Tissue bioaccumulation patterns, xenobiotic biotransformation and steroid hormone levels in Atlantic salmon (Salmo salar) fed a diet containing perfluoroactane sulfonic or perfluorooctane carboxylic acids

In the present study, groups of juvenile Atlantic salmon (Salmo salar) were fed gelatine capsules containing fish-food spiked with PFOA or PFOS (0.2 mg kg⁻¹ fish) and solvent (methanol). The capsules were given at days 0, 3 and 6. Blood, liver and whole kidney samples were collected prior to exposure (no solvent control), and at days 2, 5, 8 and 14 after exposure (Note: that day 14 after exposure is equal to 7 d recovery period). We report on the differences in the tissue bioaccumulation patterns of PFOS and PFOA, in addition to tissue and compound differences in modulation pattern of biotransformation enzyme genes. We observed that the level of PFOS and PFOA increased in the blood, liver and kidney during the exposure period. Different PFOS and PFOA bioaccumulation patterns were observed in the kidney and liver during exposure- and after the recovery periods. Particularly, after the recovery period, PFOA levels in the kidney and liver tissues were almost at the control level. On the contrary, PFOS maintained an increase with tissue-specific differences, showing a higher bioaccumulation potential (also in the blood), compared with PFOA. While PFOS and PFOA produced an apparent time-dependent increase in kidney CYP3A, CYP1A1 and GST expression, similar effects were only temporary in the liver, significantly increasing at sampling day 2. PFOA and PFOS exposure resulted in significant decreases in plasma estrone, testosterone and cortisol levels at sampling day 2, and their effects differed with 17α-methyltestostrerone showing significant decrease by PFOA (also for cholesterol) and increase by PFOS. PFOA significantly increased estrone and testosterone, and no effects were observed for cortisol, 17α-methyltestosterone and cholesterol at sampling day 5. Overall, the changes in plasma steroid hormone levels parallel changes in CYP3A mRNA levels. Given that there are no known studies that have demonstrated such tissue differences in bioaccumulation patterns with associated differences in toxicological responses in any fish species or lower vertebrate, the present findings provide some potential insights and basis for a better understanding of the possible mechanisms of PFCs toxicity that need to be studied in more detail.     

Determination of dioxin concentrations in fish and seafood samples using a highly sensitive reporter cell line, DR-EcoScreen cells

There is a strong need for the development of relatively rapid and low-cost bioassays for the determination of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), and dioxin-like polychlorinated biphenyls (dl-PCBs) in environmental and food samples. In this study, we applied a reporter gene assay using DR-EcoScreen cells (DR-cell assay), which is highly sensitive to dioxins, to the determination of PCDD/Fs and dl-PCBs in fish and seafood samples. The PCDD/Fs and dl-PCBs were extracted from homogenated samples (10 g) of 30 fish and shellfish, purified by clean-up procedure using a multilayered silica gel column and an alumina column, and applied to DR-cell assay. Interestingly, the bioanalytical equivalent (BEQ) values obtained from the DR-cell assay [<0.1 ∼ 5.4 pg BEQ g⁻¹ wet weight (ww)] were closely correlated with the toxicity equivalent (TEQ) values from conventional high-resolution gas chromatography/high-resolution mass spectrometry (HRGC-HRMS) analysis (r² = 0.912), and the slope of regression line was 0.913. Therefore, we multiplied the BEQ values from the DR-cell assay by a conversion coefficient (1.095, the reciprocal of 0.913) to approximate the TEQ values from the HRGC-HRMS analysis. Furthermore, we used this DR-cell assay to perform a prescreening test of PCDD/Fs and dl-PCBs in 16 fish and seafood samples purchased from a supermarket, revealing that a sample from the fatty flesh of a bluefin tuna exceeded 8 pg TEQ g⁻¹ ww (the European Union-tolerance limit). Taken together, these results suggest that the DR-cell assay might be applicable as a rapid and low-cost prescreening method to determine dioxin levels in fish and seafood samples.     

Evaluation of ABC efflux transporters genes expression in kidney of rainbow trout (Oncorhynchus mykiss) fed with melamine and cyanuric acid diets

Gene expression experiments were targeted in order to monitor the ABC efflux transporters, which is potentially involved in cellular detoxification/defense. Changes in expression levels of different ABC genes in kidney of Oncorhynchus mykiss fed with melamine and melamine + cyanuric acid enriched diets were recorded in both treated groups by mRNA ΔΔ_CT relative quantification method. Expression profiles of eight different ABC genes basically showed low alterations in melamine group and more consistent changes in melamine + cyanuric acid treated fish, compared with own control. In the last group ABCC2 gene over expression was the more evident alteration. These results suggest that ABC efflux system could be involved in mobilization of hydrophilic molecules in the forcing condition of chronic exposure.     

The profile of antibiotics resistance and integrons of extended-spectrum beta-lactamase producing thermotolerant coliforms isolated from the Yangtze River basin in Chongqing

The spreading of extended-spectrum β-lactamases (ESBL)-producing thermotolerant coliforms (TC) in the water environment is a threat to human health but little is known about ESBL-producing TCs in the Yangtze River. We received 319 ESBL-producing stains obtained from the Chongqing basin and we investigated antibiotic susceptibility, bla gene types and the presence of integrons and gene cassettes. 16.8% of TC isolates were ESBL-producing bacteria and bla_TEM+CTx-M was the predominant ESBL type. 65.2% of isolates contained class 1 integrons, but only 3 carried intI 2. Gene cassettes were amplified and sequenced. aadA, drfA, cmlA, sat1, aar3 and two ORF cassettes were found. In conclusion, Yangtze River is heavily polluted by ESBL-producing TC bacteria and the combined bla gene type could enhance antibiotic resistance. Class 1 integrons were widespread in ESBL-producing isolates and play an important role in multi-drug resistance. Characterization of gene cassettes could reveal the dissemination of antibiotic resistance genes.     

