HUMAN PRACTICES

"Not everything that is faced can be changed, but
nothing can be changed until it is faced."
- James Baldwin

Content


  1. Methodology
  2. Conceptualizing
  3. Objectives
  4. Solution


Human Practices aims to fully understand the problem in all its dimensions—from ethical considerations to practical solutions, from a human-centered approach to a business perspective, from the laboratory to society. It seeks to offer a sustainable, knowledge-based solution that is both practical and impactful, based on relationships and the integration of diverse viewpoints. At the same time, it recognizes the barriers, strengths, implications, and long-term consequences of our project.

By adopting a Human Prcatices approach to decision-making and then integrating the information into the project, we ensure a holistic perspective throughout all phases of our project development. This is why we have placed such a strong emphasis on this area, ensuring that every step we take is thoughtful, inclusive, and aligned with the broader societal context.


Our methodology


In order to accomplish our objectives in Human Practices, we have developed a dynamic, action-driven strategy to focus our efforts where it counts. Building on iGEM's established frameworks, we have created a closed-loop, four-phase approach that digs deep into the data and integrates ethical insights and real-time feedback from industry leaders. These four phases are Reflection, Action, Implementation, Evaluation, all of which combined conclude in a system designed to deliver real-world, applicable results again and again.



Conceptualizing the Problem


Overview and Inspiration

Growing food demand and emerging health risks make food safety a critical challenge. Aflatoxin B1 (AFB1) contamination of food, as seen in our Project Description, poses a significant health threat, including the risk of cancer from prolonged exposure. In addition, food waste, either by direct contamination during growth or by cross-contamination in transport and storage, exacerbates the problem.

To design an adequate solution to this problem, it is necessary to know it from all points of view, which is why Human Practices has set to work reflecting and searching for bibliography, talking to stakeholders, contacting society and integrating all this information with the aim of improving the quality of life of all people through synthetic biology.

The sanitary approach

First Human Practices loop

Aflatoxin B1 is not just a theoretical concern, but a serious global health threat, related to several cases of liver failure and cancer, among other things. Seeing the gravity of the problem, we came up with our first idea: creating a probiotic microorganism that captures and detects aflatoxin B1 in the intestines, allowing it to be safely expelled from the body. But to make this vision a reality, we needed real-world insights. So, our first move was clear—talk to the experts and gather critical health data.


After hitting a roadblock in accessing clinical data at Málaga’s main hospitals, Hospital Virgen de la Victoria, also known as Hospital Clínico, and Hospital Regional Universitario, bureaucratic delays hindered our efforts. We also tried to get advice from our university professor Maximino Redondo, who also works at Hospital Costa del Sol in Marbella, and also two other employees of that hospital, Paula Gómez and Marilina García. They all tried to gide us on how to get information from health services such as the oncological and toxicological areas of their hospital, but this still yielded no results, leaving us to question why so little information about aflatoxin B1 impact in helath is accessible in Spain. The main problems we faced during our investigation were:

  • Bureaucratic Hurdles: accessing clinical data in Spain proved difficult and time-consuming, rendering it unhelpful for our project's immediate needs.
  • Lack of Available Data: it appears to be very limited research and public data on aflatoxin B1 in Spain, despite its well-documented global threat.

All of this led us to investigate other possible solutions to this problem. At the end of the day, we asked ourselves if it was possible that the most efficient way to tackle this issue was not to focus all of our efforts on the halth sector, but instead, try to face it from the root cause.

We thought about the idea of creating a possible solution that addressed aflatoxin contamination from farm to fork, protecting people before it becomes a health crisis, and thus, we were ready to move forward and open our minds to think beyond our previous ideas.

