The use of pulse amplitude modulated fluorescence techniques for metal toxicity assessment in fluvial biofilms

Metal pollution in rivers is in great concern with human activities in the fluvial watershed. This thesis aims to investigate the potential use of chl-a fluorescence parameters as biomarkers of metal toxicity, and to find cause-effect relationships between metal exposures, other environmental factor (i.e. light), and functional and structural biofilm responses. This thesis demonstrates that the use of chl-a fluorescence parameters allows detect early effects on biofilms caused by zinc toxicity, both in the laboratory as in polluted rivers. In microcosm experiments, the use of chl-a fluorescence parameters allows evaluates structural changes on photosynthetic apparatus and in algal groups’ composition of biofilms long-term exposed to zinc. In order to evaluate the effects of chronic metal pollution in rivers, it is recommended the use of biofilm translocation experiments and the use of a multi-biomarker approach.

Availability and use of organic matter in stream ecosystems: the role of biofilms

La comunitat bentònica dels ecosistemes fluvials processa una gran quantitat de la matèria orgànica que arriba als rius. L'origen de les entrades de material (autòctones o al·lòctones), la seva composició química i la seva quantitat (freqüència de les entrades i concentració assolida en el riu), determinen l'estructura de la comunitat bentònica autotròfica i heterotròfica, les seves relacions tròfiques i les seves interaccions potencials (competència, sinergisme). L'objectiu d'aquesta tesi és posar de manifest la utilització de la matèria orgànica dissolta (MOD) per part dels biofilms bacterians bentònics fluvials i determinar l'eficiència del sistema fluvial en l'ús dels diferents materials que hi circulen. Amb aquesta finalitat s'han portat a terme diversos experiments, tant de camp com de laboratori, per tal de conèixer els efectes de la disponibilitat de la matèria orgànica (quantitat) i la seva qualitat (composició química i biodegradabilitat) i els efectes deguts a l'augment de temperatura de l'aigua del riu.

Distributional patterns of diatom communities in Mediterranean rivers

Aquesta tesi tracta la jerarquia i l'heterogeneïtat dels sistemes fluvials que afecten l'estructura de les comunitats bentòniques de diatomees. A nivell regional, es van buscar diferents grups de punts i les seves espècies indicadores, es va estudiar la resposta de les comunitats de diatomees als gradients ambientals, es va avaluar la utilitat de diferents índexs de diatomees i es va buscar el millor sistema de classificació per a condicions de referència. A nivell de conca, es volien definir els factors que determinen la distribució longitudinal de la diversitat de les comunitats de diatomees. Finalment, a nivell d'hàbitat es van determinar quins factors afecten les algues i els cianobacteris a aquesta escala i es va examinar la contribució relativa de l'ambient i l'espai en la distribució de la biomassa i composició d'algues i cianobacteris. Per tant, els diferents capítols d'aquesta tesi han estat desenvolupats seguint aquest esquema.

Effects of priority and emerging pollutants on river biofilms

L'activitat humana representa una de les majors causes d'entrada d'una gran varietat de substàncies en els ecosistemes fluvials. L'objectiu principal d'aquest treball es investigar els efectes que els tòxics orgànics poden exercir en els biofilms fluvials. El riu Llobregat ha estat sotmès a fortes pressions, fet que l'ha portat a uns nivells molt elevats de contaminació. En aquest estudi s'ha observat una influència dels plaguicides presents al riu en la distribució de la comunitat de diatomees, així com efectes en el biofilm a nivell funcional i estructural. Experiments amb canals experimentals han mostrat que l'herbicida diuron i el bactericida triclosan poden ocasionar una cadena d'efectes en els biofilms, incloent efectes directes i també efectes indirectes en les relacions entre els components del biofilm. Experiments amb cultius algals han mostrat que aquests tòxics, aplicats en barreja, poden tenir una major toxicitat de la prevista pels models, resultant en efectes sinèrgics.

