Team:Tec-Chihuahua/Human Practices

Erwinions

Overview

"Human Practices is the study of how your work affects the world, and how the world affects your work."
— Peter Carr, Director of Judging

Beyond the technical work on the lab, we comprehend our project exists within a world that requires us exploring aspects that involve issues such as ethics, sustainability, safety, legal framework, security, between others. These issues are complex and approached only through our human practices. Being aware of this we engaged with the communities that might be impacted by our work to invite them to shape the course of our investigation.

These social considerations were upfront in our project and integrated with the design and execution of our project, impacting in our Entrepreneurship, Wet lab, and Mathematical model work. We aimed to develop a thoughtful project with inspiring Human Practices for other teams to build upon and come up with new and creative approaches for this section, striving not only to understand how the world affects our project but also aiming to contribute to how synthetic biology is understood and practiced around the world.

Social Science Research Description

Since our Human practices involved interacting with communities to understand the social concern we are addressing, it was essential for us to approach these communities correctly by getting informed on our institution and national guidelines. This way we were able to conduct our social science research respectfully and always looking after the well-being of our cooperators.

The Mexican National Guidelines for integration and operation of the Research Ethics Committee (REC) declare the inclusion of ethical aspects as a quality indicator in social science research. The development of a bioethic culture is favorable to guarantee individual and social well-being since the discipline promotes the concepts of equity, solidarity, quality, principles of benefit, no maleficence, as well as the protection of human rights.¹

Human subjects research entails risks; this makes it necessary to guarantee ethically acceptable conditions in the production of knowledge, considering that the legal framework is not sufficient to elucidate or resolve conflicts of a moral or ethical nature that arise in scientific activity.¹

To protect our contributor’s human rights, respect their dignity and privacy, avoid any form of discrimination as well as ensure their integrity and wellness, we took into consideration the universally recognized ethical standards which are principles of benefit, no maleficence, justice and respect for autonomy.

We also made sure our procedures where according to the Mexican Federal Law on Protection of Personal Data Held by Individuals.² We requested a letter from our institution approving our social science research was done in accordance with their ethics code and values and under the stipulated in the annexed letter, we also requested a consent letter filled up by our participants where they accept we share their information on our webpage and in the Giant Jamboree's presentation. Trough this letter they also recognize our behavior and procedures were done according to the iGEM values.

References

Ruiz de Chávez, M. (2016). Guía Nacional para la integración y el funcionamiento de los Comités de Ética en investigación. 5th ed. México: Secretaría de Salud, Comisión Nacional de Bioética.
http://www.diputados.gob.mx/LeyesBiblio/pdf/LFPDPPP.pdf

Consent Letter
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Letter of Approval
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Silver Human Practices

From the first time you set foot outside the laboratory and leave scientific articles aside to interact with people related to your project, you begin to realize how crucial human practices are.

Human practices allowed us to investigate and validate the purpose, design, scope, and execution of our project. Is our product suitable for beekeepers to use? Will it disturb the innocuousness of honey? Is it capable of counteracting more diseases? Is it permissible before the law? These are some of the questions that were answered throughout our human practices chronology; we were driven to think, reflect, and act on it as our integrated human practices.

Beekeepers, beekeeping societies, international organisms, government representatives, and researchers were all essential for fortifying our research.

Government Institutions

Thanks to regulations and laws that promote safety and well-being, it is possible for a project to be fully implemented. The rapprochement with governmental institutions focused on the primary sector allowed us to consider essential legal aspects for the development and execution of our project; It was thanks to these meetings that the legal plan was formed.

June 27^th
Chihuahua, Chihuahua
Meeting with MVZ Tanús, MVZ Arroyo, and Ing. González

July 17^th
Morelos State
Video call with MVZ Juan Diego Pérez de la Rosa

June 27^th
Meeting with MVZ Ernesto Tanús Sánchez, MVZ Adolfo Arroyo, and Ing. José Luis González Miranda

Counseling about some parts of bee anatomy.
Discussion of aspects like:

Search of precise statistics on the presence of foulbrood around the world.
Important national and international normative and regulations to be considered regarding the design and execution of our project.
Difficulties and impediments regarding governmental legislations related to bees
How to ensure the peptides have arrived at the larva?

Detailed Description

For us, it was very important to receive feedback from specialists and be able to exchange opinions about our project. After the video call we had with Ernesto Tanús, we were able to keep in contact with him, since he was visiting the city of Chihuahua we took advantage of the opportunity and we scheduled a meeting that was also attended by the President of the National Association of Veterinary Doctors Specialists in Bees (ANMVEA) (MVZ Adolfo Arroyo) and a beekeeping technician from the Secretary of Agriculture, Livestock, Rural Development, Fishing and Food (SAGARPA) (Ing. José Luis González Miranda). The legal framework consultant accompanied by two active members of

human practices attended this meeting that lasted approximately 1h 30. We were looking forward to presenting our project, for them to the give us their general insights, discuss the considerations of legal aspects that involve our project, and receive counseling of the bee's anatomy.

For this date, we were in search of precise statistics on the presence of foulbrood around the world. Mr. Tanús recommended us to consult the OIE (International Office of Epizootics) where the countries report the presence of different diseases. After this meeting, we were able to consult and obtain specific data of each country to better track Fouldbrood distribution (Consult the distribution map in the detailed project description). The specialists also shared with us that the last time AFB was diagnosed in Mexico, was in 2015 and that the infected apiary was eliminated. We considered very important the fact that the causal agent was a strain with resistance to many antibiotics, reinforcing the previously mentioned need for an effective treatment alternative for these pockets of infection.

The three specialists oriented us on the several regulations and normative that we should check over in order to proceed according to the national and international law and security

Residuality: Described in the national program of control and monitoring of toxic residues in honey, carried out by SENASICA. (National Service of sanitation, innocuousness, and agri-food quality)
Regulations of the European Union regarding food safety.
Zoosanitary regulations (NOM-ZOO)
Federal Animal Health Law
Entomology protocols
Maximum limit of allowed residues in honey (After looking over we found out there is no regulation regarding AMPs; we could later validate this information in the IV State’s Beekeeping Forum with Dr. Sóstenes Rodríguez)

Since the aspects mentioned were evaluated after the meeting, we realized the need to create a legal plan where the project would really considerate and comply with all its legal and security implications.

In addition to covering the legal aspect of our project, we were able to consult with the MVZ bee specialists about the anatomy of the bee; an important aspect to consider since the nanocapsule will travel through the nurse bee until reaching the larvae. After arduous research and incorporation of the received advice in this meeting, we were able to know in detail the anatomy of the bee and integrate the important factor of it to our project design. (Consult details about the bee anatomy). When we were discussing the terms of the anatomy of the bee we both realized that some terms had different words but referred to the same part. We also had the opportunity to become aware that some very specific parts of the bee could influence the nanocapsule path that we were analyzing.

Regarding the technical part of the project, the attendees to the meeting recommended us that at the time of the experimentation part with bees, some innocuous dye could be added to make sure that the nanocapsules were reaching the larva. In addition, it was discussed how there is greater production of larvae during flowering, and in this time artificial feeding is not widely used; thus we concluded how our product would be optimal to be used before the period of flowering, so at the initiation of the greater production of larvae, if the bacteria, which mainly infects in the first days of the bee larva, attacks, the antimicrobial peptide will already be found in the medium to reach the objective.

We also had the opportunity to know a little about the perspectives and concerns of the three persons in the meeting. They perceive the problem that the regulations regarding the handling of bees and their products are not enough for reaching a strong and growing beekeeping sector, and although they have tried to promote the implementation of a NOM (Official Mexican Norm), they have seen little possibility and many impediments. They also shared with us their concern about how there exists a barrier between researchers and producers; the needs and advances sometimes do not go hand in hand and there is no adequate implementation and development.

Device where queen bees are transported

We had the privilege to share all this important talk with queen bees and worker bees! They were being transported and couldn’t stay in the car. We could get to know how the transport should be done and how queen bees were marked in the superior part of their thorax to be recognized.

The three assistants to the meeting could obtain a new approach to beekeeping when we presented them our project and the scopes of synthetic biology. They congratulated us, approved the project and put themselves at our disposal for what we could later need.

We consider this event very important in the development of our project. Here, the legal plan originated thanks to the guide of laws that we were told were essential to review; a project like ours needs to adhere to the norms and regulations of the place to be implemented. Finally, it is essential to have the points of view and recommendations

of MVZ specialists in bees regarding the experimental plan of the mathematical model that is based on the different environments of the bee anatomy.

References

Image retrieved from: Segundo a Segundo. (2018). Puerta-de-Chihuahua-Sebastian. https://www.segundoasegundo.com/puerta-de-chihuahua-sebastian/

July 17^th
Video call with MVZ Juan Diego Pérez de la Rosa

Discussion of aspects like:

Ambiguities with the data of the presence of foulbrood in Mexico.
Is it possible that a negative overaccumulation of AMP’s results in the bee?

Detailed Description

As our project was growing we sought to investigate and validate the part of regulation and monitoring of animal products; importation aspects, mobilization, exportation, confirmation of quality and innocuousness, etc. SENASICA oversees this mentioned procedures, and its regulations are for us an essential part to consider, so what a better implementation than to seek validation from a specialized governmental entity related to this area. As we have previously received different recommendations to speak with SENASICA’s (National Service of Sanitation, Innocuousness, and Agri-food Quality) chief department, we took advantage of this great opportunity and indeed contacted MVZ Juan Diego Pérez de la Rosa to present him our project and discuss new insights and considerations. He is a bee specialist, but also owns a great panorama about legal implications in the agri-food sector! The meeting took place by a video call and the legal framework consultant conducted this reunion that lasted approximately 35 minutes.

First, in the meeting, we presented our project and the scope that synthetic biology can have in the beekeeping area. Regarding the theme of foulbrood distribution, the MVZ Juan Diego Perez de la Rosa commented us that in Chihuahua two cases of American foulbrood had been registered last year. By this time in the development of our project, we had found many ambiguities with the data of presence of foulbrood in Mexico, and we were able to conclude that many times beekeepers do not report that they own infected hives because they look forward to saving money and preserving the stability of their business.

As the reunion flowed, we were also able to discuss some technical factors of our project. Sr. Juan Diego told us that AMPs were perfect for what we wanted and for keeping the innocuousness of honey. We also consulted with him a question that had been asked to us previously: When it comes to increasing the concentration of AMP’s, is it possible that a negative overaccumulation results in the bee? The MVZ analyzed and suggested that bees are exposed to many pathogens, with our product they could become more resistant; the treatment was perceived by him like a supplement that will help bees defend themselves. With so many pathogens attacking bees, we can hardly create an adverse problem, instead would fortify hives in hard times.

