耐受重金属的芘降解真菌的筛选及芘降解动力学研究

为了提高复合污染土壤修复的微生物资源的丰富度,为混合菌群修复污染土壤积累资料,利用多环芳烃-重金属双抗培养基在污染土壤中筛选得到一株对Cu和Cd有高耐受性的芘降解真菌,经分子生物学鉴定为米曲霉。探究了米曲霉对芘污染水体的降解效果及对重金属Cu和Cd的耐受程度,利用缺乏生长基质的毒性抑制动力学模型对芘单基质降解过程进行了拟合,以期为后续共代谢、固定化的研究及实际工程应用提供一定的理论支撑。结果表明:(1)米曲霉以芘为单基质代谢时,降解率为33%;(2)米曲霉对重金属Cu和Cd的耐受浓度分别为500 mg/L和50 mg/L,分别高出国家土壤重金属二级标准5倍和83倍;(3)米曲霉对单基质芘的降解符合Crridle提出的毒性抑制动力学简化模型Sc=Sc0·Tb*cX0(1-e-bt)(R2=0.9237)。芘初始浓度Sc0=80 mg/L,米曲霉投加量X0=85 mg/L时,数值拟合得到内源呼吸常数b=0.027,生物转化量Tb*c=0.2875。该米曲霉对单基质芘及重金属Cu和Cd表现出一定的降解性能及耐受性能,故可经过适当强化后作为多环芳烃-重金属污染土壤的微生物修复菌种。     

双齿围沙蚕对芘的生物可利用性研究

为近海岸沉积质中PAHs的生态风险评价及其污染生境的修复提供理论依据,以双齿围沙蚕为研究对象,以芘为暴露污染物,开展双齿围沙蚕对芘的生物可利用性研究,分别考察了芘对双齿围沙蚕抗氧化酶活性的影响以及芘在双齿围沙蚕体内的生物富集特征。结果表明,双齿围沙蚕SOD、CAT和GSH-PX活性均随诱导时间的增加而升高,且SOD活性随芘浓度的增加出现显著性抑制,CAT和GSH-PX活性则随芘浓度的增加出现显著性升高。芘在双齿围沙蚕体内的生物富集特征如下:双齿围沙蚕体内芘的蓄积量随暴露浓度的增加而增加,各处理组双齿围沙蚕体内芘的蓄积量随时间的变化均呈类凸形的二次曲线关系,在第10天达到蓄积的最大值,最大蓄积量依次为(6.18±0.62)mg·kg~(-1)、(12.37±1.23)mg·kg~(-1)和(24.76±1.51)mg·kg~(-1)。生物富集系数BCF随芘浓度的增加而减小,分别为0.0692、0.0684和0.0674。上述研究结果表明,在一定浓度范围内,双齿围沙蚕抗氧化酶活性对芘的暴露具有很好的指示作用;此外,在短期内,双齿围沙蚕对芘具有明显的富集效应。     

Distribution and sources of polycyclic aromatic hydrocarbons in Wuhan section of the Yangtze River, China

Polycyclic aromatic hydrocarbons (PAHs) are important organic contaminants with great significance for China, where coal burning is the main source of energy. In this study, concentrations, distribution between different phases, possible sources and eco-toxicological effect of PAHs of the Yangtze River were assessed. PAHs in water, suspended particulate matters (SPM) and sediment samples at seven main river sites, 23 tributary and lake sites of the Yangtze River at the Wuhan section were analyzed. The total concentrations of PAHs in the studied area ranged from 0.242 to 6.235 μg/l in waters and from 31 to 4,812 μg/kg in sediment. The average concentration of PAHs in SPM was 4,677 μg/kg, higher than that in sediment. Benzo(a)pyrene was detected only at two stations, but the concentrations were above drinking water standard. The PAHs level of the Yangtze River was similar to that of some other rivers in China but higher than some rivers in foreign countries. There existed a positive relationship between PAHs concentrations and the TOC contents in sediment. The ratio of specific PAHs indicated that PAHs mainly came from combustion process, such as coal and wood burning. PAHs may cause potential toxic effect but will not cause acute biological effects in sedimentary environment of the Wuhan section of the Yangtze River.     

表面活性剂Tween80及DOM对土壤中菲、芘解吸的影响

采用室内序批试验研究了水溶性有机物（DOM）及表面活性剂Tween80对污染土壤中菲、芘解吸行为的影响.结果表明,DOM能够增加土壤中菲、芘的解吸率,且猪粪堆肥的DOM作用效果较好;随着体系中Tween80浓度的增加,土壤中菲、芘的解吸率也明显地增加,当Tween80浓度为150 mg·L^-1时,菲、芘的解吸率分别是对照的1.7倍（菲）和6.2倍（芘）;DOM与Tween80联合作用时受Tween80浓度的影响,低浓度时联合作用效果不明显,当Tween80为150 mg·L^-1时,菲、芘的解吸率显著增加且大于两者单独作用的结果之和.试验结果还表明,同等条件下高相对分子质量DOM组分（>25000）对土壤中菲、芘的解吸作用大于低相对分子质量DOM组分（<1 000）.     

