A monoclonal antibody against Lates calcarifer vitellogenin and a competitive ELISA to evaluate vitellogenin induction after exposure to xenoestrogen

A monoclonal antibody specific to sea bass(Lates calcarifer) vitellogenin(VTG) was developed,for use as a tool for monitoring endocrine disrupting chemicals(EDCs). VTG was induced in sea bass by intramuscular injection of 17β-estradiol(E_2: 2 mg/kg) every three days. Blood was collected three days after the last injection. Plasma VTG was then purified by chromatography in hydroxyapatite and a sephacryl-S300 column. Characterizations of purified VTG were done by phospholipoglycoprotein staining on a native-PAGE with confirmation by mass spectrometry(LC-MS/MS). Antibody was raised in mice by injection of purified VTG. After monoclonal antibody production, the hybridoma clone No. 41(MAb-sea bass VTG 41)was selected and developed for quantification of VTG by competitive enzyme-linked immunosorbent assay(ELISA). The ELISA method was sensitive with a detection limit of VTG 40 ng/mL. MAb-sea bass VTG 41 was specific to VTG from E_2-treated sea bass and others EDCs(Nonylphenol, Benzo[a]pyrene and CdCl_2). Moreover, cross-reactivity was also found in E_2-treated coral grouper(Epinephelus corallicola). The ELISA method obtained from this work can be further applied for the assessment of EDCs in Thailand and Southeast Asia's aquatic environment.     

Einsatz der Hasse-Diagrammtechnik zur vergleichenden Datenanalyse von Biomarkerantworten und ?kotoxikologischen Tests

Die ?kotoxikologie mu? sich mit biologischen Systemen und ihren Antworten auf schadstoffinduzierten Stre? auseinandersetzen. Die hierzu verwendete experimentelle Technik eilt dabei den M?glichkeiten dynamischer, proze?orientierter mathematischer Modellierung weit voraus. In dieser methodisch orientierten Arbeit werden Techniken aus der mathematischen Disziplin der Verbandstheorie vorgestellt, die eine Datenanalyse unter den Aspekten vergleichender Bewertung erlauben. Wichtige Ergebnisse sind dabei

低浓度Pb2+、Cd2+对鲫鱼肝脏组织中 HSP70诱导的影响

以鲫鱼(Carassius auratus)作为实验对象,经过40 d Pb2+、Cd2+不同浓度的暴露后,运用SDS-PAGE和Western Blotting方法检测鱼肝脏组织内应激蛋白HSP70的诱导表达情况.结果表明,在实验浓度下,与对照组相比,Pb2+、Cd2+对鱼肝脏内HSP70有显著的诱导(P＜0.05),但在Cd2+浓度为0.2 mg/L有停止表达HSP70的现象,可能是由于浓度过高造成了组织病理损伤而破坏了诱导表达机制.实验还发现,在实验浓度低于国家渔业用水标准时,HSP70仍然表现为明显诱导(P＜0.05),充分说明运用分子生物学指标要比传统的环境检测指标敏感,具有对污染物早期预警的作用.     

生物标志物在全新世古气候和古环境研究中的应用进展

全新世作为最年轻的一个地质年代,其气候环境变化与人类社会发展息息相关。回顾了近年来全新世古气候和古环境研究的进展,总结了重建古气候和古环境的常用方法,重点介绍了生物标志物包括萜类化合物、长链烯酮、甘油二烷基甘油四醚、甾醇、烷基酯类、木质素和色素在该领域的应用,并提出了开发新型生物标志物、构建多指标体系,以及与地球微生物学、生物化学、微生物学等相关学科开展交叉研究等展望。     

生物标志物

生物标志物技术是常规生物检测方法的必要补充，其在环境管理，风险性评价和生态恢复方面有着较广泛的应用，该文主要介绍了相对于常规生物检定方法而言，生物标志物所具备的优点，以及常用生物标志物的类型，生物标志物在环境保护领域的应用等     

Biochemical responses in armored catfish (Pterygoplichthys anisitsi) after short-term exposure to diesel oil,pure biodiesel and biodiesel blends

Biodiesel fuel is gradually replacing petroleum-based diesel oil use. Despite the biodiesel being considered friendlier to the environment, little is known about its effects in aquatic organisms. In this work we evaluated whether biodiesel exposure can affect oxidative stress parameters and biotransformation enzymes in armored catfish (Pterygoplichthys anisitsi, Loricariidae), a South American endemic species. Thus, fish were exposed for 2 and 7 d to 0.01 mL L⁻¹ and 0.1 mL L⁻¹ of pure diesel, pure biodiesel (B100) and blends of diesel with 5% (B5) and 20% (B20) biodiesel. Lipid peroxidation (malondialdehyde) levels and the activities of the enzymes glutathione S-transferase, superoxide dismutase, catalase and glutathione peroxidase were measured in liver and gills. Also, DNA damage (8-oxo-7, 8-dihydro-2′-deoxyguanosine) levels in gills and 7-ethoxyresorufin-O-deethylase activity in liver were assessed. Pure diesel, B5 and B20 blends changed most of the enzymes tested and in some cases, B5 and B20 induced a higher enzyme activity than pure diesel. Antioxidant system activation in P. anisitsi was effective to counteract reactive oxygen species effects, since DNA damage and lipid peroxidation levels were maintained at basal levels after all treatments. However, fish gills exposed to B20 and B100 presented increased lipid peroxidation. Despite biodiesel being more biodegradable fuel that emits less greenhouse gases, the increased lipid peroxidation showed that biofuel and its blends also represent hazards to aquatic biota.     

