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

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<center><img src="https://static.igem.org/mediawiki/2018/3/33/T--Tec-Chihuahua--basis2.png" width="950px" height="auto"></center>
 
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<div class="a"><p align="justify"> Excretion of antimicrobial peptides in <i>Pichia pastoris for</i> <i>Paenibacillus larvae</i> and <i>Melissococcus plutonius</i> inhibition. The initial plan was to apply genetically modified yeasts engineered to excrete AMP’s and surfactin in a solution with hexane extract of the plant A. satureioides; surfactin has a synergistic effect with the hexane extract. This solution would be applied to the bee bread we knew larvae ingested in their diet.
 
<div class="a"><p align="justify"> Excretion of antimicrobial peptides in <i>Pichia pastoris for</i> <i>Paenibacillus larvae</i> and <i>Melissococcus plutonius</i> inhibition. The initial plan was to apply genetically modified yeasts engineered to excrete AMP’s and surfactin in a solution with hexane extract of the plant A. satureioides; surfactin has a synergistic effect with the hexane extract. This solution would be applied to the bee bread we knew larvae ingested in their diet.
 
</p></div><br>
 
</p></div><br>
 +
 
<div class="a"><p align="justify">Learned obstacles regarding the method:</p></div>
 
<div class="a"><p align="justify">Learned obstacles regarding the method:</p></div>
 
<ol class="o">
 
<ol class="o">
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<li>An overdose of yeasts would cause fermentation of the honey and an alteration in the microbiota of the bees which could be counterproductive since this would cause diseases such as Nosema and American foulbrood to arrive.</li>
 
<li>An overdose of yeasts would cause fermentation of the honey and an alteration in the microbiota of the bees which could be counterproductive since this would cause diseases such as Nosema and American foulbrood to arrive.</li>
 
</ol><br>
 
</ol><br>
 +
 
<center><img src="https://static.igem.org/mediawiki/2018/3/39/T--Tec-Chihuahua--basis3.png" width="950px" height="auto"></center><br>
 
<center><img src="https://static.igem.org/mediawiki/2018/3/39/T--Tec-Chihuahua--basis3.png" width="950px" height="auto"></center><br>
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<div class="a"><p align="justify">Production of antimicrobial peptides in Escherichia coli for Paenibacillus larvae and Melissococcus plutonius inhibition. The second plan consisted in applying the purified antimicrobial peptides, after their production in E. coli BL21 (DE3), in the bee bread we knew larvae ingested in their diet.</div></p>
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<div class="a"><p align="justify">Production of antimicrobial peptides in Escherichia coli for Paenibacillus larvae and Melissococcus plutonius inhibition. The second plan consisted in applying the purified antimicrobial peptides, after their production in E. coli BL21 (DE3), in the bee bread we knew larvae ingested in their diet.</div></p><br>
  
 +
<div class="a"><p align="justify">Learned obstacles regarding the method:</p></div>
 +
<ol class="o">
 +
<li>Bee bread is mainly stored in the hive so there is little chance that the peptides reach the larva.</li>
 +
</ol>
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<center><img src="https://static.igem.org/mediawiki/2018/8/89/T--Tec-Chihuahua--basis4.png" width="950px" height="auto"></center>
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<div class="a"><p align="justify">The third plan consisted in applying the purified peptides sprinkled as a liquid solution or as a powder on the hives.</p></div>
 +
 +
<div class="a"><p align="justify">Learned obstacles regarding the method:</p></div>
 +
<ol class="o">
 +
<li>When the product is found scattered in the hive, it can be easily affected by external factors such as air or other insects.</li>
 +
</ol>
 +
 +
<div class="a"><p align="justify">We were able to arrive at the <b>final application method</b> with the recommendation of <b>MVZ Ernesto Tanús</b> who considered that the most suitable way to apply our product would be through the liquid food beekeepers provide to the hives and that nurse bees ingest to feed the larvae subsequently.</p></div><br>
 +
 +
<h4>Experimentation plan</h4>
 +
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<div class="a"><p align="justify">We integrated three aspects discussed during our interviews within the <a href="https://2018.igem.org/Team:Tec-Chihuahua/Results">future  experimentation plan</a> of the project:</p></div>
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 +
<ol class="o">
 +
<li>Add an innocuous color dye to the nanospheres to make the track in the bee system and ensure that the product reaches the larvae. This topic was discussed during the meeting with <b>MVZ Ernesto Tanús Sánchez, MVZ Adolfo Arroyo,</b> and <b>Eng. José Luis González Miranda.</b></li>
 +
<li>Test the research on the pure Italian <i>Apis mellifera</i> spp. <i>ligustica</i>, as it is the most widespread in the world. This information was considered thanks to the video call with FAI presidential staff member <b>Lorenzo Della Morte.</b> </li>
 +
<li>Extract the hypopharyngeal glands of the nurse bees after they consume the liquid food where nanospheres were added in order to determine if there is the presence of AMP's. Consideration taken from the conversation with <b>Ph.D. David de Jong.</b></li>
 +
</ol>
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<br>
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<h4>Important considerations for the project</h4>
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<div class="a"><p align="justify"><b>MVZ Ernesto Tanús</b> considered it was essential to take into consideration the factor of the microbiota of the bee, regarding the fact that there would be AMP's supplied that can alter the natural microbiota. Regarding this same subject, <b>Ph.D. David De Jong</b> considered that our product works like the antibiotics humans use; there is the negative part that the intestinal flora will be affected, but this is in search for a greater good, attacking pathogens. The experimentation is necessary in order to be able to know what would happen with the microbiota of the bee and what consequences this situation could bring.</p></div><br>
 +
 +
<div class="a"><p align="justify">Another of the considerations of <b>Ph.D. David De Jong</b>, was the analysis on how when you give certain substance to an animal, there is a proclivity for it to produce less of this substance; risk addressed in the <a href="#1">bioethical essay</a> and mentioned in the <a href="#2">legal plan</a>.</p></div><br>
 
