复杂介质中典型溴化阻燃剂的污染诊断、归趋及生态毒理效应研究
摘要
溴化阻燃剂(Brominated flame-retardants,BFRs)是一类新型的全球性环境污染物。其产量在过去的20年间迅猛增长。当前由于其在环境、野生生物以及人体内的存在,而受到越来越多的关注。然而,关于它们在土壤中的归趋以及生态毒理效应的研究还非常有限。
为此,我们对四溴双酚A(tetrabromobisphenol A, TBBPA)和六溴环十二烷(hexabromocyclododecane, HBCD)在土壤中的归趋,TBBPA对颤蚓和小麦的生态毒理效应以及六溴环十二烷(hexabromocyclododecane, HBCD)对小麦的生态毒理效应进行了研究。结果显示：
1、土壤对TBBPA和HBCD的物理吸附导致二者在种植和未种植土壤中的回收率下降了90%。而在混合种植的情况下将近50%的HBCD被回收了,这充分说明植物种间竞争会增强HBCD的生物有效性。虽然白菜和萝卜都可以吸收TBBPA和HBCD,但是白菜吸收和富集污染物的能力要比萝卜强,并且各植物组织中HBCD的浓度约为TBBPA浓度的3.5-10.0倍。HBCD在不同介质中的分布与其异构体类型有关,a-HBCD倾向于在植物组织中分布,而y-HBCD则倾向于在土壤中。以上这些说明吸附会降低二者的生物有效性,以及人类暴露的风险,但是植物的作用会使这种风险增加。2、暴露8d后,颤蚓体内SOD的活性发生显著变化,变化趋势明显分为升高、降低、再升高3段,在0.05 mg L-1时SOD的活性被最大诱导(p<0.01),达到对照组的7.8倍,各浓度组SOD的活性均显著高于对照组,由对照组的1.5倍到对照组的7.8倍。同时CAT的活性变化则呈升高、降低、再升高、再降低4段,0.5 mg L-1时其活性达到最大值(p<0.01),除0.005 mg L-1和0.25 mg L-1浓度组CAT活性受抑制外,其余浓度组CAT的活性均高于对照组,由对照组的1.1倍到1.9倍。染毒浓度为0.25 mg L-1时,GST的活性达到最大诱导(p<0.01),其活性先缓慢上升后下降,并且各处理组GST的活性均显著高于对照组(p<0.05)。在单一浓度TBBPA污染暴露的10d时间里,颤蚓体内SOD的活性变化趋势形成了一个变形的“M”形曲线,在第3d时其活性达到最大诱导,而CAT活性的变化则表现为一个不规则的“N”形曲线,在第5d时其活性达到最大诱导,并且SOD的活性比CAT的活性受时间的影响小,相对来说更稳定。可见,SOD与GST的活性变化似乎更能反映出TBBPA对颤蚓的污染效应及其毒性作用,而SOD的活性变化似乎更为灵敏,但是二者能否作为指示TBBPA污染的生物标志物尚需进一步研究。
3、在0.002-1.0 mg·L-1暴露浓度范围内,TBBPA并未对小麦种子的发芽率产生显著影响；各TBBPA暴露浓度下小麦芽生长的比生长速率(μ)随时间的变化趋势相同,均在污染暴露16h时达到峰值。而根伸长的μ随时间的变化却略有差异,其在染毒12-20 h这一时间段内趋于稳定且随着TBBPA暴露浓度的加大这一趋势逐渐显著；0.002-0.02 mg·L-1TBBPA处理浓度对小麦芽的生长起到促进作用,暴露浓度为0.02 mg·L-1时促进作用达到最大。当染毒浓度为0.002-0.05 mg·L-1时对小麦的根伸长起到促进作用,其中染毒浓度为0.05 mg·L-1时促进作用达到顶峰。4、经过7和12天的暴露后,小麦叶片中叶绿素的含量降低,脂质过氧化含量升高。经过12天的暴露后,小麦叶片中增加的POD活性显示小麦可以通过增加POD活性的方式来保护自己。当TBBPA的浓度为0.5-50 mg kg-1时,在暴露的初始阶段,植物可以抵抗氧化胁迫,但是随着暴露时间的延长这种能力会逐渐消失。除此之外,TBBPA浓度的增大也会导致抗氧化酶防御能力的丧失经过7天的暴露后,CAT活性的升高是由SOD引起的,而经过12天的暴露后POD活性的升高就不仅是由SOD引起的。抗氧化酶活性与TBBPA的浓度间并无剂量-效应关系。POD与CAT可作为TBBPA在土壤中严重胁迫的生物标志物。
5、经过7-10天的暴露后,5-500 mg kg-1 HBCD处理组使小麦叶片中CHL的含量显著降低。小麦暴露于HBCD引起了小麦叶片和根中MDA含量的升高。随着暴露时间的延长,SOD活性显著升高。相反,小麦根中SOD活性降低了在10天的实验期间,小麦叶片和根组织中POD的活性显著升高。经过8天的暴露,0.5-50 mg kg-1 HBCD处理组,小麦叶片中CAT的活性显著升高。然而,小麦根中CAT的活性未被显著诱导。这些说明小麦具有通过构建抗氧化防御系统来适应HBCD毒性的能力。并且小麦根比叶片对HBCD的毒性敏感。增强的CAT活性应该是由SOD源诱导产生的H202引起的,而增强的POD活性则是由其他源诱导产生的H202引起的。POD在解除AOS毒性的过程中起重要作用。
Brominated flame-retardants (BFRs) are a new, diverse group of global environmental pollutants. BFR production has increased dramatically over the past 20 years. Recently, concern for this emerging class of chemicals has risen duo to their occurrence in the environment, wildlife, and people. Whereas, knowledge about their fate in siol and ecotoxicological effects on organisms (aquatic and terrestrial) is very limited.
