Accelerated sludge solubilisation under sulphate reducing conditions: the effect of hydrolytic enzymes on sludge floc size distribution and EPS composition

Extracellular polymeric substances (EPS) are the construction materials for microbial aggregates such as biofilms, flocs and sludge, and greatly contribute to the structural integrity of sludge flocs in wastewater treatment processes. The loss of integrity of the sewage sludge floc is believed to be due to enhanced hydrolysis of important structural components such as lignin, protein and cellulose in the sludge floc matrix. The mechanism of enhanced sludge floc fracture, due to the action of enzymes hydrolysing these structural components, remains a key element in our understanding of how the floc integrity in systems utilising a sulphate reducing system is compromised. A range of relatively non-specific exogenous enzymes (ß-glucosidase, cellulase, proteases: trypsin, pronase E and chymotrypsin) were added to a sulphidogenic bioreactor- (containing both sulphate reducing bacteria (SRB) and a methanogenic bacterial system) and a (control) methanogenic bioreactor sample, and the effect of these enzymes on sludge floc size (diameter) distribution and EPS composition was investigated. Sludge samples from the bioreactors were examined under bright field and differential interference contrast light microscopy. Proteolytic and glucohydrolytic activity of the enzymes were monitored using standard enzyme assaying techniques, and Bradford, Somogyi-Nelson, and total carbohydrate assays were performed to establish the composition of the EPS (after extraction with 3% (v/v) glutaraldehyde and Sephacryl S-400 size exclusion chromatography). Sludge flocs present in the sulphidogenic environment of the sulphidogenic bioreactor were found to have smaller diameters than their counterparts present in the methanogenic bioreactor. The addition of hydrolytic (i.e. proteolytic and glycohydrolytic) enzymes resulted in an increased rate of matrix hydrolysis, leading to increased rates of floc fracture and deflocculation. The presence of ß-glucosidase, cellulase, and proteases naturally residing within the sludge floc was confirmed. We propose that the addition of commercially available enzymes may be prohibitively costly on a large scale, and that the activity of the enzymes naturally residing within the floc matrix be optimised or enhanced. As the bulk of the EPS was shown to be composed of mainly polysaccharides, we propose that by increasing the activity of the naturally occurring ß-glucosidases residing within the floc matrix, the process of deflocculation may be enhanced. As sulphide has been shown to increase the activity of this very important key enzyme, we propose that this is one of the contributing factors why sludge solubilisation is accelerated under sulphate reducing conditions

Microwave Gaseous Discharges

Contains reports on six research projects.Atomic Energy Commission under Contract AT(30-1) 184

Microwave Gaseous Discharges

Contains research objectives and reports on five research projects

Risk assessment-led characterisation of the SiteChar UK North Sea site for the geological storage of CO2

Risk assessment-led characterisation of a site for the geological storage of CO2 in the UK northern North Sea was performed for the EU SiteChar research project as one of a portfolio of sites. Implementation and testing of the SiteChar project site characterisation workflow has produced a ‘dry-run’ storage permit application that is compliant with regulatory requirements. A site suitable for commercial-scale storage was characterised, compatible with current and future industrial carbon dioxide (CO2) sources in the northern UK. Pre-characterisation of the site, based on existing information acquired during hydrocarbon exploration and production, has been achieved from publicly available data. The project concept is to store captured CO2 at a rate of 5 Mt per year for 20 years in the Blake Oil Field and surrounding Captain Sandstone saline aquifer. This commercial-scale storage of 100 Mt CO2 can be achieved through a storage scenario combining injection of CO2 into the oil field and concurrent water production down-dip of the field. There would be no encroachment of supercritical phase CO2 for more than two kilometres beyond the field boundary and no adverse influence on operating hydrocarbon fields provided there is pressure management. Components of a storage permit application for the site are presented, developed as far as possible within a research project. Characterisation and technical investigations were guided by an initial assessment of perceived risks to the prospective site and a need to provide the information required for the storage permit application. The emphasis throughout was to reduce risks and uncertainty on the subsurface containment of stored CO2, particularly with respect to site technical performance, monitoring and regulatory issues, and effects on other resources. The results of selected risk assessment-led site characterisation investigations and the subsequent risk reassessments are described together with their implications for the understanding of the site. Additional investigations are identified that could further reduce risks and uncertainties, and enable progress toward a full storage permit application. Permit performance conditions are presented as SiteChar-recommended useful tools for discussion between the competent authority and operator

