Difference between revisions of "Team:NYMU-Taipei/Notebook-Lab Book-fruit flies hemolymph"

Line 586: Line 586:
 
</div>
 
</div>
  
−
<script>if (window.runOnloadHook) runOnloadHook();</script>
+
<script>if (window.runOnloadHook) runOnloadHook();</script></div>
 +
 
 +
<div class="prototypeprototypesp">
 +
<div id="wrap">
 +
<div class="fund">
 +
<h2 style="margin-top:30px; margin-bottom:10px;">Overview</h2><hr>
 +
 
 +
<p style="font-size:16px;">
 +
This section of our wiki contains the protocols for our insect experiments, including cultivation, infection rate tests, and hemolymph extraction.</p>
 +
 
 +
</div>
 +
 
 +
<div class="fund">
 +
 
 +
<h2 style="margin-top:30px; margin-bottom:10px;">Related experiment schedule & Lab note</h2><hr>
 +
<br /><h4 style="margin-top:30px; margin-bottom:10px;">Experiments</h4>
 +
<table border="1" class="tablesatisfy">
 +
　<tr>
 +
　    <td style="width:600px;">Experiments related to B. dorsalis</td>
 +
　    <td style="width:150px;">Number</td>
 +
　    <td style="width:150px;">Date</td>
 +
　</tr>
 +
　<tr>
 +
　    <td style="width:600px;">Diameter of the tunnel that allows passage to one B. dorsalis</td>
 +
　    <td style="width:150px;">1</td>
 +
　    <td style="width:150px;">8/30～9/1</td>
 +
　</tr>
 +
　<tr>
 +
　    <td style="width:600px;">Infecting B. dorsalis with non-designed M. anisopliae</td>
 +
　    <td style="width:150px;">2-1</td>
 +
　    <td style="width:150px;">9/30~10/7</td>
 +
　</tr>
 +
　<tr>
 +
　    <td style="width:600px;">infecting female B. dorsalis by copulation with infected male B. dorsalis</td>
 +
　    <td style="width:150px;">2-2</td>
 +
　    <td style="width:150px;">10/10~10/17</td>
 +
　</tr>
 +
</table>
 +
 
 +
<div class="tablesatisfysp" style="overflow-x:auto;">
 +
<table border="1">
 +
　<tr>
 +
　    <td style="width:600px;">Experiments related to B. dorsalis</td>
 +
　    <td style="width:150px;">Number</td>
 +
　    <td style="width:150px;">Date</td>
 +
　</tr>
 +
　<tr>
 +
　    <td style="width:600px;">Diameter of the tunnel that allows passage to one B. dorsalis</td>
 +
　    <td style="width:150px;">1</td>
 +
　    <td style="width:150px;">8/30～9/1</td>
 +
　</tr>
 +
　<tr>
 +
　    <td style="width:600px;">Infecting B. dorsalis with non-designed M. anisopliae</td>
 +
　    <td style="width:150px;">2-1</td>
 +
　    <td style="width:150px;">9/30~10/7</td>
 +
　</tr>
 +
　<tr>
 +
　    <td style="width:600px;">infecting female B. dorsalis by copulation with infected male B. dorsalis</td>
 +
　    <td style="width:150px;">2-2</td>
 +
　    <td style="width:150px;">10/10~10/17</td>
 +
　</tr>
 +
</table>
 +
</div>
 +
 
 +
<table border="1" class="tablesatisfy">
 +
　<tr>
 +
　    <td style="width:600px;">Experiments about hemolymph extraction</td>
 +
　    <td style="width:150px;">Number</td>
 +
　    <td style="width:150px;">Date</td>
 +
　</tr>
 +
　<tr>
 +
　    <td style="width:600px;">Blatta lateralis</td>
 +
　    <td style="width:150px;">1</td>
 +
　    <td style="width:150px;">9/20~9/30</td>
 +
　</tr>
 +
　<tr>
 +
　    <td style="width:600px;">Bactro</td>
 +
　    <td style="width:150px;">2</td>
 +
　    <td style="width:150px;">9/29~10/5</td>
 +
　</tr>
 +
　<tr>
 +
　    <td style="width:600px;">Bombyx mori</td>
 +
　    <td style="width:150px;">3</td>
 +
　    <td style="width:150px;">10/3~10/</td>
 +
　</tr>
 +
</table>
 +
 
 +
<div class="tablesatisfysp" style="overflow-x:auto;">
 +
<table border="1">
 +
　<tr>
 +
　    <td style="width:600px;">Experiments about hemolymph extraction</td>
 +
　    <td style="width:150px;">Number</td>
 +
　    <td style="width:150px;">Date</td>
 +
　</tr>
 +
　<tr>
 +
　    <td style="width:600px;">Blatta lateralis</td>
 +
　    <td style="width:150px;">1</td>
 +
　    <td style="width:150px;">9/20~9/30</td>
 +
　</tr>
 +
　<tr>
 +
　    <td style="width:600px;">Bactro</td>
 +
　    <td style="width:150px;">2</td>
 +
　    <td style="width:150px;">9/29~10/5</td>
 +
　</tr>
 +
　<tr>
 +
　    <td style="width:600px;">Bombyx mori</td>
 +
　    <td style="width:150px;">3</td>
 +
　    <td style="width:150px;">10/3~10/</td>
 +
　</tr>
 +
</table>
 +
</div>
 +
<br /><h4 style="margin-top:30px; margin-bottom:10px;"">Routine works</h4>
 +
 
