Difference between revisions of "Team:Toulouse/Design"

 
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  <div class="group center">
 
  <p class="text">
 
TrApiColi has been designed in order to take into account ethical
 
reflection, safety and ease of use for beekeepers. The use of
 
genetically modified organisms in a field, and because our project is associated with
 
edible products, underlies both applying of regulations and public interest.
 
In this context, as explain above, we searched a solution being able to isolate our engineered bacteria from
 
the environment, but allowing its growth, metabolism and gas diffusion. We found the
 
project of the iGEM Groeningen 2012 team which used the polymer TPX® in
 
order to contain their bacteria separated of the meat [1]. Thus, we
 
decided to ask them TPX® characteristics. We finally contacted the TPX® supplier Mitsui Chemicals. We obtained
 
some samples of TPX® by contacting the company MitsuiChemicals. TPX®, Polymethylpentene (4-methylpentene-1 based polyolefin, Mitsui Chemicals, Inc.), is a polymer.
 
 
 
<br>
 
<br>
 
To check if our device can be safe and feasible, several tests have been performed:
 
</p>
 
</div>
 
 
 
<ul style="font-size:15px;margin-bottom:10px;">
 
<li>Safety test: Impermeability of bacteria through the bag of TPX®
 
</li>
 
<li>Gas diffusion tests: Permeability of butyric acid and formic acid through the bag of TPX®
 
</li>
 
<li>Growth tests in TPX®: Culture of the strain <i>E. coli</i> BW25113 can grow in a TPX® bag (<i>ie.</i> in microaerobic conditions, without agitation and miming batch culture condition), as it would be in the field
 
</li>
 
<li>Bacterial survival over 15 days in microaerobic condition
 
</li>
 
<li>Carbone source test: choice of Carbon source to produce acids during 10 days
 
</li>
 
<li>Acid toxicity on <i>E. coli</i>
 
</li>
 
</ul>
 
 
 
 
 
    
 
    
 
   <div class="title">
 
   <div class="title">
  <h3>Trap Construction</h3>
+
      <a href="#main1"><h3>Content</h3></a>
 
   </div>
 
   </div>
 
    
 
    
 +
  <center> 
 +
    <div id="breadcrumb" class="clear" style="float: center;" >
 +
  <ul>
 +
        <li><a href="#trapconstruct">- Trap Device </a></li>
 +
        <li><a href="#tpx">- ApiColi containment and culture</a></li>
 +
      </ul>
 +
    </div>
 +
 +
  </center>
 +
 
   <div class="group center">
 
   <div class="group center">
 
   <p class="text">
 
   <p class="text">
Our goal is to create a solution against varroas. In order to use ApiColi to treat varroosis, we designed a trap, named TrApiColi.
+
Our goal is to create a solution against hives infestation by varroas. In order to get the best out of ApiColi in the treatment of varroosis, we designed a trap named TrApiColi. TrApiColi was thought up to take into account ethical reflection, safety and ease of use for beekeepers.
 
   </p>
 
   </p>
 
   </div>
 
   </div>
    
+
     <div class="group center">
+
   <div id = "trapconstruct"></div>
  <p class="text">
+
  <div class="title" >
Since the production of the two pathways are regulated by day light, our bacteria need to be outside of the beehive. Thus, the trap was made to be placed at the entrance of the hive, in order to prevent the entry of the mites.
+
  <h3>Trap Device</h3>
 +
  </div>
 +
      
 +
<div class="group center">
 +
<p class="text">
 +
Since the switch between the production of the two molecules is regulated by light, our bacteria need to be outside of the beehive. Thus, the trap was designed to be placed at the entrance of the hive, in order to act before infection and try to prevent the entry of as many mites as possible.
 
