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LOW-COST, HAND-HELD, AND NON-INVASIVE OPTICAL SENSOR FOR MULTIPARAMETRIC FIELD ANALYSIS OF GRAPES AND LEAVES IN VINEYARDS

Final Report Summary - PREMIVM (LOW-COST, HAND-HELD, AND NON-INVASIVE OPTICAL SENSOR FOR MULTIPARAMETRIC FIELD ANALYSIS OF GRAPES AND LEAVES IN VINEYARDS)



Executive Summary:

In August 2009 was fully implemented the reform of the Common Market Organization for wine (CR Nº479/2008, 29-04-2008). It aims at reducing the 24 million hl/year surplus of basic low quality wine, phasing out the €500 million/year spent on wine disposal subsidies and making EU wine more competitive. This reform, together with other market constraints will endanger all EU winegrowing SMEs. To survive, they will have to increase crop value, reduce the production of basic wine and convert part into premium quality. This can be achieved by implementing new and more effective field control methods.

PREMIVM (www.premivm.eu) proposes a low-cost, handheld device capable of non-invasively estimating ripeness and vigor parameters for grapes and vine plants. All this in the vineyard, by means of the innovative use of chlorophyll fluorescence and reflectance multispectral data correlated by specific mathematical models, with GPS tags for all readings. The measurement principle is based on the natural response of vegetal species to light, where specific molecules are excited at a certain wavelength and emit radiation of a different wavelength. The emission spectrum provides qualitative and quantitative data that can be used by vineyard managers to precisely control the field, reduce vineyard variability, define optimal harvesting times and increase production value. The device will provide data to precisely control the field, and increase production value up to 25% (expected yearly €5.000-10.000/ha income increase for winegrowing SMEs like SAPRUEM, QMF and PERACCIO).

With a foreseen commercial price of €2500/ unit, consortium manufacture and distribution SMEs PSI, DVC and AGRI estimate, through a strategic alliance, to reach at least 2% of the market in 5 years. The consortium provided the complementary business capabilities, commercial networks and research expertise to guarantee the technology a quick route to the market. All members were fully committed to ensure the success of the project, led by the SMEs in testing, validating, using and protecting the results outsourced to the necessary expertise in Chl-F&R, optical instrumentation, botany, IT solutions, communications and prototyping of RTDs CRIC, KIT and Czech Globe

The project was divided into research, demonstration, dissemination, exploitation and management activities by means of Work packages. After 27 months of research PREMIVM achieved all the objectives foreseen.

Project Context and Objectives:

The European wine industry is a strategic economic sector that is facing growing competition in the international market, and at the same time dealing with reduced support from the CAP. With this reality in mind, an international consortium comprised of 3 technical companies and 3 winegrowers together with 3 research groups, setup the EU FP7 Project PREMIVM in 2011.

PREMIVM proposes a solution (under the trademark WinePen) capable of noninvasively estimating sugars, chlorophyll and polyphenols in grapes. Field analyses are possible by means of the innovative use of chlorophyll fluorescence and reflectance multispectral data, correlated by specific mathematical models. The measurement principle is based on the natural response of vegetal species to light, where specific molecules are excited at a certain wavelength and emit light of a different wavelength. The emission spectrum provides qualitative and quantitative data that is transformed into useful field information to the vineyard managers.

The WinePen instrument will be a low-cost, handheld spectrometer (estimated price range of €2.500-3.500 for commercial model) with touch screen display with the capability to make wavelength and intensity readings of fluorescence and reflectance. The existing pre-commercial prototype has a memory capacity up to 8000 measurements with a battery life of 48 hours of continuous operation. The dimensions are 255 x 75 x 40mm and 300g of weight.

All field readings made with WinePen have a GPS tag, giving the user total liberty to easily extend their previously implemented sampling schemes. When uploaded to the PREMIVM vineyard management software, the data is processed into easy to interpret spatial field analysis information accessible on the Web (PC, Tablet or Smartphone). The software can be used to optimize the growth/ripening of grapes, reduce vineyard variability, define optimal harvesting time, set differential harvesting schemes, select proper grape variety/terroir combinations, micromanage local differences in the vineyard and assist integrated agriculture practices.

The results obtained with the pre-commercial prototype of WinePen, in three consecutive seasons (2011-2013), demonstrated that this device is suitable for non-invasive and fast estimation of grapevine ripening. The companies involved in the project are currently planning the next steps for the development of a commercial prototype and respective market route.

To achieve the current results, the objectives set for the PREMIVM project were:

1) Develop a Chl F&R calibration library specific for vineyards- Achieved by Month 12 and updated with field assays in the summers of 2012 and 2013

Study the relationship between chlorophyll and fruits, using fluorescence and reflectance for a better understanding of phytophysiology in vineyards.

Build an extensive spectral fluorescence and reflectance characterization library specific for vineyards, analyzing samples collected during the fruit ripening stage.

Chemically characterize all samples (like polyphenols, sugars, chlorophyll, etc.) using different reference methods.

Pair F&R data with reference data and implement a feature selection approach using a neural network methodology to create robust correlation algorithms.