Change of paradigm

Second Human Practices loop


During our second Human Practices cycle, we embraced a transformative shift in our project's paradigm. Instead of solely concentrating on the impact of aflatoxin B1 after ingestion, we researched about the need to address the problem at its root. Then we focused our efforts on the agri-food sector, including every type of company involved in the production and manufacturing of food. To effectively tackle the problem, we decided to prioritize the development of an early detection tool. The development of such tool invoved both technical and social approaches, such as:

  • Enhancing prevention through improved tracking systems.
  • Raising public awareness about the risks and solutions.

As we delved into the pressing issue of aflatoxin contamination, we uncovered a troubling reality that spans the globe. The data we have compiled paints a stark picture of AFB1 contamination rates, with South and Southeast Asia emerging as the hardest-hit regions. To know more, visit our Project Description page.


Aflatoxin contamination world wide



Among 60-80% of food crops worldwide are contaminated with aflatoxins (Eskola et al., 2020).


Specific data from Spain


The data revealed how impactful aflatoxin B1 contamination of food was worldwide, but how little it was known about it in Spain. Taking this into consideration, our team conducted a comprehensive analysis of aflatoxin B1 management practices across the agri-food chain on a local frame, revealing a wide spectrum of approaches related to the handling of this mycotoxin. We called and emailed more than thirty companies, reseiving a response from 21 of them. Here there is what we discovered from these companies:


Agriculture and Disinfection

Fresón de Palos

Recognizes aflatoxins, particularly AFB1, as a risk primarily linked to cereals and grains. Testing for AFB1 occurs only upon request or when a specific risk is identified. If contamination is detected, products are recalled following a thorough investigation.

Desur

Specializes in pest control but does not only focus on AFB1. They remove affected cereals but do not identify specific fungi or conduct AFB1 tests, leaving this responsibility to the Junta de Andalucía, the main gouvernamental organism responsible in this type of situation.


Livestock and Feed Manufacturing

Agammasur

A goat cheese producer with no routine AFB1 testing, as their supplier, DCOOP, manages it. They note that aflatoxins are a greater risk in cows than goats, and that there are no exhaustive analysis to detect AFB1 contamination in goats' milk.

Vall Companys

Actively monitors AFB1 and other mycotoxins in livestock feed using advanced detection methods like ELISA and HPLC. They strictly adhere to legal limits for AFB1 and have never exceeded them. The expenses of their analysis are among the average costs for industrial ELISA and HPLC procedures.


Storage Practices and Quality Control

Almacenes Antonio Guerrero

Conducts preventive and periodic AFB1 testing. Although they rarely handle contamination, they follow strict isolation and testing protocols when needed, guaranteeing quality and safety.

S.C. Andaluza Agrícola De Estepona

Does not perform quality control tests themselves. Their primary buyers, TROPS and Frutas Fajardo, are responsible for these procedures, but they do not have any information about how these companies manage the AFB1 detection protocols.

DCOOP

Conducts multiresidue tests for oil quality and is implementing AFB1 testing using HPLC. They outsource AFB1 tests to meet regulatory demands.

Natibero Food

Relies on prior quality checks from suppliers and does not conduct internal AFB1 tests, as the regulation does not obligate them and they do not find them necessary.


Commerce and Contaminant Control

Agrupa Algarrobo

Implements random testing and basic safety measures for storage. However, they largely depend on producer testing to guarantee proper safety meassures.

Cereales y Frutos Secos El Torcal

Conducts multiresidue testing, including AFB1, for corn and cottonseed destined for livestock feed, ensuring compliance with European Union limits.

Cereales Martín Miguel

Sends regular samples for lab testing, placing trust in the producer's quality control measures.

Frutos Secos Salen

Enforces strict quality controls from suppliers. Nevertheless, they also perform annual random AFB1 tests.

BIOLES Cooperativa

Sends annual product samples for AFB1 testing. They follow stringent protocols for handling contamination but have experienced economic losses due to contaminated products.

Cooperativa Nuestra Señora del Carmen

Produces olive oil, with DCOOP managing storage and AFB1 testing.

