洛克沙胂在土壤中的化学行为及其生物响应
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摘要
洛克沙胂(3-硝基-4-羟基苯砷酸)是一种有机胂饲料添加剂，在提高饲料利用率，对畜禽生长具有明显的促进作用的同时，还具广谱杀菌和抗球虫病的作用。故在我国畜禽养殖业中广泛使用。然而洛克沙胂在畜禽体内吸收较少，主要以药物原形随粪便排出。我国对于畜禽粪便无害化处理机制还不够完善，一般只经简单处理或不处理便作为有机肥用于农业生产，因此，洛克沙胂通过有机肥施用或随养殖场排污系统进入环境后，将可能对土壤和水质量造成不利影响，也可能对环境中的微生物、动植物乃至人类等产生生态毒性。
本研究选取作为饲料添加剂的洛克沙胂为供试污染物，采用室内模拟培养、定期采样和室内分析的方法，研究了洛克沙胂在土壤中的化学行为及其生物响应状况。主要包括洛克沙胂在土壤中的降解特性；在土壤中的吸附解吸特性以及洛克沙胂对土壤微生物活性的影响。为进一步明确洛克沙胂的环境效应,丰富有机胂化合物在土壤中的行为研究以及科学的调控洛克沙胂的生物效应提供数据参考和理论依据。主要研究结果如下：
1.对不同浓度残留的洛克沙胂进行室内模拟试验，得出洛克沙胂在土壤中的降解率随残留浓度的增大而降低，随着时间的延长而变缓，其半衰期介于42.92d~82.13d之间，参照农药在土壤中降解半衰期的等级划分标准，故洛克沙胂属于易降解有机化合物。
2.洛克沙胂在土壤中的数量变化主要由微生物降解、光降解以及土壤颗粒吸附三种途径造成，分别占减少总量的53%、12.99%和34.01%。洛克沙胂的降解与土壤温度均成正比，升高温度能够促进土壤中洛克沙胂的降解；土壤相对湿度为70%时洛克沙胂的降解作用最强，湿度过高或过低不利于洛克沙胂降解；土壤有机质的减少造成微生物活性降低从而延缓洛克沙胂的降解。
3.洛克沙胂在土壤及去除组分后土壤中的吸附和解吸过程均在24h基本达到平衡。原土壤对洛克沙胂的最大吸附量（Qm）为116.3mg·kg~(-1)，去除土壤有机质能使其对洛克沙胂的吸附量增大，而分别去除游离氧化铁和无定形氧化铁后对洛克沙胂的吸附量显著降低，其贡献率分别为~(-1)5.10%、62.79%和34.88%。洛克沙胂在土壤上的解吸存在滞后现象，滞后系数描述其解吸难易顺序为去除游离氧化铁>去除无定形氧化铁>原土壤>去除有机质，即去除游离氧化铁的土壤最难被解吸下来。
4.洛克沙胂在人工合成赤铁矿、针铁矿、水铁矿上的吸附、解吸平衡时间均为2h。人工合成铁氧化物对洛克沙胂的吸附量随初始浓度的增加而增大，铁氧化物对洛克沙胂的吸附属“L”型等温吸附线，即对低浓度洛克沙胂的亲和力较强，但吸附比例随着其浓度的增加增幅降低。
5.采用Langmuir方程对洛克沙胂在三种人工合成铁氧化物上的等温吸附结果拟合效果最佳，计算得到最大吸附量分别为23.81g·kg~(-1)、45.05g·kg~(-1)，和94.34g·kg~(-1)，是自然土壤最大吸附量的200-800倍，就三种氧化铁矿物而言，其对洛克沙胂的吸附量大小顺序为水铁矿>针铁矿>赤铁矿。由Freundlich方程所得lgKf和1/n等参数也表明水铁矿的吸附容量要强于针铁矿和赤铁矿，而吸附强度正好相反。
6.Langmuir方程对等温解吸结果拟合效果最佳，计算得到最大解吸量分别为14.90g·kg~(-1)、24.81g·kg~(-1)和31.45g·kg~(-1)。柠檬酸对氧化铁所吸附的洛克沙胂的解吸过程存在滞后现象，平均滞后系数分别为0.268，0.351和0.714，可见，赤铁矿<针铁矿<水铁矿，即水铁矿上所吸附的洛克沙胂被解吸下来的趋势最强。
7.添加不同浓度洛克沙胂能显著抑制土壤脲酶、过氧化氢酶和中性磷酸酶活性，且各处理均表现为在培养的第10天达到最大抑制作用，在培养的第10天以后，各处理酶活性均逐渐开始恢复。毒性影响程度和影响时间顺序均为中性磷酸酶>过氧化氢酶>脲酶，毒性影响时间分别为55~188.4d、50.0~154d和21.8~51.7d。
8.用米氏动力学方程推导出的模型y=c/1+bx计算得出土壤脲酶活性的ED50值为1667mg·kg~(-1)~10000mg·kg~(-1)，土壤过氧化氢酶活性的ED50值为434.8mg·kg~(-1)~1111.1mg·kg~(-1)，土壤中性磷酸酶活性的ED50值为169.5mg·kg~(-1)~588.2mg·kg~(-1)，土壤中性磷酸酶活性的ED50值最小，在一定程度上可以表征土壤受洛克沙胂的污染程度。
9.添加不同浓度的洛克沙胂能够不同程度的抑制土壤呼吸、氨化作用和硝化作用，到培养末期（27d）各处理对土壤呼吸作用的影响并不显著（p>0.05）；到培养末期（27d）低浓度(20、50、100mg·kg~(-1))的洛克沙胂对氨化和硝化作用影响不显著（p>0.05），而高浓度处理（150、250mg·kg~(-1)）仍具有显著差异。
Roxarsone (3-nitro-4-hydroxy arsenate) is an organoarsenic feed additives,which is used to improving feed utilization and animal growth stimulation,in the sametime it also have a broad-spectrum bactericidal and anti-ball insect role. Therefore, it iswidely used in livestock breeding industry in China. However, roxarsone is lessabsorption in the animal body, and excreted mainly in the drug prototype. Livestock andpoultry faeces harmless handling mechanism is not perfect in our country yet, it is usedfor agricultural production as organic fertilizer only after simply process or treatment.Consequently roxarsone come into the environment with organic fertilizer or with farmssewage system, which are likely to have adversely affected on soil and water quality,and may also have eco-toxicity on microorganisms in the environment, animals,plantsand even human.