2) Develop a Chl F&R optical system specific for vineyards - Achieved by Month 14 and continuously improved during the life-time of the project

Develop a miniaturized optical system capable of acquiring directly and non-invasively full spectrums of chlorophyll fluorescence and reflectance from fruits; investigate the optimization of excitation wavelengths for multispectral analysis and demonstrate that this analysis can provide ripeness and vigour information.

Perform outdoor active sensing measurements under any light conditions.

Perform reliable measurements at a specific distance from sample (non-invasive).

Perform simultaneous measurements of several optical signatures non-invasively, in real-time and with no preparation of sample.

To produce multiple wavelength excitations - UV and visible for fluorescence, and 350-1500 (max) nm for reflectance, and to detect fluorescence and reflectance signals in the range of 400-1500 (max) nm.

To have low energy consumption.

To evaluate the performance of the optical system in laboratory against reference methods

3) Develop a fully integrated and functional prototype - Achieved by Month 18

To integrate the final system to achieve a full working device with a final price of €2500 in mass production.

Integrated GPS module + antenna to permit geopositioning in vineyards

Easy interface with the device through a touch screen

Store all acquired data in an internal memory to store multiparametric data entries and easily interface a PC with a USB port for data transfer

Have suitable power autonomy of at least 12 hours with a Li-ion rechargeable battery

Have a compact and robust final design (< 3kg, operation at 8-55ºC; resistant to water, dust, grease and chemicals)

4) Develop the Vineyard Management software - Achieved by Month 14

develop an input data processor and storage module that automatically receives and processes the geospatial data transferred from the sensor.

To implement the correlation algorithms that through a graphical representation tools bundle will process the stored data to automatically generate field reports and statistics.

To make field tests with geopositioning and dummy data to debug the software

To have a graphical user interface to be easily and intuitively used by the winegrower.

To be able to use both PC and Pocket PC hardware platforms

5- Demonstrate the functionality of the device in the field - Achieved by Month 27-.

To use the developed prototype for a better understanding of fruit ripeness and cultivar evolution in the vineyard. The information provided by PREMIVM will be evaluated on the extent of which it can aid in understanding the dynamics of viticulture fields and provide information to assist in the micromanagement of vineyards

To perform field trials with PREMIVM in partner’s vineyards during the entire ripening stage (July, August and September +/- 1 month) and compare the performance of the prototype against the current analytical tools implemented by each winegrower.

To demonstrate how PREMIVUM can provide useful information, with a performance comparable to standard laboratory F&R techniques and equivalent to extensive and complex laboratory analysis using reference analytical methods.

6) Train SMEs, disseminate results at EU level and define the ways to protect and exploit Foreground- Achieved by Month 27

Develop a plan for participating SMEs to be trained on the use of PREMIVM, namely its functionality, operability, troubleshooting, maintenance and calibration.

PREMIVM will be disseminated through publications, conferences, fairs, workshops, paper/internet articles, and newsletters.

The plan to protect and exploit the Foreground will be formally closed. All strategies and corporative alliances will be defined within a thorough Business Plan.

Project Results:

The project results per WP can be summarized as follows:

In the first period, in WP1 the consortium completed the definition of sampling plans and laboratory protocols during the first three months of the project. KIT and CZECH GLOBE worked together on the definition of analytical methodologies to be implemented in Germany and Czech Republic. CRIC and AGRI worked together in the creation of sampling plans aligned with the analytical necessities in the workpackage and the expected performance of the final prototype. The winegrowing SMEs SAPRUEM, QMF and PERACCIO helped in this tasks and implemented specific analytical and sampling protocols to characterize and benchmark their own vineyard plots which would be used to make all field tests of the prototype during the second year.

KIT led the main efforts in the collection and analysis of samples from vineyards: a) Work in the vineyards of the Geilweilerhof Institute for Grape Breeding: specification of the measuring site, specification of the measuring times and procedures b) Measurements with single berries: Brix (sugar content), pH by means of pH indicator strips, chlorophylls and carotenoids of the berry skin, anthocyanins of the red wine berry skin, single flavonoids of the berry skin by means of curve fitting c) Measurements with grape clusters: Brix (sugar content), FTIR analysis: pH, malic and tartaric acids, glucose and fructose, nitrogen content d) Procedures in the correlation between fluorescence and reflectance measurements and the reference measurements.

A massive sampling was carried out in several vineyards in Germany, Czech Republic, Portugal and Italy. PSI assisted KIT in the characterization of the samples by using different approaches on fluorescence and reflectance techniques. The samples collected in vineyards were quantitatively analysed using standard analytical methods. The winegrowers QMF, PERACCIO and SAPRUEM also contributed at this stage, by characterizing their own vineyards to be used in the field validation of the integrated prototype scheduled for the summer of 2012. The data generated by each set of assays was used to find the best correlation models that corresponds to a given instrumental setup and methodology. By seeking the optimum instrumental specifications and work conditions (light sources, detectors, wavelengths, filters, temperature, environmental light, excitation ratios) for each correlation model through a series of combinatory experiments, this task was the precursor of the work done in WP2 on the construction of the final miniaturized ripeness transducer to be assembled in 2012. The preliminary results, conclusions, configurations and designs from WP2 and WP3, were already used to assemble a PREMIVM pre-prototype which was used in WP1. As expected the preliminary results were merely encouraging as the set-up used is not the final one.