 +
<table border="1" class="tablesatisfy">
 +
　<tr>
 +
　    <td style="width:750px;">Routine tasks of feeding Bactrocera dorsalis</td>
 +
　    <td style="width:150px;">Date</td>
 +
　</tr>
 +
　<tr>
 +
　    <td style="width:750px;">Produce & change the tube with the specific agar</td>
 +
　    <td style="width:150px;">8/24~9/20</td>
 +
　</tr>
 +
　<tr>
 +
　    <td style="width:750px;">Produce & change water gel and the specific ratio fodder</td>
 +
　    <td style="width:150px;">9/22~10/</td>
 +
　</tr>
 +
</table>
 +
 
 +
<div class="tablesatisfysp" style="overflow-x:auto;">
 +
<table border="1">
 +
　<tr>
 +
　    <td style="width:750px;">Routine tasks of feeding Bactrocera dorsalis</td>
 +
　    <td style="width:150px;">Date</td>
 +
　</tr>
 +
　<tr>
 +
　    <td style="width:750px;">Produce & change the tube with the specific agar</td>
 +
　    <td style="width:150px;">8/24~9/20</td>
 +
　</tr>
 +
　<tr>
 +
　    <td style="width:750px;">Produce & change water gel and the specific ratio fodder</td>
 +
　    <td style="width:150px;">9/22~10/</td>
 +
　</tr>
 +
</table>
 +
</div>
 +
 
 +
<table border="1" class="tablesatisfy">
 +
　<tr>
 +
　    <td style="width:750px;">Routine tasks of hemolymph extraction</td>
 +
　    <td style="width:150px;">Date</td>
 +
　</tr>
 +
　<tr>
 +
　    <td style="width:750px;">Collect the mulberry leaves to feed the silkworm</td>
 +
　    <td style="width:150px;">10/3~10/</td>
 +
　</tr>
 +
　<tr>
 +
　    <td style="width:750px;">Sterilize the related equipments</td>
 +
　    <td style="width:150px;">9/20~10/</td>
 +
　</tr>
 +
</table>
 +
 
 +
<div class="tablesatisfysp" style="overflow-x:auto;">
 +
<table border="1">
 +
　<tr>
 +
　    <td style="width:750px;">Routine tasks of hemolymph extraction</td>
 +
　    <td style="width:150px;">Date</td>
 +
　</tr>
 +
　<tr>
 +
　    <td style="width:750px;">Collect the mulberry leaves to feed the silkworm</td>
 +
　    <td style="width:150px;">10/3~10/</td>
 +
　</tr>
 +
　<tr>
 +
　    <td style="width:750px;">Sterilize the related equipments</td>
 +
　    <td style="width:150px;">9/20~10/</td>
 +
　</tr>
 +
</table>
 +
</div>
 +
 
 +
</div>
 +
 
 +
</div><br /><br />
 +
 
 +
<div class="expand">
 +
<center><p style="font-size:24px; color: #666666 !important; text-shadow: 0px 0px 1px #000000 !important; margin-bottom:100px;">
 +
<a href="#" onclick="ddaccordion.collapseall('technology'); return false"
 +
style="color: #666666 !important; text-shadow: 0px 0px 1px #000000 !important;  /*margin: 12px !important;*/  font-size: 18px !important;  text-decoration: none;">[Collapse all]</a>  |
 +
<a href="#" onclick="ddaccordion.expandall('technology'); return false"
 +
style="color: #666666 !important; text-shadow: 0px 0px 1px #000000 !important;  /*margin: 12px !important;*/  font-size: 18px !important;  text-decoration: none;">[Expand all]</a></p></center></div>
 +
 
 +
<div class="technology">8/23</div>
 +
<div class="thelanguage">
 +
<li><p style="font-size: 16px;">Oriental fruit flies sent to lab and cultivated by large transparent box to avoid oriental fruit flies flying out.</p></li>
 +
 
 +
<div class="imageimage">
 +
<div style="position:relative; width:100%;">
 +
<img src="https://static.igem.org/mediawiki/2016/2/2a/T--NYMU-Taipei--photo-OFF-related-14724253_120300000672924380_2135741640_o.jpg" width="100%" /></div>
 +
</div>
 +
 
 +
<p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology', 0); return false">[Collapse]</a></p>
 +
</div>
 +
 
 +
<div class="technology">8/24</div>
 +
<div class="thelanguage">
 +
<li><p style="font-size: 16px;">We used both of centrifuge tube and feeding box to feed oriental fruit flies for comparing and ensuring the status of oriental fruit flies that established the independent space for observing oriental fruit flies.</p></li>
 +
 
 +
<div class="imageimage">
 +
<div style="position:relative; width:100%;">
 +
<img src="https://static.igem.org/mediawiki/2016/4/42/T--NYMU-Taipei--photo-OFF-related-%E6%8B%8D%E7%85%A7.jpg" width="100%" /></div>
 +
</div>
 +
<br />
 +
<div class="imageimage">
 +
<div style="position:relative; width:48%;">
 +
<img src="https://static.igem.org/mediawiki/2016/3/3d/T--NYMU-Taipei--photo-OFF-related-14658400_120300000674554374_884455077_n.jpg" width="100%" /></div>
 +
<div style="position:relative; width:48%;">
 +
<img src="https://static.igem.org/mediawiki/2016/7/7d/T--NYMU-Taipei--photo-OFF-related-14686491_120300000678083870_2056369023_n.jpg" width="100%" /></div>
 +
</div>
 +
 
