Biological effect monitoring in dab (<Emphasis Type="Italic">Limanda limanda</Emphasis>) using gene transcript of CYP1A1 or EROD—a comparison

BACKGROUND, AIM, AND SCOPE: Gene expression analyses with real-time (RT)-polymerase chain reaction (PCR) gains importance in marine monitoring. This new technique has to be compared to the classical approaches like the well known biomarker ethoxyresorufin-O-deethylase (EROD) to test their suitability for monitoring programmes. The goal of the present study is to compare EROD activity and CYP1A1 mRNA expression in the important monitoring fish species dab (Limanda limanda) and to answer the question of whether these parameters reflect the polycyclic aromatic hydrocarbon (PAH) contamination of the fish. Further on, glyceraldehyd-3-phosphate dehydrogenase (GAPDH) was investigated as a potential housekeeping gene. MATERIALS AND METHODS: Female dab were caught in the summer of 2004 in the North Sea and in the Baltic. EROD activity was determined in liver samples by a kinetic fluorimetric assay according to a standard protocol. The gene expression of CYP1A (cytochrome P450 1A) and GAPDH were determined by means of RT-PCR. Results were compared to gonado somatic index and to the concentration of PAH metabolite 1OHPyr (1-hydroxypyrene) analysed in the bile fluids of the fish, respectively. RESULTS: Dab from all stations showed a considerable individual variation in the levels of both CYP1A mRNA and EROD. Highest mean values for CYP1A mRNA and EROD were detected in the northern part of the sampling area. In contrast, the PAH metabolite 1OHPyr was found at the highest concentration in fish caught near the German coast. CYP1A mRNA and EROD showed only a minor but significant correlation (r = 0.32, p < 0.05, n = 123). 1OHPyr in bile correlated significantly (p < 0.05) with the amount of GAPDH mRNA content in the liver. DISCUSSION: The significant but low correlation of CYP1A mRNA and EROD activity on an individual basis illustrates that these two parameters are apparently not closely linked. However, maximum EROD values correspond with maximum CYP1A mRNA concentrations when station means are regarded. Because EROD and CYP1A mRNA in dab follow different physiological principles, their application will lead to related but not identical monitoring results. This should be taken into account when future marine monitoring programmes are designed. The results also indicate that PAH are not the crucial factor for CYP1A and EROD levels in dab from the off-shore areas in the North Sea. This is remarkable because the PAH metabolism is known to be CYP1A-dependent and the widely used biomarker EROD has been recommended for monitoring PAH-related effects in fish from the North Sea. Due to a correlation between GAPDH and 1OHPyr, GAPDH was not suitable as housekeeping gene for dab. CONCLUSIONS: Neither the results from EROD nor from CYP1A1 mRNA measurements in dab reflected their exposure to PAH as measured by the PAH metabolite 1OHPyr. Thus, the question arises of whether EROD or CYP1A mRNA is a suitable biomarker at all to indicate PAH exposure in dab from the open North Sea. RECOMMENDATIONS AND PERSPECTIVES: For future biological effect monitoring, it is advisable to measure more and predominately independent parameters by RT-PCR and to incorporate more components of the detoxification system.     

Transport of simazine in unsaturated sandy soil and predictions of its leaching under hypothetical field conditions

The potential contamination of groundwater by herbicides is often controlled by processes in the vadose zone, through which herbicides travel before entering groundwater. In the vadose zone, both physical and chemical processes affect the fate and transport of herbicides, therefore it is important to represent these processes by mathematical models to predict contaminant movement. To simulate the movement of simazine, a herbicide commonly used in Chilean vineyards, batch and miscible displacement column experiments were performed on a disturbed sandy soil to quantify the primary parameters and processes of simazine transport. Chloride (Cl(-)) was used as a non-reactive tracer, and simazine as the reactive tracer. The Hydrus-1D model was used to estimate the parameters by inversion from the breakthrough curves of the columns and to evaluate the potential groundwater contamination in a sandy soil from the Casablanca Valley, Chile. The two-site, chemical non-equilibrium model was observed to best represent the experimental results of the miscible displacement experiments in laboratory soil columns. Predictions of transport under hypothetical field conditions using the same soil from the column experiments were made for 40 years by applying herbicide during the first 20 years, and then halting the application and considering different rates of groundwater recharge. For recharge rates smaller than 84 mm year(-1), the predicted concentration of simazine at a depth of 1 m is below the U.S. EPA's maximum contaminant levels (4 microg L(-1)). After eight years of application at a groundwater recharge rate of 180 mm year(-1) (approximately 50% of the annual rainfall), simazine was found to reach the groundwater (located at 1 m depth) at a higher concentration (more than 40 microg L(-1)) than the existing guidelines in the USA and Europe.     

