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.     

The influence of humidity on the emission of di-(2-ethylhexyl) phthalate (DEHP) from vinyl flooring in the emission cell “FLEC”

Asthma in children appears to be associated with both phthalate esters and dampness in buildings. An important question is whether the concentrations of phthalate esters correlate with dampness (expressed as relative humidity—RH) in indoor air. The objective was to study the influence of RH on the specific emission rate (SER) of di-(2-ethylhexyl)phthalate (DEHP) from one type of vinyl flooring in the well characterized Field and Laboratory Emission Cell (FLEC). The vinyl flooring with ca. 17% (w/w) DEHP as plasticizer was tested in 6 FLECs at 22 °C. The RH in the 6 FLECs was 10%, 30%, 50% (in triplicate) and 70%. The RH was changed after 248 d in 2 of the 50%-FLECs to 10% and 70%, and to 50% in the 10%-and 70%-FLECs. The data show that the SER of DEHP from vinyl flooring in FLECs during a 1 yr period is independent of the RH. A new physically based emission model for semivolatile organic compounds was found to be consistent with the experimental data and independent of the RH. The model helps to explain the RH results, because it appears that RH does not significantly influence any of the identified controlling mechanisms.     

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.     

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.     

Experimental assessment of bio-reduction of di-2-thylhexyl phthalate (DEHP) under aerobic thermophilic conditions

The reduction of organic contaminants in sewage sludge is of great importance for a further sludge disposal or agricultural utilization. Laboratory scale batch experiments were performed to assess the potential use of the aerobic thermophilic treatment technique to reduce the concentration of difficult to degrade organic chemicals. Di-2-ethylhexyl phthalate (DEHP) was chosen as a model representative of these chemicals. The effect of the sludge temperature and aeration rate on the reduction of DEHP concentration as well as on the reduction of the organic dry solid (oDS) was investigated. With a specific air flow rate of 16 m3/m3.h and a thermophilic temperature of 63 degrees C it was possible to achieve up to 70% reduction of the DEHP concentration and 61% of oDS within 96 hours. The maximum degradation of the oDS matter occurred within the first 24 hours of operation whereby only little oDS was degraded afterward. During the experiments the reactor content was routinely monitored for pH, COD, along with the ammonia nitrogen and orthophosphate concentrations.     

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.     

A systematic review on the adverse health effects of di-2-ethylhexyl phthalate

Di (ethylhexyl) phthalate (DEHP) is a global environmental pollutant. This study aims to systematically review the literature on health effects of exposure to DEHP including effects on reproductive health, carcinogenesis, pregnancy outcome, and respiratory system. The literature search was done through Scopus, ISI Web of Science, Google Scholar, PubMed, Medline, and the reference lists of previous review articles to identify relevant articles published to June 2016 in each subject area. The inclusion criteria were as follows: original research, cross-sectional studies, case–control studies, cohort studies, interventional studies, and review articles. Both human and animal studies were included. The search was limited to English language papers. Conference papers, editorials, and letters were not included. The systematic review was conducted and reported in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement. Overall, 152 of the 407 papers met the inclusion criteria. We provided an up-to-date comprehensive and critical assessment of both human and animal studies undertaken to explore the effects of DEHP. It revealed that in experimental studies, exposure to DEHP mainly targeted the reproductive, neurodevelopment, and respiratory systems. Human studies reported that exposure to this contaminant had carcinogenic effects and influenced neurodevelopment in early life. This systematic review underscored the adverse health effects of DEHP for pregnant women and the pediatric age group. It summarizes different response of humans and experimental animals to DEHP exposure, and some suggested underlying mechanisms.     