 +
<p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology', 1); return false">[Collapse]</a></p>
 +
</div>
 +
 
 +
<div class="technology">8/25</div>
 +
<div class="thelanguage">
 +
<li><p style="font-size: 16px;">B. dorsalis started to break through the cocoon. We conducted to put B. dorsalis to centrifuge tubes, avoiding the feeding box cannot load all oriental fruit flies. We poured the special agar to centrifuge tubes, and put centrifuges into entrance to catch oriental fruit flies. Then we sealed the centrifuge tubes by cotton. </li></p>
 +
 
 +
<div class="imageimage">
 +
<div style="position:relative; width:100%;">
 +
<img src="https://static.igem.org/mediawiki/2016/5/5c/T--NYMU-Taipei--photo-OFF-related-14697277_120300000677127274_438898518_o.jpg" width="100%" /></div>
 +
</div>
 +
 
 +
<p style="font-size: 16px;">(The team members put agar into centrifuge tubes which would be their feed.)</p>
 +
 
 +
<div class="imageimage">
 +
<div style="position:relative; width:48%;">
 +
<img src="https://static.igem.org/mediawiki/2016/e/ea/T--NYMU-Taipei--photo-OFF-related-14741005_120300000677094868_1093963562_n.jpg" width="100%" /></div>
 +
<div style="position:relative; width:48%;">
 +
<img src="https://static.igem.org/mediawiki/2016/4/40/T--NYMU-Taipei--photo-OFF-related-14696947_120300000676485877_378050777_n.jpg" width="100%" /></div>
 +
</div>
 +
 
 +
<p style="font-size: 16px;">(The left picture is that oriental fruit flies broke through cocoon; the right picture is the feeding box & centrifuge tubes where fed oriental fruit flies.)</p>
 +
 
 +
<p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology', 2); return false">[Collapse]</a></p>
 +
</div>
 +
 
 +
<div class="technology">8/30</div>
 +
<div class="thelanguage">
 +
<li><p style="font-size: 16px;">First time to Conducting Experiment 1</p></li>
 +
 
 +
<p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology', 3); return false">[Collapse]</a></p>
 +
</div>
 +
 
 +
<div class="technology">9/3</div>
 +
<div class="thelanguage">
 +
<li><p style="font-size: 16px;">Changing the centrifuge tubes where oriental fruit flies lived. (Avoid the environment being mess.) /p></li>
 +
 
 +
<p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology', 4); return false">[Collapse]</a></p>
 +
</div>
 +
 
 +
<div class="technology">9/5</div>
 +
<div class="thelanguage">
 +
<li><p style="font-size: 16px;">Changing the centrifuge tubes where oriental fruit flies lived.</p></li>
 +
 
 +
<p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology', 5); return false">[Collapse]</a></p>
 +
</div>
 +
 
 +
<div class="technology">9/6</div>
 +
<div class="thelanguage">
 +
<li><p style="font-size: 16px;">The oriental fruit flies in centrifuge tubes are dead quickly, so we conducted trouble shooting and sent the mail to professors who research oriental fruit flies.</p></li>
 +
<li><p style="font-size: 16px;">We took the trap to Beitou orchard to try to catch oriental fruit flies. </p></li>
 +
 
 +
<p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology', 6); return false">[Collapse]</a></p>
 +
</div>
 +
 
 +
<div class="technology">9/7</div>
 +
<div class="thelanguage">
 +
<li><p style="font-size: 16px;">Second time to Conducting Experiment 1.</p></li>
 +
<li><p style="font-size: 16px;">Conducting trouble shooting.</p></li>
 +
 
 +
<p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology', 7); return false">[Collapse]</a></p>
 +
</div>
 +
 
 +
<div class="technology">9/10</div>
 +
<div class="thelanguage">
 +
<li><p style="font-size: 16px;">Searching the store selling the silkworm for extracting hemolymph which is used to induce pMcl1.</p></li>
 +
 
 +
<p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology', 8); return false">[Collapse]</a></p>
 +
</div>
 +
 
 +
<div class="technology">9/13</div>
 +
<div class="thelanguage">
 +
<li><p style="font-size: 16px;">Visiting two farmers working in the orchard to ask the information about oriental fruit flies in the nature environment, and installing the trap of catching oriental fruit flies.</p></li>
 +
 
 +
<p style="font-size: 16px;">(Left picture: the team member install the trap; Right picture: the farmer and our team member took a picture)</p>
 +
 
 +
<div class="imageimage">
 +
<div style="position:relative; width:50%; margin-right:10%;">
 +
<img src="https://static.igem.org/mediawiki/2016/d/d5/T--NYMU-Taipei--photo-OFF-related-14740977_120300000687733839_596021223_n.jpg" width="100%" /></div>
 +
<div style="position:relative; width:90%; margin-top:10%;">
 +
<img src="https://static.igem.org/mediawiki/2016/7/74/T--NYMU-Taipei--photo-OFF-related-14741635_120300000673924526_1894584987_n.jpg" width="100%" /></div>
 +
</div>
 +
 
 +
<li><p style="font-size: 16px;">Sent the mail to researchers to ask for making public the content of the mail.</p></li>
 +
 
 +
<p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology', 9); return false">[Collapse]</a></p>
 +
</div>
 +
 
