英砂做滤料,在滤速0.5 m/h的条件下,进水邻苯二甲酸二(2-乙基己基)酯在5～35μg/L范围内时...