Trine Lund Hansen

Environmental assessment of solid waste systems and technologies : EASEWASTE

A new model has been developed for evaluating the overall resource consumption and environmental impacts of municipal solid waste management systems by the use of life cycle assessment. The model is named EASEWASTE (Environmental Assessment of Solid Waste Systems and Technologies) and is able to compare different waste management strategies, waste treatment methods and waste process technologies. The potential environmental impacts can be traced back to the most important processes and waste fractions that contribute to the relevant impacts. A model like EASEWASTE can be used by waste planners to optimize current waste management systems with respect to environmental achievements and by authorities to set guidelines and regulations and to evaluate different strategies for handling of waste. The waste hierarchy has for decades been governing waste management but the ranking of handling approaches may not always be the most environmentally friendly. The EASEWASTE model can identify the most environmentally sustainable solution, which may differ among waste materials and regions and can add valuable information about environmental achievements from each process in a solid waste management system.

Evaluation of environmental impacts from municipal solid waste management in the municipality of Aarhus, Denmark (EASEWASTE)

A new computer based life cycle assessment model (EASE-WASTE) was used to evaluate a municipal solid waste system with the purpose of identifying environmental benefits and disadvantages by anaerobic digestion of source-separated household waste and incineration. The most important processes that were included in the study are optical sorting and pre-treatment, anaerobic digestion with heat and power recovery, incineration with heat and power recovery, use of digested biomass on arable soils and finally, an estimated surplus consumption of plastic in order to achieve a higher quality and quantity of organic waste to the biogas plant. Results showed that there were no significant differences in most of the assessed environmental impacts for the two scenarios. However, the use of digested biomass may cause a potential toxicity impact on human health due to the heavy metal content of the organic waste. A sensitivity analysis showed that the results are sensitive to the energy recovery efficiencies, to the extra plastic consumption for waste bags and to the content of heavy metals in the waste. A model such as EASE-WASTE is very suitable for evaluating the overall environmental consequences of different waste management strategies and technologies, and can be used for most waste material fractions existing in household waste.

Life cycle modelling of environmental impacts of application of processed organic municipal solid waste on agricultural land (EASEWASTE)

A model capable of quantifying the potential environmental impacts of agricultural application of composted or anaerobically digested source-separated organic municipal solid waste (MSW) is presented. In addition to the direct impacts, the model accounts for savings by avoiding the production and use of commercial fertilizers. The model is part of a larger model, Environmental Assessment of Solid Waste Systems and Technology (EASEWASTE), developed as a decision-support model, focusing on assessment of alternative waste management options. The environmental impacts of the land application of processed organic waste are quantified by emission coefficients referring to the composition of the processed waste and related to specific crop rotation as well as soil type. The model contains several default parameters based on literature data, field experiments and modelling by the agro-ecosystem model, Daisy. All data can be modified by the user allowing application of the model to other situations. A case study including four scenarios was performed to illustrate the use of the model. One tonne of nitrogen in composted and anaerobically digested MSW was applied as fertilizer to loamy and sandy soil at a plant farm in western Denmark. Application of the processed organic waste mainly affected the environmental impact categories global warming (0.4-0.7 PE), acidification (-0.06 (saving)-1.6 PE), nutrient enrichment (-1.0 (saving)-3.1 PE), and toxicity. The main contributors to these categories were nitrous oxide formation (global warming), ammonia volatilization (acidification and nutrient enrichment), nitrate losses (nutrient enrichment and groundwater contamination), and heavy metal input to soil (toxicity potentials). The local agricultural conditions as well as the composition of the processed MSW showed large influence on the environmental impacts. A range of benefits, mainly related to improved soil quality from long-term application of the processed organic waste, could not be generally quantified with respect to the chosen life cycle assessment impact categories and were therefore not included in the model. These effects should be considered in conjunction with the results of the life cycle assessment.