Apícolas Milosi

Regularly conducts fungal and mold tests but has not detected aflatoxins, including AFB1 on their honey and pollen production.

Horticultores El Torcal

Engages in preventive testing for AFB1 in vegetables and utilizes accredited labs when Aspergillus is visually detected.

Sabor en Rama

Performs internal multiresidue testing and random batch analyses but finds widespread testing impractical due to volume. They apply proper cleaning meassures to guarantee that the olive oil production does not get conminated with AFB1 from different batches.


Transportation and Food Safety

Frío Antakira & Miafruto

These intermediaries rely on prior quality controls for the products they transport and do not conduct specific AFB1 tests.

Alitrans

Focuses on cleaning transport vehicles but does not handle temperature controls or delicate goods, representing a potential gap in food safety.


Analysis and Laboratories

Laboratorio Antakira

Has not detected AFB1 in their tests. They collaborate with specialized labs to ensure food safety.

After talking to all of these companies, we decided to visit those companies in which AFB1 has a bigger impact. Along several weeks, we went to three different towns of Málaga and visited BIOLES Cooperativa, DCOOP and Sabor en Rama. They gave us a lot of information about what they do in order to prevent AFB1 contamination of their products, the test they perform, or the negative impact this mycotoxin has had for them:


BIOLES Cooperativa

In the meeting with the company BIOLES Cooperativa, we had the opportunity to speak with Carlos Aragón, one of its founders. We discussed the strict regulations BIOLES adheres to in terms of contaminant detection. The company faces challenges with random inspections and the detection of contaminants such as aflatoxins and ochratoxins, as well as logistical issues during transportation. BIOLES also highlighted that in some cases, contaminants like aflatoxins were detected long after products had entered the market, necessitating costly recalls. We discussed how our probiotic project could help detect aflatoxins economically and quickly in products such as corn and improve the overall safety and traceability within the food production chain.


DCOOP

During our visit to DCOOP, Mª Rosario Luque Luque, head of the laboratory, Silvia López Feria, head of R&D, and José Repullo Gómez guided us through their olive oil production process, explaining how olives are processed and categorized while highlighting their rigorous quality control procedures. One of the most interesting parts was the tasting room, where a team of at least eight people evaluates oil samples anonymously to ensure the highest quality. Regarding aflatoxin management, they informed us that they currently do not perform AFB1 (aflatoxin B1) tests in-house due to a lack of appropriate equipment; instead, they send samples to external laboratories for analysis but are working on acquiring HPLC equipment for in-house testing in the future. Additionally, we learned about the impact of recent droughts on oil production and visited their cold storage area, where they analyze up to 3,000 oil samples per day. After the tour, we presented our iGEM project, AflaxOFF, and engaged in a discussion about oil varieties, preservation, and potential improvements.


Sabor en Rama

Javier, owner of Sabor en Rama, gave us a personal tour of the facilities, where he showed us the machinery used for processing olives. He explained that they handle each client's olives separately, cleaning the machines between batches to prevent contamination. We also discussed aflatoxins, which he acknowledged are present in crude olive oil, but no specific treatment is applied to remove them. Although inspections occur twice a year, multitoxin analysis is rarely requested, and oils with mycotoxins are not always removed from the market. However, he assured us that they have never had a health alert for excessive toxins, and in such cases, authorities would step in. Javier emphasized the careful tracking of olive origins by property owners, though aflatoxin testing is not performed. He also pointed out that transporters don’t monitor temperature or humidity during transport, and that phytosanitary or toxin controls on imported olives are handled by the administration.

