Final Report Summary - BIOCORIN (NEW BIOCOATING FOR CORROSION INHIBITION IN METAL SURFACES) Executive Summary:Among the different types of corrosion, Microbial Influence Corrosion (MIC) is estimated to be involved in at least 10% of the corrosion problems of structures. MIC is a very aggressive form of corrosion with many proposed mechanisms for its prevention but these solutions include biocides and solutions, not environmentally friendly. BIOCORIN project aims to develop an innovative biomimetic and eco-efficient environmental technology for inhibiting microbial induced corrosion (MIC) produced by biofouling, through the integration of microorganisms in a sol-gel coating for metal surfaces of civil engineering structures in marine and terrestrial environments such as highway bridges, gas and liquid transmission pipelines, waterways and ports, airports and railroads.During the first part of the project, the diversity of microbial communities on metal surfaces exposed to different environment was described using metagenomic analysis. In addition, microorganisms most suitable to counteract the fouling and MIC threats in civil infrastructures affected by these phenomena under different types of conditions have been identified. Samples were collected from different environments in some partners' countries. Two candidates were selected and recognized with the ability to synthesize compounds with antifouling properties. Bioassay against three representative microorganisms showed the bioactivity of two candidates against several representative microorganisms. During the second part of the project, a coating based on sol-gel technology was designed for immobilization of living microorganisms that will prevent MIC corrosion. The sol-gel system developed has been highly biocompatible with the microorganism identified, allowing to incorporate living cells and very sensitive biomolecules. In addition, the design and formulation of sol-gel coatings has been compatible with the defined metal surfaces and must show very good adhesion to the metal surfaces. The performance of BIOCORIN developments has been characterized according to their corrosion protection properties by Electrochemical Impedance Spectroscopy (EIS). EIS-test of the sol gel coating demonstrates a very good protection against corrosion. In addition to that good general corrosion results were obtained so far using microorganism inside sol-gel coating.The technology was demonstrated and validated on three case studies with different environmental conditions selected on the basis of geographical criteria to evaluate the effects of different environmental conditions in anti-MIC capabilities of the developed technology. The environmental performance of the solution was evaluated from the life cycle analysis (LCA) perspective. Preliminary results shown that the innovative BIOCORIN coating presents significant reduction (88% of CO2 emission and 91% of methane emissions) resulted in a decrease of the impact category of Global Warming Potential (GWP) compared with an epoxy coating. BIOCORIN project presents a green alternative to traditional anti-fouling compounds, proposing a biomimetic based technology that use anti-fouling microorganisms successfully identified. For the first coating solution a coating system based on a protective coating and a porous topcoat that contains of the anti-MIC microorganisms was developed. First EIS-test of the sol gel coating demonstrates a very good protection against microorganism inside sol-gel coating. LCA preliminary results have shown that BIOCORIN is a solution with low impact in the environment.Project Context and Objectives:Corrosion is the primary means by which metals deteriorate. The economic losses due to the corrosion of infrastructures and equipment's worldwide are immense. According to experts 25 to 30 percent of the annual cost of corrosion could be avoided if optimum corrosion management practices are employed. In addition, unexpected failures due to corrosion can cause important losses, not only economic but also environmental and social, affecting public safety and security of supply and requiring costly repairs.Different types of corrosion can appear in structures or systems made of metal depending on the environmental conditions and type of metals involved. Among the different types of corrosion, Microbial Influence Corrosion (MIC) refers to the influence of microorganisms on the kinetics of corrosion processes of metals caused by their adhesion to the interfaces of the metal (usually called biofilms). These biofilms are the first stage of the biofouling generation which refers to the accumulation of unwanted material on solid surfaces, most often in an aquatic environment but also in terrestrial environment with high rates of moisture. MIC is caused by bacterial microorganisms in combination with four other environmental conditions: metals (host location), nutrients, water, and oxygen (although some types of bacteria need only very small amounts of oxygen). Some of these microorganisms are capable of producing metal dissolving metabolic by-products such as sulfuric acid, and are often identified within a classification termed sulphur reduced bacteria. Considering that given the proper conditions, most alloys including steel, cast iron, copper, and even stainless steel can be susceptible to MIC corrosion causing irreversible damages on the system in just a few years with the great economic losses and inconveniences associated. MIC is a common problem in many sectors, such as infrastructures (i.e underground pipelines, culvert piping in cities and highways), building sector (building water piping systems, HVAC systems, fire sprinkler systems), transportation (i.e marine and shipping industry, aviation industry), production (i.e onshore and offshore oil and gas industries), manufacturing (i.e chemical processing industries, metal working industry) and other industries (i.e sewage handling and treatment industry). Microbial Influenced Corrosion is estimated to be involved in at least 10% of the corrosion problems of structures (Mehana et al, 2008). In the case of subterranean pipes, it is known that at least 50% of the corrosion problems come from MIC. Several solutions have been described with relative good and acceptable anti-corrosion properties. However, as they do not present any antifouling properties, they experiment rapid deterioration rates when MIC appears, reducing their performance in a short period of time, and therefore leaving the structures' surfaces unprotected against the first stages of the corrosion mainly cause by microorganism. In addition, most of the techniques have high cost and therefore their cost-performance ratios cannot be considered as a competitive alternative for MIC protection. Microbial Induced Corrosion (MIC) is a very aggressive form of corrosion with many proposed mechanisms for its prevention but as yet there is no internationally agreed mechanism against it. In the past, antifouling coatings most commonly used to protect structures (mainly in the shipping industry) from MIC corrosion were based in Tributyltin (TBT) compounds, which were banned in 2008. Other existing solutions of antifouling and biocides include compounds and techniques such as epoxy coatings, steel treatments, copper and silver ions. Nowadays, latest research has begun to focus on environmentally friendly replacements, but up to now, with low environmental performance and durability ratios. This fact has caused an urgent demand for greener, non-toxic or low-toxicity (green Anti-Fouling agents) and longer lasting antifouling compounds and technologies.BIOCORIN project aims at developing an innovative biomimetic and eco-efficient environmental technology for inhibiting microbial induced corrosion (MIC) produced by biofouling through the integration of microorganisms in a sol-gel coating for metal surfaces of civil engineering structures in marine and terrestrial environments such as highway bridges, gas and liquid transmission pipelines, waterways and ports, airports and railroads.The scientific and technical objectives that will allow achieving the main objective of the project are described as follows:1. Selection and production of the antifouling compounds to include in the Sol-gel matrix.o Identification of the microorganism most commonly present on metal surfaces for different environments.o Evaluation of those microorganisms most active in MIC corrosion among the microorganism identified.o Identification of the antifouling microorganisms most suitable for inhibiting biofilm formation and evaluation of their anti-MIC activity.o Classification of secreted antifouling compounds (AF) and evaluation of their inhibition effects.o Quantification of the optimal concentrations of the selected antifouling microorganisms.o Identification of the inhibiting microorganisms' viability in a MIC environment.o Establishment of the required conditions for inhibitors compounds to be considered in sol-gel production.o Design of the fermentation scale-up process and production of the inhibiting microorganisms' quantities required for the tests.2. Design and production of the anti-fouling coating.o Synthesis and production of sol-gel matrix compatible with the AF microorganism selected.o Integration of the selected AF microorganisms within the sol-gel coating and study of the viability of the microorganisms within it.o Design of the final coating solution considering both laboratory test and case studies' results.o Validation of the anti-corrosion coating performance and durability test at