Phytoplankton dynamic in permanent and temporary waters of Empordà Salt marshes (NE Spain)

Se ha estudiado la dinámica del fitoplancton en las lagunas costeras de Aiguamolls de l'Empordà. El fitoplancton esta sujeto principalmente al control "bottom-up", la variabilidad hidrológica y la disponibilidad de nutrientes tienen una mayor influencia en la composición y distribución de tamaños del fitoplancton, que el zooplancton. La concentración de materia orgánica disuelta es el factor ambiental más correlacionado con el crecimiento de la biomasa fitoplanctónica. Dada la proximidad entre las lagunas costeras y el mar, donde la ocurrencia de Proliferaciones de Algas Nocivas es cada vez más frecuente, se realizan un inventario general de las especies más abundantes del fitoplancton y se llevan a cabo análisis extensivos de la toxicidad. La mayoría de especies de dinoflagelados encontradas son potencialmente nocivas. Hay pocas especies en común entre el mar y las lagunas, sin embargo, existen especies productoras de PANs características de los ambientes lagunares.

Response of the benthic macroinvertebrate community to a point source in La Tordera stream (Catalonia, NE Spain)

Vam monitoritzar paràmetres físics i químics, macroinvertebrats bentònics, clorofil·la a, productors primaris i matèria orgànica durant un any (2001-2002) per examinar els efectes d'una font puntual sobre la composició taxonòmica, la estructura de la comunitat, l'organització funcional, la utilització de l'habitat i la estoquiometria al riu la Tordera (Catalunya). Aigües avall de la font puntual, concentració de nutrients, cabal i conductivitat eren majors que al tram d'aigües amunt, mentre que oxigen dissolt era menor. La densitat de macroinvertebrats era més elevada al tram d'aigües avall però la biomassa era similar als dos trams. La riquesa taxonòmica al tram de dalt era un 20% més alt que al tram de baix. Els anàlisis d'ordenació separen clarament els dos trams en el primer eix, mentre que els dos trams presentaven una pauta temporal similar en el segon eix. La similaritat entre els dos trams en composició taxonòmica, densitats i biomasses després de les crescudes d'abril i maig de 2002, indiquen que les pertorbacions del cabal poden actuar com a un mecanisme de reinici de la comunitat bentònica i jugar un paper important per a la restauració d'ecosistemes fluvials. Els dos trams presentaven una biomassa de perifiton, plantes vasculars, CPOM i FPOM similars, mentre que clorofil·la a, algues filamentoses, molses i SPOM eren majors al tram d'aigües avall. La densitat relativa de trituradors era menor sota la font puntual mentre que col·lectors i filtradors van ser afavorits. La biomassa relativa de trituradors també era menor sota la font puntual, però la biomassa de col·lectors i depredadors va augmentar. Les relacions entre densitat de grups tròfics i els seus recursos eren rarament significatives. La relació s'explicava millor amb la biomassa de macroinvertebrats. Els dos trams compartien la mateixa relació per raspadors, col·lectors i filtradors però no per trituradors i depredadors. La densitat i la biomassa de macroinvertebrats es trobaven positivament correlacionades amb la quantitat de recursos tròfics i la complexitat d'habitat, mentre que la riquesa taxonòmica es trobava negativament relacionada amb paràmetres hidràulics. La influència dels substrats inorgànics prenia menor rellevància per a la distribució dels macroinvertebrats. Els anàlisis d'ordenació mostren com les variables del microhabitat de major rellevància eren CPOM, clorofil·la a, algues filamentoses i velocitat. La cobertura de sorra només era significativa per al tram d'aigües amunt i les molses, al d'aigües avall. El número de correlacions significatives entre macroinvertebrats i les variables del microhabitat era més elevat per al tram de dalt que per al de baix, bàsicament per diferències en composició taxonòmica. La biomassa de macroinvertebrats va aportar una informació semblant a la obtinguda per la densitat. Perifiton i molses tenien uns continguts de nutrients similars en els dos trams. Els %C i %N d'algues filamentoses també eren similars en els dos trams però el %P sota la font puntual era el doble que al tram de dalt. Les relacions estoquiomètriques en CPOM, FPOM i SPOM eren considerablement menors sota la font puntual. Els continguts elementals i relacions van ser molt variables entre taxons de macroinvertebrats però no van resultar significativament diferents entre els dos trams. Dípters, tricòpters i efemeròpters presentaven una estoquiometria similar, mentre que el C i el N eren inferiors en moluscs i el P en coleòpters. Els depredadors presentaven un contingut en C i N més elevat que la resta de grups tròfics, mentre que el P era major en els filtradors. Els desequilibris elementals entre consumidors i recursos eren menors en el tram d'aigües avall. A la tardor i l'hivern la major font de nutrients va ser la BOM mentre que a la primavera i a l'estiu va ser el perifiton.

