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SYMPHONY Newsletter Integrated SYsteM based on PHOtonic microresonators and Microfluidic Components for rapid detectioN of toxins in milk and dairY products Fondazione Bruno Kessler April 2015 Newsleer, Issue 1 This project has received funding from the European Unions Seventh Framework Programme FP7-ICT-2013-10 under grant agreement No 610580. Project duraon: 1 st November 2013 - 31 st October 2016. www.symphony-project.eu The project SYMPHONY is a research and innovaon project, funded by the European Commission to find a rapid, cheap and effecve soluon to detect toxins in milk. More infos about the project and the consorum at pag. 2-3 Achievements First prototypes of Photonic sensors demonstrated to achieve Limit Of Detecon (LOD) of 0.8*10 -6 Refracve Index Units (RIU) . Find more results aſter 18 months of the project at page 4. The Milk DayExperts in milk analysis and quality, sciensts in microsystem technologies and biosensors and end users gathered in a workshop organized in Trento in February 2015, to talk about milk. More details from pag. 6. IN THIS ISSUE

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Page 1: Newsletter, Issue 1 SYMPHONY Newslettersymphony-project.eu/sites/... · SYMPHONY onsortium 3 SYMPHONY objectives SYMPHONY is a demand-driven project pursuing: Advancement of MN S

SYMPHONY Newsletter Integrated SYsteM based on PHOtonic microresonators and

Microfluidic Components for rapid detectioN of toxins in milk

and dairY products

Fondazione Bruno Kessler April 2015 Newsletter, Issue 1

This project has received funding from the European Union’s Seventh Framework Programme FP7-ICT-2013-10 under

grant agreement No 610580. Project duration: 1st November 2013 - 31st October 2016.

www.symphony-project.eu

The project SYMPHONY is a research and

innovation project, funded by

the European Commission to

find a rapid, cheap and

effective solution to detect

toxins in milk. More infos

about the project and the

consortium at pag. 2-3

Achievements First prototypes of Photonic

sensors demonstrated to

achieve Limit Of Detection

(LOD) of 0.8*10-6 Refractive

Index Units (RIU) . Find more

results after 18 months of the

project at page 4.

The “Milk Day” Experts in milk analysis and

quality, scientists in

microsystem technologies and

biosensors and end users

gathered in a workshop

organized in Trento in February

2015, to talk about milk. More

details from pag. 6.

IN THIS ISSUE

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SYMPHONY in a nutshell Integrated SYsteM based on PHOtonic

microresonators and Microfluidic Components for

rapid detectioN of toxins in milk and dairY products

I n the dairy industry, one of the most pressing needs for food quality and

safety is the timely detection of aflatoxin M1 contaminations that originate

from animal feed and are secreted into milk. This issue represents a hazard

for human health and an economic loss for the dairy industry. The available

technology for aflatoxin detection is laboratory-based, in most cases

requiring sample preparation and does not provide timely identification of

contaminants, thus fails to deliver cost-effective management of milk

quality. The entire supply chain relies on the fact that the dairy products

delivered to consumers are safe. Current best practices for Aflatoxin

management in milk in the modern dairy industry recommend aflatoxin M1

analysis to be carried out routinely for quality assurance and quality control

purposes as well as for feed-back mechanisms to farm and animal health

management, as requested by European Commission regulation (EC) No.

1881/2006. The current procedure involves the collection of samples from

collection tanks. The milk samples are analysed in a laboratory with

screening tests such as Enzyme-Linked Immunosorbent Assay (ELISA).

Contaminated milk that exceeds the 50 ng/Kg (ppt) aflatoxin limit is not

allowed in the market and has to be disposed.

T he SYMPHONY STREP project (FP7-ICT-2013-10) addresses the needs of

the dairy industry with novel microfluidic technologies and biochemistry for

sample preparation, photonic integrated sensors and compact hardware for

integration in the production chain of the dairy industry, leading toward

precision process management.

This project has received funding from the European Union’s

Seventh Framework Programme FP7-ICT-2013-10 under grant

agreement No 610580. Project duration: 1st November 2013 -

31st October 2016.

Interview with the coordinator

Dr. Leandro Lorenzelli, Head of

the MST research unit at Fonda-

zione Bruno Kessler, Trento

Could you explain what SYM-

PHONY is about?

SYMPHONY is proposing a

new generataion of high-tech

devices allowing for fast and

reliable analysis of the dairy

products quality. These would

lead to a more precise and

effective management of pro-

duction and distribution pro-

cesses.

What is SYMPHONY main

objective?