Case study on the efficacy of a lanthanum-enriched clay (Phoslock®) in controlling eutrophication in Lake Het Groene Eiland (The Netherlands)

Lake Het Groene Eiland was created in the beginning of 2008 by construction of dikes for isolating it from the surrounding 220-ha water body. This so-called claustrum of 5 ha was treated using lanthanum-modified clay (Phoslock®) to control eutrophication and mitigate cyanobacterial nuisance. Cyanobacteria chlorophyll-a were significantly lower in the claustrum than those in the reference water body, where a massive bloom developed in summer, 2008. However, PO4-P and TP did not statistically differ in these two waters. TN and NO3-N were significantly lower in the claustrum, where dense submerged macrophytes beds developed. Lanthanum concentrations were elevated after the applications of the modified clay in the claustrum, but filterable lanthanum dropped rapidly below the Dutch standard of 10.1 μg l−1. During winter, dozens of Canada geese resided at the claustrum. Geese droppings contained an average of 2 mg PO4-P g−1 dry weight and 12 mg NH3-N g−1 dry weight and might present a growing source of nutrients to the water. Constructing the claustrum enabled unrestricted bathing in subsequent three summers, as no swimming bans had to be issued due to cyanobacteria blooms. However, the role of the modified clay in this positive outcome remains unclear, and longevity of the measures questionable.

Shop Notes

Contains a report on a research project

Geology of the Edinburgh district

This Open Report describes the geology of the Edinburgh district and surrounding area, and accompanies the BGS Sheet 32E (Edinburgh) sheets, with a Bedrock edition (2003) and a Bedrock and Superficial Deposits edition (2006). The report was originally intended as a Sheet Description, but this series was discontinued before revisions and editing of the manuscript were complete. Some authors retired from BGS, and the manuscript languished in the bottom drawer for a decade. With the new BGS strategy for Maps and Models for the 21st century (BGS 2023), it was decided that the manuscript still represented a valuable contribution to the knowledge of the geology of Edinburgh district. As a result, although published in 2025, the report represents the state of knowledge and understanding of the geology as reached in the early 2000s, and the reader needs to take into account the vintage of the report. However, this report has been updated in two aspects. The first concerns the re-interpretation of the Devonian-Carboniferous boundary in the district, and indeed in the Midland Valley as a whole. This boundary was previously thought to occur within the Kinneswood formation, but work in the Scottish Borders (Marshall et al. 2018) has shown that the Devonian-Carboniferous boundary occurs at the boundary between the Kinneswood and Ballagan formations. The relevant strata can be reliably correlated to the Edinburgh district, so the chronostratigraphy of these formations has been updated in this report. Secondly, changes in society and policy related to the green transition and decarbonisation of energy sources renders part of the Applied Geology section as out-of-date. Sections on fossil fuel sections are to a degree redundant, but have been retained for historic purposes. Comments in bold are added to provide more modern context and to reflect recent policy changes. The section on geothermal energy has been extensively updated by Alison Monaghan to match current interest and directions of research. Alison Monaghan and Teddy Reeves have tracked revisions and corrections, and have extensively edited the manuscript, so that the outstanding work of the original authors Mike Browne, David Gould, Alison Monaghan and Maxine Akhurst can finally be published