Our conclusions


Addressing the Knowledge Gaps Across Key Sectors

Once we collected all the data we needed, we realized that several industries were surprisingly unaware of the serious risks associated with aflatoxin B1:

  • Disinfection and Pest Control: this sector tends to overlook fungi that produce AFB1. Often, when mold is detected in cereals, companies remove and aerate products without identifying the fungi or performing AFB1 analysis. This reactive approach highlights a critical knowledge gap and a missed opportunity for early detection and prevention.
  • Livestock Sector: many companies in this industry rely on their suppliers for AFB1 testing of livestock feed and milk, rather than conducting their own analysis. This dependency creates potential blind spots, leaving them vulnerable to contamination risks that could affect the safety of their products.
  • Agricultural Sector: a similar reactive mentality exists here, where routine AFB1 testing is only conducted when a specific threat is identified. This lack of regular monitoring, especially with imported grains, increases the chance of contaminated products slipping through the cracks.

Strengthening the Ineffective Detection Systems

The current system for detecting contaminated food, particularly AFB1, involves monitoring, analysis, and regulatory compliance. However, based on our discussions with these companies, we identified several weaknesses that undermine the effectiveness of this system:

  • Inconsistent Testing: many companies do not conduct regular AFB1 testing and only test when a threat arises, increasing the risk of contamination going undetected until it's too late.
  • Overreliance on Suppliers: both the livestock and agricultural sectors depend heavily on their suppliers for AFB1 testing, leaving gaps in accountability and oversight. This lack of direct control over testing procedures limits their ability to proactively manage risks.
  • Limited Expertise in Specific Areas: certain sectors, like disinfection and pest control, lack the expertise and focus on AFB1, resulting in inadequate detection and management of this highly toxic substance.
  • Outsourced Testing: many companies outsource their AFB1 analyses to third-party labs, which can lead to delays in detection. When testing is infrequent, contamination risks increase, as companies are left without immediate feedback.
  • Regulatory Blind Spots: some companies are either unaware of or not fully compliant with existing regulations, resulting in insufficient testing protocols and safety measures. This creates regulatory gaps that leave them exposed to significant contamination risks.

Going a step further

Third Human Practices loop


With all the information we had gathered, it was time to implement key improvements and continue developing our work in Human Practices. We needed to keep listening to those affected by aflatoxin B1 and present the advancements in our project that could benefit them.

BIOLES Cooperativa, DCOOP, and Sabor en Rama had provided valuable insights and support, so we decided to reach out to companies we had not spoken with before. With a better understanding of the food production sector, we could engage with them more effectively.

To ensure our progress was aligned with industry needs, we reconnected with companies that had guided us earlier. Our first follow-up was with DCOOP, where we explained how we had tailored the project based on their feedback. We focused on creating an engineered organism capable of producing a quick, visible response when exposed to AFB1, with a specific and fast reaction. Their positive response encouraged us to keep refining this approach.


Next, we reached out to Agroquivir, a company we initially held off contacting until our project was better developed. They emphasized the importance of a quick detection test to remain competitive in the market. The detector needed to give an almost immediate response to AFB1, as existing methods were slower and more laborious. Additionally, they highlighted the need for precision in our qualitative detection method to avoid unnecessary product isolation. Warehouses and crop reception sites, where goods from multiple sources are combined, would benefit most from this system. By using a fluorescent protein, our detector could also reduce costs, as it would not require specialized equipment for detection.

We also had a productive conversation with CETAEX, a company we met during the Transfiere forum in March. They provided valuable insights on where AFB1 contamination could occur, such as how some fungi from the Penicillium genus can contaminate fruits such as tomatoes and produce aflatoxins. Their interest in our project reaffirmed its potential to address aflatoxin contamination in different food sectors.


In order to finish our work getting information from companies, we found it important to visit new ones too. Thus, in order to conclude or final steps on the research of information and feedback, we visited Piensos Campillo, a company in Málaga that produces animal feed. They shared their experiences with AFB1, including a significant outbreak in 2013. They also mentioned their participation on a study on how ants contribute to aflatoxin contamination in cattle feed, revealing a new angle on the issue. Unexpectedly, they gifted us a sequestrant used to prevent AFB1 absorption in cattle, allowing us to conduct comparative tests.

