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Revision as of 13:30, 12 October 2018

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HUMAN PRACTICES
Intergrated Human Practices
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Rokiškio sūris
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TEAM

INTEGRATED HUMAN PRACTICES

INTRODUCTION
    PLACEHOLDER
Cheesemaker from Rokiškio sūris
  • Our project is designed to make cheese products safer. If applied, this idea would influence current cheesemakers. However, if we would like cheese makers to consider our innovation, first we need to examine their suggestions because they are experts in making cheese and so can advise us on our project design.
  • We have arranged a phone talk with Vadimas Kličius, the director of new product development in “Rokiškio sūris” (a major cheese manufacturer in Lithuania) and discussed our project idea.
  • The takeout message from the talk was that mesophilic L.lactis bacteria culture is not an optimal choice for the biosensor. The bacteria culture could not be used in making hard cheeses and some of the soft cheeses, that we are aiming for this project, do not use L.lactis in the making process. To improve our project design, we have optimised codon sequences for a thermophilic bacteria Streptococcus thermophilus.
  • Also, Vadimas mentioned that GMOs in cheese are not acceptable and that would be the main issue in applying our biosensor in any cheese making company. This is why we are modelling the isolated cell detection system so we would not need to integrate modified L.lactis to cheese products. This also inspired our discussions with the European Commission about current GMO regulations and their impact on our project.
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Rokiškio sūris
The Whole story


To get a real-world cheese maker's perspective on the industrial constraints affecting our project we spoke to Vadimas Kličius, who is the director of new product development in “Rokiškio sūris” - a major cheese manufacturer in Lithuania. Mr. Kličius introduced us to the safety measures employed in the company to keep products safe. To ensure safety, samples are taken during different stages of the cheese making process. Since tests for pathogenic bacteria (e.g. Listeria or Salmonella) cannot be run in the cheese making factory (because tests require growing potentially pathogenic cultures, which are forbidden for safety reasons), all samples are sent to external microbiology labs for analysis. It takes around 1 to 3 weeks to get the results back. Most interesting for the assessment of the economic feasibility of our alternative testing device was information on the price of this off-site testing of cheese samples. Current tests involve the following costs:

Example prices at National food and veterinary risk assessment institute of Lithuania:

  • Detection of L. monocytogenes in (26 g - 125 g of the sample) - 27 Eur ≈ £ 24 ≈ $31.8
  • Detection of L. monocytogenes by BAX Q7 method - 21 Eur ≈ £ 19 ≈ $24.7
  • Detection of the number of L. monocytogenes in food products - 12-16 Eur ≈ £ 11-14.5 ≈ $14.1-18.8
  • Detection of the number of L. monocytogenes in food products - 12-16 Eur ≈ £ 11-14.5 ≈ $14.1-18.8
  • Detection of L. monocytogenes by PCR method - 30 Eur ≈ £ 27 ≈ $35.3

We learned from Mr. Kličius that L. lactis, the bacterial species we are targeting in our iGEM project, is not necessarily used to make soft cheeses (e.g. mozzarella). The species is instead mostly used for making semi-hard cheeses, which are currently not the target of our detector device. However, according to Mr. Klicius, modified L.lactis for Listeria detection could theoretically also be integrated into the soft-cheese starter cultures without negative effects on product quality.

On the other hand, we learned that the choice of a mesophilic bacterium L.lactis might not be optimal for industrial applications of our system. Firstly, some soft cheeses (e.g. mozzarella) have a formation step where the pre-cheese mass is heated up to 59-62°C and cheese is formed. Also, the process of making the majority of hard cheeses includes a stage where cheese granules are heated up to 38-58°C to dry them and control the fermentation process. A sensor system based on L.lactis bacteria would probably lose its function or even would not survive in such temperatures. Secondly, to make mature cheeses, they are kept at 9-12°C to keep the fermentation process slow. If L.lactis bacteria are added, the maturation rate would increase because such temperature is optimal for our mesophilic bacteria to grow. As a result, cheese quality and flavour might change.

