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Bacteria detector labels for smart packaging
 
            Isaure GUERRA et Inès MOURHALI
Élèves ingénieurs 2e année
Mai 2016
Mise en ligne - Mars 2017
Avertissement
Ce mémoire d'étudiants est une première approche du sujet traité dans un temps limité.
À ce titre, il ne peut être considéré comme une étude exhaustive comportant toutes les informations
et tous les acteurs concernés.
       
  Plan  
I - Introduction
II - Bacteria detection, smart packaging & TTI
III - Smart bacteria detector labels
IV - Smart bacteria detector labels market
V - Conclusion
VI - Bibliography
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I - Introduction

Plan

   
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            Bacteria      
  Figure 1 - Bacteria  

Detection of bacteria is needed in many contexts and for different reasons. In the medical field, doctors need to know as quickly as possible if there is a bacterial contamination, but also if the tools used to operate are safe and sterilized. In the military field, detection of bacteria is used to spot quickly bacteriological attack. In the food sector, industrials have the obligation to ensure the safety of their products, and the reglementation will certainly become more stringent.

Actually, detection of bacteria is possible in a laboratory and need time to produce results. These methods are applicable to the medical, military and food sectors, which want to reduce the time required to obtain results. In consequence, each sector finds new ways of detection.

In this report, we will focus on the food industry which needs only to detect and not to quantify bacteria. To know if a product is consumable or not, industrials invented different ways like smart packaging and active packaging. Many of these packagings have the purpose to reduce food waste. Indeed, it represents nearly a third of the world production, which make 1,3 billion of tons lost by year. This food is lost or wasted since the initial production to the household consumption. It's estimated from 95 to 115 kg/year by inhabitant in Europe or in North America. The European Parliament estimates that if nothing is done within 2020, food waste could rise to 40%. Moreover, as more than one billion of people are suffering of malnutrition, the issue of food waste involves a particular ethical dimension. Food waste is mainly done at home and at the production place.

     
                Proportion of the different stages of the food chain in the production of food waste in the 27 countries of the Union      
  Figure 2 - Proportion of the different stages of the food chain
in the production of food waste in the 27 countries of the Union
[European Parliament]
 
     

Bacteria detector labels are also a way to reduce food waste, but do they have the development possibility in smart packaging? Firstly, we will study the traditional methods of detection. Then, we will explain what smart packaging is and focus on time-temperature indicators (TTI). Thirdly, we will analyse the market of TTI labels. Fourthly, we will examine detector of bacteria adapted as label. Finally, we will discuss what might be the future market for these labels.

II - Bacteria detection, smart packaging & TTI

Plan

II-1 - Bacteria detection

II-1-1 - Traditional ways of detection

Antoni van Leeuwenhoek has observed for the first time bacteria thanks to his own microscope in 1668. First detection methods of bacteria used colorant combined with microscopic analysis. In the food sector, traditional tests of detection are based on biochemical and immunological characteristics of bacterium. New developments in molecular biology have completed traditional methods in food sector: tests, now based on genetic information, are more precise than other because they are not receptive to environmental conditions. They use DNA hybridization or amplification of bacterial DNA by polymerase chain reaction. These methods are more specific and faster than older methods. Industrials can find the origin of the contamination and avoid disproportionate measure.

So now, there are three types of conventional methods.

Some of them are not expensive, but they are limited by the time needed to make the culture of bacteria and to identify bacteria. While the industry is constantly expanding, the risks of contamination are increasingly present. We have therefore identified several bacteria that have already been responsible for outbreaks, and those that will be detected in our labels.