Insect pathogens as biological control agents: back to the future

The development and use of entomopathogens as classical, conservation and augmentative biological control agents have included a number of successes and some setbacks in the past 15 years. In this forum paper we present current information on development, use and future directions of insect-specific viruses, bacteria, fungi and nematodes as components of integrated pest management strategies for control of arthropod pests of crops, forests, urban habitats, and insects of medical and veterinary importance. Insect pathogenic viruses are a fruitful source of MCAs, particularly for the control of lepidopteran pests. Most research is focused on the baculoviruses, important pathogens of some globally important pests for which control has become difficult due to either pesticide resistance or pressure to reduce pesticide residues. Baculoviruses are accepted as safe, readily mass produced, highly pathogenic and easily formulated and applied control agents. New baculovirus products are appearing in many countries and gaining an increased market share. However, the absence of a practical in vitro mass production system, generally higher production costs, limited post application persistence, slow rate of kill and high host specificity currently contribute to restricted use in pest control. Overcoming these limitations are key research areas for which progress could open up use of insect viruses to much larger markets. A small number of entomopathogenic bacteria have been commercially developed for control of insect pests. These include several Bacillus thuringiensis sub-species, Lysinibacillus (Bacillus) sphaericus, Paenibacillus spp. and Serratia entomophila. B. thuringiensis sub-species kurstaki is the most widely used for control of pest insects of crops and forests, and B. thuringiensis sub-species israelensis and L. sphaericus are the primary pathogens used for medically important pests including dipteran vectors,. These pathogens combine the advantages of chemical pesticides and microbial control agents (MCAs): they are fast acting, easy to produce at a relatively low cost, easy to formulate, have a long shelf life and allow delivery using conventional application equipment and systemics (i.e. in transgenic plants). Unlike broad spectrum chemical pesticides, B. thuringiensis toxins are selective and negative environmental impact is very limited. Of the several commercially produced MCAs, B. thuringiensis (Bt) has more than 50% of market share. Extensive research, particularly on the molecular mode of action of Bt toxins, has been conducted over the past two decades. The Bt genes used in insect-resistant transgenic crops belong to the Cry and vegetative insecticidal protein families of toxins. Bt has been highly efficacious in pest management of corn and cotton, drastically reducing the amount of broad spectrum chemical insecticides used while being safe for consumers and non-target organisms. Despite successes, the adoption of Bt crops has not been without controversy. Although there is a lack of scientific evidence regarding their detrimental effects, this controversy has created the widespread perception in some quarters that Bt crops are dangerous for the environment. In addition to discovery of more efficacious isolates and toxins, an increase in the use of Bt products and transgenes will rely on innovations in formulation, better delivery systems and ultimately, wider public acceptance of transgenic plants expressing insect-specific Bt toxins. Fungi are ubiquitous natural entomopathogens that often cause epizootics in host insects and possess many desirable traits that favor their development as MCAs. Presently, commercialized microbial pesticides based on entomopathogenic fungi largely occupy niche markets. A variety of molecular tools and technologies have recently allowed reclassification of numerous species based on phylogeny, as well as matching anamorphs (asexual forms) and teleomorphs (sexual forms) of several entomopathogenic taxa in the Phylum Ascomycota. Although these fungi have been traditionally regarded exclusively as pathogens of arthropods, recent studies have demonstrated that they occupy a great diversity of ecological niches. Entomopathogenic fungi are now known to be plant endophytes, plant disease antagonists, rhizosphere colonizers, and plant growth promoters. These newly understood attributes provide possibilities to use fungi in multiple roles. In addition to arthropod pest control, some fungal species could simultaneously suppress plant pathogens and plant parasitic nematodes as well as promote plant growth. A greater understanding of fungal ecology is needed to define their roles in nature and evaluate their limitations in biological control. More efficient mass production, formulation and delivery systems must be devised to supply an ever increasing market. More testing under field conditions is required to identify effects of biotic and abiotic factors on efficacy and persistence. Lastly, greater attention must be paid to their use within integrated pest management programs; in particular, strategies that incorporate fungi in combination with arthropod predators and parasitoids need to be defined to ensure compatibility and maximize efficacy. Entomopathogenic nematodes (EPNs) in the genera Steinernema and Heterorhabditis are potent MCAs. Substantial progress in research and application of EPNs has been made in the past decade. The number of target pests shown to be susceptible to EPNs has continued to increase. Advancements in this regard primarily have been made in soil habitats where EPNs are shielded from environmental extremes, but progress has also been made in use of nematodes in above-ground habitats owing to the development of improved protective formulations. Progress has also resulted from advancements in nematode production technology using both in vivo and in vitro systems; novel application methods such as distribution of infected host cadavers; and nematode strain improvement via enhancement and stabilization of beneficial traits. Innovative research has also yielded insights into the fundamentals of EPN biology including major advances in genomics, nematode-bacterial symbiont interactions, ecological relationships, and foraging behavior. Additional research is needed to leverage these basic findings toward direct improvements in microbial control

