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Development of a portable miniature mass spectrometer to monitor important, but technically difficult parameters in aquaculture

Final Report Summary - AQUAMMS (Development of a portable miniature mass spectrometer to monitor important, but technically difficult parameters in aquaculture)

Executive Summary:
The AquaMMS project idea was to develop an efficient, robust and portable unit made up of individual sensors for continuous online measurements of important and difficult parameters in aqueous environments. Seafood production through intensive aquaculture in water reuse and recirculating systems (WRAS and RAS) is increasing. Using recirculation technology is highly relevant in fish, shellfish and aquaponics production. A number of unwanted substances may accumulate to unsafe levels for the organisms in such systems. Thus proper water quality is a key issue to succeed.

The AquaMMS project has developed an innovative real-time online multi-sensor monitoring device for the aquaculture industry. The device uses an array of advanced approaches, including mass spectrometry and optical technologies, to measure a wide range of parameters that can affect the water quality in fish farms, more specifically in recirculation systems. The AquaMMS technology will provide immediate advanced warning of a broad range of potential pollutants that otherwise inflict chronic stress on the farmed fish resulting in disease outbreaks and/or poor product quality, which means lost profit to the fish farms. The GUI system was designed to run on a single computer with possibility for remote monitoring. The system is able to compute effects of correlated parameters as well as complex cross-value relationships. Three separate alert type pairs can be set for each sample type: Warning (high and low), Alarm (high and low) and Change rate (up and down). The system status and alert reports can be generated set thresholds break and sent by e-mails to staff members in charge.

The development of the AquaMMS technology to the aquaculture sector required further development of already patented miniature mass spectrometer technology (Q-Technologies Ltd) in conjunction with existing knowhow for specialised membrane inlet and orthogonal sensory technologies. The mass spectrometer allows qualitative and quantitative measurements of dissolved gases (e.g. O2, CO2, N2, H2S and TGP) and volatile organic compounds (e.g. CH3SCH3 and BTEX). A fluorescence-based metal ion sensor was developed and found as a proper solution to complement the MMS system monitoring inorganic metallic contaminants (e.g. Mg, Ca, Zn, Ni, Hg, Pb, Cd, Fe, Al and Cu). The developed sensors enables unique and wide-ranging capabilities allowing low concentrations detection levels <
The AquaMMS project lasted two years from October 2013 to September 2015 and was divided into several work packages (WP) covering scientific understanding, R&D work, integration, testing and validation, innovation related and dissemination activities. The AquaMMS consortium consisted of partners from UK (Q-Technologies, Anglesey Aquaculture and University of Liverpool), Ireland (Faaltech and Cork Institute of Technology), Germany (BAMO) and Norway (Telemarkrøye and Teknologisk Institutt). SMEs from UK, Ireland and Germany are the main beneficiaries and owners of project results.

Project Context and Objectives:
The AquaMMS project was created to develop a new real-time online multi-sensor monitoring device for the aquaculture industry. AquaMMS is a unique multi-sensing instrument platform offering a comprehensive measurement solution for monitoring a broad range of routine and technically difficult parameters affecting water quality. The AquaMMS integrates mass spectrometry, UV fluorescence and optical pH sensor technologies. The instrument intends to be easily operated by non-specialists.

The use of recirculation systems in aquaculture (RAS) is growing rapidly in Europe and worldwide. The use of RAS technology is very positive as it reuses a large percentage of the water, reducing water use, controlling effluents, minimising environmental pollution and at the same time achieving high fish yields. Sustainable and market oriented expansion of the aquaculture sector depend on reliable monitoring and rapid analyses of critical water quality parameters. The AquaMMS aims at providing an innovative monitoring system that is expected to supplement and be competitive to instruments already available on the market. The primary market will be land-based aquaculture industry, but other markets are also relevant including the environmental monitoring sector and the water supply industry.

Controlling the rearing environment is crucial to ensure good fish welfare in aquaculture situations. Thus, water quality monitoring of the conditions in the fish tank is essential as well as controlling the different steps of the water treatment process taking place in RAS. The AquaMMS project used an array of advanced approaches to develop a suitable combined instrument for challenging and harsh environments. The AquaMMS system will work in both fresh- and seawater conditions and the instrument enables the detection of very low concentration of important contaminants and pollutants. AquaMMS will during the project have been tested and validated in commercial fish farms producing seabass and arctic char, being marine and freshwater species, respectively.