Environmental modelling of use of treated organic waste on agricultural land: a comparison of existing models for life cycle assessment of waste systems:

Modelling of environmental impacts from the application of treated organic municipal solid waste (MSW) in agriculture differs widely between different models for environmental assessment of waste systems. In this comparative study five models were examined concerning quantification and impact assessment of environmental effects from land application of treated organic MSW: DST (Decision Support Tool, USA), IWM (Integrated Waste Management, UK), THE IFEU PROJECT (Germany), ORWARE (ORganic WAste REsearch, Sweden) and EASEWASTE (Environmental Assessment of Solid Waste Systems and Technologies, Denmark). DST and IWM are life cycle inventory (LCI) models, thus not performing actual impact assessment. The DST model includes only one water emission (biological oxygen demand) from compost leaching in the results and IWM considers only air emissions from avoided production of commercial fertilizers. THE IFEU PROJECT, ORWARE and EASEWASTE are life cycle assessment (LCA) models containing more detailed land application modules. A case study estimating the environmental impacts from land application of 1 ton of composted source sorted organic household waste was performed to compare the results from the different models and investigate the origin of any difference in type or magnitude of the results. The contributions from the LCI models were limited and did not depend on waste composition or local agricultural conditions. The three LCA models use the same overall approach for quantifying the impacts of the system. However, due to slightly different assumptions, quantification methods and environmental impact assessment, the obtained results varied clearly between the models. Furthermore, local conditions (e.g. soil type, farm type, climate and legal regulation) and waste composition strongly influenced the results of the environmental assessment.

In silico and in vitro evaluation of PCR-based assays for the detection of Bacillus anthracis chromosomal signature sequences

Bacillus anthracis, the causative agent of anthrax, is a zoonotic pathogen that is relatively common throughout the world and may cause life threatening diseases in animals and humans. There are many PCR-based assays in use for the detection of B. anthracis. While most of the developed assays rely on unique markers present on virulence plasmids pXO1 and pXO2, relatively few assays incorporate chromosomal DNA markers due to the close relatedness of B. anthracis to the B. cereus group strains. For the detection of chromosomal DNA, different genes have been used, such as BA813, rpoB, gyrA, plcR, S-layer, and prophage-lambda. Following a review of the literature, an in silico analysis of all signature sequences reported for identification of B. anthracis was conducted. Published primer and probe sequences were compared for specificity against 134 available Bacillus spp. genomes. Although many of the chromosomal targets evaluated are claimed to be specific to B. anthracis, cross-reactions with closely related B. cereus and B. thuringiensis strains were often observed. Of the 35 investigated PCR assays, only 4 were 100% specific for the B. anthracis chromosome. An interlaboratory ring trial among five European laboratories was then performed to evaluate six assays, including the WHO recommended procedures, using a collection of 90 Bacillus strains. Three assays performed adequately, yielding no false positive or negative results. All three assays target chromosomal markers located within the lambdaBa03 prophage region (PL3, BA5345, and BA5357). Detection limit was further assessed for one of these highly specific assays.

Metagenomic Detection Methods in Biopreparedness Outbreak Scenarios

In the field of diagnostic microbiology, rapid molecular methods are critically important for detecting pathogens. With rapid and accurate detection, preventive measures can be put in place early, thereby preventing loss of life and further spread of a disease. From a preparedness perspective, early detection and response are important in order to minimize the consequences. During the past 2 decades, advances in next-generation sequencing (NGS) technology have changed the playing field of molecular methods. Today, it is within reach to completely sequence the total microbiological content of a clinical sample, creating a metagenome, in a single week of laboratory work. As new technologies emerge, their dissemination and capacity building must be facilitated, and criteria for use, as well as guidelines on how to report results, must be established. This article focuses on the use of metagenomics, from sample collection to data analysis and to some extent NGS, for the detection of pathogens, the integration of the technique in outbreak response systems, and the risk-based evaluation of sample processing in routine diagnostics labs. The article covers recent advances in the field, current debate, gaps in research, and future directions. Examples of metagenomic detection, as well as possible applications of the methods, are described in various biopreparedness outbreak scenarios.

The Workshop on Animal Botulism in Europe

A workshop on animal botulism was held in Uppsala, Sweden, in June 2012. Its purpose was to explore the current status of the disease in Europe by gathering the European experts in animal botulism and to raise awareness of the disease among veterinarians and others involved in biopreparedness. Animal botulism is underreported and underdiagnosed, but an increasing number of reports, as well as the information gathered from this workshop, show that it is an emerging problem in Europe. The workshop was divided into 4 sessions: animal botulism in Europe, the bacteria behind the disease, detection and diagnostics, and European collaboration and surveillance. An electronic survey was conducted before the workshop to identify the 3 most needed discussion points, which were: prevention, preparedness and outbreak response; detection and diagnostics; and European collaboration and surveillance. The main conclusions drawn from these discussions were that there is an urgent need to replace the mouse bioassay for botulinum toxin detection with an in vitro test and that there is a need for a European network to function as a reference laboratory, which could also organize a European supply of botulinum antitoxin and vaccines. The foundation of such a network was discussed, and the proposals are presented here along with the outcome of discussions and a summary of the workshop itself.