The MVZ was very interested in the alternative we propose and said we could consider later on, the possibility of nanoencapsulating our peptides with his help. “The approach is excellent and it is a work of great quality.” He was astonished when we talked about the physiology of the bee regarding our project and he kindly also mentored us on how we should use fewer technicalities when we attended the IV State’s Beekeeping Forum, as it was approaching.

“Bees are warriors! Its immune system has to face everything”

References

Image retrieved from: México es Cultura. (2017). La Ciudad de las Flores y Sol. Cuernavaca, Morelos. https://www.mexicoescultura.com/actividad/175504/la-ciudad-de-las-flores-y-sol-cuernavaca-morelos.html#prettyPhoto

United Nations

As part of our Human Practices, we wanted to expand our horizons towards an international frame. Which are the most relevant international GMOs' protocols that apply to our project? What is a Genetically Modified Organisms Environmental Risk Assesment and how is it executed? How can our team change the way synthetic biology is understood and practiced around the world? How is it our project affected by this international frame? Trough our United Nations activities we were able to address this questions.

September 18^th
Guadalajara, Mexico
Video call with Dr. María Mercedes Roca

October 26^th
Mexico city, Mexico
Mexico Model United Nations (MEXMUN)

September 18^th
Video call with Dr. María Mercedes Roca

Dr. Mercedes invited us to collaborate with her team in the Genetically Modified Organisms Environmental Risk Assessment guide by helping in its translation to Spanish, making able for their work to reach Spanish speaking people.
Dr. Mercedes invited us to join Saúl Pizarro and her at the COP 14 as members of Youth Biotech, an international group of young leaders who are interested in the development of science communication, policy making, and biotechnology regulation.

Dr. Mercedes will present this guide at the Fourteenth meeting of the Conference of the Parties (COP 14) in Egypt.
She recommended us to base our Integrated Human Practices in the United Nations’ three dimensions for a sustainable development: Economic, Environmental and Social. On the social aspect, she recommended us to include our projects’ ethical and legal aspects.
She suggested us to talk about a complex; there is not a complete understanding nor a single reason why bees are disappearing, it might be due to various diseases, malnutrition, insecticides, is a complex of reasons. Our project won't solve the problem, but it will contribute to managing the Colony Collapse Disorder.
Is very common to tend to talk about human rights, but these are not the only rights that have to be worth, the environment should have his rights just for its intrinsic value.
She also advised us in our United Nations’ three dimensions approach.
She contacted us with Dr. Sol Ortíz García, the Director of The Intersecretarial Commission On Biosecurity Of Genetically-Modified Organisms (CIBIOGEM for its Spanish Acronym) and with Dr. Natalie Beatriz Campos also part of the Executive Secretariat of the CIBIOGEM, focusing her work on Policies and Regulations Management.

Detailed Description

Signed by 150 government leaders at the 1992 Rio Earth Summit, the Convention on Biological Diversity is dedicated to promoting sustainable development, Mexico is one of the countries that signed this Convention. The Conference of the Parties is the governing body of the Convention and advances implementation of the Convention through the decisions that take place at its periodic meetings. The Fourteenth meeting of the Conference of the Parties (COP 14) will be held in Sharm El-Sheikh, Egypt, 17^th - 29^th November 2018.¹

"MOP" stands for "Meeting of the Parties" of the protocol. This year will be COP-14 and MOP-9 of the Cartagena Protocol. Later, will be the MOP-2 of The Nagoya Protocol on Access and Benefit-sharing and the MOP-1 of The Nagoya - Kuala Lumpur Supplementary Protocol on Liability and Redress.

Biotechnology is being promoted as a better way to grow crops and produce medicines, but it has raised concerns about potential side effects on human health and the environment, including risks to biological diversity. In some countries, genetically altered agricultural products have been sold without much debate, while in others, there have been vocal protests against their use, particularly when they are sold without being identified as genetically modified. In response to these concerns, during the past two decades, two Protocols have been adopted by the Parties to the CBD. In January 2000, the Parties approved the Cartagena Protocol on Biosafety. This Protocol seeks to ensure the safe handling, transport, and use of living modified organisms (LMOs) resulting from biotechnology. To that end, the Protocol establishes procedures that allow countries to make informed decisions before agreeing to the import of LMOs into their territories.

More recently, the Parties adopted the Nagoya Protocol. It reaffirms each nation’s sovereignty over its genetic resources and calls for the fair and equitable sharing of benefits arising out of the utilization of genetic materials. By ensuring benefit-sharing, the Protocol seeks to create incentives to conserve and sustainably use biodiversity. The Nagoya Protocol entered into force on October 14^th 2014.³

One of the issues discussed in the Nagoya Protocol is the genome of a particular species may now be sequenced within a provider country and that information may be transferred digitally to a company or research entity for downloading to a DNA synthesizer.² Another significant issue discussed in the COP is if synthetic biology is different enough from modern biotechnology that it must require a different protocol and a different risk assessment, the GMOs of modern biotechnology.

The Parties to the CBD considered regulation of synthetic biology during their 12th COP meeting in October 2014. There, they agreed to take a precautionary approach to the release of any LMOs resulting from synthetic biology under the Cartagena Protocol. At the same time, the Parties called for the formation of an Ad Hoc Technical Expert Group on Synthetic Biology (AHTEG) to evaluate, among other things, whether existing arrangements constituted a comprehensive regulatory framework for addressing potential impacts from synthetic biology technologies.³

Article 15 of the Cartagena Protocol establishes that to do genetic engineering to develop or liberate a GMO, a risk assessment must be done first. The AHTEG was created to develop a risk assessment guide which was created about 8 years ago. Members from different countries that assisted to the COPs were invited to participate in this group and a risk assessment guide was started within the Cartagena protocol.

Dr. María Mercedes Roca is a professor of Biotechnology, formerly at the Monterrey Institute of Technology and Higher Education, Guadalajara campus in Mexico. She was one of the founding members of the National Biotechnology and Biosafety Committee of Honduras (CNBB for its Spanish acronym) which reviews submission for risk assessment of GMOs and advice the Honduran Government.

We were able to contact with Dr. Mercedes Roca thanks to Saúl Fernando Pizarro, a schoolmate and friend of ours who has participated in iGEM as a team member (2015), team leader (2016), and Co-chair of the Latin American Committee (2016). Dr. Mercedes is part of a group of Latin Americans unsatisfied with the rigorous risk assessment guide the AHTEG developed. Aware that in Latin America a risk assessment is needed and by taking into account the principles of biosafety in a very responsible way, this group made a guide under international confinements of risk assessment that people can follow and that countries can address, the guide is named Genetically Modified Organisms Environmental Risk Assessment Guide.

"There is a whole area of science called risk analysis; we made a guide that falls within the international parameters to assess risks internationally."

The guide her team has developed is a guide they expect servers biotechnology students like us, making it able for them to start a business or sell a product. Dr. Mercedes invited us to collaborate with her team in this guide by helping in its translation to Spanish, making able for their work to reach Spanish speaking people. We are pleased we worked for hand in hand with Saúl Pizarro in this translation.

AHTEG United Nation's guide was introduced in the thirteenth meeting of the Conference of the Parties to the Convention on Biological Diversity last year in Mexico but wasn't approved by the Parties, this year Dr. Mercedes will present her team’s alternative guide at the Fourteenth meeting of the Conference of the Parties (COP 14) in Egypt. Acquiring this guide won't have any type cost; it is an effort of a group of academics that want to give to the regulators of the countries, developers, and students an environmental risk assessment they can rely on.

Dr. Mercedes invited us to join Saúl Pizarro and her at the COP 14 as members of Youth Biotech, an international group of young leaders who are interested in the development of science communication, policy making, and biotechnology regulation. It was born on December 2016 after a group of biotech students attended the UN Biodiversity Conferences as a part of the Public Research and Regulation Initiative (PRRI).

We are truly honored for the invitation and for the opportunity to join Youth Biotech, 10 of our members are now part of this group and excited to participate and represent the community of young biotechnologists internationally.

Dr. Mercedes recommended us to base our Integrated Human Practices in the United Nations’ three dimensions for a sustainable development: Economic, Environmental and Social. On the social aspect, she recommended us to include our projects’ ethical and legal aspects. Formerly, we were planning on managing 4 dimensions, ethical, legal, social and economic.

She suggested us to talk about a complex; there is not a complete understanding nor a single reason why bees are disappearing, it might be due to various diseases, malnutrition, insecticides, is a complex of reasons. Our project won't solve the problem, but it will contribute to managing the Colony Collapse Disorder. She said is very common to tend to talk about human rights, but these are not the only rights that have to be worth, the environment should have his rights just for its intrinsic value.

Environmental

She advised us to be very emphatic about what is the loss of habitat, why is it that bees are having nutritional complications? The world is urbanizing; there are not enough plant species that bees can feed on.
We should discuss how in countries where GM crops are not cultivated, such as in Europe, bees are dying and in countries where GM crops are grown, such as Argentina, Brazil, the United States, and Mexico, some are also dying, this means there are other factors such as pesticides and especially insecticides affecting the welfare of bees.
Bees are essential to agriculture and to maintain a healthy ecosystem.

Economic

There is a need of having economic activity, Mexico needs to export for uses of its population.

Legal

Article 3 of the Mexican Biosecurity Law of GMOs establishes that all GMOs that are destined to be released commercially must be previously submitted to satisfactory tests according to risk studies and a risk assessment.

Thanks to our collaboration in translating a section of the Genetically Modified Organisms Environmental Risk Assessment Guide and reading the complete guide, we are now much involved in how to carry a risk assessment. At the end of our video call with Dr. Mercedes, she mentioned something that changed profoundly the way we perceive our career and biotechnology in general; beyond teaching molecular biology and microbiology to her students, she is aware they will become researches, entrepreneurs or both, she wants them to learn about regulations but most of all she wants them to get involved in changing them for the better.

Ultimately, Dr. Mercedes congratulated us for our project and contacted us with Dr. Sol Ortíz García, the Director of The Intersecretarial Commission On Biosecurity Of Genetically-Modified Organisms (CIBIOGEM for its Spanish Acronym) and with Dr. Natalie Beatriz Campos also part of the Executive Secretariat of the CIBIOGEM, focusing her work on Policies and Regulations Management. For more information of our participation in the translation of this guide, visit our Education and Public Engagement Section.

References

CBD Secretariat. (2018). Convention on Biological Diversity. Retrieved from: https://www.cbd.int/
Convention on Biological Diversity (2000). Sustaining life on earth. [Montreal]: Secretariat of the Convention on Biological Diversity.
Manheim, B. (2016). Regulation of synthetic biology under the Nagoya Protocol. Nature Biotechnology, 34(11), pp.1104-1105.