laboratory scale. o Design of the sol-gel coating's up-scaling process and production of the quantities required for the demonstrations. o Definition and selection of the different metal surfaces where the coating will be applied.3. Analysis of the environmental aspects of the biomimetic antifouling coating.o Quantification of the developed solution's environmental profile.o Determination of the environmental and economic impacts of the technology developed by means of LCA and LCCA analysis.o Definition of eco-innovation for the developed technology.4. Demonstration and validation of the coatingo Implementation of the coating solution in three demonstration scenarios.o Evaluation of the coating performance and identification of possible drawbacks of the technology. o Assessment of the results and validation of the technology.o Development of a technical guideline for the application and maintenance of the AF coating.5. Dissemination and exploitation of the technology developed.o Deployment of an effective dissemination plan to reach the largest target audience possible.o Identification of the market opportunities and establishment of the business strategy for the further exploitation of the project's results.Project Results:The development of the BIOCORIN project has been based on five work packages:1. Microorganism and inhibitors to be included in the sol-gel matrix2. Synthesis of a sol-gel enriched matrix for corrosion inhibition3. Environmental aspects of the biomimetic developed coating4. Demonstration5. Awareness and dissemination6. Business model and exploitation7. Project ManagementThe scientific development of different solutions to control microbial induced corrosion has been researched and demonstrated within the technical tasks of WP1, WP2, WP3 and WP4. WP1: Microorganism and inhibitors to be included in the sol-gel matrixThe aim of WP1 was to identify the microorganism performing the major role in fouling an MIC and identify antifouling microorganisms, evaluate its anti-microbial activity and design the flow chart for the scale-up process. First step was the isolation and identification of those microorganisms that play a major role in fouling and MIC corrosion for different environmental conditions. Samples were collected from: dry Mediterranean environment (Madrid, Spain), Continental Mediterranean climate (Naples, Italy), the Atlantic Ocean (Galicia, Spain) and the North Sea (Harlingen, The Netherlands).Bacterial community present on the metal surface of corroded material found in the environment was analyzed using culture independent approaches and culture dependent approaches.Initially, a bibliographical study of MIC causative microorganisms was performed. This search was focused specifically on the procedures and more suitable culture media for MIC isolation. As a result, several microorganisms belonging to bacteria, fungi and yeasts were isolated from the four received samples and were identified by means of 16S and 18S ribosomal DNA (rDNA) sequencing.These first results were the basis for the search of microorganisms with anti-MIC properties. The results obtained were compared with the public databases NCBI (National Center for Biotechnology Information) and RDP (Ribosomal Database Project) in order to identify the unknown microorganism.In addition, metagenomics analyses of the four samples were also performed in order to isolate those microorganisms unable to be identified by traditional methods. 16S rDNA gene was amplified for further diversity analyses using denaturing gradient gel electrophoresis (DGGE) and sequence identification using molecular cloning and sequencing. These first results are the basis for the search of microorganisms with anti-MIC properties.Metagenomic analysis offers a comprehensive insight into the composition and dynamics of bacterial communities living in nature. In the case of metal surfaces that are exposed in the environment or used as part of an industrial process, the bacterial communities that develop on the surface, and potentially lead to MIC, follow a case specific succession process, due to the characteristics of the metal, the physiochemical and biological parameters of the environment, time of exposure, surface protection strategies, etc.The isolation and the analysis of the microorganism community on the corroded metal surfaces have demonstrated that the high diversity of MIC causing microbial communities could be present on metal surfaces due to metal type, environmental factors and the stage of corrosion. The results are important for understanding the conditions that anti-MIC bacteria incorporated into the coating will be getting in contact with. It was demonstrated that some environments have a higher potential for the development of colonizing bacteria on metal surfaces as compared with the others. Literature checking of isolated microorganisms was done in order to identify microorganisms with potential antifouling properties. As a result of this search, two candidates (Candidate A and B) were recognized with the ability to synthesize compounds with antifouling properties. In addition, the candidates appeared in all the analyzed environments. To identify and classify the secreted antifouling compounds, extracts from Candidate A and Candidate B were analyzed by means of liquid chromatography-mass spectrometry (LC/MS) and gas chromatography-mass spectrometry (GC-MS) techniques identifying a battery of antimicrobial and antifungal compounds. In parallel, the genome of both isolated candidates was sequenced and annotated and the genes for the potential anti-MIC were identified.In order to evaluate the inhibitory capacity of the candidates, tests were performed by means of bioassay analyses against three representative microorganisms (Bacillus subtilis, Micrococus luteus ATCC 9341 and Escherichia coli ESS-22-31). Bioassay results showed (Figure 1) the bioactivity of two candidates extracts on the three tested concentrations. The inhibition zones are similar for both candidates in all tested cases. Figure 1: Antibacterial activity of anti-MIC candidates selected in the presence of the inducer extracts against E. coli ESS-22-31; M. luteus ATCC 9341 and B. subtilis. ND: Non diluted extract;Survival tests were performed in order to evaluate the viability of candidates in MIC environment. Top-agarose with candidates was inoculated over non-corroded and corroded surfaces. Different samples were taken at selected times. Sampling was made by scratching or collecting the top-agarose matrix from the steel plates and, then, isolated on agar plates to check the survival rates. The results of the viable cell count can be seen in Figure 2. In the case of not corroded steel plate control the two candidates had a similar behavior. In both cases, there was a dramatic increase in its growth in the first 3 incubation days. In the case of corroded steel plates, there was a moderate increase in the concentration of microorganism during the 60 days test period. In all cases the concentration of microorganisms equals at longer times, probably due because the top-agarose matrix dried with time. Figure 2: Survival rate of Candidate A and Candidate B on steel plates.The evaluation of the optimal growth conditions and concentration to be included in the final BIOCORIN solution was also performed. Finally we worked in the scaling up process of Anti-MIC microorganisms production at both levels upstream and downstream processing to optimize its production. WP2: synthesis of a sol-gel matrix for corrosion inhibitionThe main objectives of WP2 was to develop and produce a sol-gel matrix compatible with the anti-fouling microorganism selected, identify the physical and chemical properties of the developed coatings, to integrate the selected microorganisms, to demonstrate the durability of the developed coating and to design the flow chart for the scale-up process. Based on the different approaches that were tested within the framework of BIOCORIN two coating concepts were selected as final BIOCORIN coating designs.1) BIOCS1: B1+Biofilm is composed by a first protective layer of sol gel matrix cured at high temperature (B1) and second polymers layers (Biofilm) with encapsulated microorganisms; 2) BICOS2: B2+B2S is composed by a first protective layer of sol gel matrix cured at room temperature (B2) and second sol-gel matrix layer where the microorganisms are embedded (B2S). The selected bio-coatings B1 and B2 are based on sol-gel chemistry. The sol-gel process involves the conversion of precursors into a colloidal solution called sol, which can be further converted into a solid coating. Based on the catalyst that is used for the conversion discrete particles or linear and slightly branched polymers are preferably formed. In the case of B1 and B2 linear polymers were formed by under acidic conditions. Methyltriethoxysilane (MTEOS, MeSi(OEt)3) was used as main component. To achieve a coating with sufficient adhesion to metal substrates different functionalized silanes were tested and 3-(glycidoxypropyl)triethoxysilan (GLYEO, O(CH2)3O(CH2)3Si(OEt)3) identified as most effective. Based on the structural composition of B1 and B2 both coating systems can be used as clear coats and show good to excellent anti-corrosion properties without anti-corrosion pigments or other additives. The good protective properties are based on the formation of a dense coating. In addition, the high amount of methyl-groups in the coating generates a long lasting hydrophobization effect. A substantial difference between B1 and B2 is that the latter can be cured at room temperature