When asked about the potential of our Man-Cheester project, Mr. Klicius responded that the idea is promising but currently no one would buy a cheese-containing GMOs. He even mentioned that big companies which are buying products from “Rokiškio sūris” ask for tests which prove that products do not contain GMOs. This confirmed our earlier insights talking to artisan cheese makers, who expressed similar concerns. Thus, although interaction with potential customers indicated that they might be much more open to GMO for cheese safety testing, the commercial viability of our device would depend on a change in risk perception among cheese producers and sellers, as well as changes in the regulation of GMO in food. As a result, we had intense discussions with the European Commission regulating relevant GMO policies, but we also explored possible modifications to our device that would ensure that no live GMOs are contained in the final product - considering that GMO-derived ingredients, such as vegetarian rennet, are widely accepted in commercial cheese products.

Cheesemaker from Burt's cheese
  • We have visited a local cheese maker Claire Burt to get to know about the cheese making process and cheesemaker’s perspective about our idea.
  • We found that the commercial cheese making process is very sensitive and needs to be highly controlled.
  • Claire explained to us that the major sources of Listeria contamination are various surfaces or water used in the process. That suggested us that having a cell-free or an isolated-cell test would be more convenient than a biosensor integrated into the product.
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Visit to Burt’s Cheese
The Whole story


On the 5th of June, we were kindly invited to ‘Burt’s Cheese’ in Cheshire. The owner, Claire Burt, was generous enough to give us a full demonstration of the cheese making process where we learned the importance of pH and temperature control in cheese manufacture. With this data, we were able to amend our current cheese making protocol to fit better for a soft, white mould cheese. Our original cheese making protocol did not use any other live culture besides our E. coli but Claire advised us to include other cultures. These other cultures are used to produce flavors and/or CO2 in cheese with eyes. This meant we now needed to control for pH and temperature more strictly. For example, culture must be added at 17°C and increased slowly to 30℃ rather than simply heating to 32°C. We were also informed that it was important to check the pH of the milk is as close to 6.74 as possible for an optimal batch.

Claire Burt also gave us a good insight into the hygiene practices she must adhere to ensure her cheese is safe. For example, Claire uses an ozone producing machine in order to control phage contamination. There is also a strict hygiene checkpoint at which hairnets, lab coats and shoes are changed to minimise the introduction of outside contaminants. We discovered where and how any foodborne pathogens can enter the process. Of particular concern to Mrs. Burt was the hygiene quality of the large metal vat that the farm used to store pasteurised milk. She also stated that Listeria, when present, is often on surfaces and within the water. In addition to this, we learnt that small-scale cheese producers often only send swab samples to labs every 3 months due to the high costs associated with testing. An outbreak of Listeria can be extremely damaging within this space of time. For example, a recent L. monocytogenes outbreak in South Africa lasted 15 months, infecting over 1000 and killing over 200 people. (Blomfield, 2018)

From our visit we decided that, seeing as Listeria monocytogenes contamination was possible at the first stage of processing (the storage vat) we should integrate our biosensor into the cheese itself. Rather than developing the sensor in an external Escherichia coli bacterium, we made the milestone choice to integrate the E. coli into the cheese starter culture. In doing this, the bacterium will always be detecting L. monocytogenes, drastically reducing the time the pathogen spends in the food. Claire Burt also prompted us to research the pathogen detection market to see if we could produce our biosensor as a cheaper alternative to lab grade testing for smaller producers.

British Science Week 2018
  • We were volunteering in the British Science Week 2018 and presented at the stall "Making medicines in cells".
  • We have introduced school students to the concept of DNA replication and translation.
Conversation with Dairy Science Food Technology

In our search for sponsorship and council, we contacted Michael Mullen of ‘Dairy Science Food Technology’. On hearing the details of our project he expressed significant concern about the legality or commercial viability of our project, stating that “this has limited practical application and if I were the expert assessor I would voice my concerns. Currently, we are not legally permitted to use genetically engineered cultures in cheese making” and that it is “not practical in our current legislative environment and would be unlikely to be looked on favourably by consumers either”. As our first piece of significant negative feedback on our project, we took this criticism very seriously. On learning that the selling of a product such as ours would be prevented by legislation, we were prompted to look further into current EU laws surrounding the deliberate release of GMOs. Mr. Mullen’s concerns about the consumer opinion of our novel starter culture also worried us. We decided that we needed to deliver an outreach event at The Manchester Museum to see if Mr. Mullen’s prediction about our products lack of commercial viability was correct.