II-1-2 - Bacteria detected

Bacteria Products or locations at risk Infectious dose
(bacteria ingested)
Incubation period
Listeria monocytogenes
  • Non pasteurized milk
  • Crude meat or fish
  • Crude products
<1000 2 days to 3 weeks
Campylobacter Bowel of poultry and cattle 400-500 2-5 days
Escherichia coli
  • Undercooked ground meat products
  • Raw milk and derivatives
  • Apple juice or cider
<10 2-4 days
Salmonella
  • Milk and derivatives
  • Undercooked eggs
  • Shellfish
15-20 12-24 h
Staphylococcus aureus
  • Hospitals
  • Health facility
- -
Clostridium Canteens retain long hot food - -
Bacillus cereus Unrefrigerated food before cooking - 1-24 h

Table 1 - Most common bacteria in the concerned sectors

Escherichia coli is the most dangerous, with less than 10 bacteria needed to trigger infection.So it will be the first to be detected by labels. Most foods at risk are of animal origin. For some bacteria, the infectious dose is very small and the incubation period relatively short. This is why doctors need fast and ubiquitous detectors of bacteria. They are not yet on the market. Industrials have found other ways to ensure products security, like smart packaging and more precisely Time-Temperature Indicators (TTI).

II-2 - Smart packaging & time-temperature indicators

A smart packaging is able to give information about the content's quality to the consumer. It contains a sensor or an indicator which reacts to environmental changes and indicates them. There are many sort of smart packaging which helps to detect leak, break in the cold chain, bacteria, gas… or to follow the product point by point.

Time-temperature indicators have been developed to know if a product has been safely transported without doing some analysis. Traceability is an essential issue in the food chain. Indeed, dealers have to know the product's origin, but also if its transportation was done in good conditions. For example, if a product needs to be carried in a refrigerated truck, they want to know if there were some temperature overtakings and if the product is consumable.

II-2-1 - Thermochromic inks indicators

Thermochromic inks indicator has been developed in USA for some 15 years. In France, developments and applications are more recent. Among the different suppliers on the market, we can name:

Temptime with Fresh-Check® Time Temperature Indicators. This label shows a ring and a central pastille which is the reference. Inside the ring, a thermochromic ink is put. Initially, this ink is invisible, but In case of break of the cold chain, it blackens. This system is declined in seven versions adapted to the product. If it is needed, an explication text can be noted to help the consumer in the reading of the indications.

     
                Fresh-Check® Time Temperature Indicators      
  Figure 3 - Fresh-Check® Time Temperature Indicators
[Fresh-Check]
 
     

Hallcrest, one of the leaders in microencapsulation and in thermochromic products, offers the multiple color label: heat alert integrated into label, inner circle converts from black to red at a temperature back to black below set temperature.

     
                Multiple color label      
  Figure 4 - Multiple color label
[Hallcrest]
 
     

Topcryo, created by the French company Cryolog, is a microbiological thermal pastille which shows the impact of time and temperature by a colour change from green to red. It is easy to detect products which temperature has risen.

                Topcryo time temperature integrator       Topcryo time temperature integrator  
  Figure 5 - Topcryo
[Cryolog]
 
         

American army use this system of time-temperature to store soldier's food. Rations have to be conserved during many months and have to support a journey through the world. So the question is: after all these troubles, is food still fresh to be consumable? These indicators will allow to make the link between the quality of the rations, the conservation time and the temperature since the product's manufacturing. It's still a colored ring which surround an invisible thermochromic ink pastille. The ink will too blacken with time and the rise of the temperature.

These labels can not identify which bacteria prevent the consumption of the product. They only indicate the break of the cold chain. However, maybe the product remains consumable, which is why there is another type of label, more accurate.

Hallcrest offers an irreversible indicator, Thermostrip. It is a temperature control label which have a scale based on the property of phase change of the chemical products present in the ink. So, it is simple to know the highest temperature that the product have reached during its processing.

     
                Thermostrip      
  Figure 6 - Thermostrip
[Hallcrest]
 
     

This Thermostrip is very clear and usable in countries with differents laws, because you can see the maximum of temperature the product underwent, so it’s faster to notice that there is an overtaking of temperature.

II-2-2 - Label based on microorganism

Cryolog has also invented another solution to know if the cold chain has been broken for the alimentary and medical sectors. Traceo is a microbiological indicator of cold chain break. It is an adhesive label which indicates by a colour reaction a critical accumulation of cold chain break. This label exploit a biological reaction activated by the presence of a bacteria and become opaque. The goal is to prevent reading the barcode and the payment.