This study selected a feed additive-roxarsone as a tested pollutants, Simulatedindoor incubation experiment,regular sampling and laboratory analysis were carried outto study the roxarsone behavior in the soil and its biological response status. Includingroxarsone degradation characteristics in soil; the effect of roxarsone on soil microbialactivity in soil; the adsorption and desorption characteristics in soil in order to providethe reference data and theoretical basis for the further study of roxarsone environmentaleffects, enrich the behavior of organoarsenic compounds in the soil and the scientificregulation of theroxarsone biological effects. The main results are as follows:
1. Indoor simulation experiments has carried out on different concentrations ofroxarsone residues, it is showed that the degradation rate of roxarsone in the soildecreases with the increasing of the residual concentration,and slowing down withextension of time, its half-life ranged from42.92d to82.13d, roxarsone are easilydegradable organic compounds reference to the grading standards of pesticide in the soildegradation half-life.
2. The quantity change of roxarsone in the soil primarily by three ways: microbial degradation, light degradation and soil particles adsorption,which take up53%,12.99%and34.01%of the total decrement respectively. In the range of moisture andtemperature conditions in this experiment, the degradation of roxarsone is directproportional with soil temperature，so that the increase of temperature can promote thedegradation of roxarsone in soil; the reduction of soil organic matter result indecreasding microbial activity and bring about slowing down the roxarsonedegradation.
3.The adsorption and desorption process of roxarsone in soil and componentsremoved soil are reach balance in24h.The max adsorption quantity is116.3mg·kg~(-1)insoil, which is larger in organic matte removed soil and fewer in free iron oxides andamorphous iron oxides removed soil significantly the contribution rate is~(-1)5.10%、62.79%and34.88%.The desorption process of roxarsone which absorbed on iron oxidesby citric acid exist lag phenomenon.Hysteresis index indicates that the difficultcoefficient is free iron oxides removed soil>amorphous removed soil>contrastsoil>organic matter removed soil, so the least desorption quantity is in free iron oxidesremoved soil.