The project had several goals related to the different work packages (WP1 Design criteria, WP2 Development of interfaces and vacuum system, WP3 Development of orthogonal sensors, WP4 Development of electronic control unit, WP5 Processing of sensor data, WP6 Integration and validation of prototype, WP7 IPR, Dissemination and Training and WP8 Consortium management) which included scientific, technological and dissemination objectives. The project motivation was through the different work packages to develop an efficient, robust and portable unit made up of individual sensors for continuously online measurements of important and difficult parameters in aquaculture.

To achieve the main goals of completion of the different subsystems and the integration into one AquaMMS system the project was planned with several objectives within different work packages. The following are the main scientific and technological objectives relevant for the development work:
• Improve the understanding of water quality management issues in land-based farms.
• Enhance knowledge of the impact of pollutants and metabolic by-products in RAS and thereby define limits necessary to optimise performance and ensure animal welfare.
• Ensure design specifications for the membrane inlet and orthogonal UV sensing technology.
• Ensure design specifications for electronic control unit hardware.
• Ensure design specifications for Graphical User interface and integration of AquaMMS into current recirculation monitoring.
• To develop a novel membrane inlet (MI) methodology to enable deployment of a miniature mass spectrometer within the aquaculture environment.
• To develop a compact fluorescence sensing instrument with arrayed sensing elements complement to the MMS system extending and enhancing the detection capabilities of the device for water quality monitoring in aquaculture.
• To develop a customised electronic control unit to drive the miniature mass spectrometer, including ion source, quadrupole mass filter and detector.
• To develop the Graphic User Interface (GUI) so it presents the essential information to the fish farmer in a comprehensive manner.
• Integrate the prototype AquaMMS into a freshwater and marine commercial fish farm where it will be validated in real fish farm conditions.

The development work related to the mass spectrometer was to be based on a patent held by Q-Technologies of a MMS (miniature mass spectrometer) fabricated with hyperbolic form electrodes. In conjunction with existing knowhow for specialised membrane inlet technologies and orthogonal sensory technologies, it was permitted to develop sensors suitable for aqueous environments and with precise sensitivity to low concentrations of dissolved gases and volatile organic compounds. To develop complementary optical fluorescence sensors to detect metal ions and pH major scientific challenges had to be overcome. The toxicity to the fish of most metals increases with decreasing levels of dissolved oxygen. It was therefore regarded of great advantage to develop an instrument that can give immediate response to the content of certain metal ions in combination with other crucial parameters in aquaculture situations. To integrate the fluorescence sensing system with MMS, a plan for integrating the fluorescence sensors with MMS system need to be built and programming on the PC is required to allow users to read the fluorescence measurements of different parameters.

The measurement of the core water parameter such as pH is of constant need for the aquaculture industry. The pH level of water varies according to a variety of factors including chemical composition and the present of trace minerals. This being the case, different aquaculture species are adapted to different pH levels – the level that works for one species might not work for another. To ensure healthy aquaculture it is of high importance to continuously measure the pH of water. Recently, the optical pH sensors are possible the most frequently described group of optodes (optical sensor devices). Optical sensors have gained a lot of attention due to the fact of wide availability of the miniature photodetectors and light sources. An optical pH sensor developed and sold at the right cost as a routine consumable, lends itself readily within the sector in combination with other sensors (AquaMMS) or on its own (stand-alone sensor).

It is foreseen that the combined AquaMMS solution will be resource and cost efficient thus ensuring sustainable, profitable and competitive production by European fish farmers also in the future. The project partners in general and the SMEs in particular will after the project lifetime continue to disseminate the results to relevant industry sectors encouraging them to implement the new technology.

Project Results:
A working AquaMMS protoype was successfully developed during the project. The AquaMMS instrument is made up of individual sensors which are integrated into an efficient, robust and portable unit. Specifications of the different system components; electronic control unit (ECU), mass spectrometer (MS) system, membrane inlet (MI) and orthogonal ultraviolet (UV) sensing and graphical user interface (GUI) and communication interface. A harsh environment prototype enclosure was designed and built, and the system was tested in laboratory and commercial RAS facilities.