The label is composed of two parts:

     
                Traceo (Cryolog)      
  Figure 7 - Traceo (Cryolog)
[PakBec]
 
     

All of these labels (with thermochromic inks and based on microorganism) change of colour because of temperature changes. They indicate simply and quickly if there were temperature overtakings and so if the cold chain was break. However, they don’t indicate if bacteria grows or not.The next sort of TTI is not as simple, but it is more precise.

II-2-3 - Electronic plotter of temperature

It's a plotter which uses small recorder to store the temperature history from manufacturing to transportation. It can reveal all the temperature overtakings: the measured temperature can vary from -40°C to +85°C with a precision of 0,5°C. As the plotter is small and strong, it can be put everywhere to follow the cold chain. It can also be used as an inviolable electronic label and as a security to identify the origin, the pack...

Proges Plus produces this plotter: Thermo Button. The problem is that it’s not a label, and it will be difficult to put this in a packaging with food.

     
                Thermo Button      
  Figure 8 - Thermo Button
[Proges Plus]
 
     

II-2-4 - Fruit maturity indicator

The New Zealand group Jenkins Freshpac Systems has developed a smart label, ripeSense®, which indicates the fruit maturity stage. This concept is based on a flavour sensor integrated to the label which is glued to a rigid plastic punnet. Molecules which are released during fruit maturation, change the label colour even if it is stored in the fridge. The label turns from red to yellow passing by orange which correspond to a crunchy, firm or juicy fruit.

These packagings are used for pack of four pears and forward every peer should have one label. This concept is studied to be adapted to other products like kiwi, avocado, mango and melon. Moreover, this new packaging allows to protect fruits against unfortunates manipulations which can cause damages. So dealer can reduce the amount of waste thanks to this label. The consumer can knowingly choose his fruits without surprise.The idea is to rise fruits consumption, and to make it more practical.

     
                ripeSense®      
  Figure 9 - ripeSense®
[Jenkins Freshpac Systems]
 
     

These TTI have many advantages: the simplicity of reading, but they can also alert the consumer to the product sustainability, then consumer can choose the maturity fruit they want and reduces the food waste.

II-3 - TTI labels market

These TTI can be applicated for fresh food, dairy products, sea products, etc. In a global way, they can be applicated to the majority of packed products which are distributed chilled and then stored by the consumer in the fridge before using. Thanks to TTI, the consumer can do a better food management and reduce waste. Indeed, the limit date of consumption has a large safety margin which causes the waste of consumable products when they have been correctly transported and stored. In the contrary, some products are not consumable before this date because of bad transport conditions. These indicators will lead to more transparency and could eventually replace the limit date of consumption.

Moreover, these TTI are not a problem for food contact because they are based on the watch of an outside system, so they are not in contact with the food. This technology is ruled by the framework regulation 1935/2004/EC (27/10/2004) art. 4,  and the specific regulation 450/2009/EC (29/05/2009) in European Union. Until now, all of the new products have to be validated by the European Food Safety Authority (EFSA), which is an obstacle to a quick development.

For regulation, TTI just completes the limit date of consumption, so it is problematical to give two informations which can be contradictory. How a consumer can handle the fact that the LDC is over and the TTI indicates “fresh”? Who certifies product compliance? More fair and transparent solutions have to be found to handle this complexity in order to push forward the LDC and to reduce food waste related to LDC. Concerning the question of the responsibility, the solution could be to institute a certified organization which ensure the TTI conformity.

Main actors of this market in France.

In Europe.

In the world, industrial development is more important in Japan and USA.

The actual market for this indicator is for 95% in Business to Business. In France, Evidencia sells the fresh chip Timestrip at 15% for the food industry and at 80% for pharmaceutical industry. In 2000, Monoprix imposed on its dealers to put TTI on their fresh products. Picard had also integrated a freshness chip to his frozen food. The most enduring initiative is Carrefour, which has equipped milk cartons for 7 to 8 years. All of these French initiatives have been given up because of too many products returns. So another solution to reduce food waste has to be found. Label as detector of bacteria could be one. Indeed, this label could indicate more precisely the product's freshness and then reduce the food waste.