 +
<div class="technology">9/15</div>
 +
<div class="thelanguage">
 +
<li><p style="font-size: 16px;">Communicating with Chang Ming ecological education sericulture, asking the information of buying silkworm.</p></li>
 +
 
 +
<p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology', 10); return false">[Collapse]</a></p>
 +
</div>
 +
 
 +
<div class="technology">9/16</div>
 +
<div class="thelanguage">
 +
<li><p style="font-size: 16px;">Picking up the oriental fruit fly larval from rotten guavas and feeding them.</p></li>
 +
<li><p style="font-size: 16px;">Left picture: the team members were picking up the oriental fruit fly larval from rotten guavas; Right picture: the larval have picked up and put into another box.  Expected becoming pupae after six days. </p></li>
 +
 
 +
<div class="imageimage">
 +
<div style="position:relative; width:48%;">
 +
<img src="https://static.igem.org/mediawiki/2016/f/f9/T--NYMU-Taipei--photo-OFF-related-14699910_120300000674856561_1048345374_n.jpg" width="100%" /></div>
 +
<div style="position:relative; width:48%;">
 +
<img src="https://static.igem.org/mediawiki/2016/e/e4/T--NYMU-Taipei--photo-OFF-related-14627744_120300000676522237_1703903936_n.jpg" width="100%" /></div>
 +
</div>
 +
 
 +
<p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology', 11); return false">[Collapse]</a></p>
 +
</div>
 +
 
 +
<div class="technology">9/22</div>
 +
<div class="thelanguage">
 +
<li><p style="font-size: 16px;">For the situation that our fruit flies in the centrifuge tubes were dead quickly , so we visited Dr. Yu-Bing Huang in Taiwan Agricultural Research Institute, Applied Zoology Division, and asking the problems about the right way for feeding oriental fruit flies. We change our way of feeding oriental fruit flies. </p></li>
 +
 
 +
<p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology', 12); return false">[Collapse]</a></p>
 +
</div>
 +
 
 +
<div class="technology">9/23</div>
 +
<div class="thelanguage">
 +
<li><p style="font-size: 16px;">Producing new feeding box which is larger and ventilation, and design the experimental boxes for Experiment 2: Infecting B. dorsalis with non-designed M. anisopliae.</p></li>
 +
 
 +
<div class="imageimage">
 +
<div style="position:relative; width:48%;">
 +
<img src="https://static.igem.org/mediawiki/2016/9/91/T--NYMU-Taipei--photo-OFF-related-14697029_120300000683154255_1538186769_n.jpg" width="100%" /></div>
 +
<div style="position:relative; width:48%;">
 +
<img src="https://static.igem.org/mediawiki/2016/3/30/T--NYMU-Taipei--photo-OFF-related-14699699_120300000675475754_434414381_n.jpg" width="100%" /></div>
 +
</div>
 +
 
 +
<p style="font-size: 16px;">(Left picture: experimental boxes; Right picture: the internal space of feeding box)</p>
 +
 
 +
<p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology', 13); return false">[Collapse]</a></p>
 +
</div>
 +
 
 +
<div class="technology">9/30</div>
 +
<div class="thelanguage">
 +
<li><p style="font-size: 16px;">The new oriental fruit flies broke through cocoon in the new feeding box</p></li>
 +
 
 +
<p style="font-size: 18px;">the new way for feeding oriental fruit flies is using gauze to be the windows that could be ventilation, and we can put watered agar and artificial feed on the top of gauze, then there were no water and feed in the box which lead to the environment being mess, and they also could have enough water and feed.</p>
 +
<br />
 +
<li><p style="font-size: 16px;">Infecting B. dorsalis with non-designed M. anisopliae started to conduct.</p></li>
 +
 
 +
<div class="imageimage">
 +
<div style="position:relative; width:48%;">
 +
<img src="https://static.igem.org/mediawiki/2016/c/c5/T--NYMU-Taipei--photo-OFF-related-14696875_120300000677699020_204079543_n.jpg" width="100%" /></div>
 +
<div style="position:relative; width:48%;">
 +
<img src="https://static.igem.org/mediawiki/2016/4/46/T--NYMU-Taipei--photo-OFF-related-14696862_120300000674829370_219600743_n.jpg" width="100%" /></div>
 +
</div>
 +
<br />
 +
<div class="imageimage">
 +
<div style="position:relative; width:48%;">
 +
<img src="https://static.igem.org/mediawiki/2016/b/b9/T--NYMU-Taipei--photo-OFF-related-14686253_120300000675039507_1525612271_n.jpg" width="100%" /></div>
 +
<div style="position:relative; width:48%;">
 +
<img src="https://static.igem.org/mediawiki/2016/c/c7/T--NYMU-Taipei--photo-OFF-related-14627961_120300000679852339_2089167734_n.jpg" width="100%" /></div>
 +
</div>
 +
 
 +
<p style="font-size: 16px;">Photos shows three concentration condition and negative control environment of experiment "Infecting B. dorsalis with non-designed M. anisopliae"</p>
 +
 
 +
<p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology', 14); return false">[Collapse]</a></p>
 +
</div>
 +
 
 +
<div class="technology">10/1</div>
 +
<div class="thelanguage">
 +
<li><p style="font-size: 16px;">Keeping on Experiment 2-1 (9/30~10/7)</p></li>
 +
 
 +
<p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology', 15); return false">[Collapse]</a></p>
 +
</div>
 +
 