AquaMMS is a multi-sensor combining multiple technologies that simultaneously measure a wide range of parameters that can affect the water quality in fish farms. Real time in situ monitoring of target analytes of interest will provide a warning system to assist in farm management which will provide an increase in farm productivity. A desktop type pre-prototype system was developed as opposed to a fully portable system. The final prototype was mounted in a suitcase sized wheeled enclosure easy to transport around during transport and between tanks at the fish farm. The custom integrated system is optimised for aquaculture parameters of interest and will be suitable for ‘bench-top’ operation interfacing through a standalone computer or laptop. The integrated system should be robust and chemically inert (those elements which interact with farm water). The system should be easy to use, data and readout easy to interpret and maintenance should be minimal, the cost low and easy to implement.

The AquaMMS was developed by using an array of advanced approaches, including mass spectrometry and optical technologies, to measure a wide range of parameters that can affect the water quality in fish farms, more specifically in recirculation systems (RAS).

The different sensors developed include a mass spectrometer, UV fluorescence spectrometer and an optical pH sensor. Each device has been designed and built with a view to monitoring targeted compounds in aquaculture. The mass spectrometer is able to monitor in real time the dissolved gas composition including any volatile organic species. Metallic ion species in the water are determined by the fluorescence unit and these are both complimented by an optical pH sensor which gives the user holistic overview of water chemistry. The AquaMMS prototype incorporates all of these sensing technologies into a single unit which is suitably ruggedized to operate in a harsh environment.

The project was organised in eight work packages (WP); WP1 Design criteria, WP2 Development of interfaces and vacuum system, WP3 Development of orthogonal sensors, WP4 Development of electronic control unit, WP5 Processing of sensor data, WP6 Integration and validation of prototype, WP7 IPR, Dissemination and Training and WP8 Consortium management. Specific results were provided as presentations, posters, brochures or other reports at project meetings, exhibitions, conferences, and on the public and password protected project web site. The results from the different task works are provided to the consortium in several comprehensive deliverable reports. The main S&T results were obtained in WPs 1-6.

The description of RTD work and main results are as follows:

The objectives of WP1 “Scientific understanding” was to gather all necessary data and user requirements to make a final list of parameters and specifications to govern the developments in WPs 2-5. Background knowledge from several sources including relevant partners and potential users was gathered and the requirements made the platform for the short-list of prime parameters to be monitored by the AquaMMS technology. Available information on concentration limits of different substances affecting fish welfare was used to determine the upper and lower values to be detected and linked to alarm and display warnings. The identified requirements served as important input to the processing of sensor data and GUI development. This instrument specifications for electronic control unit (ECU), mass spectrometer (MS) system, membrane inlet (MI) and orthogonal ultraviolet (UV) sensing components was identified as well as the design and implementation of the graphical user interface (GUI) and integration of the AquaMMS with existing management systems used by fish farmers. A MMS Monitor system was specified to be responsible for collecting data from one or more MMS instruments. The MMS monitor system monitors and manages measurements from one or more instruments. The MMS instrument specifications describes the instrument with necessary electronics to operate as a unit. The main characteristics of the MMS monitor were: Running on a PC, written in Microsoft Visual C#, store data in a suitable SQL DB, XML protocol to one or more instruments. The AquaMMS design specifications ensured the development of MMS monitor so that it can integrate well into current recirculation monitoring at end-users aquaculture farms and present the essential information to the fish farmer in a comprehensive manner. The SW design was to be modular in order to prepare for future expansions and custom specific solutions.
The necessary information to produce a short-list of parameters to be measured by the AquaMMS was obtained through written communication, telephone and personal interviews. The input about most useful parameters from the fish farmer point of view was obtained from the project partners, large Atlantic salmon producers in Norway and researchers working closely with the aquaculture industry in the partner countries. More general and species specific information was based on social media networking, available literature, by talking to local environmental and fishery authorities and test laboratories. Background knowledge about relevant parameters for water quality monitoring in aquaculture was obtained via several channels to reveal the needs and requirements for water quality monitoring in aquaculture water reuse and recirculating systems (WRAS and RAS). Literature surveys, results from laboratory analyses and guidelines together with interviewing of the project partners, other industry, science, and authority representatives provided input to the report. The methodology used to collect the already documented information was based on searching available literature databases for relevant publications. In addition to scientific data bases the “grey” literature, covering papers, reports, technical notes or other documents produced and published by governmental agencies, academic institutions and other groups that are not distributed or indexed by commercial publishers, were searched or obtained through the contact network of the consortium.
Background water quality data was collected from the end-users participating in the project to document the potential concentration level of the actual parameters. The species specific tolerance levels were obtained from several documented information sources such as scientific journals, reports and presentations at meetings and conferences. The existing information of species- and stage specific tolerance limits is not complete. Some information therefore had to be generalised and evaluated as most-likely values. The obtained information was used to provide description of essential parameters and their potential challenges encountered while assessing the parameters.
The final selection of parameters was based on management options for important commercial species and recommendations by the project team. An initial comprehensive list of potential parameters that could be monitored by the AquaMMS technology was prepared and followed by a decision making process within the consortium to form the short-list of parameters required for the development of AquaMMS.
Based on input from the industry and associates a set of useful candidate parameters were identified. Ammonia nitrogen (NH3-N), nitrite (NO2-), dissolved CO2 and pH are all parameters that are considered to have great market potential if made for direct and frequent measurement that can be monitored online. Other important basic and essential parameters are oxygen, temperature and salinity. The implementation of magnesium, calcium, iron, aluminium and copper may improve the control of toxic substances in farms and facilitate immediate management decisions.