Diffusion of the synthetic pyrethroid permethrin into bed-sediments

Bed-sediments are a sink for many micro-organic contaminants in aquatic environments. The impact of toxic contaminants on benthic fauna often depends on their spatial distribution, and the fate of the parent compounds and their metabolites. The distribution of a synthetic pyrethroid, permethrin, a compound known to be toxic to aquatic invertebrates, was studied using river bed-sediments in lotic flume channels. trans/cis-Permethrin diagnostic ratios were used to quantify the photoisomerization of the trans isomer in water. Rates were affected by the presence of sediment particles and colloids when compared to distilled water alone. Two experiments in dark/light conditions with replicate channels were undertaken using natural sediment, previously contaminated with permethrin, to examine the effect of the growth of an algal biofilm at the sediment-water interface on diffusive fluxes of permethrin into the sediment. After 42 days, the bulk water was removed, allowing a fine sectioning of the sediment bed (i.e., every mm down to 5 mm and then 5-10 mm, then every 10 mm down to 50 mm). Permethrin was detected in all cases down to a depth of 5-10 mm, in agreement with estimates by the Millington and Quirk model, and measurements of concentrations in pore water produced a distribution coefficient (K-d) for each section, High K-d's were observed for the top layers, mainly as a result of high organic matter and specific surface area. Concentrations in the algal biofilm measured at the end of the experiment under light conditions, and increases in concentration in the top 1 mm of the sediment, demonstrated that algal/bacterial biofilm material was responsible for high K-d's at the sediment surface, and for the retardation of permethrin diffusion. This specific partition of permethrin to fine sediment particles and algae may enhance its threat to benthic invertebrates. In addition,the analysis of trans/cis-permethrin isomer ratios in sediment showed greater losses of trans-permethrin in the experiment under light conditions, which may have also resulted from enhanced biological activity at the sediment surface.

Presubmergence and green manure affect the transformations of nitrogen-15-labeled urea under lowland soil conditions

The effect of presubmergence and green manuring on various processes involved in [N-15]-urea transformations were studied in a growth chamber after [N-15]-urea application to floodwater. Presubmergence for 14 days increased urea hydrolysis rates and floodwater pH, resulting in higher NH3 volatilization as compared to without presubmergence. Presubmergence also increased nitrification and subsequent denitrification but lower N assimilation by floodwater algae caused higher gaseous losses. Addition of green manure maintained higher NH4+-N concentration in floodwater mainly because of lower nitrification rates but resulted in highest NH3 volatilization losses. Although green manure did not affect the KCl extractable NH4+-N from applied fertilizer, it maintained higher NH4+-N content due to its decomposition and increased mineralization of organic N. After 32 days about 36.9% (T-1), 23.9% (T-2), and 36.4% (T-3) of the applied urea N was incorporated in the pool of soil organic N in treatments. It was evident that the presubmergence has effected the recovery of applied urea N.