Chamber studies on mass-transfer of di(2-ethylhexyl)phthalate (DEHP) and di-n-butylphthalate (DnBP) from emission sources into house dust

For a number of phthalates and especially for di(2-ethylhexyl)phthalate (DEHP), surprisingly high house dust concentrations are reported in the literature. Therefore, the uptake of the most prominent compounds DEHP and di-n-butylphthalate (DnBP) from plasticized indoor materials into house dust samples of different organic content has been experimentally determined. The experiments have been performed within 45 days which is sufficient for the more volatile phthalate (DnBP) to reach equilibrium conditions. DnBP reaches considerably higher concentrations in the chamber air compared to real room measurements and, thus, also elevated dust concentrations. In contrast, the mass transfer of DEHP in the dust via the gas phase was significantly lower. However, small chamber experiments showed elevated mass transfer of DEHP in case of direct contact between emission source and sink. This aspect is experimentally determined using an plasticized PVC polymer with and without direct contact to house dust. A transfer into the dust could be observed in dependence of the initial concentration in the material. However, the results do not allow the differentiation between the two uptake mechanisms via capillary forces and contact to the material’s boundary layer. The results illustrate that the reasons for elevated DEHP concentrations in dust indoors can be traced back to direct contact of source and sink, abrasion from the source, and transport via airborne particles.     

Studies of mobility of di-iso-butyl phthalate (DiBP), di-N-butyl phthalate (DBP), and di-(2-ethyl hexyl) phthalate (DEHP) by plant foliage treatment in a closed terrestrial simulation chamber

A climate chamber was constructed for model studies of mobility and effects of chemical substances within simplified terrestrial systems. The chamber functions and its performance were tested by foliar applications of phthalates on higher plants. A low elimination rate from foliage was observed for DEHP. More than 95% of DiBP and DBP were eliminated within 15 days. 1.5 μg cm⁻² of DBP caused chlorosis on the leaves of Sinapis alba L.     

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.     

Impact of drying and composting procedures on the concentrations of 4-nonylphenols, di-(2-ethylhexyl)phthalate and polychlorinated biphenyls in anaerobically digested sewage sludge

Enhanced treatments of sewage sludge produce a more manageable product for agricultural use by stabilizing the material, removing water, and reducing the possibility of pathogen transfer. We investigated the impact of pilot-scale composting and drying of sludge on physicochemical characteristics and on the concentrations of some organic contaminants. During the 143 day composting procedure, organic matter fell 22% and moisture by half. Concentrations of 4-nonylphenols (4-NPs) fell by 88% and di-(2-ethylhexyl) phthalate (DEHP) by 60%; losses continued throughout the procedure. Losses of total polychlorinated biphenyls (PCBs) were 11%, mostly from the lower molecular weight congeners, suggesting volatilization as the most likely loss mechanism. The drying process was much shorter, 40 days, yet organic matter content decreased by 27% and moisture by 85%. Losses of 4-NPs (39%) and DEHP (22%) were less than in composting and stopped when moisture content became constant. There were no losses of PCBs. Both treatments are simple, practical procedures that reduce the volume of waste and are applicable in situ on farms. Composting would be the method of choice for reducing organic contaminants but requires much longer times than drying.     

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.     

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.     

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%。     

Production of male neonates in Daphnia magna (Cladocera, Crustacea) exposed to juvenile hormones and their analogs

We exposed the water flea Daphnia magna (Cladocera, Crustacea) to either juvenile hormone I (JH I), juvenile hormone II (JH II), or the juvenile hormone-mimicking insecticides kinoprene, hydroprene, epofenonane, or fenoxycarb. By 21-day reproduction tests, we investigated the effects on the number of neonates born per female and the offspring sex ratio. All six chemicals induced D. magna to produce male neonates; the male sex ratio of the offspring increased as the chemical concentration increased. EC50 values for production of male neonates were estimated as 400 (JH I), 410 (JH II), 190 (kinoprene), 2.9 (hydroprene), 64 (epofenonane), and 0.92 (fenoxycarb) microg/l. The number of neonates produced was reduced with all chemicals at the concentrations investigated. At the EC50 for male production, five of the six chemicals reduced the reproductive rate to less than 50%; the exception was epofenonane, which caused only a slight reduction in reproductive rate. These results were similar to those obtained for five juvenoids studied previously, one of which was studied here again. There are now 10 chemical substances--all juvenile hormones or their analogs-that are known to induce D. magna to produce male neonates. This suggests that juvenile hormone is involved in initiating male production followed by sexual reproduction in D. magna, and probably in most cladocerans that exhibit cyclic parthenogenesis.     