The objective of WP2 “Development of interfaces and vacuum system” was the development of an in-situ sampling methodology capable of operating in an aquaculture environment, development of a suitable vacuum system/housing for effective operation of the mass spectrometer and optimised coupling between the sample inlet and the mass spectrometer ion source. Sampling was achieved using a semi-permeable membrane interface which works on the principle of pervaporation. This involves the permeation of an analyte through the membrane and then its evaporation into the vapour phase. A hydrophobic membrane was used to selectively transport analytes of interest whilst resisting the transport of water through the membrane. The key aspect of this technique is that both the sample and vacuum side can flow continuously which allows real-time monitoring. This was optimized in terms of geometry, flow, temperature and the material make-up of the membrane to allow optimal sampling of the targeted analytes. In addition to the requirements set out in WP2, a membrane heating control system and a fail-safe mechanism were developed to prevent damage to delicate hardware in the unlikely event of a membrane failure.

A vacuum system was designed and built to meet the design requirements for AquaMMS. The design allowed effective mass spectrometric operation and close coupling of the membrane inlet probe to the ion source region. The bespoke design maximises analyte flux to the ion source region and considers other AquaMMS practical system requirements such as automatic venting and membrane fail-safe capabilities. The AquaMMS prototype system was calibrated, aligned and tuned using the optimal entry conditions determined using the University of Liverpool custom 3D mass spectrometer rapid simulation package.
The objective of WP3 “Development of orthogonal sensors” was to develop complementary fluorescence sensors to the MMS for detecting metal ions and pH level. The optodes of different composition of the sensing membrane were fabricated in sol-gel process. For application as a pH optode, two different pH indicator dyes were incorporated in the sensing membrane. Series of fluorescence based calibration tests were used to characterize the performance of optodes in buffers of various pH. Additionally, the low-cost electronic setup was developed to simulate the performance of the pH optodes outside the laboratory environment.

The key process chosen to develop an optical pH sensor is sol-gel. In the sol-gel process the light-sensitive dye is physically encapsulated in pores of the ‘inorganic glass’ such that the molecules are immobilized with limited leaching out. In the same time the inorganic glass is porous enough to allow the transportation of the solvent and ions into the interior. The inorganic glasses can be fabricated into desired shapes and sizes such as monoliths or thin films on the transparent substrates.

For the pH sensor for AquaMMS, a range of pH sensitive dyes were chosen and tested over the wide pH range of 5 to 9. The pH sensitive dyes were immobilized within the silica thin film made by acidic hydrolysis and condensation of tetraethyl orthosilicate (TEOS) itself or in the mixture with other silicate precursors: trimethoxy(propyl) silane (TMPS), triethoxymethylsilane (TEMS), dimethoxydimethylsilane (DMDMS), triethoxyphenylsilane (TEPhS), tetramethyl orthosilicate (TMOS), trimethoxymethylsilame (TMMS), triethoxyethylsilane (TEES) in the molar ratios of 1:1, 2:1 and 4:1, and deposited onto the glass slide through the dip-coating process. Bromocresol purple (BP) was chosen as the dye for the final sensors. The stability of the chosen sensor designs was evaluated over 900 minutes. The response was measured every 30 min at pH 6. The decrease of the emission value was less than 10% and 5% for the BP dye within TEOS and TEOS/TEMS membrane, respectively. The short-term stability was also measured for the same sensors. During the experiment the emission was recorded every 60 sec for 20min at pH 6. The results show the stable response of the sensor obtained with t90 lower than 60 sec.

