The marine environment offers both economic and scientific potential which are relatively untapped from a biotechnological point of view. These environments whilst harsh are ironically fragile and dependent on a harmonious life form balance. Exploitation of natural resources by exhaustive wild harvesting has obvious negative environmental consequences. From a European industry perspective marine organisms are a largely underutilised resource. This is not due to lack of interest but due to a lack of choice the industry faces for cost competitive, sustainable and environmentally conscientious product alternatives. Knowledge of the biotechnological potential of marine organisms together with the development of sustainable systems for their cultivation, processing and utilisation are essential. In 2010, the European Commission recognised this need and funded a collaborative RTD/SME project under the Framework 7-Knowledge Based Bio-Economy (KBBE) Theme 2 Programme 'Sustainable culture of marine microorganisms, algae and/or invertebrates for high value added products'. The scope of that project entitled 'Sustainable Production of Biologically Active Molecules of Marine Based Origin' (BAMMBO) is outlined. Although the Union is a global leader in many technologies, it faces increasing competition from traditional rivals and emerging economies alike and must therefore improve its innovation performance. For this reason innovation is placed at the heart of a European Horizon 2020 Strategy wherein the challenge is to connect economic performance to eco performance. This article provides a synopsis of the research activities of the BAMMBO project as they fit within the wider scope of sustainable environmentally conscientious marine resource exploitation for high-value biomolecules.

Polyunsaturated fatty acids (PUFAs) play an important role in human health. Due to the increased market demand, the production of PUFAs from potential alternative sources such as microalgae is receiving increased interest. The aim of this study was to perform a life cycle assessment (LCA) of the biotechnological production of eicosapentaenoic acid (EPA) from the marine diatom Phaeodactylum tricornutum, followed by the identification of avenues to improve its environmental profile. The LCA tackles two production schemes of P. tricornutum PUFAs with an EPA content of 36%: lab and pilot scales. The results at lab scale show that both the electricity requirements and the production of the extraction agent (chloroform) have significant influence on the life cycle environmental performance of microalgal EPA production. An alternative method based on hexane was proposed to replace chloroform and environmental benefits were identified. Regarding the production of EPA at pilot scale, three main environmental factors were identified: the production of the nitrogen source required for microalgae growing, the transport activities and electricity requirements. Improvement alternatives were proposed and discussed concerning: a) the use of nitrogen based fertilizers, b) the valorization of the residual algal paste as soil conditioner and, c) the anaerobic digestion of the residual algal paste for bioenergy production. Encouraging environmental benefits could be achieved if sodium nitrate was substituted by urea, calcium nitrate or ammonium nitrate, regardless the category under assessment. In contrast, minor improvement was found when valorizing the residual algal paste as mineral fertilizer, due to its overall low content in N and P. Concerning the biogas production from the anaerobic digestion, the improvement on the environmental profile was also limited due to the discrepancy between the potential energy production from the algal paste and the high electricity requirements in the culturing and extraction stages.

The current growing demand for marine resources, in particular High Value Added molecules (HVAB’s) could pose a serious threat to marine ecosystems and marine biodiversity. Instead of exploiting the natural marine resources, environmental friendly and economically sustainable ways for culturing organisms with economically interesting composition should be developed. BAMMBO addresses all key issues associated with the culture of marine organisms and will overcome these bottlenecks by designing economically sustainable and scalable culturing methodologies for industrial scale production of HVAB’s. BAMMBO will screen and identify a broad range of marine organisms (e.g. bacteria, fungi, sponges, microalgae, macroalgae and yeasts) from diverse global locations for potential as sustainable producers of HVAB’s. BAMMBO will apply various analytical methods for the extraction, purification and enrichment of targeted bioactive compounds. Moreover, a detailed life cycle analysis of the production pathways developed in the project will be undertaken to fully evaluate the sustainability of production of biologically active products from marine organisms. The EU funded FP7 project BAMMBO started in 2011. BAMMBO has brought together a multidisciplinary consortium of specialist research and SME partners. The knowledge and technologies developed during the project is transferred to relevant stakeholders in industry and the research community, as well as to policy-makers. Innovative technologies developed in the project will be demonstrated with the involvement of industry partners, and the results will be of interest not only to companies directly involved in the marine sector, but to other large scale industry players such as pharmaceutical companies with interest in added-value bioactive compounds. The laboratory

Chlorination is the most common antifouling procedure, but the search for alternatives is ongoing. Although concentrations that kill adults will also be effective against larvae, it is advisable to evaluate the toxicity of any candidate toxicant against the combatable life stage. For mussels, the earliest life stages are the most vulnerable ones and thus may require the lowest doses biocides. Since the period of larval presence is restricted to a couple of months, a pointed dosage of biocides during this period will be as effective as a continuous dosage throughout the year. This study reports on the lethal acute toxicity of sodium hypochlorite and peracetic acid to 4 h old embryos of Mytilopsis leucophaeata and Dreissena polymorpha. Chlorination was found effective against M. leucophaeata from a concentration of 0.6 mg/l onwards, even at short exposure times. Commercial peracetic acid showed to be a very good alternative in both species although the most appropriate level still has to be determined.