III - Smart bacteria detector labels

Plan

Bacteria can be detected directly or indirectly.

III-1 - Direct detection of bacteria

To detect bacteria in the medical field, scientists from the University of Sheffield use polymer paired with antibiotic which can bind to both gram negative and positive. When they encounter some bacteria, they attach to it and collapse around the bacteria. This change of shape influence the dye incorporated in the polymer which release a fluorescent signal detectable by hand-held UV lamp, within a few hours. They found that the PNIPAM polymer, modified with vancomycin (antibiotic) and containing ethidium bromide as a fluorescent dye release a strong signal in present of gram negative bacteria. Other polymer present response with gram positive bacteria such as S.aureus.

     
                PNIPAM polymer      
  Figure 10 - PNIPAM polymer
[JACS]
 
     

Moreover, scientist from CNRS- Aix-Marseille Université and CNRS- Université Paris-Sud invented another method to detect and identify living gram negative bacteria such as E.coli, Salmonella Typhimurium and Legionella Pneumophila. They put in touch bacteria with a specific sugar KDO (3-deoxy-d-manno-octulosonic acid) modified thanks to an azide function. This sugar is absorbed by the bacteria that need it to synthesize a specific polysaccharide of their cell membrane. After this absorption, a molecule which attach only with azide is injected. Then it is possible to identify and count bacteria. This method has the double advantage to be fast and reliable.

     
                E. coli marked by the incorporation of modified KDO and pairing with a fluorescent molecule      
  Figure 11 - E. coli marked by the incorporation of modified KDO
and pairing with a fluorescent molecule
[CNRS]
 
     

The biosensor developed by Radke and Alocilja for E.Coli detection works like that: bacterium is sandwiched between two antibodies, and the antibody is paired with a fluorescent probe. This technical is basically foreseen to simultaneously identify few disease causing micro-organisms. The detection limit of E. Coli in food like ground beef, apple juice or Turkey sausages is 5x104 CFU/mL. (Colony Forming Units). The major difficulty for biosensors to detect Listeria is the ligand specificity for immunosensors development. Indeed, most of antibodies developed react with most of members of the Listeria group, and some of them are not pathogenic. Moreover, these sensors don’t differentiate dead cells from living cells.

Another process uses fluorescent semiconductor quantum dots to detect Salmonella Typhimurium cells. This quantum dots are used as fluorescent labels in immunoassays to detect quantitatively the bacteria. The salmonella cells are extracted from chicken carcass thanks to a water wash and to the use of anti-salmonella antibody coated magnetic beads. After that, Salmonella reacts with the second biotin-labeled antibody. The biotins react with quantum dots coated with streptavidin which are added. The intensity of fluorescence produced by quantum dots measured help to quantify the microbial population. This method can detect a limit concentration of 103 CFU/ml.

With all these scientific discoveries, it's possible to imagine a label that can be in contact with a liquid in a transparent container, which could show the presence of bacteria by its fluorescence under UV light.

A new multiplexed paper strip can detect bacteria in water within minutes. The novel approach for selective and ultra-sensitive multiplexed detection of E. coli using a lab-on-paper test strip (bioactive paper) is based on intracellular enzyme activity. The test strip is composed of a paper support, onto which enzymes and FeCl3 were entrapped using sol–gel-derived silica inks in different zones via an ink-jet printing technique. The sample is lysed and put in contact with the paper, causing a change from colorless-to-blue and/or yellow to red-magenta. The color depends of the enzyme activated so it can indicate a precise bacteria. The limit of detection is ~5 colony-forming units (cfu) per milliliter for E. coli O157:H7 and ~20 cfu/mL for E. coli BL21, within 30 min without cell culturing. Thus, these paper test strips could be suitable for detection of viable total coliforms and pathogens in bathing water samples.