 +
<div class="technology">10/7</div>
 +
<div class="thelanguage">
 +
<li><p style="font-size: 16px;">Experiment 2-1 finished.</p></li>
 +
 
 +
<p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology', 16); return false">[Collapse]</a></p>
 +
</div>
 +
 
 +
<div class="technology">10/10</div>
 +
<div class="thelanguage">
 +
<p style="font-size: 16px;">Experiment 2-2 "infecting female B. dorsalis by copulation with infected male B. dorsalis" started to conduct.</p>
 +
 
 +
<p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology', 17); return false">[Collapse]</a></p>
 +
</div>
 +
 
 +
<div id="wrap">
 +
 
 +
<div class="fund" style="white-space: pre-wrap;">
 +
 
 +
<h2 style="margin-top:30px; margin-bottom:10px;">Q&A Section</h2><hr />
 +
 
 +
<p style="font-size:16px;">How to CULTURE oriental fruit flies?</p>
 +
<p style="font-size:16px;"> The oriental fruit flies can grow in a transparent large container and centrifuge tubes with a piece of cotton placed at the opening to prevent them from escaping while still allowing air exchange.</p>
 +
<p style="font-size:16px;"> Feeding on sucrose-peptone agar (sited by ATCC) when you choose the centrifuge tubes as their living environment; a ratio of composition below is the way to feed the fruit flies in a larger container. The larval need the other formula to grow up whether living in the centrifuge tubes or the larger transparent container. The adult feeding formula without peptone can also alive the oriental fruit flies but not breeding.</p>
 +
<p style="font-size:16px;">                Larva formula                                            Adult formula ratio
 +
Yeast extraction 35g                      Sucrose: yeast extraction: peptone
 +
Sucrose 60g                      3:1:1
 +
Oat                 120g
 +
Sodium benzoate 1g
 +
Hydrochloric acid 4.5ml, 37%
 +
Water 450ml
 +
</p>
 +
<p style="font-size:16px;"> The advantage of cultivating in large container is avoiding the tight space that oriental fruit flies can almost only walk and then causing the growth medium spoil easily. On the other hand, cultivating the fruit flies in the centrifuge tubes can easily distinguish the situation of the oriental fruit flies, for example, gender, health, quantity to take the correct actions.</p>
 +
<p style="font-size:16px;">How to prevent COMTAMINATION?</p>
 +
<li><p style="font-size:16px;"></p>Adding preservatives, for instance, sodium benzoate, and decreasing environmental humidity can decease the growth of the mold on agar.</li>
 +
<li><p style="font-size:16px;"></p>Sterilize cotton according common lab procedure or just buy pre-sterilized one.</li><br />
 +
<p style="font-size:16px;">How do you KEEP the oriental fruit flies in place during experiments?</p>
 +
<li><p style="font-size:16px;"></p>Using carbon dioxide gas to induce comatose and try your best to contain the gas within the tube and the oriental fruit flies should be unconscious after 10-20 seconds.</li>
 +
<li><p style="font-size:16px;"></p>Induce coma under low temperature.</li><br />
 +
<p style="font-size:16px;">How about the methyl eugenol? Are there any suggestions on attracting the male oriental fruit flies or the actual situation that farmer using it to prevent the oriental fruit fly pest?</p>
 +
<li><p style="font-size:16px;">It is best to complete experiments using methyl eugenol before 2:00 PM.</p></li>
 +
<li><p style="font-size:16px;">Several time experiments may cause the male oriental fruit flies became desensitized to methyl eugenol, so it's important to change the sample you test on the experiments routinely to avoid the factor to interfere the result.</p></li>
 +
<li><p style="font-size:16px;">The attraction traps putting in the orchard need to add pure and undiluted methyl eugenol to capture the oriental fruit flies. It also needs some cottons or gauzes to separate the oriental fruit flies and methyl eugenol to prevent the chain of fruit flies and eventually causing the death.</p></li>
 +
<li><p style="font-size:16px;">It's not a good idea to set the trap when (a) it's a rainy day (b) it's not the harvest season of the guava or tangerine, or the number of the fruit flies captured will not high.</p></li><br />
 +
<p style="font-size:16px;">Is the common characteristic among the hemolymph of Bactrocera dorsalis and other insects sufficiently confirmed the accuracy of the experiment result?</p><br />
 +
<p style="font-size:16px;">Can the designed M. anisopliae actually invade B. dorsalis?</p><br />
 +
 
 +
</div>
 +
 
 +
<div class="fund" style="white-space: pre;">
 +
 
 +
<h2 style="margin-top:30px; margin-bottom:10px;">Acknowledgements</h2><hr />
 +
 
 +
<p style="font-size:16px;">Beitou orchard
 +
S. J. Chen Grower
 +
</p>
 +
<p style="font-size:16px;">Beitou orchard
 +
Z. L. Liao Grower
 +
</p>
 +
<p style="font-size:16px;">Bioresource Collection and Research Center
 +
G. Y. Liou Fellow
 +
</p>
 +
<p style="font-size:16px;">Chiayi Agricultural Experiment Branch
 +
P. H. Chen Research assistant
 +
</p>
 +
<p style="font-size:16px;">Kaohsiung District Agricultural Research and Extension Station, COA, EY
 +
M. N. Tseng Research director
 +
</p>
 +
<p style="font-size:16px;">National Chung Hsing University, the Department of Entomology
 +
S. Y. Hwang Professor
 +
</p>
 +
<p style="font-size:16px;">National Chung Hsing University, the Department of Entomology
 +
Y. Y. Tseng Research assistant
 +
</p>
 +
<p style="font-size:16px;">Taiwan Agricultural Research Institute, Applied Zoology Division
 +
M. Y. Chiang Research assistant
 +
</p>
 +
<p style="font-size:16px;">Taiwan Agricultural Research Institute, Applied Zoology Division
 +
Y. B. Huang Professor
 +
/p>
 +
 