Mytilopsis leucophaeata (Conrad, 1831) is a brackish-water species with a typically high resistance to environmental changes (Verween, Vincx & Degraer, in press). The species invaded European waters in the nineteenth century (Nyst, 1835), but it was only when it became a severe fouling species in the 1990s (Rajagopal, Van der Velde & Jenner, 1995; Verween et al., 2005), clogging European industrial cooling-water systems, that it attracted the attention of the industrial and scientific communities. Knowledge of the species’ life history is almost completely lacking, both within its original habitat and its newly invaded environment (Verween et al., in press). Populations of the closely related Dreissena polymorpha, the zebra mussel, have an annual gametogenic cycle with one or more spawning events during summer and autumn and a high degree of gametogenic synchronization (Borcherding, 1991). Temperature is a key environmental factor governing the timing of both gametogenesis and spawning in dreissenid mussels (Borcherding, 1991; Fong et al., 1995; Ram, Fong & Garton, 1996; Nichols, 1996; Claxton & Mackie, 1998) and food availability has also been suggested as an important regulator (Borcherding 1991; Nichols, 1996; Ram et al., 1996). Based on data on the presence of larval M. leucophaeata in the water column (Verween et al., 2005), we hypothesized that M. leucophaeata has only one spawning period, and does not display the bimodal pattern often found in D. polymorpha. In addition, we investigated the correlation between gametogenesis and environmental variables such as temperature, salinity and food availability. Perhaps the most useful and reliable information concerning seasonal trends in gametogenesis is obtained from histological preparations of the gonads (Seed & Suchanek, 1992). Although laborious (probably the major reason for its limited use), this method can give detailed information about the entire reproductive cycle, including the actual time of spawning. The scale of gametogenic development for both male and female mussels was determined using an original arbitrary classification system, described by Seed (1969) for M. edulis. This system has been successfully applied to D. polymorpha (Borcherding, 1991) and was compared with M. leucophaeata to distinguish more easily between the classification stages. This classification method is widely used to identify broad trends of the sexual cycle but, as in any system of arbitrary classification, intermediate stages inevitably occur, resulting in some subjectivity. To make the classification as objective as possible, Seed (1969) used multiple criteria in the assessment of each stage (Table 1). The classification stages include the resting or spent condition (0), the gamete development period (Developing I–V) and the spawning period (Spawning IV–I). The mean gonad index (MGI), defining the breeding condition of any sample, was determined monthly by multiplying the number of individuals in each stage by its numerical score (0–V, deduced from the arbitrary rating of the stage) and by dividing the sum of these products by the total number of individuals in the sample. The resulting value ranges between 0, when all the individuals are spent or resting, to V, when all individuals are sexually mature. An increase in the MGI indicates a period of development in gonadal tissue, while a decrease in the MGI indicates a period of active spawning (Seed, 1975). All data originated from an industrial site in the harbour of Antwerp, along the Schelde River in Belgium (51821.370N; 4817.300E). The study area is situated in the oligohaline zone where M. leucophaeata causes fouling problems. Mussels were collected monthly from January to December 2006. The gonads were removed carefully from the surrounding tissues, fixed in Bouin’s fluid and embedded in paraffin (608C) and sections cut at 5–10 mm were stained with toluidin blue (Pearce, 1985). An average of 25 individuals was processed every month. The slides were analysed using a Leica DMLB microscope at 200 and 400 magnification. Mussel length averaged 13.58 mm+SE 0.11. Monthly differences in gametogenic stages of M. leucophaeata were tested by analysis of variance (ANOVA). Univariate Pearson correlation matrices were used to determine correlations between environmental variables and the MGI. Gametogenesis started in January with a slowly increasing trend in late winter, accelerating through spring and early summer until the highest MGI was reached in July–August (Fig. 1), which we interpreted as the main spawning period for M. leucophaeata. Data on larval abundances of M. leucophaeata (Verween et al., 2005) endorse this pattern, with larvae appearing in the water column from June onward, with maximal densities occurring in August. The development from an egg to a D-shaped larva, large enough to be monitored in this study, takes c. 3–4 weeks (Mackie & Schloesser, 1996). In early autumn (September–October), the MGI rapidly decreased towards the winter minimum. The analysis of the gonadal conditions confirmed this pattern of gametogenesis and provided further detail. The redevelopment of the spent or resting gonads started in January, with an average of 52.0% of the individuals under developing conditions, but still 32.0% resting or spent. In the following months, the percentage of resting gonads decreased to 0% in July and August. From June onwards, the development percentages decreased, and many individuals (69.6%) began to spawn. The percentage of spawning individuals reached a maximum in September, but values exceeded 50% until December. Only a small part of the individuals were already spent or resting in September (4.2%). From October onwards, a high percentage of individuals was spent (average 26.2%+SE 6.2), with few gonads being in development (average 8.3 %+ SE 2.5). The most significant feature of the annual spawning season of M. leucophaeata was its duration; the period over which more than 50% of the individuals were spawning extended over 6 months. Thus although only a single spawning period was detected, it lasted for a very long time from June to September. This echoes the pattern observed by Bamber & Taylor (2002) Correspondence: A. Verween; e-mail: annick.verween@ugent.be

