Dry and heat stress affects H2S production of Salmonella on selective plating media

The pH of Salmonella pre-enrichment media can become acidic (pH 4.0–5.0) when feeds/ingredients are incubated for 24?h. Salmonella in feed that have been stressed by heat and desiccation exhibit different pH tolerances than non-stressed cultures. Acidic conditions can result in cell injury/death and affect biochemical pathways. In this study, eight serotypes of Salmonella were grown in sterile meat and bone meal that was subjected to desiccation and heat stress. Cultures of non-stressed and stressed isolates were subsequently exposed to acidic pH from 4.0 to 7.0 in 0.5?pH increments (3 replicates/pH increment) in citrate buffer. At 6 and 24?h, serial dilutions were plated in duplicate on XLT-4 (xylose lysine tergitol-4) agar. Four serotypes showed an impaired ability to decarboxylate lysine on XLT-4. This inability to decarboxylate lysine was dependent on isolate, stress status, and incubation time. When the isolates’ ability to decarboxylate lysine was examined using biochemical tests, cultures were found to be able to decarboxylate lysine with the exception of S. Infantis. This suggests that XLT-4 contains a biochemical stressor(s) which affects the rate of decarboxylation by these Salmonella. These results suggest that acidic conditions may influence the detection and confirmation of Salmonella in feed.     

Evolution of tolerance to PCBs and susceptibility to a bacterial pathogen (Vibrio harveyi) in Atlantic killifish (Fundulus heteroclitus) from New Bedford (MA, USA) harbor

A population of the non-migratory estuarine fish Fundulus heteroclitus (Atlantic killifish) resident to New Bedford (NB), Massachusetts, USA, an urban harbor highly contaminated with polychlorinated biphenyls (PCBs), demonstrates recently evolved tolerance to some aspects of PCB toxicity. PCB toxicology, ecological theory, and some precedence supported expectations of increased susceptibility to pathogens in NB killifish. However, laboratory bacterial challenges of the marine pathogen Vibrio harveyi to wild fish throughout the reproductive season and to their mature laboratory-raised progeny demonstrated comparable survival by NB and reference killifish, and improved survival by NB males. These results are inconsistent with hypothesized trade-offs of adaptation, and suggest that evolved tolerance in NB killifish may include mechanisms that minimize the immunosuppressive effects of PCBs. Compensatory strategies of populations persisting in highly contaminated environments provide a unique perspective for understanding the long-term ecological effects of toxic chemicals.     

Integral assessment of estrogenic potentials in sediment-associated samples

Background, aim, and scope  The enzyme-linked receptor assay (ELRA) detects estrogenic and anti-estrogenic effects at the molecular level of receptor binding and is a useful tool for the integrative assessment of ecotoxicological potentials caused by hormonally active agents (HAA) and endocrine disrupting compounds (EDC). The main advantage of the ELRA is its high sample throughput and its robustness against cytotoxicity and microbial contamination. After a methodological adaptation to salinity of the ELRA, according to the first part of this study, which increased its salinity tolerance and sensitivity for 17-β-estradiol, the optimised ELRA was used to investigate 13 native sediments characterised by different levels of salinity and chemical contamination. The applicability of the ELRA for routine analysis in environmental assessment was evaluated. Salinity is often a critical factor for bioassays in ecotoxicological sediment assessment. Therefore, salinity of the samples was additionally adjusted to different levels to characterise its influence on elution and binding processes of receptor-binding substances. Materials and methods  The ELRA was carried out with the human estrogen receptor α (ER) in a 96-well microplate format using the experimental setup known from the competitive immunoassay based on ligand–protein interaction. It is an important improvement that a physiologically relevant receptor was used as a linking protein instead of an antibody. The microplates were coated with a 17-β-estradiol-BSA conjugate, and dilution series of estradiol and of native sediment samples were added and incubated with the ER. After a washing step, a biotinylated mouse anti-ER antibody was added to each well. Receptor binding to estradiol, agonistic and antagonistic receptor binding, were determined by a streptavidin-POD-biotin complex with subsequent measurement of the peroxidase activity at the wavelength of 450 nm using a commercial ELISA multiplate reader. The sediment elutriates and pore water samples of sediments were tested in a dilution series to evaluate at which dilution step the receptor-binding potential ends. In the elution process (see Section 2.1 to 2.2), a method was developed to adjust the salinity to the levels of the reference testings, which offers an appropriate option to adjust the salinity in both directions. Statistical evaluation was made with a combination of the Mann–Whitney U test and the pT-method. Results  This part of the study characterised the environmental factor ‘salinity’ for prospective applications of the ELRA. Using reference substances such as 17-β-estradiol, the ELRA showed sigmoid concentration-effect relations over a broad range from 0.05 μg/l to 100 μg/l under physiological conditions. After methodological optimisation, both sensitivity and tolerance of the assay against salinity could be significantly raised, and the ELRA became applicable under salinity conditions up to concentrations of 20.5‰. The mean relative inter-test error (n = 3) was around 11% with reference substances and below 5% for single sediments elutriates in three replicates each. For sediment testings, the pore water and different salinity-adjusted elutriates of 13 sediments were used. A clear differentiation of the receptor-binding potential could be reached by application of the pT-method. Thereby, pT-values from one to six could be assigned to the sediments, and the deviation caused by the different salinity conditions was one pT-value. The mean standard deviation in the salinity adaptation procedure of the elutriates was below 5%. Discussion  Although the ELRA has already been used for assessments of wastewater, sludge and soil, its applicability for samples to different salinity levels has not been investigated so far. Even if the ELRA is not as sensitive as the E-screen or the YES-assay, with regard to reference substances like 17-β-estradiol, it is a very useful tool for pre-screening, because it is able to integrate both estrogenic as well as anti-estrogenic receptor-binding effects. According to the results of sediment testing, and given the integrative power to detect different directions of effects, the ELRA shows sufficient sensitivity and salinity tolerance to discriminate receptor-binding potentials in environmental samples. Conclusions  The optimised ELRA assay is a fast, cost-effective, reliable and highly reproducible tool that can be used for high-throughput screening in a microplate format in detecting both estrogenic and anti-estrogenic effects. Additionally, the ELRA is robust against microbial contaminations, and is not susceptible towards cytotoxic interferences like the common cell-culture methods. The general applicability and sufficient sensitivity of the ELRA was shown in freshwater environments. Marine and brackish samples can be measured up to salinity levels of 20.5‰. Recommendations and perspectives  In view of the proven sensitivity, functionality and the fastness of the ELRA, it is recommendable to standardise the test method. At the moment, no adequate in vitro test procedure exists which is standardised to DIN or ISO levels. The E-screen and the yeast estrogen/androgen screens (YES/YAS) sometimes underlie strong cytotoxic effects, as reported in the first part of this study. Further development of an ELRA assay using human androgen receptors appears to be very promising to gain information about androgenic and anti-androgenic effects, too. This would offer a possibility to use the ELRA as a fast and reliable pre-screening tool for the detection of endocrine potentials, thus minimising time and cost-expensive animal experiments.     