   </p>
 
   </p>
 
   </div>
 
   </div>
 
    
 
    
    <div class="group center">
+
 
 +
<center>
 +
<div style="one-half;padding:10px;">
 +
<img src="https://static.igem.org/mediawiki/2015/6/63/TLSE_Device_Trap_1.png" style="width:70%;" />
 +
</div>
 +
 
 +
<p class="legend">
 +
The four different parts of our trap
 +
</p>
 +
</center>
 +
 
 +
<div class="group center">
 
   <p class="text">
 
   <p class="text">
TrApiColi is composed of four main parts:
+
TrApiColi is composed of four main parts:<br><br>
 +
<strong>1.</strong> A grid lined up with the bottom board
 +
<br><br>The bees usually enter the hive by landing on the bottom board before walking inside. Because of the alignment of the trap with the board, it will not disturb the bees' comings and goings. The holes are big enough to let the varroas fall through them, but not the bees.
 +
</p>
 +
</div> 
 +
<br><br>
 +
   
 +
<div class="group"> 
 +
<p class="text">
 +
<strong>2.</strong> A funnel, to channel all the falling varroas
 +
</p>  </div>
 +
 +
<center>
 +
<div style="one-half;padding:10px;">
 +
<img src="https://static.igem.org/mediawiki/2015/e/e2/TLSE_Device_Trap_2.jpg" style="width:40%;" />
 +
</div>
 +
 
 +
<p class="legend">
 +
The grid and the funnel
 +
</p></center>
 +
 
 +
<div class="group"> 
 +
<p class="text">
 +
<strong>3.</strong> A transparent collector, containing the bacteria confined in a special bag
 +
<br><br>It is designed like a fish bottle trap: the tube from the funnel goes on for a few centimeters inside the collector to ease the entry of the varroas in the collector while preventing them from exiting. The special bag is described in the <a href="https://2015.igem.org/Team:Toulouse/Design#tpx">"ApiColi containment and culture"</a> part below.
 +
</p>  </div>
 +
<br><br>
 +
 
 +
<div class="group"> 
 +
<p class="text">
 +
<strong>4.</strong> A roof, to protect the trap from the rain
 +
<br><br> Associated with pannels blocking the sides of the entrance, it forces the bees to walk on the grid. This channeling does not disturbs the bees.
 +
</p>
 +
</div>
 +
 
 +
  <div class="group center"> 
 +
<p class="text">
 +
<br>
 +
The dimensions of the trap allow it to adapt to almost every beehive. Indeed, most of the hive types have the exact same entrance. Thanks to that, the trap can be easily and perfectly installed on the hive by the beekeepers without drilling or cutting.
 
  </p>
 
  </p>
 
   </div>
 
   </div>
 +
  <br><br>
 +
<div class="group center"> 
 +
<p class="text">
 +
The trap was designed using Catia and then 3D printed in order to build a prototype. It is used as a demonstration device for the beekeepers and the general public. This trap could not be tested without coating because the porous plastic used for 3D printing is permeable to liquids and gases. Moreover, the modeling showed that this version of the trap is yet to be optimized to ensure a proper diffusion of our molecules, see more in <a target="_blank" href="https://2015.igem.org/Team:Toulouse/Modeling#part5"> "Modeling" </a> part.
 +
</p>  </div>
 
    
 
    
 +
  <center> <table><tbody><tr><td>
 +
<div style="one-half;padding:10px;">
 +
<img src="https://static.igem.org/mediawiki/2015/c/c2/TLSE_Device_Trap_3.jpg" style="width:80%;" />
 +
</div>
 +
</td>
 +
<td>
 +
<div style="one-half;padding:10px;">
 +
<img src="https://static.igem.org/mediawiki/2015/0/08/TLSE_Device_Trap_4.jpg" style="width:80%;" />
 +
</div>
 +
</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
</center>
 +
 +
<center>
 +
<p class="legend">
 +
The 3D printer used to construct our trap and the result of 3D printing: TrApiColi
 +
</p></center>
 
    
 
    
 
+
<div id = "tpx"></div>
 
+
 
  <div class="title">
 
  <div class="title">
   <h3>TPX Bag</h3>
+
   <h3>ApiColi containment and culture</h3>
 
   </div>
 
   </div>
    
+
 
+
   <div class="group center">
 +
  <p class="text">
 +
The use of genetically modified organisms in a field associated with edible products requires to carefully consider the issues of safety, national legislation, and public opinion. In this context, we looked for a solution that would isolate our engineered bacteria from the environment, while still allowing its growth and gas diffusion.<br>
 +
We found the solution in the <a href="https://2012.igem.org/Team:Groningen/Sticker"target="_blank">iGEM Groeningen 2012 project</a> that used a special polymer called TPX® in order to keep their bacteria from contaminating meat, their edible product.<br>
 +
TPX® is composed of Polymethylpentene, a porous polymer, and is commercialized by the company MitsuiChemicals (4-methylpentene-1 based polyolefin, Mitsui Chemicals, Inc.). In order to perform our experiments, we contacted MitsuiChemicals who offered us some samples of TPX®.
 +
</p></div>
  