Land plants and DNA barcodes: short-term and long-term goals

Land plants have had the reputation of being problematic for DNA barcoding for two general reasons: (i) the standard DNA regions used in algae, animals and fungi have exceedingly low levels of variability and (ii) the typically used land plant plastid phylogenetic markers (e.g. rbcL, trnL-F, etc.) appear to have too little variation. However, no one has assessed how well current phylogenetic resources might work in the context of identification (versus phylogeny reconstruction). In this paper, we make such an assessment, particularly with two of the markers commonly sequenced in land plant phylogenetic studies, plastid rbcL and internal transcribed spacers of the large subunits of nuclear ribosomal DNA (ITS), and find that both of these DNA regions perform well even though the data currently available in GenBank/EBI were not produced to be used as barcodes and BLAST searches are not an ideal tool for this purpose. These results bode well for the use of even more variable regions of plastid DNA (such as, for example, psbA-trnH) as barcodes, once they have been widely sequenced. In the short term, efforts to bring land plant barcoding up to the standards being used now in other organisms should make swift progress. There are two categories of DNA barcode users, scientists in fields other than taxonomy and taxonomists. For the former, the use of mitochondrial and plastid DNA, the two most easily assessed genomes, is at least in the short term a useful tool that permits them to get on with their studies, which depend on knowing roughly which species or species groups they are dealing with, but these same DNA regions have important drawbacks for use in taxonomic studies (i.e. studies designed to elucidate species limits). For these purposes, DNA markers from uniparentally (usually maternally) inherited genomes can only provide half of the story required to improve taxonomic standards being used in DNA barcoding. In the long term, we will need to develop more sophisticated barcoding tools, which would be multiple, low-copy nuclear markers with sufficient genetic variability and PCR-reliability; these would permit the detection of hybrids and permit researchers to identify the 'genetic gaps' that are useful in assessing species limits.

Sewage effluent clean-up reduces phosphorus but not phytoplankton in lowland chalk stream (River Kennet, UK) impacted by water mixing from adjacent canal

Information is provided on phosphorus in the River Kennet and the adjacent Kennet and Avon Canal in southern England to assess their interactions and the changes following phosphorus reductions in sewage treatment work (STW) effluent inputs. A step reduction in soluble reactive phosphorus (SRP) concentration within the effluent (5 to 13 fold) was observed from several STWs discharging to the river in the mid-2000s. This translated to over halving of SRP concentrations within the lower Kennet. Lower Kennet SRP concentrations change from being highest under base-flow to highest under storm-flow conditions. This represented a major shift from direct effluent inputs to a within-catchment source dominated system characteristic of the upper part to the catchment. Average SRP concentrations in the lower Kennet reduced over time towards the target for good water quality. Critically, there was no corresponding reduction in chlorophyll-a concentration, the waters remaining eutrophic when set against standards for lakes. Following the up gradient input of the main water and SRP source (Wilton Water), SRP concentrations in the canal reduced down gradient to below detection limits at times near its junction with the Kennet downstream. However, chlorophyll concentrations in the canal were in an order of magnitude higher than in the river. This probably resulted from long water residence times and higher temperatures promoting progressive algal and suspended sediment generations that consumed SRP. The canal acted as a point source for sediment, algae and total phosphorus to the river especially during the summer months when boat traffic disturbed the canal's bottom sediments and the locks were being regularly opened. The short-term dynamics of this transfer was complex. For the canal and the supply source at Wilton Water, conditions remained hypertrophic when set against standards for lakes even when SRP concentrations were extremely low.

Effects of forage source and soybean and marine algal oil supplementation on milk fatty acid composition in ewes