     
                Pathogen Sensing Paper      
  Figure 12 - Pathogen Sensing Paper : Paper strips with ink-jet printed sensing zones
can detect low levels of pathogenic or non-pathogenic bacteria. Incorporation
of an immunomagnetic separation step results in selective detection of ~25 cfu of H7:O157
bacteria in under 1 h.
[CNRS]
 
     

This detector advantages are to be rapid, sensitive, on-site detection of bacteria without a need for sophisticated equipment or skilled personnel. These advantages are extremely important in clinical settings and rapid response scenarii, as well as in resource-limited settings.The test strip can detect potentially harmful concentrations of E. coli in water quickly and simply, with much greater accuracy than existing portable technology. However, this technology can not be adapted for now as a label because it need to lyse the sample so it can not detect bacteria on food directly. However, it could maybe be adapted in label for bottle of milk or water.

III-2 - Indirect detection of bacteria

Some indicators can show the presence of bacteria by detecting another molecule associated to bacteria. The first indicator is a label detector of gas like putrescine or cadaverine which has been designed by Massachusetts Institute of Technology (MIT) researcher. This label is made of carbon nanotube coupled with metalloporphyrins (organic molecule which has a metallic central atom such as cobalt or iron). This molecule reacts with the amin like putrescine or cadaverine which are emitted when the meat rotted. The carbon nanotube is grafted and its resistance decreases. So the electric current decreases too. This variation can be detected and followed by a smartphone.

     
                Gas detector related to putrefaction      
  Figure 13 - Gas detector related to putrefaction
[Science et Avenir]
 
     

This device can be integrated to the label and read by a smartphone and its production isn't expensive.

Another detector of gas has been designed by a research team from Dublin City University (DCU). This detector is made for fish and sea products packaging and it can indicate the freshness degree of the contain. The technology uses a sensor which changes of colour from yellow to red in attendance of gas issued from decayed fish. This gas is responsible of the rotten fish smell. The change of colour is made by a ph sensitive dye. The colour variation is visible by eye. An optical drive has also been invented to do a better quantification of the colour change. The sensor can be put on an adhesive label or fixed at the outside of a bottle with filament entering the inside.

The third indicator is a barcode which has been designed by SIRA Technologies. It can indicate if the product is contaminated. The barcode's ink can detect bacteria and becomes red. Thanks to this change of colour, the barcode can’t be scanned at the checkout. Also a mention unsafe can appear to indicate to the consumer that the product is contaminated.

     
                     
  Figure 14 - Bacteria detector barcode
[Sustainable is good]
 
     

This barcode's advantage is to be readable at home, even if the product has been opened for a coupled days. Moreover, this product can’t be bought if it is contaminated so the consumer will not try to send it back to the producer if the mention unsafe appear.

IV - Smart bacteria detector labels market

Plan

As a new technology, bacteria detector labels will enter into an existing market: the smart packaging market and more precisely the TTI market.

IV-1 - Competitive analysis

On the Porter diagram, we can see the different suppliers of the actual technology. The bacteria sensor label isn't the only one which want to enter the market and its entry will be difficult and dependent on others. The evolution of the market share will depend on research and advertising campaigns lobbies of medical and food industry.

                     
  Click on picture to enlarge
Figure 15 - Porter diagram
 
     

IV-2 - Strategic diagnosis

Strength Weakness
  • Speed of analysis
  • Specificity of the detection
  • Detection for a small quantity of bacteria
  • Detectable by the consumer
  • Control of ink jet by printers
  • Cost
  • Non recyclable
  • Depending on its composition, does not meet all sanitary standards for the product
Opportunity Threat
  • Growth of packaging market, which cause a growth of labels market
  • Demographic growth and household evolution
  • Obligation to have a quick check of the detection of bacteria
  • Development of bacteria needing innovating products
  • Detectors time / temperature
  • Low cost of these sensors ITT
  • Analyses increasingly rapid laboratory and always at low cost
  • Strict sanitary standards for food contact products in and on the label
  • Smart packaging market development
  • Wholesale