by adding a titanium alkoxide derived co-binder. A description of the different systems solutions developed and researcher within the WP2 context is presented below: BIOCORIN design 1 (B1)BIOCORIN design 1 is based on the high temperature curable protective coating B1. Thermal treatment is necessary to achieve a dense and compact coating. Thermal curing in heat cabinets (80-120C) or using IR-irradiation was successfully tested as curing methods. In accordance to the low viscosity of the coating material coating blade is the most suitable application method. Based on the promising barrier properties and the demonstrated performance as anti-corrosion coating B1 can be used as corrosion protection coating. To achieve sufficient adhesion to the metal surface pretreatment with a suitable primer coating is recommended. The used primer should have a thermal stability that matches with the curing parameters of B1. A major advantage of B1 is that the excellent barrier properties can be achieved of B1 as a clear coat. That means that the binder does not have to be formulated with pigments or other additives. Figure 3: Coating design of BIOCORIN solution B1.In Figure 3 the design of the BIOCORIN solution B1 is shown. The use of a suitable primer coating is recommended. To achieve good protective properties the coating thickness of B1 should be at least 80 Microm. BIOCORIN design 2 (B2Bs2) The second BIOCORIN coating design B2Bs2 is composed of a two layer system. As base coat the room temperature curable protective coating B2 can be applied. In case that high corrosion protection is required B1 should preferably be applied. A benefit of the room temperature curable system is, that application on-site and off-site is possible. In addition this solution can be useful as rehabilitation coating. Pretreatment of the steel substrate with a suitable primer coating is only recommended if the coating is permanently immersed in an aggressive medium to avoid delamination. To avoid delamination of the base coat, commercial epoxy primers were successfully tested. The combination of B2 and the selected MO's was selected as top-coat (Bs2) in the second BIOCORIN design. B2s can be used as a two component system. Like B2, Bs2 is composed of the binder and a titanium based co-binder system. In Figure 4 BIOCORIN design 2 is shown. Figure 4: Coating design of BIOCORIN solution B2The second BIOCORIN coating is designed as an anti-MIC release coating. That means that in the particular case of a coating damage the embedded organisms are released and colonize the surface to protect the steel against microbial corrosion. In Figure 5 the anti-MIC concept of the coating is shown. Figure 5: Concept and design of an anti-MIC release coatingB2 should be applied with an overall thickness of at least 80 Microm. Excellent corrosion protection was observed if the coating thickness was between 80 and 150 Microm. Bs2 can be applied in the same thickness range. Coating experiments in the demonstrator showed that brushing the preferred application technique.Artificial BiofilmThe artificial biofilm has been developed as a top-coating solution. The coating has been developed under an alternative approach to fight biofouling and MIC using microbial cells that are found in the environment and produce different antimicrobial products. The main idea behind the artificial biofilm is to simulate the strategy of biological films that are generally present in nature and unavoidably develop on the surfaces of materials that are exposed in the environment. By applying an artificial biofilm to the surface, we can change the succession process, in which the surface is being populated by unwanted microorganisms and which leads to MIC.The artificial biofilm uses several approaches to fight early stages of fouling and attachment of MIC microorganisms simultaneously:1. Viable microorganism's cells that produce anti-microbials.2. Cell capsules with entrapped anti-microbial slow-release properties.3. Extracellular matrix components with anti-microbial and anticorrosive potential.4. Integrated multi-layer structure that prevents unwanted cells to populate the surface. This coating solution can be used in combination with a bottom coating that prevents chemical deformation of the metal surface and has also other anti-corrosive properties. The biofilm is applied in layers on the surface and forms a multilayered structure. Alternative anti-MIC solutionsAlternative solutions where explored by using developed microorganisms as an antifouling additive inside commercial binder systems. For that different incorporations on Alkyd, epoxy and polyurethane systems were evaluated according to: coating integrity (if the protective properties of the coatings remain), microorganism survival, and anti-fouling/MIC effect. Coating integrity was successfully tested in epoxy coating, polyurethane systems and in alkyd systems. Microorganism survival was also corroborate showing that the viability of the microorganism is 100% in acrylic-rust converter, 100% in Polyurethane A, 11% in Polyurethane B, 92% in Epoxy A and 100% in Epoxy B. And also evidenced anti-fouling activity on alkyd system has been proved as it is presented in Figure 6 (from left to right: control, increased microorganism concentration). Plates with alkyd system and the anti-MIC identified in the project shows a correlation between the fouling attached to the plates and the microorganism concentration in the alkyd. Figure 6: Alkyd system (from left to right: control, increased microorganism concentration)Test applied to the different solutionsElectrochemical analysis on coatingsStandard EIS corrosion test (ISO 16773)Electrochemical impedance diagrams were obtained by EIS at different immersion times. The measuring cell contained a working electrode (coating final solutions developed during BIOCORIN project) of a 1 cm2 section, immersed in 3.5% NaCl saline water or seawater samples, a graphite counter electrode and a Ag/AgCl reference electrode. The measurements were carried out over a frequency range of 1 MHz to 0.1 Hz using a 20 mV amplitude sinusoidal voltage.Data interpretationThe coating evaluation has been conducted separately following equivalent electrical circuit analysis. In this section, the physical meaning of associated electrical elements has been listed.Figure 7: Coating electrical representationIn general, impedance spectroscopy data can be fitted to an equivalent circuit when enough parameters (i.e. elements in the equivalent circuit) are used. The results may, however, be physically meaningless.Therefore it is preferable to fit the data to the most probable impedance equivalent circuits (MPEC). With respect to this strategy, the circuits are based on the process changes according to a unified degradation mechanism model based on the formation of conductive pathways.Figure 8: Common circuit elements for coating analysis.The physical meaning associated to those elements can be listed as:- Re is the electrolyte resistance and can be neglected in many cases.- CC represents the coating capacitance, where the coating film behaves as a parallel plate capacitor. For this study a constant phase element (Q) was used instead. Q allows for small deviations from ideal capacitance behaviour and is characterised by two parameters, Y0 and n. Y0 can be related to coating capacitance if n = 1, while n itself represents a deviation from ideal coating behaviour due to roughness or coating heterogeneity. Capacitance values during time can give a good estimation about water uptake of the coating specimen.- WC is the Warburg impedance and describes the direct diffusion of some ionic species through the coatings (optional to get better fit).- Rpo represents conductive paths through the coating pores. Conductive paths develop through the coating due to coating degradation. Interconnected micro-pores between the inner and outer surfaces of the coating film, filled with electrolyte solution, enable electrical conduction through pores.- CDL represents the double layer capacitance of the metal surface and follows the same rules as Cc.- RCT is the charge transfer resistance or polarization resistance of which is proportional to the corrosion intensity (Icorr), also often named as corrosion rate.