Enumeration of methanogens with a focus on fluorescence in situ hybridization

Methanogens, the members of domain Archaea are potent contributors in global warming. Being confined to the strict anaerobic environment, their direct cultivation as pure culture is quite difficult. Therefore, a range of culture-independent methods have been developed to investigate their numbers, substrate uptake patterns, and identification in complex microbial communities. Unlike other approaches, fluorescence in situ hybridization (FISH) is not only used for faster quantification and accurate identification but also to reveal the physiological properties and spatiotemporal dynamics of methanogens in their natural environment. Aside from the methodological aspects and application of FISH, this review also focuses on culture-dependent and -independent techniques employed in enumerating methanogens along with associated problems. In addition, the combination of FISH with micro-autoradiography that could also be an important tool in investigating the activities of methanogens is also discussed.     

采用大体积进样提高水样中农药检测的灵敏度

正质谱技术的进步,使得分析方法的检测限不断降低,推动了许多有关环境和食品中痕量污染物的研究.通常情况下,亚ppb级的污染物水平分析,首选串联四极杆质谱.使用WatersXevoG2-S QTof可轻松实现对10-9级别的已知和未知化合物的筛查,如果增大样品进样体积,还可进一步提高灵敏度.而仅依靠MS/MS,是无法得到未知物或大量污染物的分析结果的.结合Xevo G2-S QTof的高精确质量数准确度(3×10-6)在低浓度水平下进行筛查实验,对许多环境分析     

Use of computer models of flood inundation to facilitate communication in flood risk management

It is now widely recognised that good communication between multi-disciplinary stakeholders is central to effective flood risk management. Recent developments in Geographical Information Systems, increased availability of accurate digital terrain models from remotely sensed data sources and improved graphical computer interfaces have made the outputs from computer models of flood inundation easily accessible to the stakeholder community. As a consequence, predictions from such models are now being used routinely as a means of communication between engineers and other stakeholders in flood risk management. This paper provides a review of the modelling methods most appropriate for flood risk communication. These are one-dimensional models which are suitable for simulating flood risk at a catchment or sub-catchment scale and appropriate for communicating the impact of strategic flood management decisions and two-dimensional models which can be applied across a range of scales but, are now being regularly applied at the relatively small scale, less than 10 km², where they have the potential to inform and communicate disaster management decisions. The role of such models in communicating between modellers and non-modellers by providing a means for immediate visualisation of “the future” is discussed and illustrated by application to two case studies.     

Migration of heavy metals in soil as influenced by compost amendments

Soils contaminated with heavy metals can pose a major risk to freshwaters and food chains. In this study, the success of organic and inorganic intervention strategies to alleviate toxicity in a highly acidic soil heavily contaminated with As, Cu, Pb, and Zn was evaluated over 112 d in a mesocosm trial. Amelioration of metal toxicity was assessed by measuring changes in soil solution chemistry, metal leaching, plant growth, and foliar metal accumulation. Either green waste- or MSW-derived composts increased plant yield and rooting depth, reduced plant metal uptake, and raised the pH and nutrient status of the soil. We conclude that composts are well suited for promoting the re-vegetation of contaminated sites; however, care must be taken to ensure that very short-term leaching pulses of heavy metals induced by compost amendment are not of sufficient magnitude to cause contamination of the wider environment.     

Fluorescent dye imaging of the volume sampled by single well forced-gradient tracer tests evaluated in a laboratory-scale aquifer physical model

This study presents a new method to visualise forced-gradient tracer tests in 2-D using a laboratory-scale aquifer physical model. Experiments were designed to investigate the volume of aquifer sampled in vertical dipole flow tracer tests (DFTT) and push-pull tests (PPT), using a miniature monitoring well and straddle packer arrangement equipped with solute injection and recovery chambers. These tests have previously been used to estimate bulk aquifer hydraulic and transport properties for the evaluation of natural attenuation and other remediation approaches. Experiments were performed in a silica glass bead-filled box, using a fluorescent tracer (fluorescein) to deduce conservative solute transport paths. Digital images of fluorescein transport were captured under ultraviolet light and processed to analyse tracer plume geometry and obtain point-concentration breakthrough histories. Inorganic anion mixtures were also used to obtain conventional tracer breakthrough histories. Concentration data from the conservative tracer breakthrough curves was compared with the digital images and a well characterised numerical model. The results show that the peak tracer breakthrough response in dipole flow tracer tests samples a zone of aquifer close to the well screen, while the sampling volume of push-pull tests is limited by the length of the straddle packers used. The effective sampling volume of these single well forced-gradient tests in isotropic conditions can be estimated with simple equations. The experimental approach offers the opportunity to evaluate under controlled conditions the theoretical basis, design and performance of DFTTs and PPTs in porous media in relation to measured flow and transport properties.     