TW-80对污染土壤中菲和芘的生物降解影响

以 Ｔ Ｗ８０ 为供试物,进行了表面活性剂对污染土壤中菲和芘生物降解影响研究． 实验周期１５０ ｄ ．并分别在３０ ｄ 、６０ ｄ 和１５０ ｄ 间隔采样监测菲、芘降解率．结果表明,３０ ｄ 后,加入 Ｔ Ｗ８０ 的不同处理土壤中,菲的降解率分别达８１ ％ ～９８ ．９ ％ ,比 Ｃ Ｋ 高２２ ．０２ ％ 以上、芘的降解率分别达７１ ．９０ ％ ～８０ ．６６ ％ ,比 Ｃ Ｋ 高９ ．４６ ％ 以上．６０ ｄ 后,加入 Ｔ Ｗ８０ 的处理中,菲的降解率达８８ ．９４ ％ ～９６ ．５１ ％ 、比 Ｃ Ｋ 高１１ ．１４ ％ 以上; 芘降解率达９２ ．３９ ％～９４ ．０２ ％ ,比 Ｃ Ｋ 高２ ．２７ ％ 以上．１５０ ｄ 后,所有处理中,菲和芘的降解率均分别达到９８ ％ 和９６ ％ 以上, 与 Ｃ Ｋ 无明显差别．研究还发现, Ｔ Ｗ８０ 土壤中含有优势真菌常见青霉、蠕形青霉、淡紫青霉和顶孢头孢霉．它们可能是促进土壤中菲、芘降解的原因     

萃取－同步荧光光谱法测定大气飘尘中苯并(a)芘

提出用浓Ｈ２ＳＯ４从大气飘尘样品的环己烷提取液中一次萃取后,用同步荧光光谱测定苯并（ａ）芘的方法,选用△λ为２０ｎｍ时可获最佳效果,用浓硫酸萃取的时间仅１ｍｉｎ,苯并（ａ）芘在浓硫酸相中１ｈ内稳定,样品加标的回收率为８５％,其它多环芳烃化合物在５３０－６００ｎｍ的测定波长范围不干扰苯并（ａ）芘的测定,用乙酰化滤纸层析－荧光分光光度的统一方法作为对比对实际样品进行了测定,结果与后者的偏差在±１５％以     

Tween80对植物吸收菲和芘的影响

以水培体系模拟研究了非离子表面活性剂Tween80对黑麦草吸收菲和芘的影响.结果表明,当培养液中菲和芘的起始浓度分别为1 00mg·L-1和0 12mg·L-1时,0～105 6mg·L-1范围内,低浓度Tween80可促进根和茎叶吸收菲和芘,Tween80浓度为6 6mg·L-1时促进作用最为明显,根和茎叶中菲和芘含量、富集系数为无表面活性剂对照处理的116%～216%;而高浓度则有抑制作用.当培养液中菲和芘的起始浓度为相应Tween80浓度下的表观溶解度时,根和茎叶中菲和芘含量随Tween80浓度上升明显增大,Tween80的增溶作用有利于根和茎叶对菲和芘的吸收.     

狗尿中1-羟基芘鉴定和排出规律

经高压液相色谱分离,萤光光谱法鉴定,狗肌肉注射芘后尿中的代谢产物为1-羟基芘。分别给狗1、2、5μmol/kg芘后,尿中排出的1-羟基芘均以第24-48h期间最高;尿中排出的1-羟基芘约占注入芘的0.05％左右。     

复合固定化法固定微生物去除芘

通过"玉米秸秆吸附-包埋-交联"复合固定化方法固定多环芳烃降解菌GY2B和GP3B,提高对芘的降解性能,并对其作用过程进行初步研究.结果表明,游离态单细菌GY2B、GP3B和混合菌GY2B+GP3B的7 d降解率分别为14.0%、55.0%和73.6%,而固定化GY2B+GP3B秸秆小球在5 d内即达98.2%,去除率得到了显著的提高;对固定化微生物小球的比表面积、孔隙率和扫描电镜图分析,说明载体表面具有较高的比表面积,内部是具有大量孔隙的骨架结构,在内部生长的微生物和基质有充分的接触面积和机会;固定化GY2B+GP3B秸秆小球类似于一个吸附降解一体化的微型反应器.GY2B+GP3B混合菌芘代谢产物有菲-4-羧酸、二甲基酞酸、1-羟基-2-萘酸、1-萘酚和水杨酸,推测芘的降解是GY2B+GP3B混合菌集体代谢的综合作用的结果,GP3B的中间转化产物作为GY2B生长代谢的基质,使得芘得到更完全的降解.     

Response of bacterial communities to short-term pyrene exposure in red soil

Pyrene, a representative polycyclic aromatic hydrocarbon (PAH) compound produced mainly from incomplete combustion of fossil fuels, is hazardous to ecosystem health. However, long-term exposure studies did not detect any significant effects of pyrene on soil microorganism. In this study, short-term microcosm experiments were conducted to identify the immediate effect of pyrene on soil bacterial communities. A freshly-collected pristine red soil was spiked with pyrene at 0, 10, 100, 200, and 500 mg·kg^-1 and incubated for one day and seven days. The bacterial communities in the incubated soils were analyzed using 16S rRNA sequencing and terminal restriction fragment length polymorphism (T-RFLP) methods. The results revealed high bacterial diversity in both unspiked and pyrene-spiked soils. Only at the highest pyrene-spiking rate of 500 mg·kg^-1, two minor bacteria groups of the identified 14 most abundant bacteria groups were completely suppressed. Short-term exposure to pyrene resulted in dominance of Proteobacteria in soil, followed by Acidobacteria, Firmutes, and Bacteroidetes. Our findings showed that bacterial community structure did respond to the presence of pyrene but recovered rapidly from the perturbation. The intensity of impact and the rate of recovery showed some pyrene dosage-dependent trends. Our results revealed that different levels of pyrene may affect the bacterial community structure by suppressing or selecting certain groups of bacteria. It was also found that the bacterial community was most susceptible to pyrene within one day of the chemical addition.