- W is the Warburg impedance on the corrosion reaction. When present, the corrosion reaction is controlled by diffusion.Coating resistance and water uptake (main representatives of coating performance) are exposed bellow.Coating resistanceOne of the most important factors of protection against corrosion by barrier coatings is through their resistance to ion transport. As a guideline of the protective behaviour of coatings often the following rule is used:- > 108 ? cm2 excellent- 107 - 108 ? cm2 good- 106 - 107 ? cm2 doubtful- < 106 ? cm2 badWater uptakeBased on the capacitance data, the volume fraction of water absorbed by the coating can be calculated using a simple empirical formula derived by Brasher and Kingsbury, where ?t is the volume fraction of water absorbed by the coating, Y0(t) is the CPE (capacitance) value at any immersion time. Y0(t=0) is the CPE (capacitance) value at the beginning of the measurement and ?h is the water relative dielectric constant.Electrochemical analysis on biofilm Electrochemical impedance measurements were performed on metal surface (standard R-46 steel and SS-34 stainless steel; both Q-Lab, Germany), coated by the biofilm (multilayered structure of cells and polymers) or only by the polymers composing the biofilm (without cells). To test the effects of MIC, IMMT provided several bacterial strains (isolated from the environment) that produced different secondary metabolites (like siderophores, organic acids, other iron chelators), which would bind Fe and speed up iron dissolution process. Of all the strains examined, R10 isolate proved to be the strongest bacterium to affect iron dissolution and was used in all further experiments. Experimental procedure was carried out by immersing metal pieces coated by the biofilm, polymers or no coat (control) in polyelectrolyte, which was a bacterial growth medium that boosted the production of secondary metabolites by R10 bacteria (LBg medium). The polyelectrolyte (medium LBg) promoted the dissolution of the metal and was thus used for impedance measurements.Electrochemical impedance diagrams were obtained by EIS at different immersion times. The measuring cell contained a working electrode (different steel panels with and without artificial biofilm treatment) of a 2 cm2 section, immersed in LBg medium or LBg medium containing R10 bacteria, a graphite counter electrode and a platinium reference electrode. The measurements were carried out over a frequency range of 1 MHz to 0.01 Hz using a 20 mV amplitude sinusoidal voltage.It was found that studding the relaxation peaks from plotting Cimag Vs frequency, being where Cimaf is the imaginary part of the capacitance, Z' is the real part of the impedance ? is the frequency in radians and |Z| is the modulus of the impedance, small capacitance events generated by biofilm formation where possible to identify.Figure 9: Cimag of polymer coated stainless steel in LBg medium; control (left) and in the presence of R10 bacteria.Flow system designA laboratory pipeline system has been designed to measure the durability of a coated pipe in a constant sea water flow of 1 m3 per day, during 4 months (Figure 10). Inside this period, periodical EIS measurements were executed and reported. Figure 10: Pipe flow system design.The pipe flow system (see design on Figure 11) contains 10 different measuring connections where different BIOCORIN developments have been placed. The system contains a water deposit that can be heated-cooled to stimulate biofilm formation. Constant monitoring sensors of pH and temperature are also included. Figure 11: Pipe flow systemDurability of BIOCORIN solutionSeveral standard tests were performed to explore the durability on the final BIOCORIN solution to further evaluate the properties of the coating:o Gloss (ASTM D 523): Measurement of the gloss of the coating depending on the angle of incidence.o Impact test (ISO 6272): Drop a spherical weight on the sample from increasing heights (measures mechanical resistance to a sudden deformation.o Hardness test (ISO 1522): Pendulum damping test (measures hardness as a pendulum oscilates on its surface).o Cross cut test (ISO 2409): Visual inspection of the detached coating when two perpendicular sets of cuts in the coating are performed (measures the adhesion of the coating).o EIS Testing (ISO 16773): Monitor the impedance of the coating at different times and frequencies (gives information on the thickness and integrity of the coating when submerged to salt water).o EIS Testing (ISO 17463): Guidelines for the determination of anticorrosive properties of organic coatings by accelerated cyclic electrochemical technique.o Salt spray test (ISO 9227): Subject the sample to an artificial atmosphere.o Laboratory pipe simulation test.o Accelerated EIS.Evaluation of microorganism survival In order to check the survival rate of the microorganisms to the sol-gel components and the curing process, a test to recover the microorganisms from the cured coatings was performed. Because of sol-gel properties, the coating had to be extracted from the steel plate by scratching with a metal spatula to obtain some layer fragments. Results showed that viability remains high in the sol-gel formulation before curing, even in the presence of solvents. In the case of the curing at high temperatures, the percentage of viable microorganisms dropped to concentrations close to 1%. The results also showed that the extraction of microorganisms is not enough effective, because a high percentage of microorganisms remain trapped inside the coating because of the cross-linked structure. Molecular analysis Molecular analysis based on the characterization of microbial DNA offers a powerful tool to study the composition and dynamics of microbial communities and all the processes these are involved in. Partner IMMT has developed a bioinformatics pipeline along with DNA isolation strategy to characterize how the characteristics of microbial communities change when different protective strategies are implemented (e.g. different protective coatings are applied to the metal surface) to inhibit chemical corrosion and MIC. Characteristic communities with specific microbial representatives develop on corroded surfaces and it is crucial to know how these change after the application of a protective coating and inhibition of the corrosion process. To overcome several problems with DNA isolation from intact or corroded metal surfaces (i.e. low microbial biomass, large surface area of particles, contaminants, low pH, extremely high concentration of metal ions), IMMT has developed and optimized a protocol for extraction of intact DNA from the metal surface (presented as a stand-alone molecular biology kit, product). Implementing next-generation sequencing this DNA is successfully characterized and the composition of the microbes identified. Using this approach, IMMT has demonstrated that the application of different protective solution can influence a shift in microbial community structure or not. We have demonstrated also that in some cases the changes can happen only for some low-abundance groups or conversely that the majority of the groups are changed and that only some of them persist and become enhanced. If these groups are beneficial or if they cause damage to the surface it is a strong indication of how the protective strategy is performing.Using this approach, any type of coating on any type of material can be tested and the effects of any strategies to manipulate or change the surface properties characterized. The analyzed plates, in different degree, were subjected to degradation processes due to environmental exposure and also microbial growth giving rise to emissions of volatile organic compounds. The analysis of the results revealed that B1 and B2BS2 solution worked better in the port of Gij?n than in Naples. Acetaldehyde and acetone were detected in the plate surfaces exposed in the port of Gij?n. On the one hand, the identified ketones (acetone, 2-butanone) are products of fundamental biochemical processes such as glycolysis and the Krebs cycle for nearly all organisms. On the other hand, aldehydes (acetaldehyde, 2-methyl propanal, 2-methyl butanal) have been found to be produced by microbial metabolism. It is also well know that microorganisms are able to produce sulfur containing compounds such as methyl sulfide, dimethyl sulfide, dimethyl disulfide and trimethyl disulfide. Another detected compound in the B1 plates exposed in Naples is benzaldehyde, a compound produced by Bacillus subtilis, Bacillus simplex, Serratia marcescens, Stenotrophomonas maltophilia and Bacillus weihenstephanensis. The Artificial biofilm solution that was exposed in the marine demonstrators showed the presence of acetaldehyde and acetone like the solutions B1 and B2BS2B. Finally, the best behavior was found with the B2BS2 solution placed in the port of Gij?n due to the low number of detected compounds.WP3: Environmental aspects of the biomimetic developed coatingThe overall objective of WP3 was the establishment and quantification of the environmental eco-innovation indicators of the solution, to perform the Life Cycle Analysis (LCA) and Life Cycle Cost of the solution (LCC) and to define the advantages and disadvantages of the solution. Quantification of environmental impacts The main of the environmental tests performed were:o testing the toxicity, chemical and environmental impacts of BIOCORIN formulations;o defining the advantages and disadvantages of the solutions.For the quantification of the environmental performances, the RISE Microcosm incubator system (Figure 12) has been used in order to reproduce in laboratory the natural dynamics by simulating different environmental conditions, controlling temperature, pressure, gas exchange, light and current speed.Figure 12: RISE incubator developed by Gruppo CSASeveral Lab tests have been set up and several incubations have been performed in marine water of the three demo areas (Gulf of Naples, Mediterranean Sea; Port of Harlingen, North Sea; Port of Gijon, Atlantic Ocean) as critical environment favoring metal corrosion and biofouling generation. Dissolved oxygen and temperature were chosen as the most representative parameters to test the response of the coating at the environmental conditions. In natural environments oxygen ranges from oxic to hypoxic values and, in critical conditions, it could reach anoxic values (<1 mg/l) causing strong environmental effects/impacts. For these reasons a specific tests have been set up alternating the environment conditions (aerobic/anaerobic) for 2 months in the RISE incubator system (Figure 13). Lab tests conditions were: o Condition 1 - aerobic. Coated samples and blank samples (uncoated) have been incubated in marine water at 20 C for 60 days in oxic conditions.o Condition 2 - aerobic/anaerobic. Coated samples and blank samples (uncoated) have been incubated in marine water at 26 C