4.The roxarsone adsorption and desorption balance time are2h on hematite、geothite and ferrihydrite. The adsorption capacity of roxarsone is increased with theinitial concentration on three kinds of synthetic iron oxides. The isothermal adsorptionline is affiliated “L” model, means the affinity is stronger to low concentrationroxarsone,but the adsorption ratio growth is lower with the increasing of concentration.
5.Langmuir equations can fitting the adsorption of roxarsone on three kinds ofsynthetic iron oxides, calculated the max adsorption quantity were23.81、45.05and94.34mg·kg~(-1), which is200~800times larger than natural soil. The order ofadsorption quantity is ferrihydrite>geothite>hematite.The parameters of Freundlichequation by the lgkf and1/n also shows that the adsorption capacity of ferrihydrite isbetter than goethite and hematite, and adsorption strength just the opposite.
6. Langmuir equations can fitting the desorption of roxarsone best, the maxdesorption amount were14.90、24.81and31.45mg·kg~(-1).The desorption process of roxarsone which absorbed on iron oxides by citric acid existing lag phenomenon，average hysteresis index were0.268、0.351and0.714, so roxarsone has a strongdesorption trend on ferrihydrite.
7.Adding different concentrations of roxarsone can significantly inhibit soil urease,catalase, and neutral phosphatase activity, and the peak inhibiting ratio appeared at the10th incubation day.After the first10days of incubation,all the processing enzymeactivity gradually began to recover. The toxic effect of the extent and chronologicalorder are neutral phosphatase> catalase> urease, and time of the toxic effect was55~188.4d,50.0~154d and21.8~51.7d respectively.
8.Using the mode y=c/1+bx derived from the Michaelis-Menten kineticsequation calculat the ED50value of urease activity,catalase activity and neutralphosphatase activity:1667mg· kg~(-1)~10000mg·kg~(-1)soil,434.8mg· kg-1~1111.1mg· kg~(-1)soil169.5mg· kg-1~588.2mg· kg~(-1)soil,respectively,and the ED50valuesof neutral phosphatase activity was minimum, and to a certain extent can characterizethe soil's pollution degree of the roxarsone.
9. Adding different concentrations of roxarsone can significantly inhibit soilrespiration, amination and nitrification. The effect on soil respiration is not significant atthe end of incubation (27d). To the end of incubation (27d) the effect on soilammoniation and nitrification is not significant by low concentration of roxarsone(20、50、100mg·kg~(-1))but it is still significant difference by high density dealing(150、250mg·kg~(-1)).