The objective is to integrate

different technologies like

photonics, microfluidics and

microbiology to create a por-

table and quick tool to detect

aflatoxin in milk.

SYMPHONY Newsletter N. 1, April 2015

Grant agreement 610580

www.symphony-project.eu

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SYMPHONY Consortium

3

SYMPHONY objectives SYMPHONY is a demand-driven project pursuing:

Advancement of MNBS devices toward real-world applications by providing:

Integrated sample preparation for analytical tools in dairy industry,

Advancement in high sensitivity photonics biosensors, by using aptamers to provide selectivity to targets and improved sensor stability and hybrid integration of sources and detectors.

Improvement of milk quality assurance by making available new technologies and procedures for the detection of aflatoxin and other contamination

Improvement of dairy industry competitiveness by early detection and effec-tive corrective action against aflatoxin outbreaks and contaminants:

On field simultaneous detection of various contaminants, thanks to arrays of photonic sensors;

Analysis of raw milk and other liquid dairy products, such as yogurt, butter milk, kefir, whey;

Quality assurance at various HACCP

Improvement of food quality with positive impact on human health

Symphony facts

EU STREP project

7 partners in 5 country

3 years project

3 M€ Funding

3 private companies

3 research and academic

centers

1 dairy cooperative

www.symphony-project.eu

Follow us on

Grant agreement 610580

SYMPHONY Newsletter N. 1, April 2015

www.symphony-project.eu

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First year scientific achievements Stakeholders needs, system specifications & architecture

Stakeholder needs were surveyed by interviews and questionnaires for the definition of user requirements, specifi-

cations and the general architecture of the system.

Antibiotics were selected as best additional target to be evaluated in addition to Aflatoxin for multianalyte system.

Development of photonic sensors

SiN asymmetric Mach-Zehnder interferometers (aMZI) were developed by LioniX

SiON microring resonators (mRR) were jointly developed by FBK and UniTN

The achieved Limit Of Detection (LOD) was 0.8*10-6 Refractive Index Units (RIU) and 1.6*10-6 RIU in the case of aMZI

and mRR, respectively.

VCSEL sources and photodiodes are being integrated by means of heterogeneous assembly

Functional surfaces and analytical methods

Aflatoxin purification is based on functional surfaces based on surface coating with mussel foot protein.

Anti-aflatoxin aptamers are immobilized by FBK on the detector surface for the selective detection of aflatoxin,

with a demonstrated layer density around 1013 aflatoxin molecules per cm2

The sensor regeneration and reusability was demonstrated up to 17 regeneration cycles.

Development of microfluidic components

Microfluidic modules for fat separation and milk fractionation produced and tested showing a fat reduction compa-

rable with laboratory techniques.

Reversible Sensor packaging with integral flow cells, has been designed and fabricated.

4

Grant agreement 610580

SYMPHONY Newsletter N. 1, April 2015

www.symphony-project.eu

Micro Ring Resonators Watch the video on: http://symphony-project.eu/

node/3

Aflatoxin response of functionalized

microring resonator

Hybrid integration of microfluidics and silicon chip

Success criteria for SYMPHONY project

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5

SYMPHONY dissemination

Other events Workshop on biosensing,

Trento 30th March 2015.

http://webmagazine.unitn.it/

evento/dphys/4244/workshop

-on-biosensing

Workshop on “Smart sy-

stems for food industries”,

Trento 15th November

2013

http://www.symphony-

project.eu/node/21

Researcher’s Night 2013,

Trento

Upcoming events

21th World micromachine

summit, Berlin 10-13 May

2015

http://www.mms2015.org/

Recent Advance in Food

Analysis (RAFA 2015), Pra-

gue 3-6 November 2015.

http://www.rafa2015.eu/

index.html

3rd EOS Conference on Op-

tofluidics (EOSOF 2015)

held under the umbrella of

the World of Photonics

Congress 2015 , Munich 22-

23 June 2015.

http://www.myeos.org/

Grant agreement 610580

SYMPHONY Newsletter N. 1, April 2015

www.symphony-project.eu

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The Milk Day rationale

The Milk Day was organized

in Trento on February 4th in

the framework of the AISEM

conference to discuss rele-

vant open issues in the appli-

cation of MST in the analysis

of milk contaminations.

Can the use of Micro

System Technologies

and biosensors provide a

solution to overcome

these limitations?

What are the major fac-

tors that may jeopardize

the integration and au-

tomation of current

analysis techniques?