by alternating 3 redox conditions: i) OXIC for 7 days, ii) ANOXIC for 7 days, iii) OXIC for 45 days.The main parameters characterizing marine water environment have been monitored in the microcosms at the beginning, during and at the end of the experiments to verify significant environmental variations. Figure 13: Water characterization based on Eh, pH and rH median values calculatedIn each microcosm physico-chemical parameters (pH, redox, conductivity, dissolved oxygen, temperature) have been monitored during tests.In each sample nutrients (organic carbon, nitrogen and phosphorous compounds, biogenic silica, sulphate), inorganic pollutants (metals), organic micropollutants, microbiological parameters (bacterial count, clostrides, E. coli, total and fecal coliforms, fungi and iron bacteria) and biotoxicity have been measured. Each test included also blanks (uncoated steel) and marine water samples to compare with coated samples. Reproducibility and reliability of the results were tested.Different coating systems set by WP2, (B1: Sol-gel coating curing in the oven; B2: Sol-gel coating curing at room temperature (RT); BS2: Sol-gel coating curing at RT+Anti-MIC microorganisms; Biofilm: natural polymers + cells) have been tested in laboratory.Moreover, two commercial coating systems, each one consisting of both a primer and an antifouling ("System1" based on Cu2O and "System2" based on ZnO), were tested to compare main results and environmental effects between commercial paints and BIOCORIN formulations. The final purpose was to test the toxicity, chemical and environmental impacts of commercial anti-corrosion coatings.The main remarks of the test on commercial paints were:o A strong release of Cu associated with organic compounds (VOC) from System1 to the water column has been detected.o A strong release of Zn from System2 to the water column has been detected.o The alternating of oxic and anoxic conditions (quite common in natural environments) affected considerably the amounts of contaminants released into the water column.o These trends led to a strong biotoxicity of both coating systems.As far as BIOCORIN formulations, in Figure 14 the samples used for BS2 oxic test are shown. No rust and no delamination in water have been detected in samples at the end of the 2nd month.Figure 14: Control and BS2 samples before and after oxic testNeither biotoxicity with respect to V. fischeri, D. tertiolecta and A. salina nor critical chemical compounds were determined after 2 months.In Figure 15 the samples used for Biofilm oxic test are shown. No rust and no delamination have been detected in samples coated with Biofilm, neither in Oxic nor in Oxic/Anoxic tests carried out with the seawater from all the three demo areas. Figure 15: Control and Biofilm samples before and after oxic test.Samples coated with Biofilm released into the seawater amounts of metals significantly lower than commercial paints. Neither biotoxicity nor critical chemical compounds were determined after 2 months. The comparison between the release of key metals into the water column from Biofilm and commercial paints shows, even under Oxic/Anoxic conditions the samples coated with Biofilm release into the seawater amounts of metals always significantly lower than commercial paints. LCA and LCC The environmental and economic impact of the BIOCorin Coating Solutions (BIOCS) has been evaluated by means of a Life Cycle Assessment (LCA) and Life Cycle Costing (LCC) methodologies, respectively. In particular, two innovative solutions have been investigated, since they are the final solutions developed during the project: 1) B1+Biofilm, 2) B2+B2S3) B1+Biofilm (herein BIOCS1) is composed by a first protective layer of sol gel matrix cured at high temperature (B1) and second polymers layers (Biofilm) with encapsulated microorganisms; 4) B2+B2S (herein BIOCS2) is composed by a first protective layer of sol gel matrix cured at room temperature (B2) and second sol-gel matrix layer where the microorganisms are embedded (B2S). First, the LCA and LCC analyses have been performed on the BIOCS solutions, from raw material extraction/production to end of life (Cradle to Grave system boundary) in order to identify the most impacting (from an environmental and economic point of views) process/flow. In detail, the LCA analysis reveals that, in both solutions, the production phase is the main contributor to overall impact in the lifecycle of both of them. The LCC results of both BIOCS solutions (Table 1) demonstrate that the main difference between the two solutions investigated is the production stage associated cost, since the cost related to the other life cycle phases can be considered comparable. In detail the highest cost in BIOCS1 is provided by the production phase, while in BIOCS2 by construction phase. Table 1: LCC of BIOCS solutionsThen, these results have been compared with the environmental and economic impacts of currently available alternatives, according to a "cradle to grave" approach. Epoxy resin has been chosen as CONVentional Coating, since it represents the most common coating solution used to protect steel surface against corrosion. Kg of solution (BIOCS1, BIOCS2, CONCS) needed to cover 1 m2 of steel surface" has been chosen as functional unit. In particular, for the selected functional unit, 0,41 kg, 0,22 kg and 1,2 kg of BIOCS1, BIOCS2 and CONCS have been compared, respectively. With regard to LCA analysis, the environmental results (Figure 16) reveal that in almost all applied impact categories the BIOCS solutions have a lower impact than CONCS. In particular, BIOCS solutions present an impact 50-90% lower than CONCS in the LCA categories; it clearly appears, that the use of BIOCS may significantly reduce the environmental contribution in GWP category. Indeed BIOCS1 and BIOCS2 reduce about of 60% and 90%, respectively the greenhouse emissions in atmosphere. These results are mainly due to:o Less amount of BIOCS used to cover 1 m2 of steel surface; approximately half amount of BIOCS are used in comparison with CONCS; o Less energy consumption is requested to produce BIOCS solutions; indeed, it has been estimated that BIOCS1 and BIOCS2 reduce about of 23% and 74%, respectively, the energy consumption;o The use of several chemical materials (e.g. Propylene) NOT presented in BIOCS composition, influence the environmental results of CONCS. These chemical components are not used in the composition of the BIOCS solutions: they are substituted by natural compounds;o Less amount of BIOCS (-50%) are replaced after the coating lifetime;o The BIOCS are replaced after 7 years, two years later than CONCS. o Less amount of BIOCS (-50%) are transported and landfilled.Figure 16: LCA comparative results; kg/m2The LCC results reveal that BIOCS2 presents the best economic profile, it is approximately 50% lower than CONCS; instead the cost of BIOCS1 is comparable with CONCS (each cost is approximately 20 euro/mq).Table 2: LCC results of BIOCS and CONCS from cradle to graveConsidering the LCA and LCC results, it clearly appears that B2+B2S or BIOCS2 solution, present the best LCA/LCC-based environmental performance. Therefore, the main objective of the project consisting in the development of an innovative biomimetic and eco-efficient environmental technology for inhibiting microbial induced corrosion has been obtained.Eco-innovation Indicators The definition and quantification of the eco-indicator has been obtained from the result of the LCA and LCC performed. A cradle to grave approach has been taking into account for this analysis taking into account the raw material extraction, use of resources, production process, distribution, use and disposal of final waste (cradle to grave). Results from the demonstration activities have been used to perform the whole life cycle of the process.The results indicated that a clear reduction of environmental performance is achieved in terms of greenhouse emission, air quality and reduction of dangerous substances. Accordingly to these results, the production of BIOCORIN solutions reduces CO2 emission in 67% (BIOCS2) to 88% (BIOCS1) that means lower contribution to the global warming potential impact. A better air quality is expected taking into account the emission reduction of particulate matters, sulfur dioxide, nitrogen oxides, volatile organic compounds and aromatic hydrocarbons. Moreover, a reduction of dangerous substances is quantified as 68% to 54% less than the emitted by an epoxy coating. Marine Aquatic Eco-Toxicity Potential is focused on impacts of toxic substances on marine ecosystems and is expressed as the total emissions to the sea water and the emission of organic and inorganic compounds. The data shows that an epoxy coating emitted 12.4 mg/m2 and 20 g/m2 of organic and inorganic compound respectively while BIOCS1 and BIOCS2 emitted 8.7 mg/m2 and 1.94 mg/m2 of organic compounds and 14.0 g/m2 and 3.2 g/m2 of inorganic compounds.Waste management indicator has shown that the amount of waste is reduced in 32% for BIOCS1 and 14% for BIOCS2. This result was expected, taking into account the reduction of the use of raw material in the production process of BIOCORIN solution. Besides this, a reduction of hazardous waste management has been achieved in relation with the use of non-toxic compounds in the synthesis of BIOCORIN solution.These results and results from LCA have shown that an important improvement of the environmental performance in term of human health, ecosystem quality and climate change has been reached for BIOCORIN solution compared with epoxy coating. On the other hand, the