The fate of tetrabromobisphenol A (TBBPA) and hexabromocyclododecane (HBCD) diastereomers (α-,β-, andγ-HBCD), the ecotoxicological effects of TBBPA on tubifex (Monopylephorus limosus) and wheat(Triticum aestivum), the ecotoxicological effects of HBCD on wheat were all investigated. The results indicated that:
1. In a short-term (8 weeks) experiment, sorption to soil matrix resulted in 90% decline in recovery of TBBPA and HBCD in planted and unplanted soils. However, nearly 50% of initial HBCD recovered under the combined cabbage and radish treatment situation, which suggested that interactions between plant species might enhance the bioavailability of HBCDs. Although both plant species could uptake TBBPA and HBCDs, cabbage showed greater accumulating ability. Up to 3.5-10.0-fold higher HBCD concentrations were observed than TBBPA concentrations in all plant tissues, and the distribution of HBCDs in plant tissues for both species was diastereomer-specific. The predominance of a-HBCD in shoot tissues for both species might be attributed to diastereomer-specific translocation of HBCDs, shift in diastereomer pattern and/or selective metabolization of y-HBCD within plants.The result showed that strong sorption to soil particles reduced the bioavailability and potential of human exposure to BFRs in the soil, However, plants may increase the exposure risk by enhancing their bioavailability in the soil and by uptaking these compounds. The results also provide insight into transport mechanisms of TBBPA and HBCD diastereomers in soil-plant systems.
2. After a 8-day exposure, the SOD activity of tubifex was enhanced at first and then inhibited gradually, at last enhanced again. The highest activity of SOD (p<0.01) was examined under 0.05 mg L-1 concentration of TBBPA. And the activity of SOD was much higher than that of control(1.5 times～7.8 times more than that of control). While the CAT activity showed a tendency of induction firstly, then inhibition, then induction again and at last inhibition. The activity of CAT reached the highest value under 0.5 mg L-1 concentration of TBBPA. Furthermore, the CAT activity was higher than that of control (1.1 times～1.9 times more than that of control) except that under 0.005 mg L"1 and 0.25 mg L-1 concentration. Moreover, the highest activity of GST (p < 0.01) was observed under 0.25 mg L-1 concentration of TBBPA. The activity of GST was enhanced gradually at first and then inhibited. As the same as SOD, the activity of GST was induced significantly (p< 0.05). The changes in the SOD activity showed an "M" trend, while that in the CAT activity showed a "N" trend. And the activity of SOD is steadier than that of CAT. Thus, the changes in the activity of SOD and GST, especially SOD, can better reflect the toxic effects of pollutants on tubifex.
3. No significant effect on the germination rate of wheat was observed with the increase of TBBPA concentration. The trend of changes in the specific growth rate (μ) value of shoot growth with exposure time was similar treated with different TBBPA concentrations. And the peak of theμvalue of shoot growth was detected after the 42-hour exposure. Whereas, there were no significant differences in theμvalue of root elongation with the increase of exposure time from 38 to 46 hours as TBBPA concentration increased. The shoot growth was stimulated with the increase of TBBPA concentration from 0.002 to 0.02 mg L-1. And theμvalue of shoot growth treated with 0.02 mg L-1 of TBBPA was the highest. While the root elongation was stimulated by 0.002-0.05 mg L"1 TBBPA treatments. The 0.05 mg·L-1 TBBPA treatment significantly increased theμvalue of root elongation.