October 26^th and 27^th
MEXMUN

We developed MEXMUN edition 2018 topic “Synthetic biology as a tool for the development of biological weapons" in the First Committee on Disarmament and International Security (CDSI in Spanish) to be discussed by high school students from different states of the republic at the UN simulation.
To make sure we had developed a wholesome and reflexive topic which students can build upon, we had to look over the international mechanisms, agreements and bodies that regulate, look after and assure the responsible and safe use of synthetic biology and GMOs.

Detailed Description

MEXMUN is a student simulation of the debates that take place in the main deliberative bodies of the United Nations Organization, discussing economic, political and social issues. In this simulation, students take on the role of representatives of different countries before the UN, knowing and maintaining their national interests, having to reach the end of a resolution.

Nearly 700 secondary, preparatory and professional students from more than fifty national and international institutions participate in it. Through 20 years of history, MEXMUN has established itself as one of the largest and most elaborate conferences of its kind in Mexico, offering debate commissions in Spanish, English and French.

As part of our Education and Public Engagement activities, we developed MEXMUN edition 2018 topic “Synthetic biology as a tool for the development of biological weapons" in the First Committee on Disarmament and International Security (CDSI in Spanish) to be discussed by high school students from different states of the republic at the UN simulation. We intended to get more students interested and involved on synthetic biology issues as well as evaluate the possible risks of employing the tools this new discipline requires, tools we use in the laboratory for the construction of our product.

To make sure we had developed a wholesome and reflexive topic which students can build upon, we had to look over the international mechanisms, agreements and bodies that regulate, look after and assure the responsible and safe use of synthetic biology and GMOs.

We investigated on the Geneva Protocol, Rio de Janeiro Earth Summit, Cartagena Protocol, Convention on Biological Diversity, Biosafety-Clearance house, the Nagoya - Kuala Lumpur Supplementary Protocol and Nagoya Protocol. Through this research, we learned on biosafety matters regarding synthetic biology and we raised self-awareness on the globalization of this discipline and how it becomes more accessible every day to acquire and implement its tools and methods, for example, for biohackers. We understood the importance of biodiversity, bioethics, regulations, and dialogue as well as we raised awareness of the scopes of synthetic biology understanding the importance and the need to adhere not only to its local and national guidelines but also to the international.

Trough this research we evaluated the possible risks of releasing GMOs into the environment like for example, the risk of genetic information transferring from one GMO to a non-GMO. We acquire knowledge of an international framework directly related to our project and necessary to fully understand the importance of the next activity described in this United Nations section.

Beekeepers

The world of beekeeping is much more than what appears in texts; no research could have compared being in the field with a beekeeper sharing knowledge, getting to know his community and their needs. Our main reasons for approaching them were to be informed about Beekeeping in their region, their opinion on our project and validate technical aspects of it. We wanted to choose with their help, the optimal application method for our product into the beehives, investigate the presence of American foulbrood and European foulbrood in their regions or countries as well as current existing treatments, their benefits, and risks.

April 28^th
Cuauhtémoc, Chihuahua
Meeting with Mr. Guillermo Rodríguez

June 15^th
Santiago de Querétaro City
Meeting with Engineer Alejandro Bianchi

September 22^nd
Province of Melipilla
Videocall with Bryan Saavedra

May 26^th
Delicias, Chihuahua
Meeting with Ing. Manuel Ramírez Salcido

July 16^th
Castellón, Valencia
Valencian beekeeper interview

April 28^th
Meeting with Mr. Guillermo Rodríguez

This was our first direct approach with the beekeeper’s community!

Discussed aspects like:

Status of Foulbrood in Cuauhtémoc
Existing methods to treat Foulbrood.
SAGARPA’s restrictions regarding products applied in the hive.
General characteristics of Apis mellifera’s life

Invitation to join the association in the next monthly beekeepers’ reunion in Cuauhtémoc

Detailed Description

Viana and Ana Laura had the opportunity to travel to Cuauhtémoc, a city specialized in fruit growing; production and exportation of apples, peaches, strawberries, walnuts, among many others.¹ Thanks to Viana, who is native from this city, we were able to get to contact this beekeeper. Mr. Rodríguez is the president of the association of beekeepers in the city and we saw an enormous opportunity in talking to him. This was our very first direct approach with the beekeeper’s community! We were very excited to start getting to know them. What would they think about our ideas? Would they be interested in our project? How many things would they have to tell us? Our project was about to begin its transformation path.

This enriching talk lasted approximately 1 hour 15 minutes and different topics were discussed. We learned many general characteristics of bees' lives thanks to the great experience of the beekeeper: The different varieties of bees that exist, specific tasks of each type of bee and the joint work bees and beekeepers do to ensure pollination.

Regarding the existing methods against Foulbrood, Mr. Rodríguez mentioned how SAGARPA (National Secretary of Agriculture, Livestock, Rural Development, Fishing, and Food) does not allow the usage of chemical contaminants in beehives because wastes may end up in honey flows. Mr. Rodríguez mentioned how beekeepers sometimes use extracts of rosemary or oregano against Foulbrood, but it is always difficult for them to know what dose to add and ensure it is adequate for fighting the diseases and at the same time does not destabilize the hive. On another hand, he mentioned that the method of burning beehives results very expensive for them. Currently, in Cuauhtémoc, there is almost no presence of American Foulbrood, since, in the past, many sick hives were exposed to this burning method, then the disease decreased. On the other hand, European foulbrood is most commonly found in Cuauhtemoc.

The beekeeper emphasized how a natural product against Foulbrood is needed, so there are not left residues in the honey and he also highlighted how beekeeping varies around the world and sometimes there are specific necessities for each region.

Thanks to the sharing we made with the beekeeper of our project, he was able to have a new panorama of beekeeping, and how emerging technologies, in this case, synthetic biology, can accurately respond to specific problematics and can relegate past methods that imply many setbacks.

Mr. Rodríguez kindly invited us to present our project with the Cuauhtémoc beekeepers association and commented us about M.C. Alejandro Romo who has done different investigations on the diseases and who we would talk with later in this story.

Getting to know real experiences of the treatment methods used against our interest diseases helped us to better measure the severity of the problem and the obstacles with which beekeepers have to fight to keep their bees’ welfare.

References

Soto, A. (2017). Cuauhtémoc, región en crecimiento económico. Retrieved from http://referente.mx/@Almer/cuauhtemoc-region-en-crecimiento-economico
Image retrieved from: Visita Chihuahua. (2018). Chihuahua y sus alrededores. http://www.visita-chihuahua.com/index.php/2015-05-06-09-42-18

May 26^th
Meeting with Ing. Manuel Ramírez Salcido

Discussion of aspects like:

Importance of pollination.
Bee diseases.
Different methods applied in hives to fight diseases.
Restrictions implicated in European Union’s honey importation.
Innocuosness of the honey.
Bee bread characteristics.
Concern about the lack of young people interested in the beekeeping sector.
Bee Culture Magazine.

Detailed Description

Key team members related to the development of the mathematical model, human practices, legal management, and wet lab traveled southeast from our city to visit Salcido apiaries in Delicias; a municipality that bases its economy fundamentally on the food industry. Since we arrived with Mr. Ramírez, it was amazing how the discussion flowed and flowed, getting a meeting that end-up lasting approximately 3 hours; all were valuable minutes that taught us many relevant aspects for the development of our project.

First, Mr. Ramírez covered the topic of the importance of bees, emphasizing how they have always been considered an essential factor for environmental health and he even showed us a funny cucumber that had not been correctly pollinated; it was very small and thin.

Bee diseases were our next topic. He commented how Foulbrood

presence isn’t that strong in Mexico, however, in Europe and in the USA, this is not the case. Mr. Ramírez took advantage and included implications of other frequent bee diseases like Nosema and Varroa in the conversation. He mentioned a very important point that we had previously started to notice: Most of the serious problems in bees caused by viruses or bacteria, are not correctly detected or even known. As Mr. Rodríguez had previously commented, Mr. Ramírez mentioned how different extracts of plants are used against diseases in bees, in this case, mint and grapefruit were the examples. On another hand it was mentioned how antibiotics are not an option at all for treatment, since the European Union does not accept them, neither does accept the presence of transgenics in the honey; there are rigorous studies exercised and if there is something wrong the EU may return an entire shipment of honey back to the remitter. We discussed the issue of innocuity of honey by highlighting that when there is a new product added to the hive, it should not generate any residue at all or change the expected properties from the consumer.

After talking about our project, a question arose in Mr. Ramírez, an issue that would be asked many times since that day and that we would do everything possible to solve and validate: Our product would actually pass from the food that the nurse bee ingest all through its system until reaching the larva by means of the royal jelly? And regarding the method of application, (initially consisting in applying the final product in the bee bread), Mr. Ramírez explained to us how this bee bread is created, and the fermentation process it goes through in the bee cell.

We also had the great opportunity of listening to some of the concerns Mr. Ramírez has as a beekeeper: He thinks the beekeeping area could be greatly impulsed by receiving more support from governmental institutions and he perceives that the validation of honey innocuousness here in Mexico results complicated as there are not enough tools to endorse the required diagnostics. On another hand, he mentioned how there are no young people able to continue with the job of the current farmers, and this is an alarming situation that requires a prompt resolution.

Mr. Ramírez gave us a lot of priceless information that was essential for the development of our whole project. He was the first to comment us about the beekeeping congress that would take place in the city of Delicias in the coming months, and which we would have the great privilege to attend, he told us that by means of the different sites where he frequently researches information he had noticed another iGEM team that was working with bees, for the month of September we could establish a collaboration with this team (consult collaborations for more information) and on this same point of the media, Mr. Ramírez explained to us how he always tried to receive literature to focus and inform himself, he mentioned the magazine “Bee Culture”, where we successfully managed to make a publication about our project and synthetic biology regarding beekeeping (consult public engagement for more information).

He also shared with us the contacts of highly recommended specialists with whom we could consult about our product and with which we would have contact later: Juan Diego Pérez de la Rosa, Efraín Rocha, Felipe Esquivel Díaz.

We were able to share a new perspective on beekeeping in relation to synthetic biology with Mr. Ramírez by explaining him our project, the beekeeper mentioned how he was more than willing to help us, and he even shared some honey and bee bread with us! This meeting was an invaluable experience for us!