better use of natural resources indicator for the production of BIOCORIN solution and the reduction compared with production of epoxy coating have been calculated. The percentage of reduced energy consumption compared with epoxy coating production process is 68% and 88% for BIOCS1 and BICOS2 respectively. In addition the use of energy from renewable resources has been reduced when the whole life cycle is analyzed. However, when the production phase only is taken into account, BIOCS1 solution increase the energy from renewable resources taking into account the curing process of the coating. In addition, it is important to highlight that the energy has been calculated for the production of the coating at laboratory scale. A decrease of water consumption has been estimated as 33% (BIOCS1) to 96% (BIOCS2) compared with the epoxy. The use of non-renewable resources also decreases in comparison with epoxy coatings. Taking into account the list of compounds and elements uses as non-renewable resources it has been identified that there is not any environmental hazardous substance listed in REACH regulation candidate list.WP4: DemonstrationThe aim of WP4 is to demonstrate, monitor and validate the performance of the BIOCORIN technology in real case studies. In order to fulfill this overall goal, three demonstration sites were proposed: o Marine demonstrator in the port of Harlingeno Marin demonstrator in the port of Gij?no Marine and terrestrial demonstrator in NaplesDemonstration site Harlingen The region in which the demonstrator has been installed is the Wadden Sea, connected to the North Sea. Figure 17: Detail of the situation of the Harlingen demonstratorThe location has been chosen for its proximity to the company, as well as representing a good sample of the conditions that the coating will have to endure on the North Sea with an increased biological activity. The selected dock is a steel structure used to perform maintenance and repairs on ships. It also contains floating platforms that allows easy access to the demonstrators for their installation and posterior monitoring. The panel holder was installed in the Harlingen Shipyard and fixed to the a column with ropes and ty-raps between the high and low tide lines, as the intention was for the panels to be submerged and exposed to air alternately. Furthermore, big panel structures derived from spray application tests where half submerged on Harlingen location. Figure 18: Steel coupons (left) and steel structure (right). Port of HarlingenDemonstration site Gijon The second demonstrator was installed in the port of Gij?n in Asturias on the north of Spain. Asturias, located in the north coast of Spain, is characterized by an Atlantic climate with mild temperatures throughout the year. Gij?n is a coastal village bathed by the Atlantic Ocean. Taking several factors into consideration, a dock in the new shipside of the part was selected (Figure 19). Figure 19. Location of the demonstrator in a dock of the port of Gij?nThe demonstrator was installed in the outer new dock of the port of Gij?n. The railing had enough length to cover with different formulations produced during the project (Figure 20). The dock lacked a concrete slab, so the first step was to condition the zone in order to secure the railing to the floor. A steel framework was installed before adding the concrete to enhance the endurance of the concrete slab. Figure 20: Railing painting (left), installation (middle) and detail of the rail (right).In addition to the railing treated with the coating, several steel coupons were placed in two hanging structures. These structures could be easily raised with a pulley to take samples without damaging the whole structures. The design took into account several factors: (1) the structures were hanged at the middle of the tidal range; (2) the coupons were placed in 45? angle to homogenize the sunlight exposition and to avoid impacts with the wall of the dock that could damage the coupons; (3) the poles were produced with non-corrodible material; (4) the weight at the bottom and the lateral ropes helped avoiding lateral movements and shifts of the structures. The structure is shown in Figure 21.Figure 21. Detail of the structure with the coupons (left). Detail of the structure hanging from the railing (right).The last step of the demonstrator work was the installation of a hanging carbon fiber pole. The coupons were fixed to the pole but the use of bolts was avoided to prevent galvanic corrosion. Both structures were hanged at the middle of the tidal range using a pulley (Figure 22). Figure 22: Structures hanging from the railing at the tidal rangeDemonstration site Naples Demonstrators have been installed in an area close to Naples, Italy: Bagnoli area (Figure 23). This zone presents typical Mediterranean conditions. In fact, the sea presence influences the temperature, the humidity and the precipitations. In general, this zone is characterized by dry and warm summers and wet and mild winters. Generally, the temperatures range between 20-30 C in summer and 0-15 C in the winter. Temperatures during winter only occasionally fall below the freezing point and snow is generally seldom seen.Figure 23: (a) Pontile Nord area; (b) Location of the Terrestrial and marine demonstrator Different types of structures have been realized in Naples.Marine demonstrator: -Two steel railings. These railings present the same characteristics in terms of dimensions and shape of the old dock. Each railing has two modulus of steel. They have been realized and installed near but in different time and with different two solutions. The design of steel railing is show in Figure 24.Figure 24: Railings- Several steel coupons have been placed in a hanging structure located near and in the sea. The structure is in polycarbonate material in order to not influence the steel corrosion of the coupons (Figure 25a). Moreover, a stainless cage has been realized in order to protect the plastic structure (Figure 25b), as well as the coupons, during the winter season and to avoid to lose them and to assure that the coupons could float.Figure 25: Steel couponsTerrestrial demonstrator:Two steel pipelines [D=20 mm, L=1 m] have been positioned in the ground under the Pontile Nord (Figure 26). Before the installation of the pipelines, the soil excavation was realized. In addition, in order to monitor the corrosion caused by the soil, a system of sensors realized by VLCI has been placed on the outer side of the pipeline.Figure 26: Steel pipelinesValidation of technologies Analysis of corrosion behaviorVisual inspection of demonstratorsNaples locationAll coupons immersed in Naples location, BIOCORIN solutions and commercial products showed an advanced corrosion state. This could be due to the particularly aggressive environment.Harlingen location. Chemical corrosion and biofouling was present in all the collected coupons. B2Bs2 BIOCORIN solution was heavy damaged on steel panels but showed good integrity in stainless still coupons Nevertheless, control stainless steel panels showed biofilm formation compared with B2Bs2 coupons. Alternative solutions using BIOCORIN microorganisms were also tested in Harlingen demonstrator, showing a marked antifouling increased behavior. Gij?n locationChemical corrosion is present in many of the collected coupons. B2BS2 BIOCORIN solution was heavy damaged on steel panels but showed good integrity in stainless still coupons. EIS monitoring on demostratorsPipe system in NaplesInstalled sensors in pipe grounded in Naples location allowed to have non-destructive testing on coating integrity. Bs2 samples showed excellent corrosion resistance at initial and 24h measurements, where ideal capacitance behaviour is observed. During 4 months on the soil the impedance dropped, decreasing considerably its corrosion protection.Rails in Naples and Gij?nStructures to monitor using EIS where already visual damaged during first moths of exposure. Due to this fact, all monitoring planning using EIS was reduced significantly. Stainless steel rails presented strong delamination of BIOCORIN solution, but results on stainless steel rail showed that, even severe delamination occur, non-delaminated minor sections present good coating integrity. Significant improvements have been done during the sol-gel coating development but the properties of the developed solutions did not hold the C5M demonstrator environment (coastal and offshore areas with high salinity). That made BIOCORIN sol-gel solutions (main research of BIOCORIN project) been not suitable for antifouling/mic prevention on costal environments. Alternative approaches using commercial binder systems specially formulated with BIOCORIN developed microorganisms give promising results as antifouling technology.Application guidelineThe single most important function which can influence paint performance is the quality of the surface preparation. The importance of removing oil, grease, old coatings, rust and other surface contaminants cannot be over stressed. The level of preparation should be to ISO 8501-1 grade St2-B, C or D.Bs1: Oven curing coating applicationThe coating solution BIOCORIN "B1" can be directly applied on metallic surfaces without any reduction. If viscosity of the coating solution is too high thinner (ethanol with water content below 0.5 %) can be used to adjust the viscosity. Excessive reduction of the coating solution can affect film build, appearance, and adhesion as well as the curing behaviour. Ethanol with water content above 1% may reduce coating performance. Ratio of coating material to thinner