Characterization of a fluvial aquifer at a range of depths and scales: the Triassic St Bees Sandstone Formation, Cumbria, UK

Fluvial sedimentary successions represent porous media that host groundwater and geothermal resources. Additionally, they overlie crystalline rocks hosting nuclear waste repositories in rift settings. The permeability characteristics of an arenaceous fluvial succession, the Triassic St Bees Sandstone Formation in England (UK), are described, from core-plug to well-test scale up to ~1 km depth. Within such lithified successions, dissolution associated with the circulation of meteoric water results in increased permeability (K~10−1–100 m/day) to depths of at least 150 m below ground level (BGL) in aquifer systems that are subject to rapid groundwater circulation. Thus, contaminant transport is likely to occur at relatively high rates. In a deeper investigation (> 150 m depth), where the aquifer has not been subjected to rapid groundwater circulation, well-test-scale hydraulic conductivity is lower, decreasing from K~10−2 m/day at 150–400 m BGL to 10−3 m/day down-dip at ~1 km BGL, where the pore fluid is hypersaline. Here, pore-scale permeability becomes progressively dominant with increasing lithostatic load. Notably, this work investigates a sandstone aquifer of fluvial origin at investigation depths consistent with highly enthalpy geothermal reservoirs (~0.7–1.1 km). At such depths, intergranular flow dominates in unfaulted areas with only minor contribution by bedding plane fractures. However, extensional faults represent preferential flow pathways, due to presence of high connective open fractures. Therefore, such faults may (1) drive nuclear waste contaminants towards the highly permeable shallow (< 150 m BGL) zone of the aquifer, and (2) influence fluid recovery in geothermal fields

Accelerated sludge solubilisation under sulphate reducing conditions: the effect of hydrolytic enzymes on sludge floc size distribution and EPS composition

Extracellular polymeric substances (EPS) are the construction materials for microbial aggregates such as biofilms, flocs and sludge, and greatly contribute to the structural integrity of sludge flocs in wastewater treatment processes. The loss of integrity of the sewage sludge floc is believed to be due to enhanced hydrolysis of important structural components such as lignin, protein and cellulose in the sludge floc matrix. The mechanism of enhanced sludge floc fracture, due to the action of enzymes hydrolysing these structural components, remains a key element in our understanding of how the floc integrity in systems utilising a sulphate reducing system is compromised. A range of relatively non-specific exogenous enzymes (ß-glucosidase, cellulase, proteases: trypsin, pronase E and chymotrypsin) were added to a sulphidogenic bioreactor- (containing both sulphate reducing bacteria (SRB) and a methanogenic bacterial system) and a (control) methanogenic bioreactor sample, and the effect of these enzymes on sludge floc size (diameter) distribution and EPS composition was investigated. Sludge samples from the bioreactors were examined under bright field and differential interference contrast light microscopy. Proteolytic and glucohydrolytic activity of the enzymes were monitored using standard enzyme assaying techniques, and Bradford, Somogyi-Nelson, and total carbohydrate assays were performed to establish the composition of the EPS (after extraction with 3% (v/v) glutaraldehyde and Sephacryl S-400 size exclusion chromatography). Sludge flocs present in the sulphidogenic environment of the sulphidogenic bioreactor were found to have smaller diameters than their counterparts present in the methanogenic bioreactor. The addition of hydrolytic (i.e. proteolytic and glycohydrolytic) enzymes resulted in an increased rate of matrix hydrolysis, leading to increased rates of floc fracture and deflocculation. The presence of ß-glucosidase, cellulase, and proteases naturally residing within the sludge floc was confirmed. We propose that the addition of commercially available enzymes may be prohibitively costly on a large scale, and that the activity of the enzymes naturally residing within the floc matrix be optimised or enhanced. As the bulk of the EPS was shown to be composed of mainly polysaccharides, we propose that by increasing the activity of the naturally occurring ß-glucosidases residing within the floc matrix, the process of deflocculation may be enhanced. As sulphide has been shown to increase the activity of this very important key enzyme, we propose that this is one of the contributing factors why sludge solubilisation is accelerated under sulphate reducing conditions