4. After both 7-and 10-day exposure, reduction of the CHL content in wheat leaves could be observed. However, the changes in the CHL content with the increasing TBBPA concentration from 50 to 5000 mg kg-1 were insignificant. Increased MDA levels detected in wheat leaves at the two time intervals reflected the presence of poisoning AOS and the oxidative stress induced by TBBPA. It can be concluded from the enhancing POD activity in wheat leaves after a 12-day exposure that wheat plants had the capability to protect themselves by increasing the activity of antioxidant enzyme POD with the exposure time. Our data also showed that the plant has the capacity to counteract the oxidative stress at the first stage of stress when the concentration of TBBPA was 0.5-50 mg kg-1, but the capacity would be lost with prolonged exposure. Moreover, the defensive effect of antioxidative enzymes would be lost with the increasing TBBPA concentration. The increasing CAT activity might be induced by H2O2 produced from sources of SOD after a 7-day exposure, but the increasing POD activity was induced not only by H2O2 produced from sources of SOD after a 12-day exposure. There were no dose-response effects in the changes between the activity of antioxidant enzymes (SOD, POD and CAT) and the concentration of TBBPA. It also could be concluded that the decrease in the activity of POD and CAT could be considered as good biomarkers of serious stress by TBBPA in soil.
5.5-500 mg kg-1 HBCD treatments caused significant damage to CHL accumulation in wheat seedlings after 7-10 days of exposure. As a marker of oxidative damage, the LPO was increased in both wheat leaves and roots. SOD activity in wheat leaves was significantly (p< 0.01) increased with prolonged exposure. On the contrary, it decreased more or less in wheat roots. POD activity in both wheat leaves and roots was significantly (p< 0.01) increased by all HBCD concentrations during the 10-day experimental period.0.5-50 mg kg-1 HBCD treatments significantly (p< 0.01) increased CAT activity in wheat leaves after the 8-day exposure. However, CAT activity in wheat roots was not significantly (p> 0.05) affected by HBCD exposure. All these results demonstrated that HBCD could induce oxidative stress in terrestrial organisms and wheat plants had the capacity to tolerate oxidative stress. POD and CAT might be induced by H2O2 produced from different sources. Wheat roots were more Sensitive to HBCD than wheat leaves. Also, POD played the main role in AOS detoxification under the stress of HBCD in soil environment.
为此,我们对四溴双酚A(tetrabromobisphenol A, TBBPA)和六溴环十二烷(hexabromocyclododecane, HBCD)在土壤中的归趋,TBBPA对颤蚓和小麦的生态毒理效应以及六溴环十二烷(hexabromocyclododecane, HBCD)对小麦的生态毒理效应进行了研究。结果显示：
1、土壤对TBBPA和HBCD的物理吸附导致二者在种植和未种植土壤中的回收率下降了90%。而在混合种植的情况下将近50%的HBCD被回收了,这充分说明植物种间竞争会增强HBCD的生物有效性。虽然白菜和萝卜都可以吸收TBBPA和HBCD,但是白菜吸收和富集污染物的能力要比萝卜强,并且各植物组织中HBCD的浓度约为TBBPA浓度的3.5-10.0倍。HBCD在不同介质中的分布与其异构体类型有关,a-HBCD倾向于在植物组织中分布,而y-HBCD则倾向于在土壤中。以上这些说明吸附会降低二者的生物有效性,以及人类暴露的风险,但是植物的作用会使这种风险增加。2、暴露8d后,颤蚓体内SOD的活性发生显著变化,变化趋势明显分为升高、降低、再升高3段,在0.05 mg L-1时SOD的活性被最大诱导(p<0.01),达到对照组的7.8倍,各浓度组SOD的活性均显著高于对照组,由对照组的1.5倍到对照组的7.8倍。同时CAT的活性变化则呈升高、降低、再升高、再降低4段,0.5 mg L-1时其活性达到最大值(p<0.01),除0.005 mg L-1和0.25 mg L-1浓度组CAT活性受抑制外,其余浓度组CAT的活性均高于对照组,由对照组的1.1倍到1.9倍。染毒浓度为0.25 mg L-1时,GST的活性达到最大诱导(p<0.01),其活性先缓慢上升后下降,并且各处理组GST的活性均显著高于对照组(p<0.05)。在单一浓度TBBPA污染暴露的10d时间里,颤蚓体内SOD的活性变化趋势形成了一个变形的“M”形曲线,在第3d时其活性达到最大诱导,而CAT活性的变化则表现为一个不规则的“N”形曲线,在第5d时其活性达到最大诱导,并且SOD的活性比CAT的活性受时间的影响小,相对来说更稳定。可见,SOD与GST的活性变化似乎更能反映出TBBPA对颤蚓的污染效应及其毒性作用,而SOD的活性变化似乎更为灵敏,但是二者能否作为指示TBBPA污染的生物标志物尚需进一步研究。