Following successful lab testing, in the next step the sensors were transferred to the experimental setup for fluorescence measurements design and built in house. The setup mimics the condition in the environment regarding the sensitivity.

A LED driver circuit was designed to modulate the LED signal to the sensor. The modulation signal can be adjusted (frequency, shape) by a function generator. Sensor and sample solution were placed in a cuvette which was fixed in the cuvette holder. Optical filters were placed in front of LED and photo-detector to allow proper incident light directed to the sensor and only florescence signal went to the detector. The florescence signal detected by the photo-detector was collected by an oscilloscope and sent to PC (labview) for signal processing and analysis

In the experiment, the BP/TEOS and BP/TEOS/TEMS sensors were tested in a buffer of increasing and then decreasing pH in time. The well-defined steps were observed after every change in pH. The changes towards increasing pH values were detected every 0.5 and every 1 pH unit. On the return to pH 6, the steps of 1 pH unit were also detected. The differences in the values in the amplitude at each pH detected while going up and down in the pH scale, were observed. Due to the limitation of the LED the experimental setup could not detect the pH lower than 6 for BP/TEOS (Figure 7B) and 6.5 for BP/TEOS/TEMS. BP/TEOS sensor was tested in the sample of river water taken from Curraheen River near CIT. The sensor was immersed in the river water and kept in for 48 hours. The measurement was taken at the time of immersion (0h) and after 48h. The results were compared with the calibration plot taken on the immersion of the sensor in the buffer solution at pH 6 and pH 9.

The results demonstrated the reusability of the sensors in the long term measurement in the river water which is chemically and biologically reach. The pH of the river water measured by the optical pH BP/TEOS optode was 7.2 which is close to pH 7.1 obtained from the measurement done by commercially available electrochemical pH sensor. The biofouling effect during the experiment was not observed. Two pH sensors: BP/TEOS and BP/TEOS/TEMS have been designed, developed and fabricated. From the results of this study, the BP/TEOS optode demonstrated improved response time, reversibility and reusability compared to the BP/TEOS/TEMS pH sensor and, thus BP/TEOS probe will be used as a pH sensor for final, integrated device.

CIT also explored the use of a fluorescence based system to detect heavy metals in the sample solutions. A known concentration of Ca ions were dissolved in a buffer solution and fluorescent measurements were made to assess the limits of detection for the metal ions. Using a fluorescence quenching approach a detection limit of approx. 0.2µM can be detected.

A complete pH sensing system has been designed using the pH sensor fabricated with conventional a LED and a silicon photo-detector. In the system, a driver circuit is used to modulate the LED light directed to the pH sensor. The modulation signal is a pulsed signal with a frequency of 500 Hz and duty-cycle of 20% which is set by a function generator. The pH sensor and the sample solution (with different pH concentrations) are placed in a cuvette which is fixed in the cuvette holder. Optical filters are placed in front of LED (bandpass filter 472 ± 30 nm) and photo-detector (longpass filter ≥ 515 nm) to allow proper incident light directed to the pH sensor and only florescence signal to be collected by the detector. The florescence signal emitted from the pH sensor will be converted into a voltage signal by the photo-detector and collected by a PC (labview) through a National Instrument DAQ board (NI USB-6008) for signal processing and analysis. The voltage signal from the photo-detector has a similar shape of the modulation signal with its peak-to-peak amplitude varies for different pH concentrations in the sample solution. The function generator and the DAQ board can be replaced by a micro-controller/FPGA based circuit and a low cost and portable version of this pH sensing system can be easily achieved which will be able to operate independently of laboratory instruments for the real-time pH monitoring

A graphic user interface (GUI) is designed based on the Labview program. The GUI collects the voltage signal of the photo-detector through the National Instrument DAQ board, calculates the pk-pk amplitude of the voltage signal and converts it to pH values. As the calibration of the pH sensor is a linear curve, two points need to be entered by end users to produce the calibration curve for pH value converting. When the measurement is ongoing, the GUI is continuously displaying the pk-pk voltage amplitude from the detector and the converted pH values which are being recorded into an .lvm data file. A real-time pH value is also shown in the GUI.