Technische installaties zoals koelwatersystemen van grote industriele bedrijven gebruiken meestal water uit nabij gelegen zeeen of rivieren om hun elektrische processen af te koelen. Met dit water worden echter ook organismen opgepompt, die terecht komen in het koelwatersysteem en er eventueel problemen kunnen veroorzaken, zoals bv. verstoppingen van de warmtewisselaars. Dit proces wordt gedefinieerd als biofouling (Jenner et al., 1998). De meest efficiente en goedkope controle tegen mossel biofouling in industriele systemen is het gebruik van chlorering als biocide (IPPC, 2000), maar het onderzoek naar andere, meer specifieke en omgevingsvriendelijke controlemaatregelen is volop bezig. De brakwatermossel Mytilopsis leucophaeata (Conrad, 1831) veroorzaakt biofouling problemen in de koelwatersystemen van verschillende industriele sites in Noordwest Europa. Daarom werd de ecologie van M. leucophaeata in de haven van Antwerpen gebruikt als een instrument in de zoektocht naar betere, ecologisch verantwoorde oplossingen tegen zijn biofouling. Een grondige studie van de biologische processen van M. leucophaeata leverde waardevolle informatie op voor de ontwikkeling van controlemaatregelen tegen zijn biofouling. De larven van M. leucophaeata – het meest kwetsbare levensstadium – komen in de waterkolom voor tijdens een beperkte periode, zeer gelijkaardig tussen de verschillende jaren. Deze strikte timing toont aan dat om nieuwe biofouling te voorkomen een gerichte dosering van biociden tijdens de periode van larvale aanwezigheid even effectief zou zijn als een continue dosering gedurende het volledige jaar. Deze strategie werkt echter reactief, d.w.z. dat larven eerst gemonitord moeten worden voor actie kan worden ondernomen, resulterend in een arbeidsintensieve monitoringsstrategie die nodig blijft om de gerichte periode van biocide dosering te bepalen. Door de ontwikkeling van een voorspellend model waarin enkel temperatuur wordt gemeten kan de larvale aanwezigheid van M. leucophaeata voorspeld worden, waardoor proactief gehandeld kan worden; actie wordt ondernomen op het moment van de aankomst van larven in het systeem. We probeerden nog een stap verder te gaan. We zochten een letale combinatie van de heersende omgevingsvariabelen die het milieutechnisch mogelijk kon maken om M. leucophaeata larven te bestrijden zonder het gebruik van schadelijke chemicalien. Het bleek echter dat zelfs de allereerste levensstadia - 4 uur oude embryo’s - al opvallend resistent waren tegen abrupte veranderingen in temperatuur en saliniteit, wat erop wijst dat een volledig natuurlijke oplossing voor M. leucophaeata biofouling op dit moment nog niet realiseerbaar is.

The brackish water mussel, Mytilopsis leucophaeata, is a rapidly expanding invasive bivalve in Europe with great biofouling capacities. Being a typical brackish water species with very broad habitat preferences and environmental limits, adults are extremely tolerant to fluctuations in temperature and salinity. The life cycle of mussels however, consists of two phases: (1) from fertilization until larval settlement they are pelagic, only protected by a larval soft shell and (2) after settlement, the individuals become benthic and develop a hard mytiliform shell. The fact that adult mussels can close their protective valves is the major reason why they are important fouling species and are difficult to remove once settled. Therefore, vulnerability of different larval life stages of M. leucophaeata to temperature and salinity was investigated during standardized acute 48 h experimental tests. In addition, the survival limits of the most vulnerable larval life stage were determined at different temperature-salinity combinations. Results indicated that larval stages show a differential vulnerability: 4 h old embryos were more vulnerable to changes in temperature and salinity than 2 day old larvae. Maximal survival of 4 h old embryos was found at 22 degrees C at salinity 15. Surrounding this optimum, conditions stayed good for survival in a rather wide range: only salinities of 0 and 25 and temperatures below 10 degrees C or above 30 degrees C caused high embryonic mortality. Thus, even the most vulnerable larval stage in the life cycle of M. leucophaeata can be considered highly resistant to environmental conditions. Considering the broad environmental limits of adult as well as larval M. leucophaeata, we can expect this species to appear many brackish water bodies worldwide, with only colder regions potentially limiting its invasion success. (c) 2007 Elsevier B.V. All rights reserved.