The consortium believed that at this stage a very important step was taken as all the data gathered pointed to the feasibility of using a tool such as PREMIVM in the vitiviniculture sector. The first measuring campaign with the new PREMIVM instrument clearly demonstrated that this device was suitable for the envisaged application in non-invasive detection of grapevine ripening. The innovative use of measuring fluorescence and reflectance multispectral data was successfully shown at this stage.

In the second period, the consortium updated the sampling plans and laboratory protocols designed in 2011 to proceed with the work in WP1 (2012 and 2013 seasons). These were already defined as standard operation protocols to compose the validation plan to be used in WP6 for the demonstration of the prototype (2013 season). KIT led once more the main efforts in the collection and analysis of samples from vineyards, performed by the members of the consortium. KIT used again the vineyards belonging to the Geilweilerhof Institute for Grape Breeding.

The grape varieties used in the field experiments conducted by KIT were the same as in the first period: Riesling, Chardonnay, Cabernet Sauvignon and Blue Portuguese. Samples were collected every week, from 3 sites representative of the vineyard. A special attention was given to collect samples in different positions and sun exposure regimes. These samples were stored in tubes with methanol and sent to Czech Globe for HPLC analysis (Flavan-3-ols, Catechin + epicatechin, Hydroxy cinnamic acids, Anthocyanins, Total polyphenols). The measurements with the prototype we performed using the following protocol for each variety: 5 repetitions in the same berry, 5 berries form selected clusters and 5 berries from different clusters. Subsequently Brix analyses were made for each berry. Entire cluster were collected for FTIR analysis (density, acidity, tartaric acid, malic acid, sucrose, fructose). Other analyses performed were pigments, berry size and color.

The construction of the correlation library was centralized in PSI, who established the procedures in the correlation between fluorescence and reflectance measurements and the reference measurements. Czech Globe and PSI assisted KIT in the characterization of the samples by using different approaches on fluorescence and reflectance techniques and data processing. A massive sampling was carried out in several vineyards in Germany, Czech Republic, Portugal and Italy. The winegrowers QMF, PERACCIO and SAPRUEM made a vital contribution at this stage, as well as the company AGRI. Several prototype units were manufactured and distributed among all these partners. They all performed field readings using the prototype and collected samples that were immediately sent to Czech Globe following the sampling plan. The samples collected in vineyards were quantitatively analyzed by the laboratories of Czech Globe using standard analytical methods. The grape varieties include the ones use by KIT, plus Pinot Noir, Saint Laurent, Syrah, Pinot Blanc, Sangiovese, Colorino, Macabeu, Moscatel Galego, Moscatel de Setúbal, Alvarinho, Viosinho, Encruzado and Arinto.

The processing of analytical data obtained from berry samples (reflectance and fluorescence spectra together with chemical analyses) was primarily done using an artificial neural networks approach, enabling to model linear and non-linear relationships between a set of input and output variables. Besides the use of neural networks for non-linear correlations, specific reflectance and fluorescence ratios were studied to establish linear correlations. The results obtained with the PREMIVM prototype, in field trials in three consecutive seasons (2011-2013), demonstrated that this device is suitable for non-invasive and fast estimation of grapevine ripening. A correlation library between the measurements of the instrument and standard chemical methods was made, with excellent correlation coefficients (r2> 0.7) for all relevant ripeness parameters: Brix, Chlorophyll, Anthocyanins, Catechin, Epi-catechin, and Caftaric acid. The results clearly show the potential for this device to replace destructive chemical analysis. The fast data acquisition of the new instrument makes it superior over the time-consuming, and mostly destructive, standard chemical analysis used in today's vine growing management.

In WP2, in order to start in the first period of the project the development of the optical transducer to be integrated in the prototype, CZECH GLOBE worked closely with PSI in the identification of specific polyphenolic reporter molecules existent in grapes and the respective excitation wavelengths to gather pertinent data. They concentrated their efforts on first measuring the absorbance spectra of standards in methanol, absorbance measured on extracts from peels (all pigments extracted: polyphenols, carotenoids and chlorophylls), absorbance measured on separated peels. They conducted a swiping analysis of samples using both fluorescence and reflectance measurement. First, they identified the absorption maxima of each standard. Then, they measured their emission spectra at these excitation wavelengths and were able to identify specific wavelengths to detect important organic compounds like acids, flavonols and anthocyanins. With the identification of excitation and emission wavelengths, CZECH GLOBE and PSI started working on the measurement of fluorescence at different wavelengths for different known concentration of standards; correlate the signal with the concentration. Then started to measure absorption and fluorescence of mixture of standards (in methanol) that would mimic real concentration measured in berries (Pinot Blanc, Pinot Gris, Laurot, Malverina, Cabernet Sauvignon) and to compare the results with measurements done on real samples. With the above mentioned information they finally started developing the optical set-up. Two preliminary optical set-ups were developed, one for fluorescence and another for reflectance measurements of grape samples. These optical set-ups were then used by KIT and CZECH GLOBE to perform more analysis of samples collected in vineyards. These analyses were compared with standard laboratory methodologies. They also tackled the design of a sample head to measure leaves and grapes. To measure leaves both CZECH GLOBE and PSI already had a suitable design since they are long time experts in the analysis of this kind of samples. The real challenge lied in grapes due to their spherical geometry, for which a preliminary design was already developed and presented in the M6 meeting in Karlsruhe (Germany) in July 2011. The SMEs AGRI, DVC, SAPRUEM and PERACCIO had the opportunity to test the functionality of this measuring head and define improvement opportunities.