Table 2 - Strategic diagnosis via the SWOT matrix

One of the opportunities for these labels to develop is the reinforcement of laws concerning fresh food. Indeed, the climate change is suitable to the proliferation of bacteria and the territories enlargement where bacteria are found. With the rise of temperature, bacteria become more virulent and epidemic can start, especially in the developing countries. Moreover, with the globalisation, products are transported from one part of the world to another easily, but not always in good conditions. Industrials are now looking for news ways to secure their products. Also, laws concerning the transportation of food and the food security will certainly become more stringent. So, the request for label which can detect bacteria easily and quickly will grow. Otherwise, with the demographic growth, the demand for food will rise and the need to reduce food waste will become stronger. As food waste is mainly done at home, industrials have to find a way to inform the customer that the product is safe and label as detector of bacteria is one simple way to do it. So the request for these labels will certainly rise.

IV-3 - Future prospects

Let us imagine different scenarios – pessimistic, optimistic and neutral – for the development of bacteria detector labels.

IV-3-1 - Scenario 1 - The end of packaging

Assumptions

Bacteria detector labels arrive on the TTI market and are cheaper than time-temperature indicators... Consumers have difficulties to know which indications they can follow and how the bacteria detector works. The non-standardization of the market losts consumers. Some people willing to trust the detector which indicate the absence of bacteria are contaminated and get sick. The low reliability of the detector make consumers suspicious. Many returns of products are done because of an indication unsafe on them. Moreover, consumers want to reduce packaging in a motion to preserve environment. Sales of bulk products increase and packaging is minimized. People don’t see the interest of bacteria detector labels and think that it's a raw material waste. Idea of bacteria detector labels is ended.

Probability: 60%

IV-3-2 - Scenario 2 - Protection against epidemics for those who pay the price

Assumptions

Bacteria detector labels become mandatory for some food and medical packaging. Moreover, specifics laws put a limit on the distance that a food can travel before sale, which reduce the intermediates number and also the packaging and labels number. The global label market stagnates, or decreases, but the market of bacteria detector labels rises. However, the high cost of these labels makes difficulties to commercialize them, so only companies providing good quality food can afford to use these labels because they know that their customers will not hesitate to pay more for their safety in terms of epidemics. As a conclusion, the bacteria detector labels market stay a niche market.

Probability: 80%

IV-3-3 - Scenario 3 - Used-by date replaced by bacteria detector labels

Assumptions

Bacteria detector labels arrive on the market. Thanks to the low cost of the labels, Monoprix, Picard and Carrefour reiterate their TTI initiatives adapted to bacteria detector labels. They are well supported by consumers who are able to handle the information's difference between used-by date and detectors, thanks to good advertising. Trust in these labels grows: consumers are seduced by the possibility of knowing at any time if the product is consumable. Less returns because they know that the possibility to buy the product means that this one is safe. At home, they do a better management of fresh products which lead to a drastic reduction of food waste. Their trust in used-by date of fresh food has decreased, which leads to its abandonment.

Probability: 50%

V - Conclusion

Plan

Bacteria detector labels are not yet on the market. They could solve some problems that our society is facing now, like food waste and bacterial epidemics. However, for this technology to grow, it needs to be validated by people who will buy products packaged with this type of label.

Alarmed by the various episodes of food contamination in recent years, consumers are wary of the agri-food industry. Of course, with an awareness-raising campaign, people could understand that these labels are one solution for securing the food system. So the market of bacteria detector labels could grow and takes its place.

Moreover, we must not forget that every year research centers are developing technologies we can't imagine today, and, who knows, maybe some of them will be competing the bacteria detector labels. Their future is surely linked to the development of connected objects. Indeed, if the customer can check that the product is contaminated with his phone, he will probably be more confident and adopt these labels.

VI - Bibliography

Plan

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    SIRA Technologies Food Sentinel System Thermal Barcode for Packaging.   Sustainable is good, 4 mars 2009
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