 +
</div>
 +
</div>
 +
 
 +
<script>if (window.runOnloadHook) runOnloadHook();</script></div>
 +
 
  
 
</body>
 
</body>
 
</html>
 
</html>

Revision as of 08:53, 19 October 2016

Lab note

Lab note


Overview

This section of our wiki contains the protocols for our insect experiments, including cultivation, infection rate tests, and hemolymph extraction.

Related experiment schedule & Lab note


Experiments

　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　
Experiments related to B. dorsalisNumberDate
Diameter of the tunnel that allows passage to one B. dorsalis18/30～9/1
Infecting B. dorsalis with non-designed M. anisopliae2-19/30~10/7
infecting female B. dorsalis by copulation with infected male B. dorsalis2-210/10~10/17
　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　
Experiments related to B. dorsalisNumberDate
Diameter of the tunnel that allows passage to one B. dorsalis18/30～9/1
Infecting B. dorsalis with non-designed M. anisopliae2-19/30~10/7
infecting female B. dorsalis by copulation with infected male B. dorsalis2-210/10~10/17
　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　
Experiments about hemolymph extractionNumberDate
Blatta lateralis19/20~9/30
Bactro29/29~10/5
Bombyx mori310/3~10/
　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　
Experiments about hemolymph extractionNumberDate
Blatta lateralis19/20~9/30
Bactro29/29~10/5
Bombyx mori310/3~10/

Routine works

　 　 　 　 　 　 　 　 　 　 　 　
Routine tasks of feeding Bactrocera dorsalisDate
Produce & change the tube with the specific agar8/24~9/20
Produce & change water gel and the specific ratio fodder9/22~10/
　 　 　 　 　 　 　 　 　 　 　 　
Routine tasks of feeding Bactrocera dorsalisDate
Produce & change the tube with the specific agar8/24~9/20
Produce & change water gel and the specific ratio fodder9/22~10/
　 　 　 　 　 　 　 　 　 　 　 　
Routine tasks of hemolymph extractionDate
Collect the mulberry leaves to feed the silkworm10/3~10/
Sterilize the related equipments9/20~10/
　 　 　 　 　 　 　 　 　 　 　 　
Routine tasks of hemolymph extractionDate
Collect the mulberry leaves to feed the silkworm10/3~10/
Sterilize the related equipments9/20~10/


[Collapse all] | [Expand all]

8/23

  • Oriental fruit flies sent to lab and cultivated by large transparent box to avoid oriental fruit flies flying out.

    • [Collapse]

    8/24

  • We used both of centrifuge tube and feeding box to feed oriental fruit flies for comparing and ensuring the status of oriental fruit flies that established the independent space for observing oriental fruit flies.


    • [Collapse]

    8/25

  • B. dorsalis started to break through the cocoon. We conducted to put B. dorsalis to centrifuge tubes, avoiding the feeding box cannot load all oriental fruit flies. We poured the special agar to centrifuge tubes, and put centrifuges into entrance to catch oriental fruit flies. Then we sealed the centrifuge tubes by cotton.

    • (The team members put agar into centrifuge tubes which would be their feed.)

    (The left picture is that oriental fruit flies broke through cocoon; the right picture is the feeding box & centrifuge tubes where fed oriental fruit flies.)

    [Collapse]

    8/30

  • First time to Conducting Experiment 1

    • [Collapse]

    9/3

  • Changing the centrifuge tubes where oriental fruit flies lived. (Avoid the environment being mess.) /p>

    • [Collapse]

    9/5

  • Changing the centrifuge tubes where oriental fruit flies lived.

    • [Collapse]

    9/6

  • The oriental fruit flies in centrifuge tubes are dead quickly, so we conducted trouble shooting and sent the mail to professors who research oriental fruit flies.

  • We took the trap to Beitou orchard to try to catch oriental fruit flies.

    • [Collapse]

    9/7

  • Second time to Conducting Experiment 1.

  • Conducting trouble shooting.

    • [Collapse]

    9/10

  • Searching the store selling the silkworm for extracting hemolymph which is used to induce pMcl1.

    • [Collapse]

    9/13

  • Visiting two farmers working in the orchard to ask the information about oriental fruit flies in the nature environment, and installing the trap of catching oriental fruit flies.

    • (Left picture: the team member install the trap; Right picture: the farmer and our team member took a picture)

  • Sent the mail to researchers to ask for making public the content of the mail.

    • [Collapse]

    9/15

  • Communicating with Chang Ming ecological education sericulture, asking the information of buying silkworm.

    • [Collapse]

    9/16

  • Picking up the oriental fruit fly larval from rotten guavas and feeding them.

  • Left picture: the team members were picking up the oriental fruit fly larval from rotten guavas; Right picture: the larval have picked up and put into another box. Expected becoming pupae after six days.

    • [Collapse]

    9/22

  • For the situation that our fruit flies in the centrifuge tubes were dead quickly , so we visited Dr. Yu-Bing Huang in Taiwan Agricultural Research Institute, Applied Zoology Division, and asking the problems about the right way for feeding oriental fruit flies. We change our way of feeding oriental fruit flies.