The objective of the present studies was to determine effects of basal dietary forage source on the response of milk fatty acid composition to an oil supplement based (2:1, respectively, w/w) on soybean oil and marine algae biomass oil high in cis-9, cis-12 C18:2n − 3 and C22:6n − 3, respectively. In Study 1, Hampshire × Dorset ewes (48) were randomly assigned to one of four treatments and 12 pens in a completely randomized design blocked on the basis of lambing date and number of lambs suckled. Control rations (60:40 forage:concentrate, dry matter (DM) basis) based on alfalfa pellets (AP) or corn silage (CS) were fed from lambing. Beginning at 22 days postpartum, three pens of ewes fed AP and three pens of ewes fed CS were supplemented with oil (30 g/kg of ration DM) in place of corn meal. Average ewe DM intake (DMI) and average daily gain (ADG) were measured weekly. Milk yield and composition were measured at 42 days postpartum. DMI was lower (P<0.02) for CS and for oil, but milk yield was not affected by forage source or oil supplementation. Milk fat content was higher for oil (P<0.10) and milk protein content was higher for AP (P<0.04). Total CLA concentration (g/100 g fatty acids) increased (P<0.01) with CS and oil, and the response to oil was greater for AP (P<0.04). Similarly, total trans-C18:1 and C22:6ω−3 concentrations were higher for CS and oil, but the response to oil was greater for CS (P<0.06 and P<0.01, respectively). In Study 2, the experiment was repeated using alfalfa haylage (AH) instead of AP. The DMI decreased (P<0.05) with oil feeding, but was not affected by forage source. Milk yield was decreased by feeding oil with AH, but not by feeding oil with CS (P<0.03). Milk fat content tended to be increased by feeding oil with AH, but tended to be decreased by feeding oil with CS (P<0.08). Total CLA concentration was increased (P<0.01) for AH versus CS and by oil, and the response to oil supplementation was greater for AH (P<0.01). In contrast, total trans-C18:1 concentration was higher for CS versus AH, with a greater response to oil for CS (P<0.05). Feeding marine oil increased the C22:6ω−3 (P<0.01) concentration, and the response was greater for AH (P<0.04). To further characterize the response of milk fat composition to dietary oil in ewes, a third study used six pens of three ewes each assigned to either the control CS diet used for Study 2 or the same diet supplemented with 45 g/kg (DM basis) of the oil mixture. Feeding oil had no effect on DMI, milk yield or milk fat concentration, but again increased (P<0.001) total trans-C18:1 and C22:6ω−3 concentrations and numerically increased (114%) total CLA concentration. Milk fatty acid composition responses to supplemental vegetable and marine oils were affected by forage source. Milk trans-C18:1 concentration was higher when CS was fed in Studies 1 and 2, but the effect of forage species on CLA concentration differed between studies, which may reflect differences in diet PUFA content and consumption, as well as amounts of dietary starch and fiber consumed. Despite large increases in trans-C18:1 concentration, milk fat content was not decreased by feeding unsaturated oils to ewes, even at diet levels of 45 g/kg of ration DM.

Dietary manipulation of fatty acid composition in lamb meat and its effect on the volatile aroma compounds of grilled lamb

The effect on lamb muscle of five dietary supplements high in polyunsaturated fatty acids (PUFA) was measured. The supplements were linseed oil, fish oil, protected lipid (high in linoleic acid (C18:2 n-6) and alpha-linolenic acid (C18:3 n-3)), fish oil/marine algae (1:1), and protected lipid/marine algae (1:1). Eicosapentaenoic acid (C20:5 n-3) and docosahexaenoic acid (C22:6 n-3) were found in the highest amounts in the meat from lambs fed diets containing algae. Meat from lambs fed protected lipid had the highest levels of C18:2 n-6 and C18:3 n-3, due to the effectiveness of the protection system. In grilled meat from these animals, volatile compounds derived from n-3 fatty acids were highest in the meat from the lambs fed the fish oil/algae diet, whereas compounds derived from n-6 fatty acids were highest in the meat from the lambs fed the protected lipid diet. (C) 2004 Elsevier Ltd. All rights reserved.

Modelling fixed plant and algal dynamics in rivers: an application to the River Frome

The development of eutrophication in river systems is poorly understood given the complex relationship between fixed plants, algae, hydrodynamics, water chemistry and solar radiation. However there is a pressing need to understand the relationship between the ecological status of rivers and the controlling environmental factors to help the reasoned implementation of the Water Framework Directive and Catchment Sensitive Farming in the UK. This research aims to create a dynamic, process-based, mathematical in-stream model to simulate the growth and competition of different vegetation types (macrophytes, phytoplankton and benthic algae) in rivers. The model, applied to the River Frome (Dorset, UK), captured well the seasonality of simulated vegetation types (suspended algae, macrophytes, epiphytes, sediment biofilm). Macrophyte results showed that local knowledge is important for explaining unusual changes in biomass. Fixed algae simulations indicated the need for the more detailed representation of various herbivorous grazer groups, however this would increase the model complexity, the number of model parameters and the required observation data to better define the model. The model results also highlighted that simulating only phytoplankton is insufficient in river systems, because the majority of the suspended algae have benthic origin in short retention time rivers. Therefore, there is a need for modelling tools that link the benthic and free-floating habitats.