should not be above 1:20.It is important to apply a wet film thickness between 75 and 150 microns. This should be measured using a comb gauge. Microorganism preparation should be added at least 30 minutes before coating is applied. For the application, brush or spray application is recommended. When sprayed, it is possible to apply B1 BIOCORIN solution in one coat of up to 75 micron thickness. However, a thickness of 100 microns is recommended as this affords the optimum compromise between film build, finish and thickness control. After spraying, immediately flush out the mixed material from the static mixer and spray line (whip end) with ethanol.The applied coating should be cured after coating as soon as possible. The curing temperature should be between 80 C and 140 C. At 100 C the curing time is 30 minutes. If curing time is less than 20 minutes the applied coating can be damaged very easy and the surface may stay sticky. After this curing time corrosion protection can be decreased. Longer curing times than one hour may cause cracking.B2BS2: RT curing coating applicationThe BIOCORIN B2BS2 solution is a three component system. All components have to be mixed before the coating material can be applied on metallic surfaces. The first step requires the thorough mixing of solution A and B to build a well crosslinked material with good adhesion properties to the metal. Recommended ratio of A:B is 6:1. Mixing should be performed in a closed flask for safety reason. Both components should be stirred or shaked vigorously after mixing for at least 1-2 minutes. After this, the mixture is allowed to react for 60 minutes before solution C is added. The curing performs best if the ratio A: C is 1:0.04 (example: 1l solution A: 0.04 l solution C). Both components should be mixed vigorously for 1-2 minutes. If the two components are mixed in an open flask or container volatile components may evaporate. Microorganism preparation should be added right after solution B was added to the solution A. For the application, brush or spray application is recommended. When sprayed, it is possible to apply B2BS2 BIOCORIN solution one coat of up to 75 micron thickness. However, a thickness of 100 microns is recommended as this affords the optimum compromise between film build, finish and thickness control. After spraying, immediately flush out the mixed material from the static mixer and spray line (whip end) with ethanol.Evaporation of volatile components can have negative influence on the film formation and coating properties, appearance, and curing behaviour. After all components are mixed, the coating solution has to be used within 30 minutes. Dry touch film (75micron) is reached at 5 min. Dried to over coat is advised at 1h.Potential Impact:Potential impact: BIOCORIN project has developed different coating solutions ideas for biofouling inhibition in infrastructures sector. Although further tests are needed to generate a longer lasting coating, this innovative solution represents the first step of a more economic coating with better environmental properties compared without using organic or inorganic chemical compounds.Environmental ImpactImprovements have been reached by means of reduction of Green House Gasses emissions from the substitution of chemical compounds by natural compounds in anti-MIC and antifouling coatings, resulted in a decrease of the impact category of Global Warming Potential (GWP). The clear reduction of environmental performance is represented by the reduction of CO2 emission in 67% to 88%, for different BICORIN solutions, lower emission of particulate matters, sulfur dioxide, nitrogen oxides, volatile organic compounds and aromatic hydrocarbons. In addition, a reduction of dangerous substances has been obtained due to the use of microorganism as antifouling and anti- MIC agent into a sol -gel/biofilm matrix. These microorganisms substitute the current chemical compounds used as biocides. The reduction of dangerous substances has been quantified from 68% to 54% less than the emitted by conventional coatings using in for anti-fouling corrosion. On the other hand, marine aquatic eco-toxicity potential focus on impacts of toxic substances on marine ecosystems has been reduced by lower emission of inorganic and organic compounds of BIOCORIN solutions and because these coatings developed do not present biotoxicity against different microorganism tested. A better use of resources has been reached by reducing resource consumption by reducing the amount BIOCORIN coating applied on m2 of steel surface (0.2 to 0.4 of BIOCORIN coating can be substitute 1.2 Kg of conventional resin). A better use of resources means a decrease in waste generation. In addition, there is a reduction of energy consumption during the BIOCORIN production process, from the synthesis of sol gel/biofilm matrix to the final development of products. This reduction of energy consumption has been estimated from 68% to 88% compared with traditional coatings. Market impactBIOCORIN project has been focused on demonstrating the solution on the infrastructure sector (port facilities), water and sewage (demonstration on a sewage pipeline) sector, therefore addressing 20% of the total European market share, which would mean 246 million € in 2010 if the technology was already in place and it will suppose a market potential of 312 million € in 2015. However, BIOCORIN solution, once enhanced and demonstrated, could be easily transferred to other market sectors such as the marine or the oil/gas transmission industry, thus increasing the initial market’s share foreseen. Additionally, some of the developments like artificial biofilms and the formulation of new sol-gels can be used in other industries.A patent analysis has been perform during the project and this analysis reveals that a small number of patents (no more than 5 anti-MIC patents and less than half of them use microorganisms) worldwide are dedicated to process similar to BIOCORIN. Also, almost no patents for sol-gel technology are used in the treatment against corrosion. That states the innovation of the BIOCORIN solution and the high possibility of go into the market with patent developments.With respect to the market impact an estimation of the final cost of the developed technologies was calculated taking into account the whole life cycle. This estimation, by analyzing the whole life cycle of the solution, has been calculated as an increase of 3% in the cost of one of the BIOCORIN solution (BICOS1) and a reduction of 28% of BIOCORIN solution (BIOCS2) compared with the cost of epoxy resin (22.71 €/m2). Economic impactThe biomimetic technology developed in BIOCORIN have the capability of performing an important economic impact once totally developed. As it is clearly known, corrosion has a huge economic and environmental impact on virtually all facets of the world’s infrastructure, from highways, bridges, and buildings to oil and gas, chemical processing, and water and wastewater systems. In addition, to causing severe damage and threats to public safety, corrosion disrupts operations and requires extensive repair and replacement of failed assets. Even considering that approximately 60% of the corrosion is unavoidable and the fact that Microbial Induced Corrosion is considered to be between 10 and 50% of the total corrosion in different environments, the potential impact of this kind of technology should be considerable.Main dissemination activities The dissemination effort has been undertaken within the BIOCORIN project to ensure proper dissemination to the target stakeholders. Several communication materials have been produced (dissemination tool), with the purpose of ensuring the optimal coverage of the available communication media. In detail:o Internet website;o Pages on Twitter, Facebook and LinkedIn;o Newsletter;o Workshops;o On site visits;o Participation to promotional events;o Publication of project results. In addition, each partner has attended public external events, in order to provide presentations concerning project objectives and achievements and prepared articles addressed to specialized publications, seminars or conferences.Internet websiteThe website is the central feature for collecting information that revolves around the project and it is the reference point for the community and for the dissemination (public access) and exchange of information, documents, etc (private access). The BIOCORIN Web Portal has been continuously updated within the project lifetime, for both internal purposes (project meetings) and public events. It has been registered into the main public search engines (among whom the most important and used is represented by Google), which guarantees the best possible visibility. Public area: The content of the website has been organized into different pages, continuously updated. This part contents: o The website Home Page, providing basic information about the project such as its scope, its main aim and achieved results, and its time frame;o Project page dedicated to the description of the project and its objectives;o Partners pages dedicated to the description the presentation of the different partners;o News and events pages dedicated to the description the main news and eventso Download area page, to publicly available information and documents (e.g. project deliverables, newsletters, etc.)Private area: It is accessible only through a login process and primarily reserved to Consortium members. However, access for users willing to obtain more information on the project, or to cooperate with BIOCORIN activities, has been