 
+
<center>
<!----ON NECRIT PAS APRES--->
+
<div style="one-half;padding:10px;">
 
+
<img src="https://static.igem.org/mediawiki/2015/thumb/2/29/TLSE_TPX_bag.jpeg/433px-TLSE_TPX_bag.jpeg" style="width:40%;" />
 
+
</main>
+
 
</div>
 
</div>
<div class="wrapper row4">
+
<p class="legend">
 
+
A TPX containing 6 ml of LB medium
<div class="container clear" style="padding-top:30px;">
+
<center><p class="maintitle">  
+
References
+
 
</p></center>
 
</p></center>
  
<div class="clear">
 
<ul>
 
  
<li>
+
<div class="group center">
[1] REFERENCE 1 Vers la page Groeningen 2012
+
<p class="text">
</li>
+
As in the Groeningen 2012 project, our bacteria need to be separated from the growth medium until the beekeeper decides to use the trap. In order to do that, freeze-dryed bacteria are contained in a small polyvynil chloride bag placed inside a bigger TPX® bag. Since polyvynil chlorid is fragile, the user have to break it in order to start the growth.
 +
<br><br>
 +
To check the feasability and safety of our device, several tests have been performed:
 +
</p></div>
  
<li>
+
<ul style="font-size:15px;margin-bottom:10px;">
+
<li><a href="https://2015.igem.org/Team:Toulouse/Results#TPX"target="_blank">Growth tests in TPX®</a>: Culture of the strain <i>E. coli</i> BW25113 in a TPX® bag (<i>ie.</i> in microaerobic conditions, without agitation and miming batch culture condition), as it would be in the field
[2] REFERENCE 2 AVEC UN LIEN <a href="http://www.google.com/patents/US8647615">See more</a>
+
 
</li>
 
</li>
 +
<li><a href="https://2015.igem.org/Team:Toulouse/Results#gaz"target="_blank">Gas diffusion tests</a>: Permeability of butyric acid and formic acid through the TPX® bag
 +
</li>
 +
<li><a href="https://2015.igem.org/Team:Toulouse/Results#safety"target="_blank">Safety test</a>: Impermeability of the bag of TPX® to the bacteria
 +
</li>
 +
<li><a href="https://2015.igem.org/Team:Toulouse/Results#biolo"target="_blank">Bacterial survival</a> over 15 days in microaerobic conditions
 +
</li>
 +
<li><a href="https://2015.igem.org/Team:Toulouse/Results#carbon"target="_blank">Carbone source test</a>: choice of a carbon source to produce acids during at least 10 days
 +
</li>
 +
<li><a href="https://2015.igem.org/Team:Toulouse/Results#tox"target="_blank">Acid toxicity</a> on <i>E. coli</i>
 +
</li>
 +
</ul>
  
<li>
+
<center>
[3] REFERENCE 2 AVEC UN LIEN qui ouvre dans une nouvelle fenêtre <a href="http://www.google.com/patents/US8647615">See more</a>
+
<div class="title">
 +
<h3>READ MORE</h3>
 +
</div> </center>
 +
 
 +
<div class="group center">
 +
<div style="width:20%;">
 +
<a href="https://2015.igem.org/Team:Toulouse/Description">
 +
<div class="title">
 +
<h3>Project Description</h3>
 +
</div>
 
</a>
 
</a>
</li>
+
</div>
 +
<div style="width:20%;">
 +
<a href="https://2015.igem.org/Team:Toulouse/Description/Attract">
 +
<div class="title">
 +
<h3>Attract</h3>
 +
</div>
 +
</a>
 +
</div>
 +
<div style="width:20%;">
 +
<a href="https://2015.igem.org/Team:Toulouse/Description/Eradicate">
 +
<div class="title">
 +
<h3>Eradicate</h3>
 +
</div>
 +
</a>
 +
</div>
 +
<div style="width:20%;">
 +
<a href="https://2015.igem.org/Team:Toulouse/Description/Regulation">
 +
<div class="title">
 +
<h3>Regulation</h3>
 +
</div>
 +
</a>
 +
</div>
  