Technologies for biological containment of GM and Non-GM crops

International Perspective The development of GM technology continues to expand into increasing numbers of crops and conferred traits. Inevitably, the focus remains on the major field crops of soybean, maize, cotton, oilseed rape and potato with introduced genes conferring herbicide tolerance and/or pest resistance. Although there are comparatively few GM crops that have been commercialised to date, GM versions of 172 plant species have been grown in field trials in 31 countries. European Crops with Containment Issues Of the 20 main crops in the EU there are four for which GM varieties are commercially available (cotton, maize for animal feed and forage, and oilseed rape). Fourteen have GM varieties in field trials (bread wheat, barley, durum wheat, sunflower, oats, potatoes, sugar beet, grapes, alfalfa, olives, field peas, clover, apples, rice) and two have GM varieties still in development (rye, triticale). Many of these crops have hybridisation potential with wild and weedy relatives in the European flora (bread wheat, barley, oilseed rape, durum wheat, oats, sugar beet and grapes), with escapes (sunflower); and all have potential to cross-pollinate fields non-GM crops. Several fodder crops, forestry trees, grasses and ornamentals have varieties in field trials and these too may hybridise with wild relatives in the European flora (alfalfa, clover, lupin, silver birch, sweet chestnut, Norway spruce, Scots pine, poplar, elm, Agrostis canina, A. stolonifera, Festuca arundinacea, Lolium perenne, L. multiflorum, statice and rose). All these crops will require containment strategies to be in place if it is deemed necessary to prevent transgene movement to wild relatives and non-GM crops. Current Containment Strategies A wide variety of GM containment strategies are currently under development, with a particular focus on crops expressing pharmaceutical products. Physical containment in greenhouses and growth rooms is suitable for some crops (tomatoes, lettuce) and for research purposes. Aquatic bioreactors of some non-crop species (algae, moss, and duckweed) expressing pharmaceutical products have been adopted by some biotechnology companies. There are obvious limitations of the scale of physical containment strategies, addressed in part by the development of large underground facilities in the US and Canada. The additional resources required to grow plants underground incurs high costs that in the long term may negate any advantage of GM for commercial productioNatural genetic containment has been adopted by some companies through the selection of either non-food/feed crops (algae, moss, duckweed) as bio-pharming platforms or organisms with no wild relatives present in the local flora (safflower in the Americas). The expression of pharmaceutical products in leafy crops (tobacco, alfalfa, lettuce, spinach) enables growth and harvesting prior to and in the absence of flowering. Transgenically controlled containment strategies range in their approach and degree of development. Plastid transformation is relatively well developed but is not suited to all traits or crops and does not offer complete containment. Male sterility is well developed across a range of plants but has limitations in its application for fruit/seed bearing crops. It has been adopted in some commercial lines of oilseed rape despite not preventing escape via seed. Conditional lethality can be used to prevent flowering or seed development following the application of a chemical inducer, but requires 100% induction of the trait and sufficient application of the inducer to all plants. Equally, inducible expression of the GM trait requires equally stringent application conditions. Such a method will contain the trait but will allow the escape of a non-functioning transgene. Seed lethality (‘terminator’ technology) is the only strategy at present that prevents transgene movement via seed, but due to public opinion against the concept it has never been trialled in the field and is no longer under commercial development. Methods to control flowering and fruit development such as apomixis and cleistogamy will prevent crop-to-wild and wild-to-crop pollination, but in nature both of these strategies are complex and leaky. None of the genes controlling these traits have as yet been identified or characterised and therefore have not been transgenically introduced into crop species. Neither of these strategies will prevent transgene escape via seed and any feral apomicts that form are arguably more likely to become invasives. Transgene mitigation reduces the fitness of initial hybrids and so prevents stable introgression of transgenes into wild populations. However, it does not prevent initial formation of hybrids or spread to non-GM crops. Such strategies could be detrimental to wild populations and have not yet been demonstrated in the field. Similarly, auxotrophy prevents persistence of escapes and hybrids containing the transgene in an uncontrolled environment, but does not prevent transgene movement from the crop. Recoverable block of function, intein trans-splicing and transgene excision all use recombinases to modify the transgene in planta either to induce expression or to prevent it. All require optimal conditions and 100% accuracy to function and none have been tested under field conditions as yet. All will contain the GM trait but all will allow some non-native DNA to escape to wild populations or to non-GM crops. There are particular issues with GM trees and grasses as both are largely undomesticated, wind pollinated and perennial, thus providing many opportunities for hybridisation. Some species of both trees and grass are also capable of vegetative propagation without sexual reproduction. There are additional concerns regarding the weedy nature of many grass species and the long-term stability of GM traits across the life span of trees. Transgene stability and conferred sterility are difficult to trial in trees as most field trials are only conducted during the juvenile phase of tree growth. Bio-pharming of pharmaceutical and industrial compounds in plants Bio-pharming of pharmaceutical and industrial compounds in plants offers an attractive alternative to mammalian-based pharmaceutical and vaccine production. Several plantbased products are already on the market (Prodigene’s avidin, β-glucuronidase, trypsin generated in GM maize; Ventria’s lactoferrin generated in GM rice). Numerous products are in clinical trials (collagen, antibodies against tooth decay and non-Hodgkin’s lymphoma from tobacco; human gastric lipase, therapeutic enzymes, dietary supplements from maize; Hepatitis B and Norwalk virus vaccines from potato; rabies vaccines from spinach; dietary supplements from Arabidopsis). The initial production platforms for plant-based pharmaceuticals were selected from conventional crops, largely because an established knowledge base already existed. Tobacco and other leafy crops such as alfalfa, lettuce and spinach are widely used as leaves can be harvested and no flowering is required. Many of these crops can be grown in contained greenhouses. Potato is also widely used and can also be grown in contained conditions. The introduction of morphological markers may aid in the recognition and traceability of crops expressing pharmaceutical products. Plant cells or plant parts may be transformed and maintained in culture to produce recombinant products in a contained environment. Plant cells in suspension or in vitro, roots, root cells and guttation fluid from leaves may be engineered to secrete proteins that may be harvested in a continuous, non-destructive manner. Most strategies in this category remain developmental and have not been commercially adopted at present. Transient expression produces GM compounds from non-GM plants via the utilisation of bacterial or viral vectors. These vectors introduce the trait into specific tissues of whole plants or plant parts, but do not insert them into the heritable genome. There are some limitations of scale and the field release of such crops will require the regulation of the vector. However, several companies have several transiently expressed products in clinical and pre-clinical trials from crops raised in physical containment.