Environmental metabolites of fluoroquinolones: synthesis, fractionation and toxicological assessment of some biologically active metabolites of ciprofloxacin

Background, aim, and scope

Biowastes produced by humans and animals are routinely disposed of on land, and concern is now growing that such practices provide a pathway for fluoroquinolone (FQs) antibacterial agents and their environmental metabolites (FQEMs) to contaminate the terrestrial environment. The focus of concern is that FQs and FQEMs may accumulate in amended soils to then adversely impact on the terrestrial environment. One postulated impact is the development of a selective environment in which FQ-resistant bacteria may grow. To find evidence in support of an accumulation of antibacterial-like activity, it was first necessary to establish whether any biologically active FQEMs could be synthesized by physicochemical factors that are normally present in the environment. However, many FQEMs are not commercially available to be used as standards in such studies. FQEMs were therefore synthesized using well-defined processes. They were subsequently analyzed using spectroscopy (UV-vis) and high performance liquid chromatography with mass spectral detection. The antibacterial-like activities of fractionated FQEMs were then assessed in novel bacterial growth inhibition bioassays, and results were compared to those obtained from instrumental analyses.

Materials and Methods

Parent FQs were either exposed to sunlight or were synthesized using defined aerobic microbial (Mycobacterium gilvum or a mixed culture derived from an agricultural soil) fermentation processes. Mixtures of FQEMs derived from photo- and (intracellular) microbial processes were isolated by preparative chromatography and centrifugation techniques, respectively. Mixtures were subsequently fractionated using analytical high-performance thin layer chromatography (HPTLC), and excised analytes were tested in bioautography assays for their antibacterial-like activities. Two bacteria, Escherichia coli (E. coli) and Azospirillum brasilense (A. brasilense) were used as reporter organisms in testing FQ standards and any subtle differences between biologically active FQEMs of ciprofloxacin (CF).

Results and discussion

FQEMs produced in the photo-synthetic process had UV-vis profiles that were indistinguishable from the parent FQs, and yet mass spectral data revealed the presence of N-formylciprofloxacin (FCF). In contrast, the UV-vis profiles of FQEMs synthesized by M. gilvum and a mixed culture of microorganisms had UV-vis profiles that were similar to one another and markedly different to the parent fluoroquinolones. Mass spectral studies confirmed the presence of FCF and N-acetylciprofloxacin in both microbial ferments. In addition, a photo-FQEM (Cp 6), three M. gilvum FQEMs (Cm 5, Cm 8, and Cm 10) and a mixed culture FQEM (Cs 6) of CF and many other FQEMs of CF, norfloxacin (NF), and enrofloxacin (EF) were fractionated using HPTLC, although their identities have yet to be confirmed. Differences between bioautography results were obtained when E. coli or A. brasilense were used as reporter organisms. Parent FQs (CF and EF) and the FQEMs of CF (Cp 6, Cm 8, and Cs 6) displayed antibacterial-like activity when using E. coli as the reporter organism. In contrast, A. brasilense was insensitive to parent CF and sensitive to EF and all tested FQEMs of CF. Results are consistent with photo- and microbial processes modifying CF in different ways, with the latter changing the UV-vis chromophores. It can be inferred that a lack of detection of analytes (especially photo-FQEMs) when using UV-vis does not necessarily indicate an absence of analyte. Additionally, similarities between the UV-vis profiles of FQEMs extracted from the (monoculture) M. gilvum and the mixed culture microbial aerobic ferments are consistent with similar processes operating in both ferments. Results of HPTLC and bioautography studies revealed that mixtures of (photo- and microbial) FQEMs could be fractionated into individual components.

Conclusions

Bioactive FQEMs of ciprofloxacin, as a representative FQ, can be synthesized by photo- and microbial processes, and their detection required the use of both instrumental and bioautography analytical techniques. It is likely that such FQEMs will also be present on agricultural land that has been repeatedly amended with FQ-contaminated biosolids.

Recommendations and perspectives

The use of instrumental analytical techniques alone and especially photometric detection techniques will underestimate antibacterial-like activities of FQEMs. Moreover, the extraction technique(s) and the selected toxicological endpoint(s) require careful consideration when assessing bioactivity. It is therefore recommended that instrumental analytical techniques and several bioautography assays be performed concurrently, and bioautography assays should use a variety of reporter organisms. Two types of bacterial growth bioassays are recommended in any assessment of antibacterial-like activity derived from CF (and possibly from other FQs). A standardized E. coli bioassay should be used as a general screening procedure to facilitate intra- and inter-laboratory exchange of data. Additionally, soil-specific (region-specific) growth inhibition bioassays should be undertaken using several species of endemic soil bacteria. It is likely that the two sets of data will be useful in future risk assessment processes.     