Milk chemistry and composi-

tion, strict regulations for

milk quality, dairy industry

requirements, the large

diffusion of milk consumers

and producers pose substan-

tial challenges, but, in the

meanwhile, offer a number

of opportunities for ex-

ploiting the huge potential of

miniaturized systems and

sensors.

Presentations available on:

www.symphony-project.eu/

TheMilkDay

Milk quality testing and regulations

Milk is principally tested at food pro-

ducers for two purposes:

1) Testing for milk payment, where

main quality parameters are:

Fat

Protein

Freezing point

Somatic cell count below 400.000

Total count of bacteria below

100.000

This is typically performed off-line in

large laboratories with very large num-

ber of test per day (25000 sample/day)

with high throughput instruments like

IR spectroscopy

2) Testing for chemical residues, patho-

gens and contaminants for safety.

Since food manufacturer are responsi-

ble for their products, they are in

charge for safety enforcement; tests

are performed for legal limits of:

Chloramphenicol

Mycotoxins

Aflatoxin M1

Trichlormethan

Radionuclides

Residues

Others

While reference lab only apply reference

methods, since testing campaigns are

limited and time is not a constraint, ref-

erence tests cannot be performed on all

samples at the manufacturer site. It is

therefore possible to accredit screening

test to have a fast feedback on all sam-

ples (or a larger number) if properly cali-

brated. For this purpose, calibration solu-

tions are required, which is an art by

itself since the challenge for test kit pro-

ducers is to assure sufficient selectivity,

sensitivity, accuracy.

6

Nicola Cologna, CONCAST, Trento

CONCAST structure, includes more than 800 associated farmers, 17 dairies and the

centralized testing lab.

Dr. Cologna presented the tests performed at Concast laboratory, which, in addition

to common milk quality parameters like fat and protein content, antibiotics, etc.

also measures technical properties of milk related to cheese making (e.g. milk coag-

ulation properties, whey starter analysis, presence of spores, which are particularly

important in hard cheese making). A specific case of introduction of a new accredit-

ed test for quality is presented, where FTMIR was correlated to milk coagulation

properties in order to process the test with fast and high-throughput procedure

with respect to traditional method, which requires 30 minutes to run. This study

was conducted in collaboration with the Department of Agronomy Food Natural

resources Animals Environment of the University of Padova (Italy).

Ulrich Leist, DRRR Germany

Dr. Leist provides an overview of DRRR activities as an accredited body for proficien-

cy testing services.

Milk quality testing in Trentino

Grant agreement 610580

SYMPHONY Newsletter N. 1, April 2015

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Focus on Aflatoxin

Alberto Mattivi, Director of the Department of Hygiene and Veterinary Public Health, Health Services Provider of the

Autonomous Province of Trento.

Statistics of dairies in Trentino region highlight the particular production structure in the region, where farms are very

small and mountains huts are used as feeding grounds in the summer. Primary production in Trentino region is the high

quality traditional cheeses, which represent an interesting point for safety enforcing with respect to large companies

found in Europe and other Italian regions.

In Trentino, the official tests are performed by audits and inspections on all production aspects (veterinary, product

safety, HACCP, etc.) and according to both regulations and internal rules agreed by producers and competent authority,

which are often more restrictive than general rules. For instance screening test for aflatoxin are required to be con-

firmed by test on single producers by ELISA and eventually by reference HPLC testing if a value of aflatoxin higher than

35ng/kg is found.

In the Department experience, the majority of non-conformances are related to non-observance of internal rules of the

production plant. Interestingly, there may be local issues with particular, new contaminants as in a recent case in Trenti-

no region related to steel manufacturing plant, which resulted in local contamination with dioxins and an intense

testing campaign to identify possible contaminated products before they reached the market. This would open the field

for new screening test if technically available. The cost per test to identify dioxins is quite high (1300-1400€/sample for

reference test, 500-600€/test for screening) with reduced number of molecules identified. Other similar examples of

specific needs may be found.

7

Milk and health in the province of Trento

“Aflatoxins are naturally occurring mycotoxins that are produced by Aspergillus flavus and Aspergillus parasiticus, species of fungi. The name, aflatoxin, was created around 1960 after the discovery that the source of "Turkey 'X' di-sease" was Aspergillus flavus toxins. Aflatoxins are toxic and among the most carcinogenic substances known. After entering the body, aflatoxins may be metabolized by the liver to a reactive epoxide intermediate” “or hydroxylated to become the less harmful aflatoxin M1 (Wikipedia)

Grant agreement 610580

SYMPHONY Newsletter N. 1, April 2015

A SEM image of an Aspergillus

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Veronica Lattanzio, Institute of Sciences of Food Production, National Research Council of Italy (ISPA-CNR) .