During the visit, Piensos Campillo introduced us to the lateral flow assay kits they use for AFB1 detection, which are made by ProGnosis. We decided to contact ProGnosis for kits to verify the effectiveness of our engineered organism. After learning about our project, ProGnosis generously agreed to sponsor us by providing the kits for free. This partnership was pivotal in advancing our experimental work, giving us the tools needed to perform more accurate and consistent assays, in line with industry standards. This collaboration underscored how our engagement with companies had a direct and positive impact on our project’s development.


Days of reflection


Between cycle and cycle, we carried out reflection days. In those reflection days, we took a step back to carefully consider the broader implications of AflaxOFF, acknowledging the risks, potential inequalities, environmental concerns, and economic impact our project might have. All of this work was carried out in the margin of our Ethical code and Values, which have been developed in parallel with the development of our project.

One of the most significant issues we recognized was the unequal burden of aflatoxin contamination across the world. Regions like Latin America and Africa are disproportionately affected, while Europe faces fewer challenges in this area. We realized that our initial idea of developing a probiotic might inadvertently widen the gap between developed and underdeveloped regions due to accessibility issues. In response, we thought that alternatives like preventing contamination directly in the field or creating a more affordable, freeze-dried version of the probiotic could ensure that it could reach the people who need it the most.

We also reflected on the environmental and economic consequences of aflatoxin contamination. With between 60 to 80% of global agricultural food production affected each year, we understood the scale of the problem and the economic losses it causes, particularly for farmers and food producers. While AflaxOFF has the potential to address some of these challenges by providing a simple detection method, we recognize that implementing such a solution is a huge undertaking. It was clear that we need to collaborate with organizations capable of large-scale production to reduce costs and ensure that our solution is accessible, especially in the most vulnerable areas.

We are aware of the regulatory challenges surrounding GMOs too, especially when it comes to human applications. We understand that this could limit the reach of our project in certain regions. However, we’re exploring alternatives, such as using processed fragments of GMOs already dead to bind aflatoxins, and are committed to following strict safety protocols to minimize any risks to the environment. We recognize that these are complex challenges, and we are learning as we go, always keeping in mind the need for responsible innovation.

Throughout our journey, we have embraced scenario planning and used SWOT analysis to explore different outcomes and pathways for our project. By reflecting on our strengths, addressing weaknesses, and anticipating challenges, we’ve been able to refine our ideas and adapt them thoughtfully. This process has been key in shaping the project’s evolution, allowing us to move forward with a clearer, more purposeful vision.


We also reflected abot the Sustainable Development Goals on how our project could be helpful to accomplish them. We then decided that AflaxOFF fit on the following categories:

Target 2.1.2: Prevalence of moderate or severe food insecurity, assessing how many people have significant difficulties in accessing food. We aim to avoid wastage of food with aflatoxin levels above EU standards, contributing to food security.

Target 3.4: Reduce premature mortality from non-communicable diseases by one-third through prevention and treatment, and to promote mental health and well-being. Our project would help prevent aflatoxin poisoning, which is associated with liver cancer.

Target 12.3.1: Halve global per capita food waste at retail and consumer level and reduce food losses in production and supply chains. AflaxOFF contributes to preventing the waste of food with aflatoxin concentrations above the European regulations.


Defining the objectives



Reflection-led Action

Our interactions with the real world have helped us sharpen our strategic focus, underscoring the pressing need for innovative solutions to combat AFB1 contamination. This is a major issue not only in the agricultural and agrarian sectors within our country, but also a significant health concern in regions outside our borders.

Guided by industry feedback and public perception, we have set the following key objectives to address this challenge:



This organism should act as a biological safeguard against AFB1 ingestion, allowing the toxin to be safely expelled from the body through feces. Beyond its immediate health benefits, this solution has broader implications for the food industry, as it can function as a rapid detection system for AFB1 contamination early in the supply chain. By identifying contaminated products before they reach consumers or livestock, we can reduce the risks of AFB1 exposure, protect animal products from contamination, and improve overall food safety.