本研究选取作为饲料添加剂的洛克沙胂为供试污染物，采用室内模拟培养、定期采样和室内分析的方法，研究了洛克沙胂在土壤中的化学行为及其生物响应状况。主要包括洛克沙胂在土壤中的降解特性；在土壤中的吸附解吸特性以及洛克沙胂对土壤微生物活性的影响。为进一步明确洛克沙胂的环境效应,丰富有机胂化合物在土壤中的行为研究以及科学的调控洛克沙胂的生物效应提供数据参考和理论依据。主要研究结果如下：
1.对不同浓度残留的洛克沙胂进行室内模拟试验，得出洛克沙胂在土壤中的降解率随残留浓度的增大而降低，随着时间的延长而变缓，其半衰期介于42.92d~82.13d之间，参照农药在土壤中降解半衰期的等级划分标准，故洛克沙胂属于易降解有机化合物。
2.洛克沙胂在土壤中的数量变化主要由微生物降解、光降解以及土壤颗粒吸附三种途径造成，分别占减少总量的53%、12.99%和34.01%。洛克沙胂的降解与土壤温度均成正比，升高温度能够促进土壤中洛克沙胂的降解；土壤相对湿度为70%时洛克沙胂的降解作用最强，湿度过高或过低不利于洛克沙胂降解；土壤有机质的减少造成微生物活性降低从而延缓洛克沙胂的降解。
3.洛克沙胂在土壤及去除组分后土壤中的吸附和解吸过程均在24h基本达到平衡。原土壤对洛克沙胂的最大吸附量（Qm）为116.3mg·kg~(-1)，去除土壤有机质能使其对洛克沙胂的吸附量增大，而分别去除游离氧化铁和无定形氧化铁后对洛克沙胂的吸附量显著降低，其贡献率分别为~(-1)5.10%、62.79%和34.88%。洛克沙胂在土壤上的解吸存在滞后现象，滞后系数描述其解吸难易顺序为去除游离氧化铁>去除无定形氧化铁>原土壤>去除有机质，即去除游离氧化铁的土壤最难被解吸下来。
4.洛克沙胂在人工合成赤铁矿、针铁矿、水铁矿上的吸附、解吸平衡时间均为2h。人工合成铁氧化物对洛克沙胂的吸附量随初始浓度的增加而增大，铁氧化物对洛克沙胂的吸附属“L”型等温吸附线，即对低浓度洛克沙胂的亲和力较强，但吸附比例随着其浓度的增加增幅降低。
5.采用Langmuir方程对洛克沙胂在三种人工合成铁氧化物上的等温吸附结果拟合效果最佳，计算得到最大吸附量分别为23.81g·kg~(-1)、45.05g·kg~(-1)，和94.34g·kg~(-1)，是自然土壤最大吸附量的200-800倍，就三种氧化铁矿物而言，其对洛克沙胂的吸附量大小顺序为水铁矿>针铁矿>赤铁矿。由Freundlich方程所得lgKf和1/n等参数也表明水铁矿的吸附容量要强于针铁矿和赤铁矿，而吸附强度正好相反。
6.Langmuir方程对等温解吸结果拟合效果最佳，计算得到最大解吸量分别为14.90g·kg~(-1)、24.81g·kg~(-1)和31.45g·kg~(-1)。柠檬酸对氧化铁所吸附的洛克沙胂的解吸过程存在滞后现象，平均滞后系数分别为0.268，0.351和0.714，可见，赤铁矿<针铁矿<水铁矿，即水铁矿上所吸附的洛克沙胂被解吸下来的趋势最强。
7.添加不同浓度洛克沙胂能显著抑制土壤脲酶、过氧化氢酶和中性磷酸酶活性，且各处理均表现为在培养的第10天达到最大抑制作用，在培养的第10天以后，各处理酶活性均逐渐开始恢复。毒性影响程度和影响时间顺序均为中性磷酸酶>过氧化氢酶>脲酶，毒性影响时间分别为55~188.4d、50.0~154d和21.8~51.7d。
8.用米氏动力学方程推导出的模型y=c/1+bx计算得出土壤脲酶活性的ED50值为1667mg·kg~(-1)~10000mg·kg~(-1)，土壤过氧化氢酶活性的ED50值为434.8mg·kg~(-1)~1111.1mg·kg~(-1)，土壤中性磷酸酶活性的ED50值为169.5mg·kg~(-1)~588.2mg·kg~(-1)，土壤中性磷酸酶活性的ED50值最小，在一定程度上可以表征土壤受洛克沙胂的污染程度。
9.添加不同浓度的洛克沙胂能够不同程度的抑制土壤呼吸、氨化作用和硝化作用，到培养末期（27d）各处理对土壤呼吸作用的影响并不显著（p>0.05）；到培养末期（27d）低浓度(20、50、100mg·kg~(-1))的洛克沙胂对氨化和硝化作用影响不显著（p>0.05），而高浓度处理（150、250mg·kg~(-1)）仍具有显著差异。
Roxarsone (3-nitro-4-hydroxy arsenate) is an organoarsenic feed additives,which is used to improving feed utilization and animal growth stimulation,in the sametime it also have a broad-spectrum bactericidal and anti-ball insect role. Therefore, it iswidely used in livestock breeding industry in China. However, roxarsone is lessabsorption in the animal body, and excreted mainly in the drug prototype. Livestock andpoultry faeces harmless handling mechanism is not perfect in our country yet, it is usedfor agricultural production as organic fertilizer only after simply process or treatment.Consequently roxarsone come into the environment with organic fertilizer or with farmssewage system, which are likely to have adversely affected on soil and water quality,and may also have eco-toxicity on microorganisms in the environment, animals,plantsand even human.