Mytilopsis leucophaeata is a biofouling bivalve causing major problems in the cooling water system of BASF, Antwerp NV, Belgium, a large water-using industrial facility. This study aimed to develop a statistical model to predict the response of M. leucophaeata larvae to environmental conditions in estuarine ecosystems. Multiple logistic regression, taking into account temporal autocorrelation, was applied on a large dataset allowing the prediction of the probability of occurrence of M. leucophaeata larvae at BASF NV as a response to the environmental variables. The final model made it possible to predict larval presence in the water column solely by monitoring water temperature. The results from subsampling indicated that the model was stable. The model was tested with 2005 data, demonstrating a 98% precise prediction of the occurrence of M. leucophaeata larvae in the water column, with a sensitivity of 100% and a specificity of 97%, even though autumn 2005 was exceptionally warm, which led to an extended presence of the larvae.

For the first time, growth of Mytilopsis leucophaeata, an important European fouling species, was investigated. By means of growth cages, individual shell growth of three cohorts, with, respectively, initial shell lengths of < or =5 mm, 10 mm and 15 mm, was monitored in the harbour of Antwerp, Belgium, during 2003 - 2004. M. leucophaeata followed an oscillatory growth pattern with a single summer growing period per year (May to August). Growth decreased during wintertime, but never ceased completely. M. leucophaeata has an average growth rate of < 3-6 mm year- 1. Temperature was found to be the main environmental factor affecting growth. The von Bertalanffy growth function was used to model growth of individuals < or =5 mm, resulting in Linfinity = 16.7 mm and K= 0.56. Based on a combination of growth of all three cohorts, the hypothetical growth of an average individual mussel could be modelled over a 5-year period, resulting in a maximum length > 19 mm with a growth rate of 0.41. Its longevity (more than 5 years) and the positive effect of higher water temperatures on growth, combined with its high resistance to chlorination, provides M. leucophaeata with a high potential for severe and long-lasting biofouling

* Corresponding author Abstract A population of Rangia cuneata (G.B. Sowerby I, 1831), an estuarine bivalve, has been recorded in the harbour of Antwerp, Belgium. This species is new to the European brackish water fauna. After initially finding only a few small individuals in August 2005, R. cuneata was encountered frequently in the pipes of the cooling water system of an industrial plant from February 2006 onwards. Before this present record, R. cuneata was only known from the Gulf of Mexico and the Atlantic coast of North America.

Mytilopsis leucophaeata Conrad, 1831, the Brackish Water Mussel, is a mytiliform bivalve (Mollusca, Bivalvia, Veneroida, Dreissenidae), which produces strong byssus to attach to hard substrates. Mytilopsis leucophaeata is a typical estuarine species, and thus resistant to a wide range of oligo- to mesohaline conditions (1). The species originates from the southern coast of the U.S. to Tampico, Mexico (2). In 1835, it was first detected in Europe, in the harbour of Antwerp (3). After a period of apparent absence, M. leucophaeata is currently found along the coast of the North Sea from Germany into France and recently in Great Britain (4). Ballast water discharges from ships were identified as a major vector in the transfer of nuisance aquatic species, such as M. leucophaeata, from one area of the world to another. The fact that the species was not detected in Belgian waters over more than 50 years does not necessarily indicate the absence of M. leucophaeata along the European coast. Because of the morphological resemblance with the closely related Dreissena polymorpha, the Zebra Mussel, species-confusion may have arisen. When M. leucophaeata became an economic problem in the nineties as an important industrial fouler, attention was brought back to this relatively unknown species. Any surface exposed to untreated water provides an opportunity for the settlement and subsequent growth of organisms. Because of the high temperature and the constant supply of food and oxygen, cooling water systems are an ideal habitat for M. leucophaeata. Given these perfect conditions, settlement occurs readily and growth can be rapid until it causes fouling at the heat exchangers and the tubes in the conduits and finally leads to the failure of the operational systems. This phenomenon is known as biofouling (5). Of all organisms causing fouling in cooling systems, mussels are known to cause the most serious problems (6).