Public Engagement session in Manchester Museum
  • We hosted a small stand in The Manchester Museum as part of ‘University of Manchester Community Fest’.
  • We introduced members of a general public to our project idea and asked if they would eat a cheese that contains GMOs if the purpose of GMOs is to detect the pathogenic bacteria. The majority of people showed an interest to our approach and supported the idea saying that they would not mind GM bacteria in cheese if it is safe to eat.
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Public Engagement event in Manchester Museum
The Whole story

We hosted a small stand in ‘The Manchester Museum’ as part of ‘University of Manchester Community Fest’. As this was an event aimed at families, we had a colouring activity for children. We asked the children to colour what they think ‘good’ and ‘bad’ bacteria would look like. While the children were engaged with this activity we asked parents questions about their views on GMO containing cheese. These questions were:

  • Do you like cheese?
  • Is there anything specific about it you enjoy?
  • Do you have any concerns about cheeses? (Particularly to do with health, disease and contamination)
  • Do you take any steps to minimise your concerns about cheeses?
  • What are your feelings regarding genetic modification?
  • What would convince you (or not) to eat GM food?
  • How would you feel about using genetic modification technology to make cheese safer?

These questions yielded a broad range of responses that are summarised below:

These questions yielded a broad range of responses that are summarised below:

Do you like cheese? Is there anything specific about it you particularly enjoy?

43 people responded to this question, with 35 stating that they enjoy cheese products regularly because they like the taste. 4 people stated that they do not like cheese, again due to its taste. 2 respondents stated their indifference to cheese. There was also one lactose intolerant individual and one vegan.

Do you have any concerns about cheese? (Particularly to do with health disease and contamination)

37 people responded to this question. 22 stated that they had no concerns about eating cheese. 7 were only concerned if they were pregnant. 3 respondents expressed dietary concerns, with 2 further expressing concerns specifically to do with cholesterol levels. 1 respondent each voiced concerns about handmade cheeses only, Listeria contamination and concern only for children.

Do you take steps to minimise your concerns about cheese?

Of the 11 people that responded to this question, 3 took no actions. 3 attempt to limit the quantity they eat. 2 keep it refrigerated. 2 did not eat soft cheeses when pregnant. 1 did not buy handmade cheeses.

What are your feelings regarding genetic modification?

We gained 27 responses to this question which are summarised below:

I try to Avoid GM Products x3
I agree with using GM for health and safety purposes x3
I do not know enough to make an informed choice x3
I do not care if food is GM or not x2
I do not agree with using GM crops for profit but using them for health and safety measures is useful
I am interested in how the laws differ around the world
They seem a good solution to many problems
I disagree with using any GMOs
I do not know anything about GMOs
Is it safe to ‘play’ with bacteria
I am scared of GM
I know that plants are modified to give a better yield
I would not eat GM food
I prefer natural products
I do not mind trying GM products but I would not give them to my child
I feel uncomfortable but would change my mind if they were cheaper

These responses informed us that much of the public are pro-GM if the technology is being used to benefit health, but are less supportive it is simply being used to reduce costs of production. There was less strong opposition than we predicted.

What would convince you to (or not) eat our GM food?

Lower Price of a product x7
If they are better educated on how the technology works x5
The use of a well known ‘good’ bacteria rather than E. coli x4
If the taste remains the same x3
Nothing could convince me x2
A full explanation of how the GMO interacts with the body
If pregnant, will the GMO interact with the fetus?
I would only eat the product if I was pregnant
I would eat the cheese if the GM organisms could be removed before consumption
I would eat the cheese if the system could be put in goats cheese rather than other soft cheese
If there are more people eating such products and they were more common

A significant number of people were supportive of our product if it were not sold at a higher price. Another common trend was a requirement of better education on GM technology to be confident in consuming our product. Possibly the most impactful comment was the trend that people are more happy to consume a GM bacteria that is commensal and well known as it removes much of the uncertainty. E. coli is commonly associated with disease in the public and media so a switch to a ‘good’ bacteria would be preferred. From these data we decided to integrate our system into the already present Lactococcus lactis rather than E. coli in order avoid the public fear of E. coli.

How would you feel about using genetic modification technology to make cheese safer? (Sticker chart for children and adults) (0=Would not eat such a cheese. 10=Would eat such a cheese as normal)

News Feed analysis

As part of our Human Practices, we analysed both recent news from around the world and slightly older news around historical Listeria outbreaks and adapted aspects of our project as a consequence of that analysis.
We found that:

  • Listeria has an effect on trade, both directly (via restrictions on import from affected countries) and indirectly (due to loss of consumer trust and reduced demand for goods).
  • It is key that food testing be decentralised to avoid a single point of failure and therefore our device should use a reporter that needs no equipment to interpret.
  • It is currently impractical to test each packet of food if contamination is suspected. Our system should be higher resolution by reporting Listeria contamination only when present and able to prevent this food waste.

