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.     

湘潭锰矿重金属环境安全及植物耐性研究

采集了湘潭锰矿红旗分矿开采区、沙圹村恢复区的代表性当季蔬菜(莴笋叶Fruticicolidae、小白菜Brassica chinensis、香葱Allium schoenoprasum、空心菜Ipomoea aquatica)、废弃区的优势植物(商陆Phytolacca acinosa、野茼蒿Crassocephalum crepidioides、苍耳Xanthium sibiricum)和3个研究区的土壤,通过原子吸收分光光度法分析了Mn、Pb、Zn含量。结果表明:开采区蔬菜Mn含量(8.3~84.5 mg/kg)明显高于恢复区(2.7~55.6 mg/kg),开采区和恢复区蔬菜都明显受到Pb污染(0.6~33mg/kg),蔬菜Zn含量范围为1.9~6.5mg/kg;3个研究区域土壤重金属均明显超标,最严重的是Pb污染(1 993.5~2 213.5mg/kg)。商陆、野茼蒿和苍耳中重金属含量差异较大,对重金属的耐性强,其中商陆表现出最好的耐性与长势。研究结论对锰矿土地合理利用以及矿区土壤重金属治理提供一定的科学依据。     

铜尾矿区9种优势植物体内重金属和氮磷含量研究

以杨山冲铜尾矿废弃地人工覆土上生长的9种优势植物为研究对象,探讨不同植物对尾矿基质中重金属和氮磷的吸收特性以及与基质理化性质之间的关联。土壤样品采集时沿基质垂直方向分为4层：A层（覆土层0~5 cm）、B层（覆土层5~20 cm）、C层（尾砂层20~25 cm）、D层（尾砂层25~40 cm）,植物样品采集时草本植物分地上和地下2部分、木本植物分根、茎和叶3部分采集。结果表明：随着9种植物生长,A层和B层基质理化性质得到明显的改善。主要表现在A层中w（总氮）、w（氨氮）、w（硝氮）、w（有效磷）和w（有机质）显著高于C、D层（P〈0.05）,B层中w（有效态Cu）、w（有效态Zn）显著低于A、C、D层（P〈0.05）。9种优势植物对Cu、Pb、Zn的吸收和积累能力具有明显差异性,其中草本植物以根系积累为主,木本植物以叶片积累为主。草本植物对尾矿基质表现出较强的适应性和耐性,其中毛叶荩草（Arthraxon prionodes）根系中Cu、Pb、Zn的富集量均最大,耐性表现最突出,是较为理想的铜尾矿基质的修复物种。另外,本研究还发现,9种植物体内氮磷质量分数也各不相同,其中木本植物叶中氮磷质量分数均大于根、茎。木本植物体内w（总氮）与w（总磷）、w（Cu）、w（Zn）相关性较好,草本植物体内w（Cu）、w（Pb）、w（Zn）之间相关性较好,表明彼此之间存在一定的促进作用。     

利用细菌表面展示技术构建镉耐受性的重组根瘤菌

通过细菌表面展示功能基因InaXN、猴金属硫蛋白α亚基四聚体MT4α和两种启动子PnifH及Plac元件,分别构建两个重组质粒pTNIM和pBLIM.进一步通过三亲本杂交,将重组质粒分别导入费氏中华根瘤菌HN01,分别构建获得诱导表达型重组菌HN01(pTNIM)和组成表达型重组菌HN01(pBLIM).在培养条件下比较测定重组菌和出发菌对镉的吸附能力和耐受性,结果表明,组成表达型重组菌HN01(pBLIM)在上述两个方面的能力得到明显增强,对镉的吸附富集能力提高了2倍.研究还发现重组根瘤菌也具有很好的静态吸附效果.盆栽实验结果表明,接种诱导表达型重组菌HN01(pTNIM)的大豆根瘤和根中Cd2+的富集量均明显高于野生菌株HN01.本实验为后续探索重组根瘤菌与宿主植物共生体系在Cd2+污染土壤修复中的应用前景奠定了基础.     

Influence of arsenic stress on synthesis and localization of low-molecular-weight thiols in Pteris vittata

The roles of low-molecular-weight thiols (LMWTs), such as glutathione and phytochelatins, in arsenic (As) tolerance and hyperaccumulation in Pteris vittata an As-hyperaccumulator fern remain to be better understood. This study aimed to thoroughly characterize LMWT synthesis in P. vittata to understand the roles played by LMWTs in As tolerance and hyperaccumulation. LMWT synthesis in P. vittata was induced directly by As, and not by As-mediated oxidative stress. Expression of PvECS2, one of the putative genes of γ-glutamylcysteine synthetase (γECS), increases in P. vittata shoots at 48 h after the onset of As exposure, almost corresponding to the increase in the concentrations of γ-glutamylcysteine and glutathione. Furthermore, localization of As showed similar trends to those of LMWTs in fronds at both whole-frond and cellular levels. This study thus indicates the specific contribution of LMWTs to As tolerance in P. vittata. γECS may be responsible for the As-induced enhancement of LMWT synthesis.     

Ammonium and nitrate tolerance in lichens

Since lichens lack roots and take up water, solutes and gases over the entire thallus surface, these organisms respond more sensitively to changes in atmospheric purity than vascular plants. After centuries where effects of sulphur dioxide and acidity were in the focus of research on atmospheric chemistry and lichens, recently the globally increased levels of ammonia and nitrate increasingly affect lichen vegetation and gave rise to intense research on the tolerance of lichens to nitrogen pollution. The present paper discusses the main findings on the uptake of ammonia and nitrate in the lichen symbiosis and to the tolerance of lichens to eutrophication. Ammonia and nitrate are both efficiently taken up under ambient conditions. The tolerance to high nitrogen levels depends, among others, on the capability of the photobiont to provide sufficient amounts of carbon skeletons for ammonia assimilation. Lowly productive lichens are apparently predisposed to be sensitive to excess nitrogen.     

活性污泥的耐盐驯化培养

以某污水处理厂二沉池好氧污泥为接种污泥,采用逐步提高盐度和稳定盐度2种方法对活性污泥进行耐盐性驯化培养,考察驯化结果表明,前一种方法更有利于耐盐菌的培养。对比不同盐度情况下各项指标的去除效果得出:本实验污泥适宜盐度为1%。使用稳定进水盐度的方法,出水指标及各指标的去除率均低于逐步提高盐度法,且镜检结果表明大量微生物死亡。     

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.     

Constitutional tolerance to heavy metals of a fiber crop, ramie (Boehmeria nivea), and its potential usage

It is observed that ramie (Boehmeria nivea), an economic fiber crop, can establish and colonize metal-contaminated sites in China. Metal tolerance and accumulation by ramie originating from 13 metal-contaminated and 4 “clean” sites in China were compared under field and hydroponic conditions. All selected populations and germplasms displayed good growth performance under diverse metal-contaminated habitats; while growth responses, metal accumulation and tolerance were similar among the 8 populations and 2 germplasms when exposed to solutions containing elevated As, Cd, Pb, or Zn in the laboratory. These revealed that ramie possesses a certain degree of constitutional metal tolerance. To our knowledge, this is the first report of constitutional metal tolerance possessed by a fiber crop. Ramie can be considered as a good candidate for both fiber production and phytoremediation of sites contaminated by multi-metals, as it accumulates relative low metal concentrations, but possesses both high biomass and high economic value.