</ul>
+
<div style="width:20%;">
 +
<a href="https://2015.igem.org/Team:Toulouse/Results">
 +
<div class="title">
 +
<h3>Results</h3>
 +
</div>
 +
</a>
 +
</div>
 
</div>
 
</div>
  
  
<div class="clear">
+
<!----ON NECRIT PAS APRES--->
  
</div>
 
<br>
 
</div>
 
  
 +
</main>
 
</div>
 
</div>
 +
 
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Latest revision as of 22:16, 18 September 2015

iGEM Toulouse 2015

Device


Our goal is to create a solution against hives infestation by varroas. In order to get the best out of ApiColi in the treatment of varroosis, we designed a trap named TrApiColi. TrApiColi was thought up to take into account ethical reflection, safety and ease of use for beekeepers.

Trap Device

Since the switch between the production of the two molecules is regulated by light, our bacteria need to be outside of the beehive. Thus, the trap was designed to be placed at the entrance of the hive, in order to act before infection and try to prevent the entry of as many mites as possible.

The four different parts of our trap

TrApiColi is composed of four main parts:

1. A grid lined up with the bottom board

The bees usually enter the hive by landing on the bottom board before walking inside. Because of the alignment of the trap with the board, it will not disturb the bees' comings and goings. The holes are big enough to let the varroas fall through them, but not the bees.



2. A funnel, to channel all the falling varroas

The grid and the funnel

3. A transparent collector, containing the bacteria confined in a special bag

It is designed like a fish bottle trap: the tube from the funnel goes on for a few centimeters inside the collector to ease the entry of the varroas in the collector while preventing them from exiting. The special bag is described in the "ApiColi containment and culture" part below.



4. A roof, to protect the trap from the rain

Associated with pannels blocking the sides of the entrance, it forces the bees to walk on the grid. This channeling does not disturbs the bees.


The dimensions of the trap allow it to adapt to almost every beehive. Indeed, most of the hive types have the exact same entrance. Thanks to that, the trap can be easily and perfectly installed on the hive by the beekeepers without drilling or cutting.



The trap was designed using Catia and then 3D printed in order to build a prototype. It is used as a demonstration device for the beekeepers and the general public. This trap could not be tested without coating because the porous plastic used for 3D printing is permeable to liquids and gases. Moreover, the modeling showed that this version of the trap is yet to be optimized to ensure a proper diffusion of our molecules, see more in "Modeling" part.

The 3D printer used to construct our trap and the result of 3D printing: TrApiColi

ApiColi containment and culture

The use of genetically modified organisms in a field associated with edible products requires to carefully consider the issues of safety, national legislation, and public opinion. In this context, we looked for a solution that would isolate our engineered bacteria from the environment, while still allowing its growth and gas diffusion.
We found the solution in the iGEM Groeningen 2012 project that used a special polymer called TPX® in order to keep their bacteria from contaminating meat, their edible product.
TPX® is composed of Polymethylpentene, a porous polymer, and is commercialized by the company MitsuiChemicals (4-methylpentene-1 based polyolefin, Mitsui Chemicals, Inc.). In order to perform our experiments, we contacted MitsuiChemicals who offered us some samples of TPX®.

A TPX containing 6 ml of LB medium

As in the Groeningen 2012 project, our bacteria need to be separated from the growth medium until the beekeeper decides to use the trap. In order to do that, freeze-dryed bacteria are contained in a small polyvynil chloride bag placed inside a bigger TPX® bag. Since polyvynil chlorid is fragile, the user have to break it in order to start the growth.

To check the feasability and safety of our device, several tests have been performed:

  • Growth tests in TPX®: Culture of the strain E. coli BW25113 in a TPX® bag (ie. in microaerobic conditions, without agitation and miming batch culture condition), as it would be in the field
  • Gas diffusion tests: Permeability of butyric acid and formic acid through the TPX® bag
  • Safety test: Impermeability of the bag of TPX® to the bacteria
  • Bacterial survival over 15 days in microaerobic conditions
  • Carbone source test: choice of a carbon source to produce acids during at least 10 days
  • Acid toxicity on E. coli

READ MORE