To be able to demonstrate fluorescence and pH measurements through the data harvesting and GUI application, a shortcut has been chosen, and raw measurements of light intensity from the fluorescent sensor and pH sensor prototypes are read by “Iota lookalike” software instead of being directly available in Q-Tech’s Iota server itself. From the user’s perspective, this workaround is transparent but two applications will have to be installed instead of one.

Data are available through the ThorLabs optical powermeter in fluorescence sensor case and through a custom electronic circuit (combined with an NI-DAQ 6008) for the pH sensor. The “Q-Tech lookalike” software will then serve the requests from AquaMMSmonitor through a telnet interface. The latter reads un-calibrated raw data from the instrument and those raw data samples can also be processed via the lookup table feature available in AquaMMSmonitor to convert them into a concentration or pH value.

A fluorescence sensing system for pH has been developed which was built by using the optical pH sensor previously developed with optical and electronic components. A labview based graphic user interface (GUI) was designed and a calibration method was developed for the pH measuring.

A plan for integrating the fluorescence sensors with MMS system has been described. Programming has been done on the PC that allows user to read the fluorescence measurements of different parameters through Q-Tech’s Iota server.

The objective of WP4 “Development of electronic control unit” involved the design, build and testing of a customised electronic control unit (ECU) (hardware and firmware) tuned to drive a portable membrane inlet mass spectrometer (including ion source, mass analyser and detector). This includes the ECU and quadrupole array design and assembly, integration of the ion source, mass analyser and detector onto a vacuum flange, and initial testing of the customised, portable mass spectrometer in the aquaculture environment. This was completed and demonstrated with the membrane inlet and vacuum system developed as part of WP2. In addition the application of an axial magnetic field to the body of the instrument was optimised to increase ion transmission leading to an overall order of magnitude increase in sensitivity. Such an increase in ion transmission is otherwise unachievable with a conventional device (without sacrificing other performance criteria). The electronic control unit and hardware developed were extensively tested at the University of Liverpool and a working membrane inlet mass spectrometer system was demonstrated for the detection of volatile organics in water.
The AquaMMS instrument was integrated into a sufficiently robust enclosure which aided portability and was designed to be robust and prevent water ingress. As part of the work package, this functionality was also demonstrated onsite at Anglesey Aquaculture. The system was used to monitor dissolved gases and organic compounds at different stages of the water treatment cycle at various points in time at Anglesey aquaculture, highlighting the capability and benefits of online in situ monitoring.
The objective of WP5 “Processing of sensor data” was mainly to develop software for harvest, process, evaluate information, and present the essential information to the user of AquaMMS in a comprehensive manner. The MMS Monitor system was developed to be responsible for collecting data from one or more MMS instruments, and to present the status of the system on a screen. The GUI was one part of this system. The MMS monitor controls and monitor its connected MMS instruments as well as showing the measured data in a suitable manner for the user. At a glance, it is possible to see the overall status of water quality, and react if necessary. The MMS monitor will run on a windows PC, alone or together with other farm systems. Microsoft development tools were used for the development.
The product design specification including full details on design and software structure of the AquaMMS system was elaborated under this work package. A system architecture which consists of a framework for data, communication and interchange format was developed. The main objective was to specify the design of the computer system using one or more AquaMMS instruments for assuring good water quality in a fish farm or other relevant installations. The system harvest data from the instruments and processes them to the necessary level. The specification also included the protocol used for data harvest and instrument control. The specifications could be modified during the integration phase of the project if it should be impossible or not feasible to fulfil them.
The product design specifications and requirements for testing served as a guide for the development of the AquaMMS data processing system. The GUI system was designed to run on a single computer with possibility for remote monitoring. The system is able to compute effects of correlated parameters as well as complex cross-value relationships. Three separate alert type pairs can be set for each sample type: Warning (high and low), Alarm (high and low) and Change rate (up and down). The system status and alert reports can be generated set thresholds break and sent by e-mails to people in charge of the monitoring. By the time of the completion of the work package the system had been extensively tested with the Q-Technologies server running an instrument simulator for the MMS instrument. The program was also successfully integrated, tested and demonstrated with the pre-prototype of the MMS instrument.
The user alarms and data presentation system includes product design description, system functionality with test and test reports, data processing and alarm software (SW) and product data package, SW and SW source.