The Probability of a Horizontal Gene Transfer from Roundup Ready® Soybean to Root Symbiotic Bacteria: A Risk Assessment Study on the GSF Lysimeter Station

The gene transfer from glyphosate tolerant soybean to Bradyrhizobium japonicum was evaluated in a free-air lysimeter experiment under natural conditions and increasing selection pressure, to monitor for the probability of horizontal gene transfer (HGT). A large volume lysimeter study that offers conditions comparable to normal farming was conducted in 2004 and 2005 with Roundup Ready® (RR) soybean and Roundup® application according to agricultural practice. Analysis of nodules showed, as expected, the presence of the transgenic 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS). However, in bacteroids that were isolated from nodules and then cultivated for several rounds in the presence of high levels of glyphosate, the EPSPS gene could no longer be detected. This indicates no stable HGT transfer of the whole EPSPS gene under field conditions.     

污水处理厂生物气溶胶抗生素抗性污染特征

为探究污水处理厂生物气溶胶抗生素抗性基因（ARGs）污染特征,在济南市某污水处理厂采用宏基因组测序技术对厂界内及周边生物气溶胶样本及污水或污泥样本进行分析.结果表明,相比于上风向,厂界内和下风向生物气溶胶具有更多的ARGs亚型种类数和更高的总相对丰度.厂界内与上风向生物气溶胶ARGs组成存在显著的差异性,差异度为47.57%;而厂界内与下风向生物气溶胶ARGs组成的差异性不显著,且差异度下降至33.98%.上风向背景空气和污水或污泥均是厂界内生物气溶胶ARGs的重要来源,两者总的源的贡献大于63.92%.共检测到43种ARGs亚型（8种ARGs主型）在至少一处污水处理单元极易负载于生物气溶胶颗粒逸出.本研究可为污水处理厂生物气溶胶抗生素抗性污染的风险评估和控制提供理论依据.     

2种烷基多环芳烃对仿刺参CYP450和p53基因表达的影响研究

海洋环境中多环芳烃类(PAHs)主要来源于海洋溢油事故以及沿海石油化工企业的废水排放,国内外大量研究发现海洋中的多环芳烃对海洋生物造成了潜在的生态风险。为了揭示不同浓度多环芳烃类污染物对海参的生态毒理效应,将仿刺参(Apostichopus japonicus)分别暴露于不同浓度的2种烷基多环芳烃3-甲基菲(5、10、100μg·L~(-1))和2-甲基蒽(5、10、50μg·L~(-1))中,检测暴露3 d、7 d和14 d后,3-甲基菲和2-甲基蒽胁迫下仿刺参CYP450和p53基因的相对表达量。结果表明,3-甲基菲和2-甲基蒽胁迫下,仿刺参CYP450和p53基因的表达均对毒物产生了不同程度的响应。与对照组相比,3-甲基菲各处理组对仿刺参CYP450和p53基因的表达均产生显著的抑制作用(P0.05); 2-甲基蒽各处理组对仿刺参CYP450和p53基因的表达影响作用不同,暴露7 d后,2-甲基蒽各处理组对仿刺参CYP450基因的表达表现出抑制作用,对p53基因的表达表现出诱导作用。相同浓度与时间胁迫下,2-甲基蒽对仿刺参CYP450和p53基因表达的影响比3-甲基菲的影响大。上述研究结果表明,3-甲基菲和2-甲基蒽均可不同程度影响仿刺参CYP450和p53基因的表达,且与3-甲基菲相比,2-甲基蒽对仿刺参CYP450和p53基因表达的影响较明显。上述结果为多环芳烃类污染物对仿刺参的生物毒性评价提供了基础数据。     

集约化柑橘种植抑制土壤磷循环微生物活性

微生物是调节土壤磷循环的关键驱动力.阐明土壤解磷菌的微生物矿化过程对于提高植物养分吸收率和作物产量具有重要意义.通过测定柑橘园与毗邻的自然林地土壤编码碱性磷酸酶基因（phoD）丰度、解磷细菌群落多样性和土壤无机磷组分,探究柑橘种植对土壤微生物获取磷策略的影响机制.结果表明,柑橘种植导致土壤pH下降,土壤有效磷累积,ω（有效磷）平均值高达112 mg·kg^-1,显著高于毗邻的自然林地（3.7 mg·kg^-1）.柑橘种植也会影响土壤磷素组成,柑橘土壤含有较高的可溶态磷（CaCl₂-P）、柠檬酸提取态磷（Citrate-P）和矿物结合态磷（HCl-P）.自然林地土壤各磷组分均显著低于柑橘土壤,而phoD基因丰度和碱性磷酸酶活性显著高于柑橘土壤.高通量测序结果表明,柑橘土壤解磷细菌Shannon指数（4.61）显著低于自然林地（5.35）,群落结构也有别于自然林地.柑橘种植改变了土壤解磷菌的群落组成,自然林地变形菌门的相对丰度显著低于柑橘土壤.土壤有效磷含量与碱性磷酸酶活性呈显著负相关,表明土壤高磷累积抑制土壤解磷细菌的活性.柑橘种植改变了土壤微生物对磷的获取策略,在柑橘园中,土壤微生物主要依赖外源磷,而自然林地土壤微生物主要以微生物分泌碱性磷酸酶矿化有机磷来获取磷的方式满足其生长需求.