Including Public Perspectives in Industrial Biotechnology and the Biobased Economy

Industrial (“white”) biotechnology promises to contribute to a more sustainable future. Compared to current production processes, cases have been identified where industrial biotechnology can decrease the amount of energy and raw materials used to make products and also reduce the amount of emissions and waste produced during production. However, switching from products based on chemical production processes and fossil fuels towards “biobased” products is at present not necessarily economically viable. This is especially true for bulk products, for example ethanol production from biomass. Therefore, scientists are also turning to genetic modification as a means to develop organisms that can produce at lower costs. These include not only micro-organisms, but also organisms used in agriculture for food and feed.The use of genetic modification for “deliberate release” purposes, in particular, has met great opposition in Europe. Many industrial biotechnology applications may, due to their scale, entail deliberate releases of GM organisms. Thus, the biobased economy brings back a familiar question; is it ethically justifiable, and acceptable to citizens, to expose the environment and society to the risks associated with GM, in order to protect that same environment and to sustain our affluent way of life? For a successful innovation towards a biobased economy, its proponents, especially producers, need to take into account (take responsibility for) such issues when developing new products and processes. These issues, and how scientists can interact with citizens about them in a timely way, are further explored in projects at Delft University and Leiden University, also in collaboration with Utrecht University.     

Incorporating dormancy in dynamic microbial community models

Biogeochemical activity in natural and engineered systems depends on the abundances, functional capabilities and physiological states of the indigenous microorganisms. Typically, only a fraction of the microbial population is active at any given time. As environmental conditions change, previously active microorganisms may switch to an inactive or dormant state, while dormant ones may become active. Here, we present an extended modeling concept for the growth and decay of microorganisms that explicitly accounts for their ability to switch between active and dormant states. The equations describing the switching between physiological states are implemented into a biogeochemical reaction simulator. The model was used to reproduce published data from two laboratory experiments in which microorganisms were subjected to intermittent substrate supply or reactivated after a prolonged period of starvation. Parameter values obtained from the simulation of these experiments were used for subsequent sensitivity analyses and for the simulation of hypothetical scenarios. Results for hypothetical microbial communities consisting of two competing species exposed to periodic feeding imply that, under certain conditions, an effective dormancy-reactivation strategy may have a competitive advantage over a fast growth strategy. That is, organisms that can switch rapidly in response to fluctuations in external conditions may outcompete fast-growing organisms. Furthermore, certain combinations of growth and dormancy strategies may lead to the long-term coexistence of the two competing species. Overall, the simulated population dynamics show that dormancy is an important feature of microbial communities, which can lead to complex responses to environmental fluctuations.     

Detrimental effects of the isopod, <Emphasis Type="Italic">Edotia doellojuradoi,</Emphasis> on gill morphology and host condition of the mussel, <Emphasis Type="Italic">Mytilus edulis platensis</Emphasis>

There are many reported associations between mussels and other invertebrates, such as pea crabs, polychaetes, turbellarians and copepods, which live in their mantle cavities. The boundary between commensalism and parasitism is often indistinguishable because of insufficient knowledge or because the interaction is variable. Preliminary evidence led to a closer examination of the relationship between the mussel, Mytilus edulis platensis, and an isopod, Edotia doellojuradoi, previously described as commensalism. Monthly intertidal samples of mussels were taken from September 2004 to August 2005 at Caleta Cordova Norte (45°43′S, 67°22′W) in southern Argentina and assessed for the prevalence and abundance levels of isopods. Mussels with and without isopods were measured, examined for evidence of gill damage and their condition (soft tissue dry weight) was determined. The overall isopod prevalence in mussels was 57.9% and infestation varied with mussel length, with maximum occurence at 30.2 mm (medium-sized mussels). Experimental evidence indicated that the position of the isopod inside the mussel depended on the feeding activity of the mussel. Female isopods were observed grasping the ventral food groove of the gill demibranchs and feeding on the mucous food strand produced by the mussel. Juveniles and males were observed clustered together on the dorsal side of the single female in each occupied mussel, suggesting extended maternal care. Gill damage was observed in 58.2% of mussels at the Argentine site and was significantly associated with isopod occurrence. Categorical regression analysis showed that the most important factor associated with the degree of gill lesions was the number of male and juvenile isopods per mussel, followed by the length of female isopods and the developmental stage of juveniles. Conversely, the degree of gill damage decreased with increasing mussel length. Overall, E. doellojuradoi had a significant effect on mussel condition throughout the year, with low flesh weight in mussels with isopods, except during the austral summer and early autumn. In contrast to previous studies, which concluded that the isopod was a commensal, the present study clearly demonstrates that E. doellojuradoi is a parasite of M. edulis platensis. Other symbiotic interactions formerly classified as commensal might not be innocuous on further investigation, especially if samples are taken at multiple sites and at different times of the year.     