During the second period, Czech Globe and PSI continued to work in the integration of the optical transducer and checked its performance in laboratory conditions. Czech Globe and PSI identified the main polyphenols varying their concentration during the ripening process and thus influencing the quality of the berries. They measured their content in berries of 4 varieties and correlated their contents with optical signals of the berries. Then they designed the hardware of the optical transducer of the PREMIVM prototype and successfully correlated the results from prototype with those of a standard fluorescence spectrometer.

The vineyard chosen to support the activities under WP2 belongs to Winberg Winery Ltd, located in Mikulov, about 180km east from Nové Hrady. All field actions were planned and performed with the supportive role of SAPRUEM and PERACCIO as winegrowing experts. Berries from 4 varieties were collected (Chardonnay, Riesling, Pinot Noir and Saint Laurent), 4 berries from one cluster (sun exposed, shade, top, and bottom), 6 sampling points per hectare. Samples were collected from August till October when the winegrowers harvested the berries. The samples were subjected to chemical analyses: sugar content, acidity, pH, berry size, polyphenols content spectroscopically and using HPLC (catechin, epicatechin, caftaric acid, resveratrol, malvidine-3-Glc).

The berries were scanned by using a flat-bed scanner. Each grape berry was scanned together with ruler (sliding guage) to determine the berry diameter. and determine color of the berries. Reflectance measurements were done for each collected berry, using an Optical Spectrometer. Fluorescence measurements were done for each collected berry, and emission spectra were taken using a commercial spectrofluorimeter. Based on the laboratory measurements, the PREMIVM optical transducer was designed with the help of PSI and with DVC in a consultative role.

In WP3, during the first period CRIC worked on the development of a custom-dimensioned casing to house the optical platform, to ensure high-strength, dust proof, splash proof, and chemical stability and temperature resistance properties. This casing had a specific internal configuration to allow the installation of all necessary components: touch screen, printed circuit boards, GPS, battery and all the remaining minor accessories plus the optical set-up developed by CZECH GLOBE and PSI in WP2. The prototype had a GPS module implemented, with a compact design and low power consumption. The module implemented would allow that while making a reading in the vineyard, the positioning through lock onto the satellite signals could be made rapidly even in areas with dense canopy removing the need for an external antenna. To store all acquired data in the field, CRIC equipped the prototype with an internal memory , so that the end users could save all multiparametric data entries with time stamp and GPS tag and later on dump all the measurements to the PC easily by means of a USB port. The prototype was equipped with a Li-ion battery to allow maintaining the device switched on for at least 12 hours. The prototype was equipped with a Resistive touch screen to allow to the user to interface with PREMIUM. The resistive touch screen is composed of two transparent and rigid layers forming a "sandwich" structure which have an internal resistive layer. This resistive system consists of a normal glass panel that is covered with a conductive and a resistive metallic layer. These two layers are held apart by spacers, and a scratch-resistant layer is placed on the top of the whole setup. An electrical current runs through the two layers while the screen is switched on. When a user touches the screen, the two layers make contact in an exact spot. A first interface software specifically for winegrowers was designed. SMEs AGRI, DVC, SAPREUM and PERACCIO analyzed its architecture and considered it to be suitable for winegrowers' needs.

In de second period, after the development of housing and accessories and respective component integration, a system test was conducted to confirm the smooth running of the GPS module and the measurement storage effectiveness. Each time one measurement was done, the PREMIVM device was able to store the measurement result, including the geopositioning points where the measurement was done. The storage of the measurement data values were done to specific folders. The graphical representation of the signal of each measurement could be done and easily represented with the PREMIVM device, without need of a PC to export the measurement data values, and check the results in situ. File reports were generated with all measurements done, including in every measure the name, number of measure, time, date, latitude, longitude and the result of the measure.

Field tests were conducted thanks to the collaboration of CRIC with "Cooperativa Agricola de Barberà de la Conca" (Spain) who gave us access to their fields. Field trials were planned with the help and supervision of SAPRUEM, PERACCIO and QMF. As the most important issue regarding this test was to verify if the GPS coordinates acquired by the device and the measurement storage were correct, the test was done by starting the first measurements in the edge of the vineyard, and making the following measurements approximately each 10 meters in alternative rows. Approximately 103 measurements were made covering the vineyard. Once the data concerning the measurements were dumped, it was demonstrated that the measurement process was done with good performance. To demonstrate that the GPS coordinates obtained in the measurements are correct, each point was represented in a map. In the generated map it was observable that the coordinates correspond to the locations where the measurements were taken.