Utilization and degradation of imazaquin by a naturally occurring isolate of Arthrobacter crystallopoietes

A species of bacteria that is capable of utilizing imazaquin as the sole carbon source was isolated from soil with repeated imazaquin applications, and was identified as Arthrobacter crystallopoietes (designated as strain “WWX-1”). This isolate degrades imazaquin as high as 200 μg ml⁻¹, and the estimated dissipation half-lives increased from 1.51 d for the treatment at 50 μg ml⁻¹ to 4.75 d for 200 μg ml⁻¹. Optimal growth of WWX-1 in mineral salt medium with 50 μg ml⁻¹ imazaquin was obtained at 35 °C and a pH of 5.0. Growth of WWX-1 was also observed in mineral salt medium with the addition of other imidazolinone herbicides such as imazethapyr and imazapyr, but not with different classes of herbicides such as metsulfuron-methyl. Two imazaquin metabolites were detected, and spectral analysis with HPLC–MS, ¹H NMR, and IR revealed one metabolite with a molecular weight (MW) of 199 as quinoline-2,3-dicarboxylic anhydride. We propose that A. crystallopoietes (WWX-1) could serve as an efficient biodegradation system for remediation of water and soils that are heavily contaminated with imazaquin or other structurally similar chemicals.     

A survey of antennal responses by five species of coniferophagous bark beetles (Coleoptera: Scolytidae) to bark volatiles of six species of angiosperm trees

Summary. Using Porapak Q traps, we collected the bark volatiles of six angiosperm trees native to British Columbia: black cottonwood, Populus trichocarpa Torr. & A. Gray (Salicaceae), trembling aspen, P. tremuloides Michx. (Salicaceae), paper birch, Betula papyrifera Marsh. (Betulaceae), bigleaf maple, Acer macrophyllum Pursh (Aceraceae), red alder, Alnus rubra Bong. (Betulaceae), and Sitka alder, A. viridis ssp. sinuata (Regel) á. L?ve & D. L?ve (Betulaceae). Utilising coupled gas chromatographic-electroantennographic detection analysis, the captured volatiles were assayed for antennal responses in five species of coniferophagous bark beetles (Coleoptera: Scolytidae), sympatric with most or all of the angiosperm trees: the Douglas-fir beetle, Dendroctonus pseudotsugae Hopkins, the mountain pine beetle, D. ponderosae Hopkins, the spruce beetle, D. rufipennis (Kirby), the western balsam bark beetle, Dryocoetes confusus Swaine, and the pine engraver, Ips pini (Say). The identities of 25 antennally-active compounds were determined by coupled gas chromatographic-mass spectroscopic analysis, and co-chromatographic comparisons with authentic chemicals. The compounds identified were: hexanal, (E)-2-hexenal, (Z)-3-hexen-1-ol, 1-hexanol, heptanal, α-pinene, frontalin, benzaldehyde, β-pinene, 2-hydroxycyclohexanone, 3-carene, limonene, β-phellandrene, benzyl alcohol, (E)-ocimene, salicylaldehyde, conophthorin, guaiacol, nonanal, methyl salicylate, 4-allylanisole, decanal, thymol methyl ether, (E)-nerolidol, and dendrolasin. A number of these compounds are known semiochemicals that are active in the behaviour of other organisms, including bark beetles, suggesting a high degree of semiochemical parsimony. Antennally-active compounds ranged from seven in A. viridis to 17 in P. trichocarpa. The fewest number of compounds (9) were detected by I. pini and the largest number (24) were detected by D. pseudotsugae. Six compounds excited the antennae of all five species of bark beetles. The large number of antennally-active compounds detected in common by numerous bark beetles and present in common in numerous nonhost trees supports the hypothesis of olfaction-based recognition and avoidance of nonhost angiosperm trees during the process of host selection by coniferophagous bark beetles. Received 13 December 1999; accepted 14 March 2000     

洪泛区林地的开发对鱼类栖息地面积的影响模拟

洪泛区的开发利用,必然会导致洪泛区林地糙率的改变。河漫滩也是河流洪泛区的一部分。洪泛区糙率的减小,会使河水流速增大,从而减小鱼类可用栖息地面积。因此,在洪水发生时,为了给鱼类提供避险场所,保护有效栖息地面积,需研究洪泛区林地的开发对鱼类的影响。利用River 2D软件模拟在一定频率的洪水条件下,不同的河漫滩糙率对应的鱼类栖息地加权可用面积(WUA),分析了糙率的改变对河流鱼类栖息地的影响。结果表明:随着河漫滩糙率的降低,加权可用面积(WUA)逐渐减小,河漫滩范围内WUA的变化幅度比较大,河漫滩的特性对鱼类栖息地面积的影响很大。洪泛区植被的砍伐,会使鱼类栖息地面积减小。因此,应防止洪泛区的过度开发,保护河流生态环境。     