The objective of WP6 “Integration and validation of protoype” was to proof the concept of AquaMMS in real life environments. The AquaMMS prototype was to be tested and validated at two commercial fish farms, one in UK and one in Norway. The test sites represented modern land-based aquaculture recirculating systems (RAS) of different scale and location. The AquaMMS prototype was functionally tested at the facilities of the two end-user partners of the project: Anglesey Aquaculture producing seabass and Telemarkrøye producing arctic/alpine char. During testing and monitoring the measurement data were logged continuously. The TeamViewer software programme was used for remotely observation of the instrument performance and of the development of selected parameters in the fish tank waters.
The instrument was tested over time under real conditions in their marine and freshwater recirculation systems, respectively. Changes in the water quality and the levels of the different parameters measured occurred regularly in association with feeding and handling of the fish. The consumption of oxygen increased during heavy feeding activity or if the fish were stressed. The addition of feed created detectable variations of O2 and CO2. During sampling of fish for grading and weighing, some handling causing stress was unavoidable. Changes during these events could also be detected by AquaMMS. Diurnal variations in temperature and light conditions were other events that resulted in water quality changes. Random water samples were analysed and compared with the AquaMMS results. During the functional field testing of the AquaMMS the staff at the two fish farms had the opportunity to follow the use of the instrument onsite. Demonstration of the instrument was also made in the laboratory and in connection with management board meetings ensuring that the industrial partners could experience the performance of the instrument and get hands-on experience. The great potential of AquaMMS and its applications was discussed with the industrial partners.

By monitoring the water quality at various stages of the treatment cycle – various processes could be assessed and evaluated. In particular, testing was completed in situ pre- and post- biofilter stage, and pre- and post- CO2 filtering. Typically, water at these stages is not monitored nor tested. There are serious issues with sample integrity when seeking to take samples (particularly with dissolved gas measurements) and then send for analysis at external laboratories. The measurements taken from various points in the treatment cycle were in keeping with general practice and confirmed by external laboratory reports. Our results were of particular interest in enabling the end user to better understand the performance of key water treatment processes such as the effectiveness of the biofilter and CO2 filter. For example, it is difficult to asses the performance of the CO2 stripper by visual inspection. By monitoring the levels of dissolved CO2 pre- and post- this process it was determined that the stripper was not operating at maximum efficiency and that some maintenance was required.

Potential Impact:
Technology and innovation
The use of RAS in aquaculture is growing rapidly in Europe and world-wide. Sustainable and market oriented expansion of the aquaculture sector depend on reliable monitoring and rapid analyses of critical water quality parameters. The AquaMMS will provide an innovative monitoring system which can supplement and be competitive to instruments already available on the market.

Metal and other element analyses often require that water samples are sent to accredited laboratory. This procedure may be costly and time consuming to the farmers and it will be beneficial if such analyses can be made on-site. The implementation of magnesium, calcium, iron, aluminium and copper may improve the control of toxic substances in farms and facilitate immediate management decisions.

A broader platform technology has been developed here. The ability to form optically active layers on glass substrates open other avenues for R&D and product development. In the case of AquaMMS the layer sensed pH levels in the 5-8 range. Using different dyes pH range in either the more acidic or basic region can be measured and for next generation sensors multi layer systems present the possibility to measure a very broad pH range in a single system. Outside of pH measurements layers could potentially be functionalised to interact with metallic ions, specific bacteria or nutrients or to detect contaminant products in waste systems. This gives the potential for moving into areas such as life sciences, medical devices and the food industry.

Impact for the participating SMEs
The industry identified candidate parameters as ammonia nitrogen (NH3-N), nitrite (NO2-), dissolved CO2 and pH to be parameters that are considered to have great market potential if made for direct and frequent measurement that can be monitored online. Other important basic and essential parameters are oxygen, temperature and salinity. Implementing these parameters in the AquaMMS analyte package, either by own technology or by integrating existing off- the-shelf sensors, will make the AquaMMS even more attractive. A prerequisite is that the sensor probes can be kept clean from biofouling for a long time in water.