Bee Bread

References

Image retrieved from: Administración. (2018). ¡Conócenos! http://municipiodelicias.com/

June 15^th
Meeting with Engineer Alejandro Bianchi

As we were speaking with him about our project, he mentioned the importance of the Varroa mite, a parasite that sucks the hemolymph of bees, the equivalent to blood in this arthropods, weakening their immune system and making them susceptible to other diseases. He ensured that enhancing the natural defenses of bees would have a positive response since healthy bees are a lot less susceptible to diseases.
He recommend us to expand our horizons to consider our product as a product that can also be applied when there is the presence of Varroa.

Urbanization is a problem bees are facing directly.
The Secretariat of Agriculture, Livestock, Rural Development, Fisheries and Food (Spanish: SAGARPA) is the governmental institution responsible for inspecting hives.

Detailed Description

Engineer Alejandro Bianchi studied Agronomy at the Monterrey Institute of Technology and Higher Education and has been dedicated to beekeeping since 30 years ago. Currently, he has a company called Kolmena Yothë that offers bee products for health and beauty purposes, his company also provides apitherapy treatments, complementing them with Bach flowers, Schuessler salts and homeopathy.

As we were speaking with him about our project, he mentioned the importance of the Varroa mite, a parasite that sucks the hemolymph of bees, the equivalent to blood in this arthropods, weakening their immune system and making them susceptible to other diseases. He ensured that enhancing the natural defenses of bees would have a positive response since healthy bees are a lot less susceptible to diseases.

He mentioned the Varroa mite as part of the named “colony collapse disorder”, baptized by this name in the US, and described it to be a phenomenon where bees leave their beehives. Perhaps he announced foulbrood are not a current problem in his state, Varroa is, so he recommends us to expand our horizons to consider our product as a product that can also be applied when there is the presence of Varroa.

“If the bees are strong they can’t get sick, but how will they be strong if there are not enough flowers?” He mentioned urbanization is a problem bees are facing directly, and they are dying because of it. As flowering and bee population lower, he went from collecting a bucket of honey per box to a bucket every 40 boxes.

He said the actual treatments for American foulbrood are formic acid, oxalic acid, thymol (thyme), among others treatments

that don’t pollute the environment. Previously, fluvalinates were used to treat this diseases, however, they stopped using them since they are carcinogenic and pollute the environment. The Secretariat of Agriculture, Livestock, Rural Development, Fisheries and Food (Spanish: SAGARPA) is the governmental institution responsible of inspecting their hives, it allows under certain requirements the use of antibiotics for treating foulbrood but has to avoid products that pollute the environment and honey. He also shared with us the treatment he uses to fight Varroa which consists in an essential oils mix and stressed it is important not to use the same treatment for a long time since the pathogen can develop a resistance towards it.

We asked him if he could share with us certain regulations or standards that had to be followed to ensure the honey’s quality, he mentioned the "Good livestock practices".Discussing other issues affecting beekeeping in his state, another serious problem in Queretaro is bee theft, as beekeepers visit their bees every 8 or 15 days, they are the target for many people that take advantage of this situation. Engineer Bianchi assists to the Beekeeper's society meetings, and he sees there are not young beekeepers to take the hard work of beekeeping, he sees that as another issue.

Finally, after approximately an hour of conversating, he shared us his book of memories from the 25th International Apicultural Update Congress which was held in Queretaro last year and invited us to visit his beehives as well as the facilities of his company.

July 16^th
Valencian beekeeper interview

Received information on aspects like:

Status of foulbrood in Valencia.
Treatment used against foulbrood in Valencia.
Beekeeper's opinion about our solution proposal.

Detailed Description

Thanks to our colleagues from Valencia_UPV, we received written feedback from a Valencian beekeeper and information about the current status of foulbrood in his province. Our Valencian colleagues interviewed the beekeeper for us, thank you Valencia_UPV team! (LINK collaborations) Given that our interest diseases have a global presence, it is essential for us to make contact with beekeepers from all around the world; knowing how they face illnesses, the used treatment methods, their effectiveness and consequences, and the beekeeper's concerns and opinions about our solution proposal in order to also integrate them into the development of our project.

The interviewed beekeeper has spent approximately 70 years dedicating himself to the invaluable work of beekeeping and he shared with us some already experienced foulbrood implications that are lived in his province. In Valencia the treatment against foulbrood consist in the following:

First, the combs with the infected brood are replaced by healthy brood and then antibiotic with powdered sugar, is applied. The adult bees, when they realize the larvae are dead, clean the cell. The beekeeper ensures that with this treatment he has managed to heal all his hives, and has not suffered any loss from the disease, the drawback for this method is that it contaminates pollen, so it would no longer be suitable for human consumption, and when this treatment is not applied on time, beekeepers get to have very large losses of money and even the total loss of their hives (between 20 to 100 hives per beekeeper). Regarding this information, we could realize that antibiotics indeed are currently used and although they are effective in treating the disease, the innocuity of the honey is always altered resulting in market issues.

Once the beekeeper heard about our project and learned about biotechnological alternatives that seek to solve beekeeping problems, he considered it an adequate proposal which does not contaminate the bee products and he was expectant before the results of the project to evaluate if it is actually an effective method.

September 22^nd
Videocall with Bryan Saavedra

Discussion of aspects like:

Presence of bee diseases in Chile and its implications.
Agricultural and Livestock Service normative.
Can our product help against varroa?
How bee’s longevity varies depending on different production stages.

Detailed Description

Keeping on with our international validation on the presence of foulbrood, its implications, and our project insights, we were able to contact a Chilean beekeeper thanks to our colleagues from UChile (LINK collaborations). Bryan Saavedra dedicates to honey production, pollination, and queen bee breeding and exportation. He has dedicated more than a decade of his life to beekeeping and he counts with more than 1000 beehives.

“Approximately 13% of the Chilean working population is engaged in agriculture, and agricultural products represent about 4% of the gross domestic product (GDP). The fruit sector is very important and includes crops such as grapes, melons, apples, and peaches.¹”

After UChile team sent us the beekeeper’s contact we were able to get in touch with him and schedule a video-call. During the meeting, that lasted approximately 35 minutes, we were able to discuss the presence of bee diseases in Chile, its implications and some activities of Chilean agencies in charge of supporting the development of agriculture, forests, and livestock.

Foulbrood occur in Chile as foci of infection, once an infection is detected, which most likely originated

from contaminated equipment, the SAG (Agricultural and Livestock Service) proceeds to burn and then bury the infected hives; the decision of burning the beehives is quite drastic but it is the only effective way to take care of the rest of the Chilean apiculture. The last outbreak of foulbrood the beekeeper noticed about, was in last February; normally the infection comes from Argentina, which is where the disease most often occurs. In Chile, they mostly struggle with varroa and nosema and for purposes of the SAG and exportations, the varroa mite is treated with organic products, that result inefficient and expensive. Antibiotics are also used against Varroa, getting back to the resistance problem. Regarding this topic, the beekeeper wondered if our product would help against varroa, and we could tell him how one of the most important consequences of varroa is that the immune system is weakened; providing the bee with new immunity components (AMP’s) will greatly help it, so the answer is yes! Our product can help bees get fortified when they are suffering from varroa.

The beekeeper also shared with us the main routes through which the disease is transmitted from Argentina to Chile, how the bee's life duration varies depending on the production stage and also shared with us details about SAC normative.

Regarding the discussed topic on how bee’s longevity varies depending on different production stages, we explained the beekeeper another approach of our project: When the bee uses less energy in the production of antimicrobial peptides it is possible that its longevity increases.²

Bryan Saavedra did not know about synthetic biology and we gave him an approach to this area and how it had so many reaches, one of them being beekeeping. While explaining our project we absent-mindedly use a somewhat technical word that is usually complicated for the community in general. A peptide. The beekeeper, as in other times of the conversation, didn't hesitate to ask us and he attentively listened to us while we explained to him what we were referring to; he was always intrigued by our project.

References

Economía de Chile: agricultura y pesca de Chile. (2018). Retrieved from http://www.voyagesphotosmanu.com/economia_chile.html
Turcatto, A. P., Lourenço, A. P., & De Jong, D. (2017). Propolis consumption ramps up the immune response in honey bees infected with bacteria. Apidologie, 49(3), 287–296. doi:10.1007/s13592-017-0553-z
Image retrieved from: Melipilla Chile: Ubicación, mapa y todo lo que desconoce. (n.d.). Retrieved from http://conociendochile.com/c-region-metropolitana-de-santiago/melipilla-chile/

Specialists and Investigators

Our project considers particular parts as the immune system of the bee, as well as bigger picture aspects like how beekeepers provide the liquid food to their hives; the required panorama is vast, and we sought to make fit each small component of the project in the life of bees and beekeepers. Researchers who have been surrounded by the beekeeping world for years were the indicated to guide us in external aspects regarding the beekeepers' field of expertise.

June 7^th
Mexico, City
Video call with MVZ Ernesto Tanús Sánchez

August 25^th
São Paulo, Brazil
Video call with Ph.D. David De Jong

June 15^th
Cuauhtémoc, Chihuahua
Center for Research in Food and Development A.C. (CIAD) visit

October 2^nd
Guelph, Canada
Video conference with Dr. Ernesto Guzmán Novoa

June 7^th
Video call with MVZ Ernesto Tanús Sánchez

The current burning of beehives threatens the environment and the loss of its resources, which is why a viable alternative to avoid such burning is necessary.
He clarified that the word of a beekeeper is not in doubt; however, it might be hard for us to obtain real data regarding the presence of American foulbrood as it is a mandatory complaint issue, so there is distrust of the producer.

Dr. Tanus suggested liquid application (in the form of syrup) which beekeepers could feed their hives. This syrup (sweetened water) could contain the peptides in it and be fed directly to the box. This way, nurse bees would feed on here. Since Africanized bees appeared, he claims, it is better to have interior feeding to avoid foragers. This method would also avoid product loss from environmental conditions such as the wind or sun.
We mentioned the possibility of adding the solution cell by cell, but he said we must think of the beekeeper. This just isn’t scalable for 3,000 or 4,000 boxes. We realized here that the easier the application, the more attractive the product.
We should consider the effect our product would have on the bee's usual microbiome.
He put us in contact with MSc Alejandro Romo, MSc Juan Diego Perez de la Rosa, and Dr. Alfonso Arroyo.
We received an invitation to the XXVI International Congress for Beekeeping Updates in 2019.

Detailed Description

After receiving feedback from beekeepers, we looked into bees’ anatomy and behavior in a healthy environment. We were looking to enrich our investigation with the point of view of a veterinary expert. Dr. Tanus provided us with an hour and a half of his time to listen to our work. He began by introducing the arrival of foulbrood in Mexico and mentioning the National Program for Honey Harmlessness and Quality, which appeals to European standards (since the EU is Mexican honey’s largest purchaser). Among these standards, Mexico asks honey producers to create antibiotic-free products.