Effects of thiobencarb herbicide to an alga (Nannochloris oculata) and the cladoceran (Daphnia magna)

Chronic toxicity studies were conducted with an algae (Nannochloris oculata) and the cladoceran (Daphnia magna) to determine their relative sensitivities to the thiocarbamate herbicide thiobencarb (S-4-chlorobenzyl diethylthiocarbamate). Most of the algal populations were initially affected by exposure to the herbicide. Thiobencarb concentrations higher than 0.5 mg/L significantly reduced algal densities after 24-h exposure. The 24-h static EC50 in D. magna was 3.01 mg/L. The sublethal effects of 0.3, 0.37, 0.5, 0.75, and 1.5 mg/L of thiobencarb concentrations on the survival, reproduction, and growth of D. magna were monitored for 21 days. The parameters used to determined the effect of the herbicide on D. magna were mean total young per female; mean brood size; days to first brood; intrinsic rate of natural increase (r); growth; and survival. Reproduction was significantly reduced at thiobencarb concentrations of 0.30 mg/L and higher while survival was affected after exposure to 0.75 and 1.5 mg/L of the pesticide. The r value decreased with increasing concentrations of thiobencarb. Growth, as measured by body length, was depressed significantly after exposure to all herbicide concentrations tested.     

Predicting bioconcentration factors (BCFs) of polychlorinated bornane (Toxaphene) congeners in fish and comparison with bioaccumulation factors (BAFs) in biota from the aquatic environment.

Polychlorinated bornanes, the main components of Toxaphene, are bioconcentrated in aquatic organisms to a high extent. However, up to this time no bioconcentration tests with individual chlorinated bornanes in aquatic organisms have been performed. Therefore, the bioconcentration factors (BCFs) of seven selected persistent chlorinated bornane congeners which are regularly found in aquatic organisms, were predicted from their n-octanol/water partition coefficients (log Kow). Furthermore, these BCF values were compared with the measured bioaccumulation factors (BAFs) in zooplankton and different fish species from the aquatic environment.     

Acute toxicity tests with Daphnia magna, Americamysis bahia, Chironomus riparius and Gammarus pulex and implications of new EU requirements for the aquatic effect assessment of insecticides

Threshold concentrations for treatment related effects of 31 insecticides, as derived from aquatic micro-/mesocosm tests, were used to calibrate the predictive value of the European Tier-1 acute effect assessment on basis of laboratory toxicity tests with Daphnia magna, Chironomus spp., Americamysis bahia and Gammarus pulex. The acute Tier-1 effect assessment on basis of Daphnia (EC(50)/100) overall was protective for organophosphates, carbamates and most pyrethroids but not for neonicotinoids and the majority of insect growth regulators (IGRs) in the database. By including the 28-day water-spiked Chironomus riparius test, the effect assessment improves but selecting the lowest value on basis of the 48-h Daphnia test (EC50/100) and the 28-day Chironomus test (NOEC/10) is not fully protective for 4 out of 23 insecticide cases. An assessment on basis of G. pulex (EC(50)/100) is sufficiently protective for 15 out of 19 insecticide cases. The Tier-1 procedure on basis of acute toxicity data (EC(50)/100) for the combination of Daphnia and A. bahia and/or Chironomus (new EU dossier requirements currently under discussion) overall is protective to pulsed insecticide exposures in micro-/mesocosms. For IGRs that affect moulting, the effect assessment on basis of the 48-h Chironomus test (EC(50)/100) may not always be protective enough to replace that of the water-spiked 28-day C. riparius test (NOEC/10) because of latency of effects.     

Assessment of the impact of the emission of certain organochlorine compounds on the aquatic environment: Part I: Monochlorophenols and 2,4-dichlorophenol

The consequences of contamination of the aquatic environment by chlorophenols (2-chlorophenol, 3-chlorophenol, 4-chlorophenol and 2,4-dichlorophenol), have been evaluated by extensive review of the available scientific data. The chlorophenols generally exert moderate toxic effects to mammalian and aquatic life, although toxicity to fish upon long-term exposure may be considerable, as has been found for 2,4-dichlorophenol. Persistence is low when adapted microflora is present, capable of biodegrading these compounds, but may become moderate to high depending on the environmental conditions. This is most pronounced for 3-chlorophenol. Bioaccumulation is expected to be low. A striking feature of these chlorophenols is their strong organoleptic effect.