Designing a solution


In order to conclude the work we have done, we have designed a implementation strategy that takes a comprehensive, dual approach aimed at improving both public health and food safety.

Two important elements of our design are the use of a visible reporter, a red fluorescent protein triggered by the presence of AFB1, and its implementation on a probiotic chassis, Sacharomyces cerevisiae var. boulardii, to use a single engineered organism to carry out both approaches. These decision were made deliberately to align with our project goals and better meet public needs. It is also important to aknowledge that the use of the boulardii variant has been reviewed biblographically, but would serve as a future implementation to optimize the project. By employing this strategic approach, we ensure that the groundwork already laid is effectively implemented, while also meeting the immediate needs of the communities affected by aflatoxin B1 exposure.


Our innovative design endows yeast with novel capabilities, making it applicable across various sectors. This versatility allows the yeast to tackle aflatoxin contamination from different angles and offers a potential solution to this global threat. Additionally, the highly adaptable signaling platform we have developed could be capable in the future of detecting not only AFB1, but other harmful substances as well. This multi-purpose functionality transforms the yeast into a powerful tool that can serve as a detector, diagnostic agent, and even a therapeutic option in specific contexts.

In its primary function, the yeast acts as an early-warning system, detecting AFB1 contamination in food products before they reach consumers. This has significant implications for the agricultural sector, where it can prevent the distribution of contaminated food shipments, reducing health risks and minimizing economic losses associated with aflatoxin exposure.

Furthermore, our future approach relies on its application when administered as a probiotic. In this case, the yeast would offer a dual-action benefit: it would detect AFB1 within the digestive system and actively sequester the toxin in the intestine. This prevents the absorption of AFB1 following the ingestion of contaminated food, offering an in vivo solution that not only protects human health but also demonstrates the powerful potential of biological tools in food safety and public health. Through this multi-faceted strategy, we address both immediate and long-term challenges posed by AFB1 contamination, ensuring a safer food supply and healthier population.

It has been thanks to the development of our ideas, and through our close engagement with communities affected by aflatoxin B1 contamination that we have come to realize that our original concept may be better suited as a future phase of our project. While our initial vision remains crucial, the insights and feedback gathered during our Human Practices cycles have allowed us to refine our approach and prioritize more immediate solutions that address today's pressing needs. By listening to the concerns of stakeholders and understanding the real-world impact of AFB1, we have been able to reorganize our efforts and create a project that not only meets current demands but also paves the way for future innovation. Our work now strikes a balance between addressing urgent challenges and remaining adaptable to what lies ahead, ensuring a lasting and meaningful contribution to public health and food safety.

References


Rodrigues, I., & Naehrer, K. (2012). A three-year survey on the worldwide occurrence of mycotoxins in feedstuffs and feed. Toxins, 4(9), 663-675.

Rodrigues, I., & Naehrer, K. (2012). Prevalence of mycotoxins in feedstuffs and feed surveyed worldwide in 2009 and 2010. Phytopathologia Mediterranea, 175-192.

Anjum, M. A., Khan, S. H., Sahota, A. W., & Sardar, R. (2012). Assessment of aflatoxin B1 in commercial poultry feed and feed ingredients. J Anim Plant Sci, 22(2), 268-272.

Eskola, M., Kos, G., Elliott, C. T., Hajšlová, J., Mayar, S., & Krska, R. (2020). Worldwide contamination of food-crops with mycotoxins: Validity of the widely cited 'FAO estimate' of 25. Critical reviews in food science and nutrition, 60(16), 2773–2789.

Tarazona, A., Gómez, J. V., Mateo, F., Jiménez, M., & Mateo, E. M. (2021). Potential health risk associated with mycotoxins in oat grains consumed in Spain. Toxins, 13(6), 421.