Monitoring anthropogenic sewage pollution on mangrove creeks in southern Mozambique: a test of Palaemon concinnus Dana, 1852 (Palaemonidae) as a biological indicator

Tropical coastal ecosystems, such as mangroves, have a great ecological and socioeconomic importance for adjacent systems and local populations, but intensive environmental impact monitoring is still lacking, mainly in East Africa. This study evaluated the potential anthropogenic disturbance on Palaemon concinnus population structure and fitness. Palaemon concinnus populations from one peri-urban (domestic sewage impacted) and two pristine mangrove creeks were studied by sampling nearly 100 shrimps per location every 15 days for 12 months. The shrimps at the peri-urban location were larger, experienced longer reproductive periods, presented higher proportion of ovigerous females and better embryo quality when compared with shrimps inhabiting pristine locations. Physiological indices (RNA/DNA ratio) were similar between shrimps at pristine and peri-urban mangroves. However, a higher level of parasitation by a Bopyridae isopod, Pseudione elongata indicated some degree of stress on the host at the peri-urban mangrove, with potential effects on the host population dynamics.     

Amendment application in a multi-contaminated mine soil: effects on soil enzymatic activities and ecotoxicological characteristics

Several amendments were tested on soils obtained from an arsenopyrite mine, further planted with Arrhenatherum elatius and Festuca curvifolia, in order to assess their ability to improve soil's ecotoxicological characteristics. The properties used to assess the effects were: soil enzymatic activities (dehydrogenase, β-glucosidase, acid phosphatase, urease, protease and cellulase), terrestrial bioassays (Eisenia fetida mortality and avoidance behaviour), and aquatic bioassays using a soil leachate (Daphnia magna immobilisation and Vibrio fischeri bioluminescence inhibition). The treatment with FeSO₄ 1 % w/w was able to reduce extractable As in soil, but increased the extractable Cu, Mn and Zn concentrations, as a consequence of the decrease in soil pH, in relation to the unamended soil, from 5.0 to 3.4, respectively. As a consequence, this treatment had a detrimental effect in some of the soil enzymatic activities (e.g. dehydrogenase, acid phosphatase, urease and cellulase), did not allow plant growth, induced E. fetida mortality in the highest concentration tested (100 % w/w), and its soil leachate was very toxic towards D. magna and V. fischeri. The combined application of FeSO₄ 1 % w/w with other treatments (e.g. CaCO₃ 1 % w/w and paper mill 1 % w/w) allowed a decrease in extractable As and metals, and a soil pH value closer to neutrality. As a consequence, dehydrogenase activity, plant growth and some of the bioassays identified those as better soil treatments to this type of multi-contaminated soil.     

Cyprodinil retention on mixtures of soil and solid wastes from wineries. Effects of waste dose and ageing

In spite of its wide-world economic relevance, wine production generates a huge amount of waste that threatens the environment. A batch experiment was designed to assess the effect of the amendment of an agricultural soil with two winery wastes (perlite and bentonite wastes) in the immobilization of cyprodinil. Waste addition (0, 10, 20, 40, and 80 Mg ha^?1) and different times of incubation of soil-waste mixtures (1, 30, and 120 days) were tested. The addition of wastes improved the soil’s ability to immobilize cyprodinil, which was significantly correlated to total C content in soil-waste mixtures. Longer incubation times decreased the cyprodinil sorption possibly due to the mineralization of organic matter but also as a consequence of the high pH values reached after bentonite waste addition (up to 10.0). Cyprodinil desorption increased as the amount of waste added to soil, and the incubation time increased. The use of these winery wastes contributes to a more sustainable agriculture preventing fungicide mobilization to groundwater.     

Localisation and quantification of elements within seeds of Cd/Zn hyperaccumulator Thlaspi praecox by micro-PIXE

Cd, Zn and Pb accumulation, spatial distribution within seeds and germinating seedlings, and seeds fitness of metal hyperaccumulating Thlaspi praecox were investigated in order to gain more knowledge on plant reproductive success at metal polluted sites. The seeds contained up to 1351 microg g-1 (dry weight) of Cd, 121 microg g-1 of Zn and 17 microg g-1 of Pb. Seed fitness was negatively influenced by seed Cd hyperaccumulation. Nevertheless, the viability of seeds was decreased by maximally 20%, indicating very efficient tolerance of the plant embryos to Cd. Localisation by micro-PIXE revealed preferential storage of most elements in the embryonic axis. Cd and Zn were preferentially localised in the epidermis of cotyledons. The restriction of seed Pb and Zn uptake and hyperaccumulation of Cd, accompanied by partitioning of Cd in the epidermal tissues of cotyledons, may enable the survival of T. praecox embryos and seedlings in Cd polluted environments.     