also ensured. The private area represents the document repository of the project, in which all relevant documents and information have been stored.Pages on Twitter, Facebook and LinkedIn;A BIOCORIN Facebook has been created, as well as a LinkedIn group and a Twitter account. The main objective has been to illustrate the project results in informal communication level. These pages are continuously updated within the project lifetime.NewsletterTwo newsletters have been submitted during the project. They described the project progresses related to the activities performed in the respective considered period. Both the newsletters have been distributed, in electronic format (PDF format), to the members of the Target Groups. The newsletters were also made available on the webpage as download. Also, paper copies of each issue have been prepared and distributed during the workshops and other relevant public events.WorkshopsTwo workshops have been organized during the project: o A national workshop at month 24 of the project: The national workshop (26th February 2014) was the first of workshops associated with the project. The workshop was hosted by STRESS, partner of the BIOCORIN project in the ACEN conference room. It was mainly dedicated to the progresses related to the activities performed in the two years of the project; moreover a discussion among representatives of key stakeholders, among the audience, and among the members of the project has been provided. In fact, national, European stakeholders and experts related to the field of BIOCORIN project (e.g. chemicals, biologists, engineers, students, etc) have participated in this workshop. o An European workshop at month 40 of the project: The European workshop (30th July 2015) was organized in Amsterdam. It was mainly dedicated to show the advantages of Biocorin solution developed within the project and the progresses related to the activities performed. All the partners presented the different topics and the different solutions developed in BIOCORIN. In addition, some experts in MIC, coatings, fouling participated as speakers. Moreover were invited European stakeholders, coating industry and experts related to the field (e.g. chemicals, biologists, engineers, etc). A discussion among representatives of key stakeholders, among the audience, and among the members of the project has been provided. Several dissemination materials were distributed.Networking with other projectsACCIONA keeps contact with FP7 projects: ? BYEFOULING: Low-toxic cost-efficient environment-friendly antifouling materials.? SEAFRONT: Innovative antifouling materials for maritime applications. ? MICSIPE: Microbiologically Induced Corrosion of Steel Structures in Port Environment: improving prediction and diagnosis of ALWC.? AERTOS.ERA-NET: Low Cost Corrosion Protection for Offshore Wind Turbines and Structures LCCP-OW.? NANOMAR: Nanocontainer-Based Active Coatings for Maritime Applications.? STEELCOAT: Development of Green Anticorrosion Coatings for Steel Protection Based on Environmentally Friendly Nanoparticles and Conducting Polymers.ACCIONA and Gruppo CSA participated in the "Satellite meeting on novel antifouling strategies" held on 5/06/2015 in Santiago de Compostela (Spain) and organized by BYEFOULING project. Furthermore, scientific coordinator of Byefouling project and one partner of Seafront project participated at the BIOCORIN Final Workshop in Amsterdam. On site visitsDuring the third period several visits to the laboratories and on real cases to show the progress in BIOCORIN project have been organized.ACCIONA has organized visits to their laboratories to show the progress in BIOCORIN project to the following visitors:? Australian Trade Commission (AUSTRADE).? MBA Program from Cox School of Business (Texas, USA).? Port Authority of Balearic Islands (Ministry of Public Works and Transport, Spain).? Port Authority of Valencia (Ministry of Public Works and Transport, Spain).In addition, visits to the Naples and Port of Gij?n demonstrators were done. Exploitation results Within the BIOCORIN project, a number of exploitable results have been achieved and the list is described below: ProductsThe main BICORIN products are: 1. Green biological anti-fouling coating additive2. Formulation of sol-gel coating able to include anti-MIC microorganism3. Sol-gel coating curable at high temperatures with anti-anticorrosion properties 4. Artificial biofilm as anti-fouling solution5. BIOCOR Gel: Eco-efficient environmental BioCoating with anti-MIC properties These products carry innovative protocols and usages can be patentable and its level of market potential is dictated by the degree of technological innovation that carries. The biocoatings form a strong and unique patentable platform of innovative and chemical formulations with multiple industrial uses, within the antifouling or anti-corrosion or other industrial sectors. The Sol-Gel global patent pool is expanding in the past few years and we are projecting this trend to continue showing an increasing industrial investment interest.ServicesServices have been developed within BIOCORIN Project: 1. Innovative corrosion monitoring: This service will include Innovative techniques including corrosion monitoring simulating a flow system, analysis of the electrochemical behavior of biofilms and monitor onsite corrosion. In addition, the design of specific sensors can be offer in this service.2. Service to compliance with REACH biocide and LCA and LCCA evaluation service for coating and biological process: Quantification of environmental impacts and test required by REACH biocide. 3. Formulation for new alternatives. This service will include the formulation of new coating, proving the viability and efficiency of anti-MIC/fouling bacterial in different coating systems., providing an ad hoc solution development. List of Websites:Website: www.biocorin.euVideos: Videos are available at https://www.youtube.com/results?search_query=biocorin ACCIONA Infraestructuras S.A.(Madrid Spain)Role in the project: Larrge Enterprise, Technical and Admin/Financial Coordinator of the Project. ACCIONA leads WP4 for demonstration activities to implement the technology in the port of Gij?n. ACCIONA was in charge of the evaluation of anti-MIC, its integration in the sol-gel coating and evaluation of its viability in the sol-gel coating and the environment and definition of eco-innovation indicators Contact details: Project Coordinator: Mrs. Edith Guedella Bustamante e-mail: edith.guedella.bustamante@acciona.comPhone: P: + 34 91 791 20 20Dr. Roc?o Barros Garc?a e-mail:rocio.barros.garcia.EXT@acciona.com Dr. Emilio Blas Galindoe-mail:emilio.blas.galindo@acciona.com Dr. Miguel Angel Paris e-mail: miguelangel.paris.torres@acciona.comBioprosperity Symbouloi Epikheironepe (Athens Greece)Role in the project: SME, Responsible partner for the development and successfully perform the business and exploitation plan. WP6 leader Contact details: Dr. Alex Pavlou e-mail: apavlou@bioprosperity.comIngg. F.&.R. Girardi Costruzioni Civili ed Industriali S.P.A. (Naples, Italy)Role in the project: SME, GIRARDI was In charge of demonstration site in Naples Contact detail: Mr. Domenico Giuliano e-mail: domenico.giuliano@girardispa.comGruppo CSA SPA (Rimini, Italy)Role in the project: SME, Gruppo CSA was in charge of WP3 to analyse the environmental aspects of the technology. Test the environmental feasibility of BIOCORIN technology by simulating a microcosms environment Contact detail: Dr. Gabriele Matteucci e-mailgmatteucci@csaricerche.comInstitut za mikrobiolo?ke znanosti in tenhologije d.o.o. (IFB) (terminated on M25) (Ljubljana, Slovenia)Role in the project: SME, IFB was involved in the metagenomic analysis of baceteria in metal surfaces TerminatedAsociaci?n de Investigaci?n INBIOTEC, Instituto de Biotecnolog?a de Le?n (Le?n, Spain)Role in the project: Research Center, INBIOTEC was in charge of WP1 for the isolation, characterization and production of Anti- Microbial induced corrosion (MIC) microorganisms, for its later integration into the coatings to protect the metallic structures against the corrosion process. INBIOTEC leaded the isolation, identification and characterization of MIC and anti-MIC microorganisms Contact details: Dr. Ricardo Vicente Ull?ne-mail: rvicu@unileon.esSviluppo Tecnologie e Ricerca per L'edilizia Sismicamente Sicura ed Ecosostenibile Scarl (STRES) (Naples, Italy)Role in the project: SME, STRES is in charge of WP5, dissemination activities of the project. Will also participate in LCA and LCCA of the developed technology. Contact details: Mr. Carmine Pascale e-mail: carmine.pascale@stress-scarl.itTechnische Universitaet Bergakademie Freiberg (TU BAF) (Freiberg, Germnay)Role in the project: University, TU BAF is in charge of WP2 related with the synthesis of the sol-gel matrix. Leading the formulation, coating application and-anti corrosion properties Contact details: Dr. Edwin Kroke e-mail: Edwin.Kroke@chemie.tu-freiberg.deVan Loon Chemical Innovations BV (VLCI) (Amstedam, Netherlands) Role in the project: SME, VLCI In charge of demonstration site of Harlingen. VLCI participated in durability test and corrosion test of the BIOCORIN solutions Contact details: Mr. Sander van Loone-mail: s.vanloon@vlci.bizIn?titut za metagenomiko in mikrobne tehnologije d.o.o. (IMMT) (Ljubljana, Slovenia)Role in the project: SME, IMMT was in charge of the artificial biofilm development and in the metagenomic analysis of the bacterial communities developing on raw metal and coating surfaces Contact details: Dr. Tomas Rijaveke-mail: tomaz.rijavec@immt.eu