The Institute of Sciences of Food Production (ISPA-CNR) is a centre of excellence, worldwide renowned, acting in the fields

of scientific research, innovation and technology transfer aimed to improve safety and quality of agro-food products

Dr. Lattanzio presents the work of ISPA institute on rapid

detection of aflatoxin M1 in milk, and an overview of ana-

lytical challenges and validation aspects.

Rapid test methods for measuring Aflatoxin M1 (AFM1) in

milk are available either as commercial kits or as research

methods.

Enzyme-linked immunosorbent assays (ELISA), lateral flow

tests, immunoaffinity columns coupled with fluorimetric

assay are common formats in the current market. The

main purpose of screening methods is to detect the pres-

ence of a contaminant at level of interest allowing rapid

decision making, i.e. in 5 to 20 minutes with respect to

about 4 hours/samples in reference methods.

European Union has set a maximum permitted limit of

0.050 µg/kg for AFM1 in milk. Besides the high sensitivity

required for AFM1 detection in milk, the major analytical

challenge when developing screening tests is to make

them reliable and robust, in particular by reducing the false

negatives, for laboratory and in-field use. This means to

cope with differences from matrix to matrix, environmen-

tal conditions, operator skills, lot-to-lot reproducibility.

Recent efforts of the European Union, for establishing

practical guidelines for the generation of fit-for-purposes

performance parameters for screening methods for myco-

toxins in foods, resulted in the Regulation 519/2014/EC.

The EC validation scheme and its practical application to

evaluate performances of a commercial kit for AFM1 de-

tection in milk were presented and discussed. It is common

experience that fat removal can change the calibration

curve of a method and sample treatment/clean-up is need-

ed in order to improve the analytical accuracy of the sys-

tem. Dilution is already a commonly used sample prepara-

tion method.

Rapid Detection of Aflatoxin M1 in Milk

8

Grant agreement 610580

SYMPHONY Newsletter N. 1, April 2015

www.symphony-project.eu

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Mark Whatton, Technical Projects Manager at QuadraChem Laboratories Ltd, UK

QCL provide rapid analytical testing solutions for a wide range of industries across the UK. Using extensive experience QCL

applies advanced technologies to the improvement of industrial processing, quality control and environmental monitoring.

Dr. Whatton discussed in his presentation an opinion on why Micro Systems Technologies, while promising exciting and novel ways to monitor the quality of food and beverages, are difficult to translate in commercial reality.

There are many reasons why promising systems fail to reach the market, but is it possible to understand and mitigate these risks early on? The presenta-tion described the target needs and wants required to hit the market, as evaluated in the development of the Symphony project:

Innovation aims need to work with commercialisation aims, since com-mercial successes are those that provide clear benefits to those who will use them. These benefits can only be known by understanding the tar-get market.

Selection of technology and design should target specific requirements from end-users.

Development phase should go from a proof-of-concept into a pre-production prototype. An industrial partner focusing on instrument pre-production prototype will understand and work to maximize the major attraction of the innovation, and bring it to final users in such a way it is clearly perceived.

Andrea Adami, FBK-Technical manager of the SYMPHONY project.

“Working out what benefits are for a project AFTER the research and innovation is too late”

Research & Business: Transitioning Innovation to Commercial Success

9

Dr. Adami presents a critical review of advantages of microfluidic devices in food analysis. Among different peculiarities

of microfluidics, the major points of interest are mainly the integration of multiple functions in an integrated system, the

faster mass and heat transfer in small channels, the possibility to create a laminar flow, multiple parallel testing and high

surface interaction. On the

other hand, small samples typi-

cally used in microfluidics are

not a key issue in food analysis,

since sampling large volumes

are often required for sample

representativeness and typical-

ly low cost of the sample. Tech-

niques, concepts and devices of

interest in food analysis include

capillary electrophoresis, micro

PCR, immunoassay, particle

and drop separation and multi-

plexing techniques.

MST for food quality and safety

Grant agreement 610580

SYMPHONY Newsletter N. 1, April 2015

www.symphony-project.eu

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Project Coordinator: Dr. Leandro Lorenzelli Tel. +39 0461 314 455 e-mail [email protected] Technical Manager: Dr. Andrea Adami Tel. +39 0461 314 497 e-mail [email protected] FBK - "Bruno Kessler" Foundation Center for Materials and Microsystems Via Sommarive, 18 38123 Trento

Project Website: http://www.symphony-project.eu/ To subscribe/unsuscribe write an e-mail to: [email protected]