Cd-tolerant Suillus luteus: A fungal insurance for pines exposed to Cd

Soil metal pollution can trigger evolutionary adaptation in soil-borne organisms. An in vitro screening test showed cadmium adaptation in populations of Suillus luteus (L.: Fr.) Roussel, an ectomycorrhizal fungus of pine trees. Cadmium stress was subsequently investigated in Scots pine (Pinus sylvestris L.) seedlings inoculated with a Cd-tolerant S. luteus, isolated from a heavy metal contaminated site, and compared to plants inoculated with a Cd-sensitive isolate from a non-polluted area. A dose-response experiment with mycorrhizal pines showed better plant protection by a Cd-adapted fungus: more fungal biomass and a higher nutrient uptake at high Cd exposure. In addition, less Cd was transferred to aboveground plant parts. Because of the key role of the ectomycorrhizal symbiosis for tree fitness, the evolution of Cd tolerance in an ectomycorrhizal partner such as S. luteus can be of major importance for the establishment of pine forests on Cd-contaminated soils.     

聚(丙烯酸-co-丙烯酰胺)/膨润土/腐植酸钠三维网络凝胶吸附剂对Cd2+吸附性能研究

制备了一种聚(丙烯酸-co-丙烯酰胺)/膨润土/腐植酸钠三维网络凝胶吸附剂,重点考察了吸附剂对Cd2+吸附的pH依赖性、吸附动力学和吸附等温线.结果表明,Cd2+溶液pH值对吸附容量有较大影响.在pH=8、吸附时间30 min、Cd2+溶液初始浓度0.02 mol/L和吸附剂用量0.10 g的条件下,吸附剂对Cd2+的饱和吸附量可达294.7 mg/g.与膨润土相比,三维网络凝胶吸附剂具有更高的吸附容量和更快的吸附速率.     

土壤中镉、锌离子的单一及竞争吸附研究

采用等温吸附法探讨了重金属镉、锌在青岛地区3种主要土壤(棕壤、潮土和砂浆黑土)中的吸附特征.结果表明,在质量浓度为20～500 mg/L的单一重金属等温吸附试验中,棕壤和潮上对Cd~(2+)、Zn~(2+)的吸附量相近,而黑土对Cd~(2+)、Zn~(2+)的吸附量明显高于棕壤和潮土.Cd~(2+)、Zn~(2+)的双组分竞争吸附表明,3种上壤对Cd~(2+)、Zn~(2+)的吸附量与其单独存在下的情形相比都有所下降.质量浓度分别为20～500mg/L的两种重金属元索等馈竞争吸附时,这3种土壤对Cd2+的吸附量比对Zn~(2+)的大.非等量竞争吸附试验的结果表明,3种土壤中,500mg/L的Zn~(2+)对100 mg/L的Cd~(2+)吸附量的影响均比同浓度的Cd~(2+)对Zn~(2+)的影响大.用Langmuir和Freundlich方程对这两种重金属离子的等温吸附试验数据进行拟合,均呈极显著相关关系.     