On American Foulbrood, he called the larvae’s first 24 hours “crucial”, since it is the time their defenses are at their lowest. He remarked that there aren´t effective remedies to these diseases, only preventive methods.

He said that bee colony arson was frowned upon and claimed that wood should not be used indiscriminately. When one suspects spores are present, it is imperative to change hives and flame the boxes by blowtorch, this is sterilization.

On our own account, we found NOM--SEMARNAT/SAGARPA-2007 which sets parameters under which it is legal to start fires on forestry, agricultural, and cattle land. It allows fires for the control of bacterial diseases, after meeting several requirements.

He mentioned foulbrood incidence has declined since 2008 thanks to the Good Honey Production Practices manual which recommends changing boxes and preventive strategies for beekeepers. He mentioned MSc Juan Diego Perez de la Rosa, Head of the Department of Genomic Sequencing at the National Center for Animal Health Verification Services, who has cultured Paenibacillus larvae for identification, from samples taken in 17 different states of the Mexican Republic.

Tanus stated that it is impossible to say that there is no presence of foulbrood in Mexico without first conducting an epizootiology study in the country. He clarified that the word of a beekeeper is not in doubt; however, it might be hard to obtain real data as it is a mandatory complaint issue, so there is distrust of the producer. He shared some characteristics that differentiate both foulbrood. He also commented on the incidence of these diseases throughout the seasons.

Dr. Tanus suggested liquid application (in the form of syrup) which beekeepers could feed their hives. This syrup (sweetened water) could contain the peptides in it and be fed directly to the box. This way, nurse bees would feed on here. Since Africanized bees appeared, he claims, it is better to have interior feeding to avoid foragers. This method would also avoid product loss from environmental conditions such as the wind or sun.

He reassured us that peptides would get to the brood, regardless of the application and only the time it takes might change. Peptides cannot arrive directly since larvae do not feed on their own: on their first 3 days, larvae are fed jelly, and with bee bread the following three. He also gave us a first draft of the path the peptides follow: from the digestive system to the hemolymph and finally to the hypopharyngeal glands in the head where they produce royal jelly.

Adult bees are not affected by P. larvae. In fact, in their digestive system, there is an anatomical structure called proventriculus and can close when the time comes to purify the food. He shared with us a new fact about hygienic behavior. As brood infected with AFB begin metamorphosis, proteolytic enzymes from P. larvae degrade fibers and reduce them to pure viscose biomass. Bees organs in their antennas can sense this decomposition; cleaner bees then break through the capped cell, remove the corpse, and clean the cell. All colonies of beehives exhibit this behavior being in some more marked than in others.

We mentioned the possibility of adding the solution cell by cell, but he said we must think of the beekeeper. This just isn’t scalable for 3,000 or 4,000 boxes. We realized here that the easier the application, the more attractive the product. This made us think of secondary factors that influence our products viability, other than its antimicrobial ability. This is further discussed in our business model. He noted that if the product is applied to cure it is susceptible to apply directly in the cells as mentioned above since in an apiary of 20 to 30 hives, this disease can be initiated in 2 or 3 of them, however, it should be applied as a preventive measure throughout the apiary.

He mentioned that we should consider the effect our product would have on the bee's usual microbiome. He asked us whether our product was a remedy or a preventive measure. We answered that the design wasn't enough to pick one or the other, and it might be both.

Finally, he recommended articles and contacts for further information. He put us in contact with MSc Alejandro Romo, MSc Juan Diego Perez de la Rosa, and Dr. Alfonso Arroyo.

He congratulated us for our work; he found our efforts and interest fascinating, especially when we put it to work to help the beekeeper community up close. He encouraged us to keep at it and invited us to the XXVI International Congress for Beekeeping Updates in 2019.

The chat was enlightening, and gave us valuable information for our research, hard to find in books or articles. He also recommended a website to check for information (OIE) which had epidemiological information on both foulbroods. We are still awaiting the 2018 update of foulbrood presence across the world.

We greatly thank Ernesto Tanus for his time and disposition, especially by giving us faith and encouragement to keep going and make this a reality.

June 15^th
Center for Research in Food and Development A.C. (CIAD) visit

In CIAD they have worked in investigations with Paenibacillus larvae and Melissococcus plutonius; they gave us advice on how to manage their in vitro growth.
We had the opportunity to interact with beehives, get to know how beekeepers work with them and how they feed them.
We were able to learn about in vitro bee breeding techniques.
We discussed the bioethical aspect of working with bees in the laboratory.

Detailed Description

The main objective of the realization of this visit was to get to know and evaluate the ideal conditions with which M. plutonius and P. larvae grow and how are they handled in vitro. On another hand, we looked forward to knowing the practices of in vitro bee breeding.

Four members of the team had the opportunity to travel to Cuauhtémoc and attend this visit where we invested approximately four hours, first in the laboratory and then in the field. M.S. Alejandro Romo Chacón (who has participated in more than 20 scientific dissemination articles related to bees or agriculture) welcomed us to the facilities and once we entered the laboratory he began to explain us about the bees’ life cycle and about some of the products and instruments beekeepers use to handle hives. Later, we were able to meet M.S. Adrián Ponce de León, who is researching about foulbrood bacteria, and along with M.S. Romo, he explained us about procedures and safety considerations in order to manage the in vitro growth of P. larvae and M. plutonius.

The day of our visit, M.S. Romo and M.S. Ponce de León planned to go to the bee field and carry out bee larvae transferences, for its following in vitro breeding. This activity is one of the parts that implies our project once we reach the experimentation stage with the insects and M.S. Romo and M.S. Ponce de León attentively invited us to accompany them to the field, they even lent us suits and veils to interact directly with the bees! Previously we had consulted a protocol that showed us all the details about the in vitro breeding of bees and by observing the techniques used by Ponce de León for the transference, we were able to validate and somehow, to begin with, the practice of this activity.

Interacting with the bees and being able to observe the composition of the bee boxes, the way in which the different foods were applied and the way in which beekeepers approached the bees were all key elements that allowed us to define the optimum application method of our product and helped us ensure its appropriate integration regarding each characteristic of the hive organization and the work done by beekeepers.

When returning to the CIAD facilities we had the opportunity to discuss the use of bees in the laboratory, how many bees were used? What was this number based on? Were the three R’s considered? The reality is that in Mexico there does not exist such regulations and the bioethical part of working with bees in the laboratory is quite unattended. We found this situation somewhat unsettling and it led us to perform a more extensive analysis of this topic. (consult the bioethical essay in).

M.S. Romo and M.S. Ponce de León could broaden their panorama on the research that is currently being done with bees and

their diseases thanks to the sharing we made with them of our project and the allusion that synthetic biology is its fundamental part.

Thanks to this visit we were able to have our first real approach with bees, we knew the reality of working with them and working with the two strains according to a specialized laboratory that has already experienced the proper biosafety considerations. This visit gave us different data that is very useful for the whole experimental plan of our project, and besides that, we lived a little of what beekeepers live every day, like traveling inside the truck with many bees that stayed inside from the field and even experiencing some bee-stings. We were also able to capture amazing photos of the hives!

References

Image retrieved from: Grupo Caresner S. de R.L de C.V. (2018). Museo Menonita. https://www.mexicodestinos.com/chihuahua/tours/museo-menonita.html

August 25^th
Video call with Ph.D. David De Jong

The application method of our product is feasible, but we should make sure bees could digest them. An excellent measure he suggested us was to eventually plan to extract the hypopharyngeal glands in bees and determine if our AMPs are entering these glands.
He believes okay for us to supply our peptides and then try to see if they help bees resist foulbrood, as for now, he thinks it's an excellent first step if we can produce some material and prove it to work on the laboratory

If we did get to test within larvae, we asked him which would be an optimum method to find out if our peptides are getting to the developing brood.
Based on his advice we can:

Consider two scenarios, feeding the larvae in vitro, where we won't be making use of a nurse bee or the second scenario where the nurse bee feeds the larvae.
See if our peptides get into the worker jelly; we can look at the hypopharyngeal glands and take a worker jelly sample from the cells where larvae are swirling.
Finally, compare to a control.

Regarding the incidence of American or European foulbrood, he mentioned he has worked in the US for control of these two diseases, the percentage was always around one percent, sometimes two. In Brazil, it's not established, they have few European foulbrood cases and there is no presence of American foulbrood, by the other hand, in Argentina, this last has been a big problem.
In Mexico, we have the presence of Varroa, he mentioned that's a concern because is probably affecting the bee's immune system, another reason to be concerned about the AMPs levels.
We asked him if it's legal to burn beehives on Brazil, he answered it’s not for someone who is not a beekeeper, the fireman is only supposed to do it if it’s a matter of environmental protection. He told us about when he was a US bee inspector, what they would do is when they find the disease, they would burn the colonies, he explained to us the methodology they followed.
In Brazil they have no regulation affecting research with honey bees, the only regulation is involving bees in the wild.
He offered to contact us to speak with Erica Weinstein Teixeira, she was a student of him, she set up a laboratory and it’s a good bee pathologist. He also offered to contact us with more researchers in his lab in case we needed to.

Detailed Description

P.hD. David de Jong is a professor of genetics and entomology at the University of São Paulo, Ribeirão Preto School of Medicine (specialist in pathology, biology, and management of Africanised honey bees); he is also technical Editor of an online scientific journal: Genetics and Molecular Research (GMR).

Our team leader, human practices leader, and our ethics, legal and mathematical model representatives were present in this 40-minute call to approach issues regarding their respective divisions. David the Jong is a researcher that has written in many papers we had previously found and read on bees; we thank MVZ Ernesto Tanús for contacting us with him. First off, he was curious if we were planning to test our peptides on the bee larvae or in the colony, so far, the idea was to test them against a bacterium within a culture in the lab, we were working on the time to see if we could test them in vivo on the larvae but we were not quite sure yet. He said that was a good first step and recommend us to emphasize that on our plan.

He was curious whether we know if it is feasible for beekeepers to eventually use our product, if it could be a costly production or if we have an idea of how much these peptides were going to cost in a final presentation. We mentioned him experimental procedures were still missing to solve these issues, like determining how much a bacterium can produce within a particular time and with given resources to give a better approximation of the cost.

He mentioned using AMPs in food for bees was newly and thinks it's feasible for us to test in culture and then determine which combination of AMPs works best to inhibit the bacteria. Before our phone call, we sent him our detailed project description to which he recommended us to add more details on our justification as well as mention the importance of honey, money

involved in its exportation and beekeeping in Mexico. He said we should also emphasize the impact of this diseases on the whole colony and beekeeping, “the larvae are interesting, but not so important concerning the application".