The Transcriptional Heat Shock Response of Salmonella Typhimurium Shows Hysteresis and Heated Cells Show Increased Resistance to Heat and Acid Stress

We investigated if the transcriptional response of Salmonella Typhimurium to temperature and acid variations was hysteretic, i.e. whether the transcriptional regulation caused by environmental stimuli showed memory and remained after the stimuli ceased. The transcriptional activity of non-replicating stationary phase cells of S. Typhimurium caused by the exposure to 45°C and to pH 5 for 30 min was monitored by microarray hybridizations at the end of the treatment period as well as immediately and 30 minutes after conditions were set back to their initial values, 25°C and pH 7. One hundred and two out of 120 up-regulated genes during the heat shock remained up-regulated 30 minutes after the temperature was set back to 25°C, while only 86 out of 293 down regulated genes remained down regulated 30 minutes after the heat shock ceased. Thus, the majority of the induced genes exhibited hysteresis, i.e., they remained up-regulated after the environmental stress ceased. At 25°C the transcriptional regulation of genes encoding for heat shock proteins was determined by the previous environment. Gene networks constructed with up-regulated genes were significantly more modular than those of down-regulated genes, implying that down-regulation was significantly less synchronized than up-regulation. The hysteretic transcriptional response to heat shock was accompanied by higher resistance to inactivation at 50°C as well as cross-resistance to inactivation at pH 3; however, growth rates and lag times at 43°C and at pH 4.5 were not affected. The exposure to pH 5 only caused up-regulation of 12 genes and this response was neither hysteretic nor accompanied of increased resistance to inactivation conditions. Cellular memory at the transcriptional level may represent a mechanism of adaptation to the environment and a deterministic source of variability in gene regulation.

Evaluation of Direct 16S rDNA Sequencing as a Metagenomics-based Approach to Screening Bacteria in Bottled Water

Deliberate or accidental contamination of food, feed, and water supplies poses a threat to human health worldwide. A rapid and sensitive detection technique that could replace the current labor-intensive and time-consuming culture-based methods is highly desirable. In addition to species-specific assays, such as PCR, there is a need for generic methods to screen for unknown pathogenic microorganisms in samples. This work presents a metagenomics-based direct-sequencing approach for detecting unknown microorganisms, using Bacillus cereus (as a model organism for B. anthracis) in bottled water as an example. Total DNA extraction and 16S rDNA gene sequencing were used in combination with principle component analysis and multicurve resolution to study detection level and possibility for identification. Results showed a detection level of 10(5) to 10(6) CFU/L. Using this method, it was possible to separate 2 B. cereus strains by the principal component plot, despite the close sequence resemblance. A linear correlation between the artificial contamination level and the relative amount of the Bacillus artificial contaminant in the metagenome was observed, and a relative amount value above 0.5 confirmed the presence of Bacillus. The analysis also revealed that background flora in the bottled water varied between the different water types that were included in the study. This method has the potential to be adapted to other biological matrices and bacterial pathogens for fast screening of unknown bacterial threats in outbreak situations.

Animal Botulism Outcomes in the AniBioThreat Project

Botulism disease in both humans and animals is a worldwide concern. Botulinum neurotoxins produced by Clostridium botulinum and other Clostridium species are the most potent biological substances known and are responsible for flaccid paralysis leading to a high mortality rate. Clostridium botulinum and botulinum neurotoxins are considered potential weapons for bioterrorism and have been included in the Australia Group List of Biological Agents. In 2010 the European Commission (DG Justice, Freedom and Security) funded a 3-year project named AniBioThreat to improve the EUs capacity to counter animal bioterrorism threats. A detection portfolio with screening methods for botulism agents and incidents was needed to improve tracking and tracing of accidental and deliberate contamination of the feed and food chain with botulinum neurotoxins and other Clostridia. The complexity of this threat required acquiring new genetic information to better understand the diversity of these Clostridia and develop detection methods targeting both highly specific genetic markers of these Clostridia and the neurotoxins they are able to produce. Several European institutes participating in the AniBioThreat project collaborated on this program to achieve these objectives. Their scientific developments are discussed here.