3种镉超富集植物毛状根体系对镉胁迫响应的比较

近年来,转基因毛状根组织被越来越多地应用于重金属和有机污染物的植物修复技术研究中,已成为进行污染物毒性响应机制研究的便捷的实验室工具。为了探究龙葵、油菜、芥菜3种镉(cadmium,Cd)超富集植物对Cd毒性胁迫响应的差异,以诱导出的3种植物毛状根为研究材料,从毛状根的生长状态、富集Cd的能力、根组织细胞的凋亡程度和抗氧化酶活性等方面进行了探讨。结果表明:Cd浓度为0~50μmol·L-1时,龙葵、油菜、芥菜毛状根受Cd毒害的影响都不明显;Cd浓度为75~100μmol·L-1时,龙葵、油菜、芥菜毛状根均表现出对Cd胁迫的防御响应。在较高的Cd浓度(100μmol·L-1)下,龙葵毛状根的生物量受Cd毒害的影响最小,芥菜次之,油菜受影响最大;同时龙葵毛状根富集的Cd含量最高(745.0μg·g-1),芥菜次之(681.4μg·g-1),油菜最差(505.2μg·g-1)。龙葵、油菜、芥菜毛状根在Cd胁迫下的细胞凋亡水平均随Cd浓度的升高而升高,当Cd浓度为100μmol·L-1时,龙葵毛状根比油菜和芥菜毛状根的细胞凋亡程度均低。同时3种植物毛状根在不同浓度Cd处理下抗氧化酶活性的变化有一定差异。从上述结果综合来看,龙葵毛状根受Cd毒害的影响最小、富集Cd的能力最好,是进一步开展Cd超富集植物转基因改造研究的较好的实验室载体。     

菲和镉单一及复合污染条件下在毛蚶体内的富集动力学研究

采用室内半静态双箱动力学模型实验,研究了菲和镉单一及复合污染条件下在毛蚶(Anadara subcrenata)体内的生物富集,通过对富集与释放过程中毛蚶体内菲和Cd的富集量进行非线性曲线拟合,获得了菲和Cd单一及复合污染条件下在毛蚶体内的吸收速率常数k1、释放速率常数k2、生物富集因子BCF、生物半衰期B1/2和平衡状态下最大富集量CA max等动力学参数。实验结果表明,菲和Cd在实验前期富集速率较高,8 d以后富集速率减缓,释放阶段与富集阶段相似。毛蚶对菲的BCF值为37.80,远大于Cd的BCF值13.12,且生物半衰期时间更长,菲更容易在生物体内富集。菲和Cd联合暴露条件下,在毛蚶体内的CA max和BCF值均大于单一作用,说明二者同时暴露时,毛蚶对菲和镉的吸收富集均有所增强。实验模型拟合度较好,输出值和实测值之间无显著性差异,拟合方程和拟合参数可信。     

化学沉淀法强化常规工艺应急去除水中的镉

在常规工艺基础上,通过投加氢氧化钠,实验进行了应急去除重金属镉的研究。实验结果表明,该方法能有效去除饮用水水源的镉,效果稳定,可进行应急处理。对pH、镉初始浓度和混凝剂投加量3个影响因素的灰色关联分析表明,3个因素对镉去除效果影响的大小排序为:滤后水pH>混凝剂投加量>镉初始浓度。在水源镉突发污染时,在原有常规水处理工艺基础上,通过控制滤后水pH可实现对重金属镉的去除,pH的控制值与水源水质有关。     

多孔性Mn3O4的制备及其对水中镉的吸附

采用硫酸锰水解-氧化两步法制备了多孔性四氧化三锰,以X射线衍射、扫描电镜和氮吸附比表面仪对其进行了表征,研究了该多孔性Mn3O4在不同的溶液pH值、吸附时间、投加量和温度下对水中Cd(Ⅱ)的吸附行为,并测定了吸附过程的动力学和热力学参数。实验结果表明,该法合成的四氧化三锰属四方体心结构,一次晶粒尺寸为45.8 nm,颗粒间存在大量的孔洞,BET法比表面积为40.27 m2/g。在293 K,pH 5.5,吸附12 h时达到平衡,饱和吸附量为12.8 mg/g。该吸附过程符合准二级反应动力学模型,其反应的吸附活化能为2.743 kJ/mol,吸附等温线较好地符合Langmuir等温方程,吸附焓变和熵变均为正值,自由能变为负值,说明该吸附过程为自发的吸热过程。