At this point of this third work package, it was demonstrated that the implementation of the main tasks, like the geopositioning module and the internal memory storage, also the component integration, worked according to specifications and with good performance. All individual components and accessories of the prototype were developed and tested, and ready to be integrated into a final prototype inWP5.

In WP4, during the first period CRIC together with QMF, and with the input from SAPRUEM and PERACCIO, defined the specifications for the Graphical User Interface and Data Base Architecture: - The Decision Support Software should store automatically the data downloaded from the device, storing and ordering it according to the date and the position of the readings;

- The software should generate a map of the vineyard so that the producer can visually see what is happening (for example using a color code to represent the phenolic maturity in different locations of the vineyard);
- The software should be capable of generating easy to understand statistical analysis. For example the producer could ask to draw a graph to see the evolution of phenolic maturity in given a area of your vineyard;
- The software should enable the possibility to access it through internet. For example he producer could have one of his/her employees taking readings in the field. This person would then plug the device to a computer and the data would download automatically with a single click. Using internet the SME owner can follow the behavior of the vineyard anywhere in the world.

CRIC presented the preliminary architecture of the Premivm Management Software in the M6 meeting, where all partners had the opportunity to discuss in detail and make a final approval of all components. The software was being developed to satisfy the need of different types of users, from low level (individual small producers), to medium level (producers with large vineyards) and finally an advanced level (Large groups or associations of SME producers). The Use Cases and Navigation Maps defined would lead to a series of standard functionalities like log, register, edition, errors, statistics, predictions, actions, among others. The central station would use Mapnik or Google Maps to pinpoint and obtain maps of the vineyards of each winegrower. On top of these maps, a series of visual overlapping layers will provide easy to interpret GIS information on different conditions like phenolic ripeness of the grapes. The User Page Mockup discussed in the consortium already presented a window with the vineyard map of QMF as well as technical details of the field as area, rows, grape varieties, etc and a simulation of working orders for actions to be implemented in the field.

In the second period, after designing the Graphical User Interface, the Business Logic, and the DAO layer in the first period, CRIC worked together with AGRI, DVC, PSI, QMF and SAPRUEM on the Data Base architecture and the final Release of the Software applications.

The central station of Premivm manages the application and interacts with user nodes. The user nodes are clients that interact with the server through a web browser. For this type of nodes the system provides the following functionalities: Management of users; Management of vineyards, blocks and work orders; Generation of vineyard status over a map; Management of user resources; Generation of statistics; Allow user customization; Upload data from devices. The different technologies selected to develop the PREMIVM central station software, include: Java, Spring, hibernate, MySQL, Tomcat, Ajax, Javascript, JQuery and Google Maps.

According with the requirements the application is divided in two different modules: a) Web application: This module provides the web based interaction with the user; b) Data Processing Module: This module processes the raw data and generate an interpolated heat map using this processed data. Additionally an external geographic information system service will be used in order to represent the data collected from the devices.

The main functionalities of the software include User Customization, Vineyard and Block registration, Work order generation, Data uploading and interpolated map generation and Statistics.

User Customization: The user can customize variables related with the description of the vineyard blocks and also with the generation of work and block orders: Activities, Row orientation, Soil type, Grape variety. The user can also create crews (work teams) assign roles to crew members and insert new members in a crew. It is necessary that the user had already included at least a crew and a role to insert new members.

Vineyard and Block registration: This is easily done by using graphical tools that allows the user to paint a shape over the map to define the vineyard bounds. The web page that allows the vineyard registration contains an embedded map that is requested by using a connection to Google maps server. Initially the map is configured to be centered and zoomed over Europe territory in a Satellite view. The user has the option to find the desired vineyard and define the bounds generating a polygon over the map with editable vertex that allows the user to fit the shape to the vineyard as accurately as he/she wants. The user can divide the vineyard into blocks using the same drawing tool.

Work order generation: The user can generate work orders and block orders. The generation is possible thanks a dynamic form in the “Work Orders section” where the user generate a work order that is a composition of block orders. It is necessary that the user has already registered a vineyard with its blocks, a crew with its members and some possible activities to assign. The table “incomplete work orders” will show to the user the work order status that will be modified as block orders become finished.

Data uploading and interpolated map generation: The data recollected by the user must be uploaded to the server that allocates the Premivm central station. This data will be used to generate interpolated maps that will help the user to interpret graphically the information obtained by the device.

Statistics: The user will be able to represent correlated data stored in database accessing the “Statistics” section. The inputs needed to represent the data are the range of time, the block/blocks of vineyard to study and the type of data.

WP5 was executed entirely in the second period of the project. Czech Globe and PSI worked on the final design and integration of the prototype, with the support of the winegrowing SMEs QMF, SAPRUEM and PERACCIO. With the designed optical head, composed of different excitation sources, it is possible to measure Reflectance and Fluorescence.