Regarding the application method of our product, we were considering putting nanocapsules containing our AMPs within the bee’s liquid food substitute. He said it’s feasible, but we would want to make sure bees could digest them. An excellent measure he suggested us was to eventually plan to extract the hypopharyngeal glands in bees and determine if our AMPs are entering these glands.

He believes okay for us to supply our peptides and then try to see if they help bees resist foulbrood, as for now, he thinks it's an excellent first step if we can produce some material and prove it to work on the laboratory, he mentioned that's a paper already. He emphasized is not easy to have real larvae on the laboratory.

We were wondering his opinion whether our AMPs could affect the larvae negatively in their development or worker bees in general. He compared or product to a human antibiotic, it kills the disease, but it also eliminates part of its microbiota, he thinks we could mention it, but we would have to test these possibilities experimentally and see what happens. He stated antibiotic treatments available now for foulbrood not always work, and they could contaminate the honey. Regarding the possibility of our AMPs getting into the honey, he thinks there are going to be low levels of our peptides, and they are going to be on the larvae, he doesn't think it's going to be a problem our AMPs getting to the honey.

If we did get to test within larvae, we asked him which would be an optimum method to find out if our peptides are getting to the developing brood.

Consider two scenarios, feeding the larvae in vitro, where we won't be making use of a nurse bee or the second scenario where the nurse bee feeds the larvae.
See if our peptides get into the worker jelly; we can look at the hypopharyngeal glands and take a worker jelly sample from the cells where larvae are swirling.
Finally, compare to a control

If we want to see if our AMPs are getting into the food nurse bees are feeding the larvae in the colony, we would have to use a colony or a mini colony, which is more complicated, he said.

"Many people have been working on ways to protect the bees, they have different systems they are using, but none of these experiments involve all these different steps you are needing because it's very ambitious and requires much time". He thinks if we can get the peptides off our bacterium and then show that they inhibit P. larvae and M. plutonius, we can talk about the next steps on a plan.

We were wondering if our peptides could activate some biochemical signal or have the larvae reject them. He said if an animal is given something it produces, the tendency is this animal will produce less. He recommended us to see if naturally, bee AMPs get into their food, it could be that maybe if bees have available more AMPs, they disactivate the bacteria, he wasn't sure about it.

When nanoencapsulated medicine is done, there is the risk that it can get accumulated somewhere within the organism since honey bees live within a month, he doesn't deem that is something we should worry about, yet he would say the fat body, that's the place bees tend to accumulate.

Regarding the incidence of American or European foulbrood, he mentioned he has worked in the US for control of these two diseases, the percentage was always around one percent, sometimes two. In Brazil, it's not established, they have few European foulbrood cases and there is no presence of American foulbrood, by the other hand, in Argentina, it has been a big problem. He was there to diagnose the first official finding of it, they had many thousands of colines dying, since then, some of their people went to Brazil to learn bee genetics and the incidence was very much reduced.

In Mexico, we have the presence of Varroa, he mentioned that's a concern because is probably affecting the bee's immune system, another reason to be concerned about the AMPs levels. If we get to see if nurse bee AMPs get to the food of larvae, that is an interesting result because it helps researches understand how bees can resist diseases. Beyond the work, we are proposing, or as part of it, he thinks is interesting to have that information.

As we continue talking about foulbrood, we asked him if it's legal to burn beehives on Brazil, he answered it’s not for someone who is not a beekeeper, the fireman is only supposed to do it if it’s a matter of environmental protection. He has burned a few beehives in a demonstration of how they control the disease when it is found, normally the government would burn beehives is there's a new problem or disease, but they are supposed to pay the beekeeper. He told us about when he was a US bee inspector, what they would do is when they find the disease, they would burn the colonies, quarantine the apiary, and tell the beekeeper he had five days to challenge, after that period, colonies that had symptoms would be burned.

What he would do is he would burn the frames and the columns on the bees and then crush the boxes so the beekeeper couldn't put bees in it anymore, nowadays, gasoline is used. The reason he did that is he wanted to make sure all the honey was destroyed because the honey stores were contaminated with the spores of American foulbrood and if the honey wasn't destroyed, all the apiaries around would get infected. Another way the disease is spread between beekeepers is through the places where honey is processed, they bring honey from different beekeepers or different countries and process it, if it is contaminated and bees get access to it, the disease is spread in the surroundings. Another way of spreading is when a beekeeper wants to buy or sell beehives or equipment that is contaminated because they don't see the symptoms and the person who is selling or buying, doesn't know there is a contamination.

We were curious if there were any biotechnical regulations that apply to bees in Brazil and the US regarding experimental work on them for example, for eradicating diseases. In Brazil they have no regulation affecting research with honey bees, the only regulation is involving bees in the wild. For experimental work, for instance, he is on the ethics committee of the hospital, he mentioned for human research a permission is needed as for animal research, but there are no insects’ permissions.

He offered to contact us to speak with Erica Weinstein Teixeira, she was a student of him, she set up a laboratory and it’s a good bee pathologist. Erika has worked with viruses and diagnosis and said she probably knows about AMPs. He also offered to contact us with more researchers in his lab in case we needed to.

Ultimately, he mentioned American foulbrood was not a problem in Brazil, but in Argentina is. He sent us some papers that could be useful to our research and said it was a good project. We are thankful for his help and for his willingness to keep supporting us.

October 2^nd
Video conference with Dr. Ernesto Guzmán Novoa

To the best of his knowledge, no one has worked with bee’s antimicrobial peptide genes to treat diseases in these arthropods.
There might be some questions we won’t be able to answer until we get into bee experimentation, but we should consider possible risks such as the addition of external AMPs to the colony atrophying the natural production of AMPs in bees, as well as the possibility of weakening their microbiota.

He emphasized that it is not an easy project to develop as working with American and European foulbrood of honey bees is extremely complicated because of the specialized infrastructure, techniques and studies required to work with honey bee brood diseases.
Regarding American and European foulbrood disease presence in Canada, he stated European foulbrood predominates over American foulbrood in Canada.
There are no regulations in Canada concerning experimentation on bees, yet, he doesn't consider there should be since regulating it would hinder and inhibit research with them.

Detailed Description

Dr. Ernesto Guzmán is a professor and head of the Honey Bee Research Centre at the University of Guelph; he received a DVM degree from the National Autonomous University of Mexico and a MSc and Ph.D. in entomology from the University of California, Davis. Dr. Guzmán has advised or co-advised the theses of more than 50 DVM, MSc, and Ph.D. students. Research in his lab focuses on three main areas: genetics, behavior and parasitic mites of honey bees, identifying factors that affect honey bee health.

On October 2, we met with Dr. Guzmán on a video conference to speak about our project; he referred to it as something new since to the best of his knowledge, no one has worked with bee’s antimicrobial peptide genes to treat diseases in these arthropods. He mentioned there might be some questions we won’t be able to answer until we get into bee experimentation, but we should consider possible risks such as the addition of external AMPs to the colony atrophying the natural production of AMPs in bees, as well as the possibility of weakening their microbiota.

He emphasized that it is not an easy project to develop as working with American

and European foulbrood of honey bees is extremely complicated because of the specialized infrastructure, techniques and studies required to work with honey bee brood diseases. It can take months or even years to standardize the protocols needed, so he was curious if we were planning to continue working on this project after the competition.

Regarding American and European foulbrood disease presence in Canada, he stated European foulbrood predominates over American foulbrood in Canada, but in Mexico, American foulbrood is a severe problem.

Finally, we were curious whether there were any regulations regarding experimentation on bees in his country, to which he answered there are none in Canada concerning experimentation on them, yet, he doesn't consider there should be since regulating it would hinder and inhibit research with them. Ultimately, he wished us luck and referred to our project as good and novel.

Beekeeping Societies

We had the great privilege of being able to get closer to complete and organized associations that show how beekeepers gather, collaborate and strengthen beekeeping. Dialogue is fundamental in a society and what a better way to join this dialogue than by discussing in these reunions, listening to the concerns or contributions expressed by the societies and building-up new group insights.

May 5^th
Cuauhtémoc, Chihuahua
Monthly beekeepers’ association meeting

July 27^th, 28^th and 29^th
Delicias City, Chihuahua
IV State Beekeeping Forum

July 19^th
Rome, Italy
Video call with FAI presidential staff member Lorenzo Della Morte

May 5^th
Monthly beekeepers’ association meeting

Until that meeting, our project consisted of the production of antimicrobial peptides in Pichia pastoris for exclusively the inhibition of the pathogen Paenibacillus larvae. The intention was to apply these yeasts in a solution with hexane extract of the plant Achyrocline satureioides in the presence also of surfactins, the surfactin would have a synergistic effect with the hexane extract. This solution would be applied to bee bread, which we knew, the larvae ingested in their diet.
They said there is not much research in the beekeeping sector, so it seems excellent that we intend to carry out scientific research in this field and try different combinations of peptides thus ensuring the best combination.

From this meeting, we set out to investigate the export requirements of honey, which is the main product exported by beekeepers, and how our genetically modified yeasts would affect or play a role in these requirements.
Through their feedback, we evaluated the risks of using yeasts in our product and the risks of introducing GMOs in what is known as "harmless honey".
They emphasized that an overdose of yeasts would cause a fermentation of the honey and an alteration in the microbiota of the bees which could be counterproductive since this would cause diseases such as American foulbrood to arrive, they recommended us to be very careful with the dose and consider the addition of a fermentation inhibitor in our formula.
Concerning the final product being a dilution, they commented that this was difficult to apply since, if our product was to be applied frequently, it would be laborious to have to spray every three days, for example, 2,000 boxes of bees.
In their city, the foulbrood that represents a problem is the European.
Finally, they congratulated us for our research and stressed the importance of feedback between researcher and producer since this way we can collaborate in a more effective way, guaranteeing a bidirectional communication.
MSc Alejandro Romo offered us to experiment with his bees in his laboratory at Cuauhtémoc.
After this meeting, our project changed to the production of antimicrobial peptides in Escherichia coli for the inhibition of Paenibacillus larvae and Melissococcus plutonius

Detailed Description

We thank Mr. Guillermo Rodríguez for inviting us to the monthly meeting of the Cuauhtemoc's beekeeping society where we had our first approach with the beekeeping community. The society consisted of approximately 33 beekeepers and researchers who gave us a warm welcome, listened to our project, and gave us their opinion on particular details that we had to consider or improve. They were happy that young students of other disciplines were interested in their work and bees, we explained to them the reason it was important for us to talk with them and why we had chosen to work with bees in our project, something that made them curious.