Removal of famoxadone, fluquinconazole and trifloxystrobin residues in red wines: effects of clarification and filtration processes

The effects of six clarification agents [egg albumin, blood albumin, bentonite + gelatine, charcoal, polyvinylpolypyrrolidine (PVPP) and silica gel] on the removal of residues of three fungicides (famoxadone, fluquinconazole and trifloxystrobin) applied directly to a racked red wine, elaborated from Monastrell variety grapes from the D.O. Region of Jumilla (Murcia, Spain) were studied. The clarified wines were filtered with 0.45 microm nylon filters to determine the influence of this winemaking process in the disappearance of fungicide residues. Analytical determination of fluquinconazole and trifloxystrobin was performed by gas chromatography with electron captor detector (ECD), while that of famoxadone using an HPLC equipped with a diode array detector (DAD). Generally, trifloxystrobin is the fungicide that is the lowest persistent one in wines, except in the egg albumin study whereas, the most persistent one is fluquinconazole. The elimination depends on the nature of the active ingredient, though the water stability in the presence of light within it has more influence than the solubility and polarity of the product itself. The most effective clarifying agents were the charcoal and PVPP. The silica gel and bentonite plus gelatine were not enough to reduce considerably the residual contents in the wine clarified with them. In general terms, filtration is not an effective step in the elimination of wine residues. The greatest removal after filtration is obtained in wines clarified with egg albumine and bentonite plus gelatine, and the lowest in those clarified with PVPP.     

Trait-mediated effects on flowers: artificial spiders deceive pollinators and decrease plant fitness

Although predators can affect foraging behaviors of floral visitors, rarely is it known if these top-down effects of predators may cascade to plant fitness through trait-mediated interactions. In this study we manipulated artificial crab spiders on flowers of Rubus rosifolius to test the effects of predation risk on flower-visiting insects and strength of trait-mediated indirect effects to plant fitness. In addition, we tested which predator traits (e.g., forelimbs, abdomen) are recognized and avoided by pollinators. Total visitation rate was higher for control flowers than for flowers with an artificial crab spider. In addition, flowers with a sphere (simulating a spider abdomen) were more frequently visited than those with forelimbs or the entire spider model. Furthermore, the presence of artificial spiders decreased individual seed set by 42% and fruit biomass by 50%. Our findings indicate that pollinators, mostly bees, recognize and avoid flowers with predation risk; forelimbs seem to be the predator trait recognized and avoided by hymenopterans. Additionally, predator avoidance by pollinators resulted in pollen limitation, thereby affecting some components of plant fitness (fruit biomass and seed number). Because most pollinator species that recognized predation risk visited many other plant species, trait-mediated indirect effects of spiders cascading down to plant fitness may be a common phenomenon in the Atlantic rainforest ecosystem.     

Insights into the pathophysiology of twin–twin transfusion syndrome

Twin–twin transfusion syndrome (TTTS) is attributed to trans-anastomotic transfusion between twins. Anastomoses are ubiquitous in monochorionic (MC) placentae, yet TTTS develops in only 15%. Although ex vivo and in vivo studies fail to identify a unique anastomotic signature, TTTS placentae are typically associated with an imbalance in unidirectional arteriovenous anastomoses with absent bidirectional anastomoses. Doppler detection of an artery-artery anastomosis reduces the chance of TTTS, whereas, in those that develop the disease, it improves stage-independent survival. Selective laser is often curative, but an increasingly recognized risk of persistent or reverse TTTS may be attributable to atypical arteriovenous anastomoses not identifiable from the chorionic plate. Simple dysvolaemia fails to explain several phenotypic features, including haematological concordancy, recipient hypertension, and reversibly absent end diastolic flow in the donor. The renin-angiotensin system is upregulated in the donor and downregulated in the recipient's kidneys, while paradoxically raised renin levels in the recipient may contribute to raised afterload along with endothelin. Although research is limited in humans by therapy and the lack of a suitable experimental model, further studies of placental and vascular pathophysiology may not only refine current treatment modalities but may also, in addition, suggest further avenues for downstream management such as genetic predisposition testing or pharmacological intervention. Copyright © 2005 John Wiley & Sons, Ltd.