The optical transducer is connected to a sample holder, as it: i) protects internal optics and electronics; ii) can be easily cleaned; iii) can be used for liquid samples in future. The sample holder was designed to follow the requirement to keep samples as much as possible at same position without the need of removing berries from the clusters.

Electronics controlling the PREMIVM instrument is based on an ARM7 microprocessor. Other components of the instruments are: a) high capacity flash memory; b) GPS receiver; c) real time clock; d) integrated high capacity Li-ION battery with USB charging capability. User interface was designed with a graphical color display and touch screen.

The firmware of the device includes several practical features to enable the user to interface with the hardware. It is composed of several menus: 1) Device: i) Calibrate – calibration can be run any time by pressing Device->Calibrate after inserting appropriate standard in the clip. Also includes how often should calibration be performed. 2) Data: i) Browse – displays data browse dialog box; ii) Erase – use this menu to erase internal data memory; iii) Memory info – displays information on used memory. 3) Settings: i) Time: Set the actual time and date (all data files are stored by time and date signature). To change time, touch on one of the values and adjust it using the arrows. 4) Miscellaneous: i) Backlight intensity – move slider to adjust backlight intensity; ii) Backlight time-out – move slider to adjust backlight time-out (time of inactivity required before backlight will dim out to save battery life).

The performance of the prototype was checked by comparison with a laboratory spectrofluorometer. During the duration of the PREMIVM project duration, representatives of the main polyphenol groups varying in their concentration during ripening process, and thus influencing the quality of the berries, were identified. Their fluorescence properties and wavelengths were specified, in which their concentration in berries can be measured.

In red varieties the main polyphenols that can be optically detected are anthocyanins that can be measured by means of reflectance and flavan-3-ols measured by means of fluorescence. In white varieties, the main polyphenols are usually flaval-3-ols and hydroxycinnamic acids (HCA), both measured by means of fluorescence. Since anthocyanins can be measured by standard procedure using reflectance, the development effort was focused on fluorescence.

To compare performance of the PREMIVM prototype with a standard laboratory spectrofluorometer, 4 red and 2 white berry varieties were tested. Berries from each variety were measured in parallel with the PREMIVM prototype and the spectrofluorometer. Then the polyphenols were extracted and measured by HPLC. The correlations between the performance of PREMIVM prototype and the standard laboratory spectrofluorometer were found to be quite satisfactory (r2 > 0.7).

In WP6, a sampling plan was designed, in order to have all partners of the consortium working with one single field protocol during the DEMO phase.

The RTDs Czech Globe and KIT proposed to measure and take samples 1x week (e.g. each Tuesday), during the last 3 weeks before harvest 2x week (e.g. Mondays and Thursdays). According to the plan initially presented by CRIC in the first period of the project, Czech Globe and KIT proposed to have approximately 6-10 sampling places/hectare for each variety. From each place 1 sample should be taken. One sample means 3-4 berries that will be firstly measured by PREMIVM instrument, then plucked and placed into the plastic tube with methanol (will be delivered to partners).

The partners measured the representative berries from clusters of each variety using the PREMIVM prototype (3-5 berries). Then each berry was detached and placed into a sample holder in such a way that the top of the berry would be oriented toward the device. Measurements were taken repeatedly 3-5x. Then the berries were placed in a plastic cube with methanol (the berries should be immersed in the methanol), and the tube was shaken. Next the tubes were labeled and placed in dark storage before being sent to the laboratories of Czech Globe for HPLC analysis. CzechGlobe and KIT additionally discriminated berries from sun and shade side of the cluster. For each variety and sampling position, two berries from sun exposed part and two berries from shadow part were inspected.

As already mentioned, before being collected and transported to the laboratory, each berry was measured in the field using the PREMIVM prototype. After each field trial, the end-users would upload in the PREMIVM software the measurements taken with GPS tags. The measured varieties were: 1) KIT: 4 varieties (Riesling, Chardonnay, Cabernet Sauvignon, Blue Portuguese); 2) CzechGlobe: 4 varieties(Riesling, Chardonnay, Pinot Noir, Saint Laurent); 3) PERACCIO: 2 varieties (Sangiovese, Colorino); 4) SAPRUEM: 2 varieties (Riesling+ Pinot Blanc); 5) QMF: 1 variety (Syrah); 6) AGRI: 7 varieties (Macabeu, Moscatel Galego, Moscatel de Setúbal, Alvarinho, Viosinho, Encruzado and Arinto).

During the execution of the field trials in Portugal, Germany, Czech Republic and Italy, the feedback of the end-users was critical to improve hardware, firmware and software of the PREMIVM prototype. The improvements made in software included: 1) Export data to Excel file; 2) Black list unwanted data; 3) Option graph + table statistics; 4) Create field notes; 5) Vineyard boundary edition. The improvements in firmware included: 1) constant updates of correlation models (more than 120 models were generated and tested); 2) 24 hours time format; 3) Synchronize time with PC; 4) Automatic installation of device driver; 5) Firmware updates uploaded from server. The improvements made at hardware level included: 1) Design a new light-cover for the measuring head of the prototype, usable for field measurements in grape clusters (grapes are not removed from clusters).