The AquaMMS technology will be relevant for use by other than aquaculturists. For instance, it is obvious that a method for direct measurement of the toxic unionized ammonia (NH3) would provide benefits when considering the challenges encountered using current methods which only estimates NH3. It is therefore recommended that AquaMMS should investigate approaches to obtain direct NH3 and other measurements. The market for such technology would probably be large, within wastewater treatment sector, aquaculture, and environmental monitoring of natural waters.

Several potential markets for exploitation of the AquaMMS technology exists. The monitoring of water quality is essential in aquaculture as well as in a row of other industrial sectors. Sectors of application may be aquaculture, aquariums, aquatic environmental consultancies, aquaculture academic R&D sites, municipal water supply & sanitation, environmental regulators, food & beverage processing, pharmaceutical, agriculture & horticulture, paper, mining, power generation and more.

Contribution to the development of a sustainable aquaculture industry sector
Increasing seafood production through intensification in aquaculture recirculating systems (RAS) has inherent risks additional to those that are already well understood by the aquaculture sector such as pH decline, rising carbon dioxide concentrations, phosphate and nitrate levels. By reducing the volume of new water replacement, the farmer can reduce the cost of water pumping and temperature control, whereas reduced daily water replacement may lead to accumulating levels of pollutants unless they are specifically monitored and removed before the fish welfare is challenged. With the AquaMMS instrument installed, the fish farm can monitor the crucial parameters continuously and an alarm can be triggered if a pollutant increases beyond a set level, giving the farmer time to take correct management decisions like dilution with new water, employment of a specific water treatment, changing feed sources etc. In this way, efficient use of resources is ensured.

As with most industries the need to control costs more tightly and to be more efficient is a concern in the aquaculture sector. In dealing with livestock this is of greater concern as unknown factors and small changes in environmental conditions can have a very dramatic effect on the health and wellbeing of stock. This is of particular concern when the stock is high value and in those cases more readily affected by such environmental changes.

The AquaMMS system has the potential to allow the aquaculture industry to increase its productivity and efficiency in a variety of different ways. Firstly the ability to remote monitor changes in the conditions and environment of the stock should lead to faster response times when dealing with issues. It also allows multiple sites to be monitored at any given time increasing the efficiency of the process and allowing staff to be more dedicated to more subtle aspects of the particular process. This in turn will also potentially allow the business to increase its stock levels and productivity thus helping to encourage growth and sustainability.

Secondly working in the aquaculture industry can present some significant health and safety risks. This is not a typical environment and hazards are more pronounced, working outdoors in wet and windy weather, handling heavy equipment and working in dark hours all present challenges. The technology presented has the capability to reduce the exposure of the worker to these hazards and only requiring the individual to work in these conditions from the point of view of monitoring when absolutely necessary.

AquaMMS technology will impact and improve the EU fish production industry and the European environment receiving its effluent by providing:
• A technology suited to all land based marine and freshwater farms and hatcheries
• Improvements in quality of farmed seafood by reducing risk of contamination
• Enhanced production through reduction of mortality and potential disease outbreaks due to chronic pollutant exposure
• Reduced farm production costs associated with water quality monitoring methods
• Improved monitoring of effluent quality and reduced environmental impact

Wider societal implications
Aquaculture may create much sought after jobs for men and women in coastal and rural areas.
Spin-off activities....
Increased activity in locally and regionally in Europe
Impact on people health and quality of life

Exploitation of results and dissemination activities
The exploitable results planned and derived from the AquaMMS project were:
1. Generated scientific knowledge and design specifications
2. Interfaces and vacuum system
3. Optical pH sensor Electronics and housing
4. Orthogonal sensors
5. Graphic User Interface, data interchange and storage SW

The specific sensor developed (result no. 3) can be adapted to work in alternative areas of research. It can also be potentially adapted to work in a fibre optic based detection system, allowing for measurements to be made in tightly controlled areas or areas with restricted access. The associated signal measurement and processing electronics are also widely adaptable. The same system can be used no matter what actual parameter is being measured. Also the associated scaling of the unit can be applied across other measurement systems where multiple parameters may be measured simultaneously or in a times or gated fashion.

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