Histopathological and biochemical alterations of the earthworm (Lumbricus Terrestris) as biomarker of soil pollution along Porsuk River Basin (Turkey)

This study investigated biomarker responses of the earthworm Lumbricus terrestris in order to evaluate the soil pollution along Porsuk River Basin. Samples consisted of animals from six sites that are agricultural regions and a forested control. Histopathological and biochemical alterations were examined. Significant histopathological alterations were observed in animals from three of the sampling sites. There was an enlargement of epithelial cell lining, mucus cell hyperplasia and increase in mucus secretion. Circular and longitudinal muscles lost their structural integrity. Chloragogenous tissue was dilated and vocuolized. Necrosis was observed in the cells and tissues of some affected worms. A load of heavy metals in tissues of animals was determined. Heavy metals were found to be accumulated particularly in longitudinal muscles of animals. CAT activity was found to be increased in animals from three of the experimental sites. GST activity was also increased in five sites while it was stable in one site. The results have shown that animals from locations particularly that are close to urbanized and industrialized regions were seriously affected from the soil pollution around the basin. These results are reflecting the biological effects of soil pollution around Porsuk River Basin on the indicator organism L. terrestris and constitute an early warning of ecological change in relation to human health.     

Anchoring novel molecular biomarker responses to traditional responses in fish exposed to environmental contamination

The responses of Dicentrarchus labrax and Liza aurata to aquatic pollution were assessed in a contaminated coastal lagoon, using both traditional and novel biomarkers combined. DNA damage, assessed by comet assay, was higher in both fish species from the contaminated sites, whereas levels of cytochrome P450 1A1 gene expression were not significantly altered. The liver histopathological analysis also revealed significant lesions in fish from contaminated sites. Alterations in ras and xpf genes were analysed and additional pollutant-responsive genes were identified. While no alterations were found in ras gene, a downregulation of xpf gene was observed in D. labrax from a contaminated site. Suppression subtractive hybridization applied to D. labrax collected at a contaminated site, revealed altered expression in genes involved in energy metabolism, immune system activity and antioxidant response. The approach and results reported herein demonstrate the utility of anchoring traditional biomarker responses alongside novel biomarker responses.     

Using a physiologically based pharmacokinetic model to link urinary biomarker concentrations to dietary exposure of perchlorate

Exposure to perchlorate is widespread in the United States and many studies have attempted to character the perchlorate exposure by estimating the average daily intakes of perchlorate. These approaches provided population-based estimates, but did not provide individual-level exposure estimates. Until recently, exposure activity database such as CSFII, TDS and NHANES become available and provide opportunities to evaluate the individual-level exposure to chemical using exposure surveillance dataset. In this study, we use perchlorate as an example to investigate the usefulness of urinary biomarker data for predicting exposures at the individual level. Specifically, two analyses were conducted: (1) using data from a controlled human study to examine the ability of a physiologically based pharmacokinetic (PBPK) model to predict perchlorate concentrations in single-spot and cumulative urine samples; and (2) using biomarker data from a population-based study and a PBPK model to demonstrate the challenges in linking urinary biomarker concentrations to intake doses for individuals. Results showed that the modeling approach was able to characterize the distribution of biomarker concentrations at the population level, but predicting the exposure-biomarker relationship for individuals was much more difficult. The type of information needed to reduce the uncertainty in estimating intake doses, for individuals, based on biomarker measurements is discussed.     

Cadmium-induced DNA damage and mutations in Arabidopsis plantlet shoots identified by DNA fingerprinting

Random amplified polymorphic DNA (RAPD) test is a feasible method to evaluate the toxicity of environmental pollutants on vegetal organisms. Herein, Arabidopsis thaliana (Arabidopsis) plantlets following Cadmium (Cd) treatment for 26 d were screened for DNA genetic alterations by DNA fingerprinting. Four primers amplified 20-23 mutated RAPD fragments in 0.125-3.0 mg L⁻¹ Cd-treated Arabidopsis plantlets, respectively. Cloning and sequencing analysis of eight randomly selected mutated fragments revealed 99-100% homology with the genes of VARICOSE-Related, SLEEPY1 F-box, 40S ribosomal protein S3, phosphoglucomutase, and noncoding regions in Arabidopsis genome correspondingly. The results show the ability of RAPD analysis to detect significant genetic alterations in Cd-exposed seedlings. Although the exact functional importance of the other mutated bands is unknown, the presence of mutated loci in Cd-treated seedlings, prior to the onset of significant physiological effects, suggests that these altered loci are the early events in Cd-treated Arabidopsis seedlings and would greatly improve environmental risk assessment.