The main conclusions drawn from the demonstration of the prototype in the field were: 1- The prototype has an excellent performance, with high repeatability of results and a very low standard deviation, even when a berry is measured in different positions; 2- The values given by all the prototype units tested (non-destructive analysis) are consistent and of the same order of magnitude as the results achieved when using standard analytical methods (destructive analysis).

While the field trials were being tackled, the SME partner DVC developed a Smartphone version of the PREMIVM software with the collaboration of CRIC. The PREMIVM mobile web application allows users to browse through the data and perform different operations such as registering users, visualizing vineyards/blocks, consulting work orders, modifying data types, showing statistics etc. The PREMIVM mobile web application was developed to be used on different types of smart phones and tablets. The application was designed for a comfort usage on a various types of devices and it automatically adapts to user's screen. Currently it supports Android, iOs and Windows Phone's browsers.

Potential Impact:

During the first period of the project, RTD to SME technology transfer took place at the quarterly project meetings, where RTD activities for the period are presented and discussed. A training plan was outlined. According to the agreed plan, the periodic consortium meetings would be used to carry out technology transfer sessions where RTD partners transfer the generated foreground to SMEs. SMEs also contributed to the technology transfer sessions by exposing their background to aid in the definition of alternative and complementary development paths to achieve the intended foreground. Partners submitted a draft version of the Plan for the Use and Dissemination of Foreground (PUDF) describing dissemination results to date and planned dissemination activities. The PREMIVM website was produced and available online at http://www.premivm.eu since M2. A leaflet for use at Trade Fairs and other dissemination events was designed as well as a poster, providing an attractive marketing image which would help disseminating the results to a wider audience during the second part of the project. Technical and End User SMEs undertook discussions in periodic project meetings regarding the exploitation potential of the technology. Technical SME partners AGRI, PSI and DVC outlined an exploitation plan which would continue to be refined during the next months of the project.

The training plan designed in the first period was executed to ensure that the lead-user and end-user SMEs assimilated the project results including the technical, socio-economic and commercial benefits of PREMIVM. Tutorial for the use of hardware, firmware and software were produced within the PREMIVM project to assist in the training and technology transfer actions.

In the second period of the project, the Exploitation Manager AGRI, with the assistance from CRIC, coordinated all the major training/ technology transfer actions between RTDs and SMEs, under WP7. Together they defined the material to be used, training programme and arranged for its production. The first step in the Training Plan was to continuously promote technology transfer during the whole duration of the project (joint R&D efforts, meetings, sharing of documentation and information), particularly between RTDs and lead-user SMEs PSI, DVC and AGRI. Specifically oriented and extended training actions were organized to be performed together with all SMEs.

The SMEs were first trained in two introductory sessions. The first one was in a meeting held in Brno (Czech Republic), organized by the SME partner PSI in June 2012. The second one was in meeting held in Wehlen (Germany), organized by the SME partner SAPRUEM in December 2012. The RTDs and PSI made the first knowledge transfer sessions concerning the prototype and its operation, as well as the web-application software.

After the first introductory session, several prototype units were produced and given to the SME winegrowers QMF, PERACCIO and SAPRUEM. During the summer of 2012, each of these SMEs made field trials using the prototypes in their own vineyards. Their feedback was paramount for the success of the second introductory session, where the consortium had the opportunity to discuss the performance of the current prototype. Several improvements were suggested from the point of view of the end-user. In the second introductory session, the SMEs were trained on the use of the PREMIVM web-application software. The RTD CRIC was in charge of this with the support of the SME AGRI. In April 2013 the consortium met in a special session in Brussels, organized by the SME DVC. In this technology transfer session the whole consortium made a complete training on the use of all the partial results of the project. Finally in July 2013, the consortium met in a final joint training session, including field trials in vineyards. This meeting took place in Lisbon (Portugal) and was hosted by the SME partner AGRI. In this event all partners used several prototypes simultaneously, conducting assays both indoors and outdoors.

The public website was updated regularly updated and improved during the second part of the project, including information about the dissemination events, media material produced by the project partners, scientific posters, links to interviews, etc. It has a restricted area for the access and storage of technical information by the consortium partners. The PREMIVM website is available online at http://www.premivm.eu since M3.

All SME partners disseminated and contributed to the design of the exploitation plan for the project-results. Every partner was responsible for dissemination in his own country. Country-overlapped or European-wide activities were carried out together by the project consortium. SMEs were primarily responsible for dissemination in trade fairs, trade magazines and to business contacts, etc. RTDs could also participate in events of this nature. RTDs performed dissemination to the scientific community, subject to approval by the exploitation board. All scientific publications include partner names (with consent), reference to the PREMIVM project, and reference to EU funding. SMEs could also participate in events of this nature. Several dissemination materials were produced: web page, logo, flyers, posters, press notes, articles and videos.

The SMEs led by the Exploitation Manager continuously performed activates under Knowledge Management, IP Protection and Exploitation Potential. The discussion on the Exploitation and Use of the Foreground with the SMEs is initiated.