Until that meeting, our project consisted of the production of antimicrobial peptides in Pichia pastorisfor exclusively the inhibition of the pathogen Paenibacillus larvae. We planned on using yeasts due to their high capacity to carry out post-translational modifications, similar to the protein folding mechanisms of bees and their ability to secrete heterologous proteins to the environment, in this case, our antimicrobial peptides. The intention was to apply these yeasts in a solution with hexane extract of the plant Achyrocline satureioides which its efficiency against P. larvae has already been shown previouslysup>2, in presence also of surfactins, the surfactin would have a synergistic effect with the hexane extract. Previous studies on B. subtilis have shown that surfactin, a lipopeptide produced by the bacteria, inhibits the pathogens of honey bees, including P. larvae, A. apis and Nosema ceranae.¹ This solution would be applied to bee bread, which we knew, the larvae ingested in their diet.

Through their feedback, we evaluated the risks of using yeasts in our product and the risks of introducing GMOs in what is known as "harmless honey". They emphasized that an overdose of yeasts would cause a fermentation of the honey and an alteration in the microbiota of the bees which could be counterproductive since this would cause diseases such as American foulbrood to arrive, they recommended us to be very careful with the dose and consider the addition of a fermentation inhibitor in our formula. From this meeting, we set out to investigate the export requirements of honey, which is the main product exported by beekeepers, and how our genetically modified yeasts would affect or play a role in these requirements.

One issue we were asked to review was whether the hexane extract of A.satureioides oxidized because if so, it loses its qualities. Currently, a treatment that they use to fight diseases in bees is the oregano essential oil which they have already found to be oxidized. After further investigation, we verified that certainly, the extract that we wanted to add is oxidized by exposure to air. Another treatment that beekeepers use to fight diseases, is the sodium bicarbonate which, when it is spread in the hive, causes the bees to begin a cleaning protocol with which they take that bicarbonate out of their hive taking away the dirt present.

Thanks to this meeting, we set out to investigate the export requirements of honey, which is the main product exported by beekeepers, and how our genetically modified yeasts would affect or play a role in these requirements. They mentioned that they do not usually have much time to try different combinations of treatments and find out the optimal combination. They said there is not much research in the beekeeping sector, so it seems excellent that we intend to carry out scientific research in this field and try different combinations of peptides thus ensuring the best combination.

They also told us about other bee pathogens, Nosema apis and Varroa destructor. They commented that, if effective, our product is excellent, however, they also mentioned that in general there are many good products and at a reasonable price, however, they are challenging to work with, occasional applications are less complicated than regular ones. They look for a practical alternative, cheap and that doesn't consume time. Concerning the final product being a dilution, they commented that this was difficult to apply since, if our product was to be applied frequently, it would be laborious to have to spray every three days, for example, 2,000 boxes of bees.

Subsequently, they mentioned that, in their city, the foulbrood that represents a problem is the European, which occurs when bees have little accessibility to pollen because of the little flowering throughout the year. Previously we had read about this disease; however, it was from this meeting that we investigated more about it and after validating the theoretical efficacy of our antimicrobial peptides of interest against this disease, we decided to address it as part of the pathogens to combat with our product.

Finally, they congratulated us for our research and stressed the importance of feedback between researcher and producer since in this way we can collaborate in a more effective way, guaranteeing a bidirectional communication. They invited us to continue attending their and MSc Alejandro Romo, a researcher at the Center for Food Research and Development (For its acronym in Spanish: CIAD), offered to do the experimentation with bees from our research in his laboratory in Cuauhtémoc, the same laboratory that we visited one month later.

After this meeting, our project changed to production of antimicrobial peptides in Escherichia coli for the inhibition of Paenibacillus larvae and Melissococcus plutonius

References

Wu, M., Sugimura, Y., Iwata, K., Takaya, N., Takamatsu, D., Kobayashi, M., … Yoshiyama, M. (2014). Inhibitory effect of gut bacteria from the Japanese honey bee, Apis cerana japonica, against Melissococcus plutonius, the causal agent of European foulbrood disease. Journal of Insect Science, 14, 129. http://doi.org/10.1093/jis/14.1.129
Chanchao, C., Stoić, M., Yoshiyama, M. and Kimura, K. (2018). Review on Apiary Management and Antimicrobial Compounds as Alternative Prevention or Treatment for Honeybee Diseases. OnLine Journal of Biological Sciences, 18(1), pp.46-61.

July 19^th
Video call with FAI presidential staff member Lorenzo Della Morte

Discussion of aspects like:

Foulbrood incidence rate in Italy.
Treatment methods used in Italy to treat American foulbrood and European foulbrood.
How legislative tools, regarding beekeeping, are often perceived as repressive.
FAI insights about our solution proposal.

Detailed Description

Looking forward to expanding our international reach, we encountered the possibility to contact the Italian Federation of Beekeepers (FAI). Successfully, after exchanging some emails we were able to schedule a virtual meeting with the FAI presidential staff member Lorenzo Della Morte. We were very excited to have the opportunity to share and also nurture our project with insights, concerns and regulations that came from the city that hosts the headquarters of many important multinational food companies and agencies, such as the United Nations Food and Agriculture Organization (FAO), the International Fund for Agricultural Development (IFAD) and The World Food Programme (WFP). Eight members of the team could attend this important reunion that lasted approximately 40 minutes.

FAI President Raffaele Cirone and his staff member Lorenzo Della Morte shared with us very invaluable information about how foulbrood is experienced in Italy:

Foulbrood incidence rate in Italy usually has a seasonal trend with the largest number of registered infections during the second semester of the year (July-December). Regarding the impact and incidence of these two diseases on Italian beekeeping, FAI Italian Federation of Beekeepers has consulted the National Centre of Reference on bee diseases, who elaborated an official data reporting 51 complaints of colony infections from 7 on the 20 Italian Regions, for the year 2017. The number of infected and then destroyed colonies is a few tens of units compared to the 1.300.000 colonies, which constitute the Italian beekeeping heritage.

From the FAI point of view, the data is not to be considered statistically reliable both for the low number of registered cases and for the missing complaints. In fact, in order to avoid the Veterinarian Police controls, many beekeepers often don’t report the infection and directly destroy the colonies in secret. According to FAI estimates the incidence rate of foulbrood can’t be lower than the 10-20% of the Italian beekeeping heritage in total.

The treatment methods used in Italy to treat AFB and EFB are the following:

For AFB:

Must be compulsorily notifiable to the Health Authority. It involves serious restrictive measures in accordance with the Veterinarian Police Regulation (DPR 8/2/1954, n.320). Apart from the Beekeeping Best Practices (BBP) there are no official and authorized control methodologies so that the infected colonies must be burned. Furthermore, the apiary is subject to health constraints until the recovery plan has been ascertained. All the beekeeper’s instruments must be disinfected and sterilized.
For EFB:

Must be compulsorily notifiable to the Health Authority. It involves serious restrictive measures in accordance with the Veterinarian Police Regulation (DPR 8/2/1954, n.320). Differently, from the American foulbrood (AFB) regulation, the European foulbrood (EFB), has a less restricted regulation. In fact, the infected colonies must be distinguished in two different clusters related to their clinical picture:

Severely compromised colonies: In this case, the approach is the same as the one for AFB and it lies in destroying both the colony and the hive and in disinfecting and sterilizing the beekeeper’s instruments.
Not compromised colonies but with clinical signs: In this case, the “swarming technique” is carried out to overcome the disease. It consists of:

Removing the infected combs from the hive.
Replacing the queen bee.
Adding adult bees from external healthy colonies.
Feeding bees with an appropriate nutrition.
Further practices suggested by the veterinarian in charge.

The apiary will be seized for at least 9 days and until the positive outcome of the treatment, in order to assess the efficacy of the “swarming technique”. Anyway, also, in this case, the beekeeper can ask the destruction of all the affected colonies to the veterinarian authority.

In both the cases AFB and EFB, the use of antibiotics is not allowed in Italy because of its ineffectiveness and to prevent the spread of antibiotic-resistant bacterial strains.

FAI emphasized a very important point that seems not only to apply in Italy but throughout the whole world: there are no different solutions to AFB and EFB diseases these days and the legislative tools are often perceived as repressive, so many beekeepers don’t notify the infection in their apiary, destroying the infected colonies and concealing the fact. This is one of the most relevant problems for the health and statistical assessment studies.

After Mr. Della Morte learned about our project and the scopes of synthetic biology regarding beekeeping, he considered our proposal as a possibility of prevention method against the diseases. Our product would help beekeepers do not have to lose all their beehives and save money.

After the video call, Mr. Raffaele Cirone and his staff reflected on our project information and kindly sent us their final insights:

The Italian Federation of Beekeepers believes that AMPABEE Project is a new and innovative approach to such incurable diseases as AFB and EFB. Preventing the spread of these bacteria and increasing the immune system of bee larvae is an important goal to achieve in the health bee risk prevent. In particular, nowadays the only preventing tools are the Beekeeping Best Practices (BBP) which depends on the personal application beekeepers carry out. In this respect, FAI considers essential to test this research on the pure Italian Apis mellifera ligustica, as it is the most widespread in the world. In this regard, FAI will collaborate and provide the necessary support, remaining at iGEM team Tec-Chihuahua disposal in case of need for further information for the research project and programme. FAI is totally sure that such a research could find a spreading interest in the beekeeping world at different scales. FAI can support the product commercialization thanks to the FAI’s Service Company and the FAI trade partnerships covering the entire national territory.

July 27^th, 28^th, and 29^th
IV State Beekeeping Forum

The forum was held in the Chihuahuan Desert Museum and had an approximate of 340 attendees from the state including beekeepers from the Cuauhtemoc’s beekeeping society, attendees from other states and from the country of Chile.
We had the opportunity to speak with diverse beekeeping companies.
Beekeepers spoke about the current contamination that exists in honey, the main problems are antibiotics, sulfonamides are still used in many parts of the country.
We were the closure presentation of the event. Juan Carlos Coronado, a Chilean beekeeper we met at the forum, invited us to visit his apiaries.