    • [Collapse]

    9/23

  • Producing new feeding box which is larger and ventilation, and design the experimental boxes for Experiment 2: Infecting B. dorsalis with non-designed M. anisopliae.

    • (Left picture: experimental boxes; Right picture: the internal space of feeding box)

    [Collapse]

    9/30

  • The new oriental fruit flies broke through cocoon in the new feeding box

    • the new way for feeding oriental fruit flies is using gauze to be the windows that could be ventilation, and we can put watered agar and artificial feed on the top of gauze, then there were no water and feed in the box which lead to the environment being mess, and they also could have enough water and feed.


  • Infecting B. dorsalis with non-designed M. anisopliae started to conduct.


    • Photos shows three concentration condition and negative control environment of experiment "Infecting B. dorsalis with non-designed M. anisopliae"

    [Collapse]

    10/1

  • Keeping on Experiment 2-1 (9/30~10/7)

    • [Collapse]

    10/7

  • Experiment 2-1 finished.

    • [Collapse]

    10/10

    Experiment 2-2 "infecting female B. dorsalis by copulation with infected male B. dorsalis" started to conduct.

    [Collapse]

    Q&A Section

    How to CULTURE oriental fruit flies?

    The oriental fruit flies can grow in a transparent large container and centrifuge tubes with a piece of cotton placed at the opening to prevent them from escaping while still allowing air exchange.

    Feeding on sucrose-peptone agar (sited by ATCC) when you choose the centrifuge tubes as their living environment; a ratio of composition below is the way to feed the fruit flies in a larger container. The larval need the other formula to grow up whether living in the centrifuge tubes or the larger transparent container. The adult feeding formula without peptone can also alive the oriental fruit flies but not breeding.

    Larva formula Adult formula ratio Yeast extraction 35g Sucrose: yeast extraction: peptone Sucrose 60g 3:1:1 Oat 120g Sodium benzoate 1g Hydrochloric acid 4.5ml, 37% Water 450ml

    The advantage of cultivating in large container is avoiding the tight space that oriental fruit flies can almost only walk and then causing the growth medium spoil easily. On the other hand, cultivating the fruit flies in the centrifuge tubes can easily distinguish the situation of the oriental fruit flies, for example, gender, health, quantity to take the correct actions.

    How to prevent COMTAMINATION?

  • Adding preservatives, for instance, sodium benzoate, and decreasing environmental humidity can decease the growth of the mold on agar.

  • Sterilize cotton according common lab procedure or just buy pre-sterilized one.

    • How do you KEEP the oriental fruit flies in place during experiments?

  • Using carbon dioxide gas to induce comatose and try your best to contain the gas within the tube and the oriental fruit flies should be unconscious after 10-20 seconds.

  • Induce coma under low temperature.

    • How about the methyl eugenol? Are there any suggestions on attracting the male oriental fruit flies or the actual situation that farmer using it to prevent the oriental fruit fly pest?

  • It is best to complete experiments using methyl eugenol before 2:00 PM.

  • Several time experiments may cause the male oriental fruit flies became desensitized to methyl eugenol, so it's important to change the sample you test on the experiments routinely to avoid the factor to interfere the result.

  • The attraction traps putting in the orchard need to add pure and undiluted methyl eugenol to capture the oriental fruit flies. It also needs some cottons or gauzes to separate the oriental fruit flies and methyl eugenol to prevent the chain of fruit flies and eventually causing the death.

  • It's not a good idea to set the trap when (a) it's a rainy day (b) it's not the harvest season of the guava or tangerine, or the number of the fruit flies captured will not high.


    • Is the common characteristic among the hemolymph of Bactrocera dorsalis and other insects sufficiently confirmed the accuracy of the experiment result?


    Can the designed M. anisopliae actually invade B. dorsalis?


    Acknowledgements

    Beitou orchard S. J. Chen Grower

    Beitou orchard Z. L. Liao Grower

    Bioresource Collection and Research Center G. Y. Liou Fellow

    Chiayi Agricultural Experiment Branch P. H. Chen Research assistant

    Kaohsiung District Agricultural Research and Extension Station, COA, EY M. N. Tseng Research director

    National Chung Hsing University, the Department of Entomology S. Y. Hwang Professor

    National Chung Hsing University, the Department of Entomology Y. Y. Tseng Research assistant

    Taiwan Agricultural Research Institute, Applied Zoology Division M. Y. Chiang Research assistant

    Taiwan Agricultural Research Institute, Applied Zoology Division Y. B. Huang Professor /p>

    Overview

    This section of our wiki contains the protocols for our insect experiments, including cultivation, infection rate tests, and hemolymph extraction.