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News Feed
The Whole story

21st December 2017
Errington's Dunsyre Blue cheese recalled over Listeria concerns
Read more

Blue cheese recalled due to possible Listeria contamination calls into question our use of a blue protein as a reporter. We could use a red colour, as it is naturally associated with danger and easy to produce with many well-characterized proteins (mRFP etc.). However some cheesemakers are attracting their customers by producing cheeses of different colours (e.g. red, orange etc.) and this would reduce the possible cheeses our product could be used for as, unlike with a blue reporter, a red reporter will not stand out very well against a red background. We could also use a smell producing biobrick to ensure that, even if the colour is not visible in all cheeses, the consumer will still be alerted to the contamination. However, cheeses are known for their strong smell and there is the possibility that a smell that would normally be considered unpleasant could be unintentionally delicious when combined with the normal cheese aroma. Ultimately, we need to revisit our design and consider if it is feasible to add a smell producing module to the device and if so, what smell should we use. We also need to re-evaluate our reporter design and potentially switch to a different coloured protein that will show up better against typical cheese colours without looking like the blue mould on a blue cheese.

8th March 2018
Why the listeriosis outbreak is 'just the tip of the iceberg' - by Alet Janse van Rensburg
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This piece highlights that the recent Listeriosis outbreak in South Africa reveals issues with general food safety for the urban poor. Processed meats are a key protein source for many people and these foods are generally produced in large factories operated by one of a few large companies. This centralised food production means that if contamination occurs, it affects many people at once. A possible solution to this general problem would be taking some of the control over testing away from the company and ensuring that in the future there is less reliance on a single point of failure for food safety testing. This means we will need to ensure that our project does not require any expensive equipment (fluorimeter etc.) or training as this would mean that people would be forced to cede control back to larger organisations, who may have a vested interest in presenting their food as safer than it actually is.

11th May 2018
Egypt just banned South African imports because of listeriosis - by Phillip de Wet
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Egypt has placed new restrictions on imports from South Africa. “[Egypt] wants assurances that SA can test for Listeria and provide accompanying paperwork.” - Phillip de Wet. This shows that testing for Listeria is still a significant barrier to international trade and an improvement in this area would help reduce Listeria's economic impact. This is especially important for highly perishable goods such as cheese, fruit or fresh meat as any delay in testing can reduce the time the consumer has to consume the goods before they go bad. It will, therefore, be important for us to model our device to ensure that it works fast enough to be useful, but also is not overly sensitive as this risks false positives which would lead to unnecessary destruction of food.

10th July 2018
Woolworths is recalling rice over fears of Listeria- by James de Villiers
Read more
23rd July 2018
TOTAL RECALL Listeria outbreak – what frozen vegetables and foods have been recalled over listeriosis concerns?- by Guy Birchall and Hollie Borland
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Indicates that products that are not cooked before eating, such as soft cheeses, sweet corn or fresh fruit, are an important vector for Listeria, but that current testing is inadequate: because testing each batch or pack is not feasible using food external methods, we need to focus on making the food itself become the sensor. In this case, large recalls are not needed because the consumer can see for themselves whether or not their specific packet is contaminated. This raises particular difficulty with licensing and safety accreditation and we will need to investigate the regulatory framework around GM organisms used inside food rather than the more established framework of GM organisms being used for food production (e.g. GM rennet production).

12th September 2018
Brexit could free UK farmers from Europe’s stringent GMO regulations - by Justin Cremer
Read more
14th September 2018
'We take a science-based approach to GM regulation': UK to consider relaxing gene editing ban post Brexit - by Katy Askew
Read more

These articles raise an important question: should we use Brexit as an opportunity to move away from the EU’s position on GMOs? Many countries in the world have much less stringent regulations for GM food and relaxing the UK’s restrictions on the use of these products may be essential for forming new trading relationships post-Brexit. Relaxing the law with regard to GM foods would be good for our project as it would be illegal to use in food under current EU law. However, serious reservations about GM food and food standards (post-Brexit) remain amongst both the general public and members of parliament. A greater analysis of GMO regulation post–Brexit is warranted as it may have profound effects on our projects viability and on the synthetic biology industry in general.