This study selected a feed additive-roxarsone as a tested pollutants, Simulatedindoor incubation experiment,regular sampling and laboratory analysis were carried outto study the roxarsone behavior in the soil and its biological response status. Includingroxarsone degradation characteristics in soil; the effect of roxarsone on soil microbialactivity in soil; the adsorption and desorption characteristics in soil in order to providethe reference data and theoretical basis for the further study of roxarsone environmentaleffects, enrich the behavior of organoarsenic compounds in the soil and the scientificregulation of theroxarsone biological effects. The main results are as follows:
1. Indoor simulation experiments has carried out on different concentrations ofroxarsone residues, it is showed that the degradation rate of roxarsone in the soildecreases with the increasing of the residual concentration,and slowing down withextension of time, its half-life ranged from42.92d to82.13d, roxarsone are easilydegradable organic compounds reference to the grading standards of pesticide in the soildegradation half-life.
2. The quantity change of roxarsone in the soil primarily by three ways: microbial degradation, light degradation and soil particles adsorption,which take up53%,12.99%and34.01%of the total decrement respectively. In the range of moisture andtemperature conditions in this experiment, the degradation of roxarsone is directproportional with soil temperature，so that the increase of temperature can promote thedegradation of roxarsone in soil; the reduction of soil organic matter result indecreasding microbial activity and bring about slowing down the roxarsonedegradation.
3.The adsorption and desorption process of roxarsone in soil and componentsremoved soil are reach balance in24h.The max adsorption quantity is116.3mg·kg~(-1)insoil, which is larger in organic matte removed soil and fewer in free iron oxides andamorphous iron oxides removed soil significantly the contribution rate is~(-1)5.10%、62.79%and34.88%.The desorption process of roxarsone which absorbed on iron oxidesby citric acid exist lag phenomenon.Hysteresis index indicates that the difficultcoefficient is free iron oxides removed soil>amorphous removed soil>contrastsoil>organic matter removed soil, so the least desorption quantity is in free iron oxidesremoved soil.
4.The roxarsone adsorption and desorption balance time are2h on hematite、geothite and ferrihydrite. The adsorption capacity of roxarsone is increased with theinitial concentration on three kinds of synthetic iron oxides. The isothermal adsorptionline is affiliated “L” model, means the affinity is stronger to low concentrationroxarsone,but the adsorption ratio growth is lower with the increasing of concentration.
5.Langmuir equations can fitting the adsorption of roxarsone on three kinds ofsynthetic iron oxides, calculated the max adsorption quantity were23.81、45.05and94.34mg·kg~(-1), which is200~800times larger than natural soil. The order ofadsorption quantity is ferrihydrite>geothite>hematite.The parameters of Freundlichequation by the lgkf and1/n also shows that the adsorption capacity of ferrihydrite isbetter than goethite and hematite, and adsorption strength just the opposite.
6. Langmuir equations can fitting the desorption of roxarsone best, the maxdesorption amount were14.90、24.81and31.45mg·kg~(-1).The desorption process of roxarsone which absorbed on iron oxides by citric acid existing lag phenomenon，average hysteresis index were0.268、0.351and0.714, so roxarsone has a strongdesorption trend on ferrihydrite.
7.Adding different concentrations of roxarsone can significantly inhibit soil urease,catalase, and neutral phosphatase activity, and the peak inhibiting ratio appeared at the10th incubation day.After the first10days of incubation,all the processing enzymeactivity gradually began to recover. The toxic effect of the extent and chronologicalorder are neutral phosphatase> catalase> urease, and time of the toxic effect was55~188.4d,50.0~154d and21.8~51.7d respectively.
8.Using the mode y=c/1+bx derived from the Michaelis-Menten kineticsequation calculat the ED50value of urease activity,catalase activity and neutralphosphatase activity:1667mg· kg~(-1)~10000mg·kg~(-1)soil,434.8mg· kg-1~1111.1mg· kg~(-1)soil169.5mg· kg-1~588.2mg· kg~(-1)soil,respectively,and the ED50valuesof neutral phosphatase activity was minimum, and to a certain extent can characterizethe soil's pollution degree of the roxarsone.
9. Adding different concentrations of roxarsone can significantly inhibit soilrespiration, amination and nitrification. The effect on soil respiration is not significant atthe end of incubation (27d). To the end of incubation (27d) the effect on soilammoniation and nitrification is not significant by low concentration of roxarsone(20、50、100mg·kg~(-1))but it is still significant difference by high density dealing(150、250mg·kg~(-1)).
引文
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