改性粉煤灰合成沸石对甲基橙的吸附动力学

以粉煤灰(FA)为原料,采用水热晶化一步法制备了Na P1型沸石(ZFA),对合成产物的结构进行了表征,并采用阳离子表面活性剂十六烷基三甲基溴化铵(CTAB)对其进行改性。通过静态吸附实验,研究了改性后Na P1型沸石(MZFA)对水溶液中甲基橙的吸附特性,从动力学角度探讨了吸附机理。结果表明,在所研究的浓度条件下,改性Na P1型沸石对甲基橙的吸附符合Langmuir等温吸附方程,在25℃时,静态饱和吸附量(Qm)为64.76 mg/g。动力学分析表明,改性Na P1型沸石对溶液中甲基橙的吸附符合准二级动力学模型。     

600 MW机组低压转子叶片裂纹分析

针对某台600 MW亚临界参数汽轮机低压转子的末三级叶片(2Cr12NiMo1W1V钢)出汽侧裂纹进行了宏观形貌观察、化学成分分析、金相组织分析、室温冲击韧性试验、硬度试验,并对断口进行了扫描电镜及能谱分析。结果表明：此次开裂的低压转子叶片,是由于其化学成分Mo元素含量低于标准要求,导致材料的KISCC降低,以及硬度偏低造成裂纹扩展所需的外界启动应力降低,进而在运行过程中拉应力和湿蒸汽的氧化腐蚀共同作用下,发生了应力腐蚀开裂。     

苹果角质层的组分特征及其对甲萘酚的吸附作用

为能够准确预测有机污染物在植物角质层上的吸附行为,用化学方法(脱蜡、皂化、酸解)分离得到7种苹果角质层组分,探讨了角质组分对甲萘酚的吸附特征及其与结构组成之间的关系.结果表明.苹果角质层由44.7%蜡质、34.6%角质单体、13.6%糖类、7.5%生物高聚物(cutan)组成,脱蜡、皂化脱角质单体后角质组分的极性指数((O N)/C)增大,而酸解脱糖后(O N)/C则减小.角质层组分对甲萘酚的等温吸附曲线均呈非线性(N=0.551～0.918),而高浓度时的等温吸附曲线则呈线性,表明植物角质与有机物的相互作用除了分配作用之外,还存在其它的特殊作用.苹果角质层(ACI)对甲萘酚的吸附系数(Knc)为442,脱蜡、脱糖后角质组分的KDC.值增大,而脱角质单体后Kuc值则大大减小,表明角质单体是主要的吸附介质;蜡质、角质单体、糖类组分对角质层吸附行为的影响存在交互作用.角质组分的Koc值随其(O N)/C变化出现"峰值",意味着甲萘酚与角质组分之间的相互作用存在最佳的极性匹配.     

维生素B1制药废水的铁炭微电解-混凝预处理工艺

针对维生素B1制药废水高有机物浓度、高悬浮物、色度高、难降解的特点,采用铁炭微电解-混凝工艺对其进行预处理,效果良好。铁炭微电解-混凝工艺的最佳运行条件为:进水pH值为4,铁炭比为1∶1,曝气量为0.2m3/h的情况下停留时间80min。出水CODCr浓度平均为1600mg/L,去除率约为79%,色度去除率为85%,出水达到了(GB8978-1996)二级排放标准。     

Growth factors, kinetics and biodegradation mechanism associated with Pseudomonas nitroreducens TX1 grown on octylphenol polyethoxylates

The growth properties and biodegradation mechanism of a Gram-negative bacterium, Pseudomonas nitroreducens TX1 that was able to grow on branched octylphenol polyethoxylates (OPEO(n), average n=9.5) as the sole carbon source over a wide concentration range (1-100,000 mgl(-1)) were studied. Analysis of growth factors indicated the highest specific growth rate (micro) of 0.53 h(-1) was obtained at an initial concentration of 5,000 mgl(-1) OPEO(n). An optimal C/N ratio of 12 was obtained for (NH(4))(2)SO(4) as the nitrogen source in a cultivated medium at pH 7. The kinetic analysis demonstrated that bacterial growth and OPEO(n) degradation followed the Monod equation and were based on a substrate concentration inhibition model and pseudo-first-order reaction, respectively. The substrate inhibition coefficient was over 18,000 mgl(-1) and this indicates that the strain has an ability to sustain growth at high concentrations of OPEO(n) and use it as the sole carbon source under such a stress condition. Furthermore, LC-MS analysis showed that the biodegradation mechanism of dodecyl octaethoxylate (AEO8) by P. nitroreducens TX1 was the sequential cleavage of the ethoxylate chain.