The main results as exploitable knowledge and exploitable products, are: 1) PREMIVM system as a whole (patent/trade secret)- Hand-held, multi-parametric and non-invasive analyzer of grapes and vine leaves; 2) New optical fluorescence/reflectance transducer (patent/trade secret)- Optical system capable of acquiring directly and non-invasively full spectrums of chlorophyll fluorescence and reflectance from fruits and leaves; 3) Correlation library (trade secret)- Extensive spectral fluorescence and reflectance characterization library which establishes the relationship between chlorophyll and nitrogen content in vine leaves and grape ripening, extending knowledge in the application of fluorescence and reflectance principles for a better understanding of plant and fruit physiology in vineyards; 4) Decision-making software (copyright)- Data processor and storage module that automatically receives and processes geospatial data transferred from a sensor. It has implemented correlation algorithms that, through a graphical representation tools bundle, process the stored data to automatically generate field reports and statistics; PREVIM app (copyright)- Smart-phone application to access primary functionalities of the decision-making software using portable terminals (mobiles and tablets).

In terms of IP Protection, the SMEs planned a combined strategy, effectively covering all results: patent, trade secret, copyright and trademark. After an extensive technology watch effort continuously made by the Exploitation Manager Carlos Lopes and his team at AGRI, the consortium partners concluded that with the situation of competitors and the positive results achieved by PREMIVM, the potential commercial exploitation of the results is viable. The PREMIVM business model is based on the strengths of the consortium which covers all of the expertises needed to commercialize the system. The commercial route envisaged for the exploitation of the results will be preferentially done via consortium members.

Partners PSI, DVC and AGRI will lead the production and initial distribution of the new technology using the Viticulture Management Services commercialized by AGRI. Partners SAPRUEM, PERACIO and QMF may assume a tactic role in the marketing of PREMIVM, particularly in their native countries. AGRI will use its solid position in the EU vitiviniculture market to rapidly coordinate the distribution of the PREMIVM system. Due to their privileged geographical position and vast experience in EU sales, PSI and DVC will assist AGRI. Due to their commercial experience in the wine sector, SAPRUEM, PERACIO and QMF can also promote the solution, acting as prescribers, representatives and/or distributors;

The prototype was successfully integrated and tested in different vineyards in Europe. The final unit is a low-cost, handheld spectrometer with touch screen display, and the capability to make wavelength and intensity readings of fluorescence and reflectance. It has a memory capacity up to 8000 measurements with a battery life of 48 hours in continuous operation. Its dimensions are 255 x 75 x 40mm, and it has a weight of only 300g. The results obtained with the pre-competitive prototype in three consecutive seasons (2011-2013), demonstrated that this device is suitable for non-invasive and fast estimation of grapevine ripening. A correlation library between the measurements of the instrument and standard chemical methods was made, with correlation factors superior to 0.75 in relevant ripeness factors. The field demonstration results clearly showed the potential for this device to replace destructive chemical analysis. The fast data acquisition of the new instrument makes it superior over the time-consuming, and mostly destructive, standard chemical analysis used in today's vine growing management.

Europe’s primary sector has in vitiviniculture one of its dominant areas, which is currently suffering strong market and regulatory pressures. The production of fewer grapes of a higher quality is the only way for the EU wine sector to maintain economic feasibility, a fact supported by the example of Portugal, which produces six times less wine than Spain and generates identical income. In today’s global market of well-informed consumers the price of a wine is dictated by its quality, not by its Protected Designation of Origin (PDO).

Current mechanisms and methodologies to control the vineyard have proven to be ineffective. On the other hand, the new field instruments fighting for a competitive position in the market do not offer the complete cost effective analytical package solution sought by winegrowers. PREMIVM implies a fixed investment of €2.500 on a hand-held multiparametric device and supporting vineyard management software. Unlike current hand-held instruments, PREMIVM will permit a straightforward interpretation of the data collected in field, by the use of a tailor-made software. This can process the accumulated data and present it numerically with statistical tools and visually though digital vineyard maps, where the evolution of fruits and plants can be presented in a very comprehensive way.

The consortium estimates that the implementation of PREMIVM will allow end-user winegrowing companies to cost-effectively increase the quality of their grapes at least one level higher (low: €300/ton, medium: €600/ton, high: €1.200/ton) which will allow the production of wines of a superior segment. Using PREMIVM, SMEs estimate the expected increase of gross retail value of vitiviniculture production at up to 40% over a five-year period by means of differential harvesting and increased average berry quality. Other mid-term estimated financial benefits, derived from the implementation of integrated agriculture practices, include reduction of laboratory analysis up to and potentially the reduction of pesticides and fertilizers.

The expected market demand for this new application shall benefit technological consortium partners PSI, DVC and AGRI as lead-users and suppliers of the PREMIVM components and finished product. With an estimated EU market penetration of 2% in five years, the partners expect to jointly have a yearly Return of Investment of 86% within 5 years. This value takes into account the budget requested for the project, costs to reach time-to-market, and production/distribution costs.

List of Websites:

http://www.premivm.eu