3、在0.002-1.0 mg·L-1暴露浓度范围内,TBBPA并未对小麦种子的发芽率产生显著影响；各TBBPA暴露浓度下小麦芽生长的比生长速率(μ)随时间的变化趋势相同,均在污染暴露16h时达到峰值。而根伸长的μ随时间的变化却略有差异,其在染毒12-20 h这一时间段内趋于稳定且随着TBBPA暴露浓度的加大这一趋势逐渐显著；0.002-0.02 mg·L-1TBBPA处理浓度对小麦芽的生长起到促进作用,暴露浓度为0.02 mg·L-1时促进作用达到最大。当染毒浓度为0.002-0.05 mg·L-1时对小麦的根伸长起到促进作用,其中染毒浓度为0.05 mg·L-1时促进作用达到顶峰。4、经过7和12天的暴露后,小麦叶片中叶绿素的含量降低,脂质过氧化含量升高。经过12天的暴露后,小麦叶片中增加的POD活性显示小麦可以通过增加POD活性的方式来保护自己。当TBBPA的浓度为0.5-50 mg kg-1时,在暴露的初始阶段,植物可以抵抗氧化胁迫,但是随着暴露时间的延长这种能力会逐渐消失。除此之外,TBBPA浓度的增大也会导致抗氧化酶防御能力的丧失经过7天的暴露后,CAT活性的升高是由SOD引起的,而经过12天的暴露后POD活性的升高就不仅是由SOD引起的。抗氧化酶活性与TBBPA的浓度间并无剂量-效应关系。POD与CAT可作为TBBPA在土壤中严重胁迫的生物标志物。
5、经过7-10天的暴露后,5-500 mg kg-1 HBCD处理组使小麦叶片中CHL的含量显著降低。小麦暴露于HBCD引起了小麦叶片和根中MDA含量的升高。随着暴露时间的延长,SOD活性显著升高。相反,小麦根中SOD活性降低了在10天的实验期间,小麦叶片和根组织中POD的活性显著升高。经过8天的暴露,0.5-50 mg kg-1 HBCD处理组,小麦叶片中CAT的活性显著升高。然而,小麦根中CAT的活性未被显著诱导。这些说明小麦具有通过构建抗氧化防御系统来适应HBCD毒性的能力。并且小麦根比叶片对HBCD的毒性敏感。增强的CAT活性应该是由SOD源诱导产生的H202引起的,而增强的POD活性则是由其他源诱导产生的H202引起的。POD在解除AOS毒性的过程中起重要作用。
Brominated flame-retardants (BFRs) are a new, diverse group of global environmental pollutants. BFR production has increased dramatically over the past 20 years. Recently, concern for this emerging class of chemicals has risen duo to their occurrence in the environment, wildlife, and people. Whereas, knowledge about their fate in siol and ecotoxicological effects on organisms (aquatic and terrestrial) is very limited.
The fate of tetrabromobisphenol A (TBBPA) and hexabromocyclododecane (HBCD) diastereomers (α-,β-, andγ-HBCD), the ecotoxicological effects of TBBPA on tubifex (Monopylephorus limosus) and wheat(Triticum aestivum), the ecotoxicological effects of HBCD on wheat were all investigated. The results indicated that:
1. In a short-term (8 weeks) experiment, sorption to soil matrix resulted in 90% decline in recovery of TBBPA and HBCD in planted and unplanted soils. However, nearly 50% of initial HBCD recovered under the combined cabbage and radish treatment situation, which suggested that interactions between plant species might enhance the bioavailability of HBCDs. Although both plant species could uptake TBBPA and HBCDs, cabbage showed greater accumulating ability. Up to 3.5-10.0-fold higher HBCD concentrations were observed than TBBPA concentrations in all plant tissues, and the distribution of HBCDs in plant tissues for both species was diastereomer-specific. The predominance of a-HBCD in shoot tissues for both species might be attributed to diastereomer-specific translocation of HBCDs, shift in diastereomer pattern and/or selective metabolization of y-HBCD within plants.The result showed that strong sorption to soil particles reduced the bioavailability and potential of human exposure to BFRs in the soil, However, plants may increase the exposure risk by enhancing their bioavailability in the soil and by uptaking these compounds. The results also provide insight into transport mechanisms of TBBPA and HBCD diastereomers in soil-plant systems.