Sediment fingerprinting as an environmental forensics tool explaining cyanobacteria blooms in lakes

Cyanobacteria (blue-green algae) blooms in water bodies present serious public health issues with attendant economic and ecological impacts. Llyn Tegid (Lake Bala) is an important conservation and amenity asset within Snowdonia National Park, Wales which since the mid-1990s has experienced multiple toxic cyanobacteria blooms threatening the ecology and tourism-dependent local economy. Multiple working hypotheses explain the emergence of this problem, including climate change, land management linked to increased nutrient flux, hydromorphological alterations or changing trophic structure - any of which may operate individually or cumulatively to impair lake function. This paper reports the findings of a sedimentfingerprinting study using dated lake cores to explore the linkages between catchment and lake management practices and the emergence of the algal blooms problem. Since 1900 AD lake bed sedimentation rates have varied from 0.06 to 1.07 g cm−2 yr−1, with a pronounced acceleration since the early 1980s. Geochemical analysis revealed increases in the concentrations of total phosphorus (TP), calcium and heavy metals such as zinc and lead consistent with eutrophication and a rising pollution burden, particularly since the late 1970s. An uncertainty-inclusive sedimentfingerprinting approach was used to apportion the relative fluxes from the major catchment land cover types of improved pasture, rough grazing, forestry and channel banks. This showed improved pasture and channel banks are the dominant diffuse sources of sediment in the catchment, though forestry sources were important historically. Conversion of rough grazing to improved grassland, coupled with intensified land management and year-round livestock grazing, is concluded to provide the principal source of rising TP levels. Lake Habitat Survey and particle size analysis of lake cores demonstrate the hydromorphological impact of the River Dee Regulation Scheme, which controls water level and periodically diverts flow into Llyn Tegid from the adjacent Afon Tryweryn catchment. This hydromorphological impact has also been most pronounced since the late 1970s. It is concluded that an integrated approach combining land management to reduce agricultural runoff allied to improved water level regulation enabling recovery of littoral macrophytes offers the greatest chance halting the on-going cyanobacteria issue in Llyn Tegid.