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Revision as of 16:08, 17 October 2018

Erwinions

Gold Integrated Human Practices

Our integrated human practices are the result of each and every single human practices activity that we carried out throughout the iGEM season. We truly believe our work, in every section of our project, including wet lab, has significantly changed towards the implementation of our relevant communities feedback. Since the first day we engaged with beekeepers, our project has been shaping and improving towards an effective, responsible, safe and ethical work.


After the video call we had with Maria Mercedes Roca, Ph.D, where emphasis was placed on the three pillars of the UN for sustainable development, we saw the importance of these three areas and decided to exploit them in our integrated human practices: the economic, social and environmental area of our project, as well as a section showing how the design and execution of it has been, molded thanks to our silver human practices.



Project Basis

The foundation of the project was evolving, improving and solidifying thanks to the interactions we experienced in our human practices. There were fundamental changes such as the production chassis of the peptides and the application method of the product, as well as recommendations for the experimental plan and insights about the risk factors we should consider.

Excretion of antimicrobial peptides in Pichia pastoris for Paenibacillus larvae and Melissococcus plutonius inhibition. The initial plan was to apply genetically modified yeasts engineered to excrete AMP’s and surfactin in a solution with hexane extract of the plant A. satureioides; surfactin has a synergistic effect with the hexane extract. This solution would be applied to the bee bread we knew larvae ingested in their diet.


Learned obstacles regarding the method:

  1. Presence of genetically modified yeats would complicate the exportation process of honey to the European Union increasing the risks of having the product rejected.
  2. The hexane extract of A.satureioides is oxidized by exposure to air; therefore, it may lose its efficiency.
  3. An overdose of yeasts would cause fermentation of the honey and an alteration in the microbiota of the bees which could be counterproductive since this would cause diseases such as Nosema and American foulbrood to arrive.


Production of antimicrobial peptides in Escherichia coli for Paenibacillus larvae and Melissococcus plutonius inhibition. The second plan consisted in applying the purified antimicrobial peptides, after their production in E. coli BL21 (DE3), in the bee bread we knew larvae ingested in their diet.


Learned obstacles regarding the method:

  1. Bee bread is mainly stored in the hive so there is little chance that the peptides reach the larva.

The third plan consisted in applying the purified peptides sprinkled as a liquid solution or as a powder on the hives.

Learned obstacles regarding the method:

  1. When the product is found scattered in the hive, it can be easily affected by external factors such as air or other insects.

We were able to arrive at the final application method with the recommendation of MVZ Ernesto Tanús who considered that the most suitable way to apply our product would be through the liquid food beekeepers provide to the hives and that nurse bees ingest to feed the larvae subsequently.


Experimentation plan

We integrated three aspects discussed during our interviews within the future experimentation plan of the project:

  1. Add an innocuous color dye to the nanospheres to make the track in the bee system and ensure that the product reaches the larvae. This topic was discussed during the meeting with MVZ Ernesto Tanús Sánchez, MVZ Adolfo Arroyo, and Eng. José Luis González Miranda.
  2. Test the research on the pure Italian Apis mellifera spp. ligustica, as it is the most widespread in the world. This information was considered thanks to the video call with FAI presidential staff member Lorenzo Della Morte.
  3. Extract the hypopharyngeal glands of the nurse bees after they consume the liquid food where nanospheres were added in order to determine if there is the presence of AMP's. Consideration taken from the conversation with Ph.D. David de Jong.

Important considerations for the project

MVZ Ernesto Tanús considered it was essential to take into consideration the factor of the microbiota of the bee, regarding the fact that there would be AMP's supplied that can alter the natural microbiota. Regarding this same subject, Ph.D. David De Jong considered that our product works like the antibiotics humans use; there is the negative part that the intestinal flora will be affected, but this is in search for a greater good, attacking pathogens. The experimentation is necessary in order to be able to know what would happen with the microbiota of the bee and what consequences this situation could bring.


Another of the considerations of Ph.D. David De Jong, was the analysis on how when you give certain substance to an animal, there is a proclivity for it to produce less of this substance; risk addressed in the bioethical essay and mentioned in the legal plan.



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