2. After a 8-day exposure, the SOD activity of tubifex was enhanced at first and then inhibited gradually, at last enhanced again. The highest activity of SOD (p<0.01) was examined under 0.05 mg L-1 concentration of TBBPA. And the activity of SOD was much higher than that of control(1.5 times～7.8 times more than that of control). While the CAT activity showed a tendency of induction firstly, then inhibition, then induction again and at last inhibition. The activity of CAT reached the highest value under 0.5 mg L-1 concentration of TBBPA. Furthermore, the CAT activity was higher than that of control (1.1 times～1.9 times more than that of control) except that under 0.005 mg L"1 and 0.25 mg L-1 concentration. Moreover, the highest activity of GST (p < 0.01) was observed under 0.25 mg L-1 concentration of TBBPA. The activity of GST was enhanced gradually at first and then inhibited. As the same as SOD, the activity of GST was induced significantly (p< 0.05). The changes in the SOD activity showed an "M" trend, while that in the CAT activity showed a "N" trend. And the activity of SOD is steadier than that of CAT. Thus, the changes in the activity of SOD and GST, especially SOD, can better reflect the toxic effects of pollutants on tubifex.
3. No significant effect on the germination rate of wheat was observed with the increase of TBBPA concentration. The trend of changes in the specific growth rate (μ) value of shoot growth with exposure time was similar treated with different TBBPA concentrations. And the peak of theμvalue of shoot growth was detected after the 42-hour exposure. Whereas, there were no significant differences in theμvalue of root elongation with the increase of exposure time from 38 to 46 hours as TBBPA concentration increased. The shoot growth was stimulated with the increase of TBBPA concentration from 0.002 to 0.02 mg L-1. And theμvalue of shoot growth treated with 0.02 mg L-1 of TBBPA was the highest. While the root elongation was stimulated by 0.002-0.05 mg L"1 TBBPA treatments. The 0.05 mg·L-1 TBBPA treatment significantly increased theμvalue of root elongation.
4. After both 7-and 10-day exposure, reduction of the CHL content in wheat leaves could be observed. However, the changes in the CHL content with the increasing TBBPA concentration from 50 to 5000 mg kg-1 were insignificant. Increased MDA levels detected in wheat leaves at the two time intervals reflected the presence of poisoning AOS and the oxidative stress induced by TBBPA. It can be concluded from the enhancing POD activity in wheat leaves after a 12-day exposure that wheat plants had the capability to protect themselves by increasing the activity of antioxidant enzyme POD with the exposure time. Our data also showed that the plant has the capacity to counteract the oxidative stress at the first stage of stress when the concentration of TBBPA was 0.5-50 mg kg-1, but the capacity would be lost with prolonged exposure. Moreover, the defensive effect of antioxidative enzymes would be lost with the increasing TBBPA concentration. The increasing CAT activity might be induced by H2O2 produced from sources of SOD after a 7-day exposure, but the increasing POD activity was induced not only by H2O2 produced from sources of SOD after a 12-day exposure. There were no dose-response effects in the changes between the activity of antioxidant enzymes (SOD, POD and CAT) and the concentration of TBBPA. It also could be concluded that the decrease in the activity of POD and CAT could be considered as good biomarkers of serious stress by TBBPA in soil.
5.5-500 mg kg-1 HBCD treatments caused significant damage to CHL accumulation in wheat seedlings after 7-10 days of exposure. As a marker of oxidative damage, the LPO was increased in both wheat leaves and roots. SOD activity in wheat leaves was significantly (p< 0.01) increased with prolonged exposure. On the contrary, it decreased more or less in wheat roots. POD activity in both wheat leaves and roots was significantly (p< 0.01) increased by all HBCD concentrations during the 10-day experimental period.0.5-50 mg kg-1 HBCD treatments significantly (p< 0.01) increased CAT activity in wheat leaves after the 8-day exposure. However, CAT activity in wheat roots was not significantly (p> 0.05) affected by HBCD exposure. All these results demonstrated that HBCD could induce oxidative stress in terrestrial organisms and wheat plants had the capacity to tolerate oxidative stress. POD and CAT might be induced by H2O2 produced from different sources. Wheat roots were more Sensitive to HBCD than wheat leaves. Also, POD played the main role in AOS detoxification under the stress of HBCD in soil environment.
引文
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