Passive drainage and biofiltration of landfill gas: results of Australian field trial

A field scale trial was undertaken at a landfill site in Sydney, Australia (2004-2008), to investigate passive drainage and biofiltration of landfill gas as a means of managing landfill gas emissions from low to moderate gas generation landfill sites. The objective of the trial was to evaluate the effectiveness of a passive landfill gas drainage and biofiltration system at treating landfill gas under field conditions, and to identify and evaluate the factors that affect the behaviour and performance of the system.The trial results showed that passively aerated biofilters operating in a temperate climate can effectively oxidise methane in landfill gas, and demonstrated that maximum methane oxidation efficiencies greater than 90% and average oxidation efficiencies greater than 50% were achieved over the 4 years of operation. The trial results also showed that landfill gas loading was the primary factor that determined the behaviour and performance of the passively aerated biofilters. The landfill gas loading rate was found to control the diffusion of atmospheric oxygen into the biofilter media, limiting the microbial methane oxidation process. The temperature and moisture conditions within the biofilter were found to be affected by local climatic conditions and were also found to affect the behaviour and performance of the biofilter, but to a lesser degree than the landfill gas loading.     

Modelling of post-fragmentation waste stream processing within UK shredder facilities

With the introduction of producer responsibility legislation within the UK (i.e., waste electrical and electronic equipment directive and end-of-life vehicles directive), specific recycling and recovery targets have been imposed to improve the sustainability of end-of-life products. With the introduction of these targets, and the increased investment in post-fragmentation facilities, automated material separation technologies are playing an integral role within the UK's end-of-life waste management strategy. Post-fragmentation facilities utilise a range of purification technologies that target certain material attributes (e.g., density, magnetism, volume) to isolate materials from the shredded waste stream. High ferrous prices have historically meant that UK facilities have been primarily interested in recovering iron and steel, establishing processing routes that are very effective at removing these material types, but as a consequence are extremely rigid and inflexible. With the proliferation of more exotic materials within end-of-life products, combined with more stringent recycling targets, there is therefore a need to optimise the current waste reclamation processes to better realise effort-to-value returns. This paper provides a background as to the current post-fragmentation processing adopted within the UK, and describes the development of a post-fragmentation modelling approach, capable of simulating the value-added processing that a piece of automated separation equipment can have on a fragmented waste stream. These include the modelling of the inefficiencies of the technology, the effects of material entanglement on separation, determination of typical material sizing and an appreciation for compositional value. The implementation of this approach within a software decision-support system is described, before the limitations, calibration and further validation of the approach are discussed.     

Validation of a passive atmospheric deposition sampler for polybrominated diphenyl ethers

An atmospheric deposition sampler was validated with respect to polybrominated diphenyl ethers (PBDEs), a compound group that is widely used as flame retardants in many types of consumer products. The deposition sampler consists of an adsorption cartridge that is connected to a glass funnel. Extraction tests with spiked cartridges using soxhlet extraction with acetone revealed recoveries of >80% for all of the investigated PBDEs. Once adsorbed, PBDEs are stable, as proven by extractions of spiked cartridges that were stored outdoors and collected after different periods of time, up to 84 days. High recoveries indicate that degradation of adsorbed PBDEs does not play a role under realistic field conditions. Bulk deposition rates of PBDEs were determined in a field test with 3 replicates, and a possible breakthrough of target compounds was assessed in the field using a second adsorption cartridge in series. No breakthrough of target compounds could be observed within a sampling period of 61 days, and a bulk deposition rate of approximately 1 ng m(-2) day(-1) for the sum of all analysed PBDEs was measured. The highest deposition rates were measured for octa-brominated congeners, followed by BDEs 99, 183, 153, and 47. Overall, the sampler was successfully validated regarding the atmospheric deposition of PBDEs.     

Distributing power? Community energy projects' experiences of planning,policy and incumbents in the devolved nations of Scotland and Wales

Community-owned energy projects are viewed as compelling contributors towards renewable energy targets. They contribute to curtailing the use of carbon intensive energy sources, consequently aiding mitigation of climate change, and can contribute towards a sustainable, localised economy. The success and expansion of the sector varies. However, there is a lack of knowledge regarding the development of the sector between sub-state nations. This qualitative, comparative study looks at the sector in the sub-state, devolved nations of Wales and Scotland within the UK. Through a series of in-depth interviews with community energy practitioners in four case study sites in Scotland and Wales, this study shows how policy and governance practices can influence the sector and those working at grassroots level. The study shows a disparity in confidence and outlook for the sector, based on the perceived (in)effective governance in each devolved nation.