Utilisation of phosphorus nutrient content in industrial scale plasmid DNA production: a waste minimisation study

Human gene therapy is currently seeing an increase in the use of plasmid (pDNA)-based vectors as a preferred choice of vehicle for delivery of the therapeutic gene into the body. In this regard, the environmental impacts of the waste streams from the possible industrial scale manufacture of pDNA require more detailed assessment. In this study, an initial assessment was made of the nutrient phosphorus (P) inputs to four fermentation processes recommended for the industrial scale production of pDNA for the purposes of gene therapy. Phosphorus inputs to each of the four selected fermentation processes ranged from approximately 60 mg l^?1 up to 3000 mg l^?1 in the fresh media. However, the spent media waste from each of the processes exhibited only a minor reduction in the phosphorus concentrations indicating minimal uptake of P by the microorganisms. This unutilised excess level of phosphorus nutrient within the waste streams poses a strong potential for environmental impact. Waste minimisation studies were undertaken on one model fermentation process with the aim of reducing unnecessary phosphorus input. An optimised media containing a 98% reduction in added P to the media was developed. This phosphorus-minimised media had little quantitative effect upon cell biomass produced and no effect upon the quantity or quality of pDNA produced, relative to the control media. The reduction in P requirement results in an overall cost savings of 12% per fermentation batch, would simplify subsequent wastewater treatment and would contribute to slowing the depletion rate of a valuable, finite natural resource.     

The water vulnerability of metro and megacities: An investigation of structural determinants

Urban areas are becoming increasingly subject to and vulnerable to water‐related natural disasters. Urban areas are a kind of socio‐ecological system wherein the human development dynamics co‐evolve with the natural dynamics. Most of the literature is focused on the impact of natural disasters on human development; we evaluate the impacts of human development on natural disasters and present an analysis of this phenomenon in the context of megacities. The approach is exploratory and begins with the construction of a database on the 595 existing megacities in the world. Multifactor analysis is then used to determine the main characteristics of these megacities. Finally, three structural components (maturity, anthropization and centrality) are identified and then correlated with data on water‐related hazards, distinguishing groups of cities according to their structure and factors of vulnerability to water‐related risks.     

How democratic innovations realise democratic goods. Two case studies of area committees in the Netherlands

Participatory planning is becoming increasingly integral to governance. Numerous planning innovations are developed which aim to increase democratic legitimacy and improve decision making. This paper critically reflects on a typical Dutch innovation: the area committee. Based on two individual case studies, we investigate whether area committees realise democratic legitimacy in existing planning practices. Analytically, we focus on four democratic goods: inclusiveness, popular control, considered judgement and transparency. Based on the interdependencies between area committees and government structures we discuss the potential and dilemmas for the area committee to contribute to the democratic legitimacy of environmental policy and rural development.     

Comparison of phosphorus forms in wet and dried animal manures by solution phosphorus-31 nuclear magnetic resonance spectroscopy and enzymatic hydrolysis

Both enzymatic hydrolysis and solution (31)P nuclear magnetic resonance (NMR) spectroscopy have been used to characterize P compounds in animal manures. In this study, we comparatively investigated P forms in 0.25 M NaOH/0.05 M EDTA extracts of dairy and poultry manures by the two methods. For the dairy manure, enzymatic hydrolysis revealed that the majority of extracted P was inorganic P (56%), with 10% phytate-like P, 9% simple monoester P, 6% polynucleotide-like P, and 18% non-hydrolyzable P. Similar results were obtained by NMR spectroscopy, which showed that inorganic P was the major P fraction (64-73%), followed by 6% phytic acid, 14 to 22% other monoesters, and 7% phosphodiesters. In the poultry manure, enzymatic hydrolysis showed that inorganic P was the largest fraction (71%), followed by 15% phytate-like P and 1% other monoesters, and 3% polynucleotide-like P. NMR spectroscopy revealed that orthophosphate was 51 to 63% of extracted P, phytic acid 24 to 33%, other phosphomonoesters 6 to 12%, and phospholipids and DNA 2% each. Drying process increased orthophosphate (8.4% of total P) in dairy manure, but decreased orthophosphate (13.3% of total P) in poultry manure, suggesting that drying treatment caused the hydrolysis of some organic P to orthophosphate in dairy manure, but less recovery of orthophosphate in poultry manure. Comparison of these data indicates that the distribution patterns of major P forms in animal manure determined by the two methods were similar. Researchers can utilize the method that best fits their specific research goals or use both methods to obtain a full spectrum of manure P characterization.     

Efficacy of Vegetated Buffers in Preventing Transport of Fecal Coliform Bacteria from Pasturelands

An experimental study was conducted in Tillamook, Oregon, USA, to quantify the effectiveness of edge-of-field vegetated buffers for reducing transport of fecal coliform bacteria (FCB) from agricultural fields amended with dairy cow manure. Installation of vegetated buffers on loamy soils dramatically reduced the bacterial contamination of runoff water from manure-treated pasturelands, but the size of the vegetated buffer was not an important determinant of bacterial removal efficiency. Only 10% of the runoff samples collected from treatment cells having vegetated buffers exhibited FCB concentrations >200 colony forming units (cfu)/100 mL (a common water quality standard value), and the median concentration for all cells containing vegetated buffers was only 6 cfu/100 mL. The presence of a vegetated buffer of any size, from 1 to 25 m, generally reduced the median FCB concentration in runoff by more than 99%. Results for FCB load calculations were similar. Our results suggest that where substantial FCB contamination of runoff occurs from manure-treated pasturelands, it might be disproportionately associated with specific field or management conditions, such as the presence of soils that exhibit low water infiltration and generate larger volumes of runoff or the absence of a vegetated buffer. Buffer size regulations that do not consider such differences might not be efficient or effective in reducing bacterial contamination of runoff.