Engineer Sóstenes Rafael Rodríguez Dehaibes, CEO of COMVERMIEL and national and international speaker in beekeeping pathology topics in Mexico, Brazil, Uruguay, Argentina, Venezuela, and Colombia. Mr. Sóstenes shared with us some of the studies made for honey exportation. Honey needs to meet specific criteria concerning quality, labeling, residues of treatments for bees and pollutants from the environment. He stressed that for antimicrobial peptides, studies had not been done yet.
A research done in 2017 has documented a dramatic decline in average airborne insect biomass of 76% (up to 82% in midsummer) in just 27 years for protected nature areas in Germany. This considerably exceeds the estimated decline of 58% in global abundance of wild vertebrates over a 42-year period to 2012. Their results demonstrate that recently reported declines in several taxa such as butterflies, wild bees and moths, are in parallel with a severe loss of total aerial insect biomass, suggesting that it is not only the vulnerable species but the flying insect community as a whole, that has been decimated over the last few decades.2
An issue Mexican beekeepers are facing are transgenic crops, there is the presence of transgenic pollen in bees' honey. In 2009 Germany banned MON810 maize from cultivation for agricultural purposes, even though the EU has approved it for release into the environment.³ Harmless honey for exportation to Germany is not allowed to have presence of MON810 maize which is cultivated here in Mexico.
In general, beekeepers told us we were doing impressive research and saw huge potential in our work as a real solution for the foulbrood problem as well as a possible treatment for Nosema and Varroa.
On the last day of the event, we visited apiaries with visitors from Mexico city, Chile, Cuauhtemoc city and other states of the republic. It was an incredible experience for us.

Detailed Description

We are truly grateful with Ernesto Tanús Sánchez, Manuel Ramírez Salcido, Juan Diego Pérez de la Rosa and Felipe Esquivel Díaz president of the Delicia’s beekeepers society in behalf of such, for inviting us as speakers at the IV State Beekeeping Forum, a forum organized by the Delicia’s Beekeepers society in coordination with the city council and the state government. The experience itself in addition to the willingness and kindness of beekeepers, government representatives, and beekeeping companies to contribute to our project, made of this event a valuable approach to understanding how the world affects our project and how our project affects the world.

The forum was held in the Chihuahuan Desert Museum and had an approximate of 340 attendees from the state including beekeepers from the Cuauhtemoc’s beekeeping society, attendees from other states and from the country of Chile. The event intended to update the sector, discuss existing options for increasing productivity and encourage beekeepers to continue making products derived from honey.

It was important for us to get involved with our relevant communities seeking stimulating dialogues and bi-directional learning. We engaged conversation with Engineer Sóstenes Rafael Rodríguez Dehaibes, CEO of COMVERMIEL and national and international speaker in beekeeping pathology topics in Mexico, Brazil, Uruguay, Argentina, Venezuela, and Colombia. Mr. Sóstenes shared with us some of the studies made for honey exportation and stressed the importance of surveillance plans for this product, which means it must be defined which parameters should be analyzed and at which intervals. This way, there is a minimization of risks, and the cost-benefit is optimized.

Honey needs to meet specific criteria concerning quality, labeling, residues of treatments for bees and pollutants from the environment. It is imperative that honey is processed protecting its nutritional components. Some examples of residues evaluated in honey exportation are sulfonamides, streptomycin, tetracyclines, chloramphenicol, nitrofurans, and phenol pesticides.¹ Above all, the analysis of residues requires sensitive methods. He stressed that for antimicrobial peptides, studies had not been done yet.

We brought two posters to the forum, one of them was the

first draft of our Giant Jamborees' poster and the other one was an explanation about what is synthetic biology and its applications. We were visited in our posters by the state delegate of the Secretariat of Agriculture, Livestock, Rural Development, Fisheries and Food (For its acronym in Spanish: SAGARPA), Isaac Zepeda Romero, who congratulated us for our work and encouraged us to continue working hard. Also, we reunited once again with MVZ Ernesto Tanús who introduced us to other more beekeepers, Dr. José Luis Reyes Carrillo was also present.

During this forum, we talked with many beekeepers, between the topics we were discussing they mentioned all wild pollinators are scarce. They told us in Germany when researchers went to the field looking for bumblebees, they did not find any, they realized that their cars had no insects that had hit their windows on the road, there is a degree of desertification of insects.

A research done in 2017 has documented a dramatic decline in average airborne insect biomass of 76% (up to 82% in midsummer) in just 27 years for protected nature areas in Germany. This considerably exceeds the estimated decline of 58% in global abundance of wild vertebrates over a 42-year period to 2012. Their results demonstrate that recently reported declines in several taxa such as butterflies, wild bees and moths, are in parallel with a severe loss of total aerial insect biomass, suggesting that it is not only the vulnerable species but the flying insect community as a whole, that has been decimated over the last few decades.²

Beekeepers also spoke about the current contamination that exists in honey, the main problems are antibiotics, sulfonamides are still used in many parts of the country. The mentioned honey that is not exported is because it is contaminated with antibiotics. Another issue Mexican beekeepers are facing are transgenic crops, there is the presence of transgenic pollen in bees' honey. In 2009 Germany banned MON810 maize from cultivation for agricultural purposes, even though the EU has approved it for release into the environment.³Harmless honey for exportation to Germany is not allowed to have presence of MON810 maize which is cultivated here in Mexico.

In the forum, we had the opportunity to speak with diverse beekeeping companies, Api ofertas, APIMAT, X-nox Inovación and Salcido apiarios, who showed us some of their beekeeping products and also treatments they market for managing Varroa. We participated in a raffle organized by Api Ofertas and we are so excited we won a Beekeeper suit we will show it to our iGEM friends in Boston!

On the first day of the forum, the Delicia’s beekeeping association invited us to join them in a dinner, at this dinner we spoke with Mr. Rodolfo Contreras Martínez and Mr. Rodolfo Contreras Lozano, two beekeepers from Chile owners of San Martín Apiaries. We spoke about pollination, the

different types of honey and their exportation. We especially thank Delicias' beekeeping society for their hospitality by offering us to stay the night in the hotel Los Cedros at Delicias city where other guests of the forum were also staying.

We were the closure presentation of the event. Juan Carlos Coronado, a Chilean beekeeper we met at the forum, invited us to visit his apiaries. In general, beekeepers told us we were doing impressive research and saw huge potential in our work as a real solution for the foulbrood problem as well as a possible treatment for Nosema and Varroa. Other presentations at the forum included topics such as pollination, neonicotinoids in Beekeeping, the honey market, between others.

On the last day of the event, we visited apiaries with visitors from Mexico city, Chile, Cuauhtemoc city and other states of the republic. We went to visit two apiaries, one of them was exclusively for breeding queen bees and the other one was for honey production. We were taught how to differentiate between the different types of bees, for example between drone bees, queen bees, and worker bees. We saw larvae in their cells and visualized a worm that parasites bees. It was an incredible experience for us.

Trough the three days, we applied surveys to beekeepers to obtain more information on foulbrood in the country. Beekeepers from a variety of ages and states answered these surveys. We had a total of 16 answered surveys. We confirmed there had been the presence of American and European foulbrood in Mexico in the last seven years with an increase in the previous 3 years. Terramycin is the most used treatment to combat this diseases. When we asked how much would they be willing to invest for a product that combats and prevents this both diseases and in general strengthens the immune system of bees, 50% of the surveyed said they would pay the necessary.

More information about our attendance to this forum has been addressed in our Education and Public Engagement section.

Survey Format
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References

Intertek (n.d.). Miel y Productos Apícolas. [online] Intertek.com.mx. Available at: http://www.intertek.com.mx/servicios/miel/
Hallmann CA, Sorg M, Jongejans E, Siepel H, Hofland N, Schwan H, et al. (2017) More than 75 percent decline over 27 years in total flying insect biomass in protected areas. PLoS ONE 12(10): e0185809. https://doi.org/10.1371/journal.pone.0185809
Edith, P. (2015). Restrictions on Genetically Modified Organisms: Germany | Law Library of Congress. [online] Library of Congress. Available at: https://www.loc.gov/law/help/restrictions-on-gmos/germany.php

Team:Tec-Chihuahua/Human Practices

Overview

Social Science Research Description

References

Consent Letter▼

Letter of Approval▼

Silver Human Practices

Government Institutions

June 27thMeeting with MVZ Ernesto Tanús Sánchez, MVZ Adolfo Arroyo, and Ing. José Luis González Miranda

Detailed Description

References

July 17thVideo call with MVZ Juan Diego Pérez de la Rosa

Detailed Description

References

United Nations

September 18thVideo call with Dr. María Mercedes Roca

Detailed Description

Environmental

Economic

Legal

References

October 26th and 27thMEXMUN

Detailed Description

Beekeepers

April 28thMeeting with Mr. Guillermo Rodríguez

Detailed Description

References

May 26thMeeting with Ing. Manuel Ramírez Salcido

Detailed Description

References

June 15thMeeting with Engineer Alejandro Bianchi

Detailed Description

July 16thValencian beekeeper interview

Detailed Description

September 22ndVideocall with Bryan Saavedra

Detailed Description

References

Specialists and Investigators

June 7thVideo call with MVZ Ernesto Tanús Sánchez

Detailed Description

June 15thCenter for Research in Food and Development A.C. (CIAD) visit

Detailed Description

References

August 25thVideo call with Ph.D. David De Jong

Detailed Description

October 2ndVideo conference with Dr. Ernesto Guzmán Novoa

Detailed Description

Beekeeping Societies

May 5thMonthly beekeepers’ association meeting

Detailed Description

References

July 19thVideo call with FAI presidential staff member Lorenzo Della Morte

Detailed Description

July 27th, 28th, and 29thIV State Beekeeping Forum

Detailed Description

Survey Format▼

References

Gold Integrated Human Practices

Project Basis ∇

Project Basis

Experimentation plan

Important considerations for the project

Social ∇

Economic ∇

Environmental ∇

Environmental

References

Economic

References

Social

Ethical

Bioethics▼

Legal

Bioethics▼

References

Follow us on social media:

We thank our sponsors:

Consent Letter
▼

Letter of Approval
▼

June 27^th
Meeting with MVZ Ernesto Tanús Sánchez, MVZ Adolfo Arroyo, and Ing. José Luis González Miranda

July 17^th
Video call with MVZ Juan Diego Pérez de la Rosa

September 18^th
Video call with Dr. María Mercedes Roca

October 26^th and 27^th
MEXMUN

April 28^th
Meeting with Mr. Guillermo Rodríguez

May 26^th
Meeting with Ing. Manuel Ramírez Salcido

June 15^th
Meeting with Engineer Alejandro Bianchi

July 16^th
Valencian beekeeper interview

September 22^nd
Videocall with Bryan Saavedra

June 7^th
Video call with MVZ Ernesto Tanús Sánchez

June 15^th
Center for Research in Food and Development A.C. (CIAD) visit

August 25^th
Video call with Ph.D. David De Jong

October 2^nd
Video conference with Dr. Ernesto Guzmán Novoa

May 5^th
Monthly beekeepers’ association meeting

July 19^th
Video call with FAI presidential staff member Lorenzo Della Morte

July 27^th, 28^th, and 29^th
IV State Beekeeping Forum

Survey Format
▼

Bioethics
▼

Bioethics
▼

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sponsors: