Difference between revisions of "Team:Tec-Chihuahua/test"

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.

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.

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 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.

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.

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.

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!

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.

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.

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.

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.