    Related experiment schedule & Lab note


    Experiments

    　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　
    Experiments related to B. dorsalisNumberDate
    Diameter of the tunnel that allows passage to one B. dorsalis18/30～9/1
    Infecting B. dorsalis with non-designed M. anisopliae2-19/30~10/7
    infecting female B. dorsalis by copulation with infected male B. dorsalis2-210/10~10/17
    　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　
    Experiments related to B. dorsalisNumberDate
    Diameter of the tunnel that allows passage to one B. dorsalis18/30～9/1
    Infecting B. dorsalis with non-designed M. anisopliae2-19/30~10/7
    infecting female B. dorsalis by copulation with infected male B. dorsalis2-210/10~10/17
    　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　
    Experiments about hemolymph extractionNumberDate
    Blatta lateralis19/20~9/30
    Bactro29/29~10/5
    Bombyx mori310/3~10/
    　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　 　
    Experiments about hemolymph extractionNumberDate
    Blatta lateralis19/20~9/30
    Bactro29/29~10/5
    Bombyx mori310/3~10/

    Routine works

    　 　 　 　 　 　 　 　 　 　 　 　
    Routine tasks of feeding Bactrocera dorsalisDate
    Produce & change the tube with the specific agar8/24~9/20
    Produce & change water gel and the specific ratio fodder9/22~10/
    　 　 　 　 　 　 　 　 　 　 　 　
    Routine tasks of feeding Bactrocera dorsalisDate
    Produce & change the tube with the specific agar8/24~9/20
    Produce & change water gel and the specific ratio fodder9/22~10/
    　 　 　 　 　 　 　 　 　 　 　 　
    Routine tasks of hemolymph extractionDate
    Collect the mulberry leaves to feed the silkworm10/3~10/
    Sterilize the related equipments9/20~10/
    　 　 　 　 　 　 　 　 　 　 　 　
    Routine tasks of hemolymph extractionDate
    Collect the mulberry leaves to feed the silkworm10/3~10/
    Sterilize the related equipments9/20~10/


    [Collapse all] | [Expand all]

    8/23

  • Oriental fruit flies sent to lab and cultivated by large transparent box to avoid oriental fruit flies flying out.

    • [Collapse]

    8/24

  • We used both of centrifuge tube and feeding box to feed oriental fruit flies for comparing and ensuring the status of oriental fruit flies that established the independent space for observing oriental fruit flies.


    • [Collapse]

    8/25

  • B. dorsalis started to break through the cocoon. We conducted to put B. dorsalis to centrifuge tubes, avoiding the feeding box cannot load all oriental fruit flies. We poured the special agar to centrifuge tubes, and put centrifuges into entrance to catch oriental fruit flies. Then we sealed the centrifuge tubes by cotton.

    • (The team members put agar into centrifuge tubes which would be their feed.)

    (The left picture is that oriental fruit flies broke through cocoon; the right picture is the feeding box & centrifuge tubes where fed oriental fruit flies.)

    [Collapse]

    8/30

  • First time to Conducting Experiment 1

    • [Collapse]

    9/3

  • Changing the centrifuge tubes where oriental fruit flies lived. (Avoid the environment being mess.) /p>

    • [Collapse]

    9/5

  • Changing the centrifuge tubes where oriental fruit flies lived.

    • [Collapse]

    9/6

  • The oriental fruit flies in centrifuge tubes are dead quickly, so we conducted trouble shooting and sent the mail to professors who research oriental fruit flies.

  • We took the trap to Beitou orchard to try to catch oriental fruit flies.

    • [Collapse]

    9/7

  • Second time to Conducting Experiment 1.

  • Conducting trouble shooting.

    • [Collapse]

    9/10

  • Searching the store selling the silkworm for extracting hemolymph which is used to induce pMcl1.

    • [Collapse]

    9/13

  • Visiting two farmers working in the orchard to ask the information about oriental fruit flies in the nature environment, and installing the trap of catching oriental fruit flies.

    • (Left picture: the team member install the trap; Right picture: the farmer and our team member took a picture)

  • Sent the mail to researchers to ask for making public the content of the mail.

    • [Collapse]

    9/15

  • Communicating with Chang Ming ecological education sericulture, asking the information of buying silkworm.

    • [Collapse]

    9/16

  • Picking up the oriental fruit fly larval from rotten guavas and feeding them.

  • Left picture: the team members were picking up the oriental fruit fly larval from rotten guavas; Right picture: the larval have picked up and put into another box. Expected becoming pupae after six days.

    • [Collapse]

    9/22

  • For the situation that our fruit flies in the centrifuge tubes were dead quickly , so we visited Dr. Yu-Bing Huang in Taiwan Agricultural Research Institute, Applied Zoology Division, and asking the problems about the right way for feeding oriental fruit flies. We change our way of feeding oriental fruit flies.

    • [Collapse]

    9/23

  • Producing new feeding box which is larger and ventilation, and design the experimental boxes for Experiment 2: Infecting B. dorsalis with non-designed M. anisopliae.

    • (Left picture: experimental boxes; Right picture: the internal space of feeding box)

    [Collapse]

    9/30

  • The new oriental fruit flies broke through cocoon in the new feeding box

    • the new way for feeding oriental fruit flies is using gauze to be the windows that could be ventilation, and we can put watered agar and artificial feed on the top of gauze, then there were no water and feed in the box which lead to the environment being mess, and they also could have enough water and feed.


  • Infecting B. dorsalis with non-designed M. anisopliae started to conduct.


    • Photos shows three concentration condition and negative control environment of experiment "Infecting B. dorsalis with non-designed M. anisopliae"

    [Collapse]

    10/1

  • Keeping on Experiment 2-1 (9/30~10/7)

    • [Collapse]

    10/7

  • Experiment 2-1 finished.

    • [Collapse]

    10/10

    Experiment 2-2 "infecting female B. dorsalis by copulation with infected male B. dorsalis" started to conduct.

    [Collapse]

    Q&A Section

    How to CULTURE oriental fruit flies?

    The oriental fruit flies can grow in a transparent large container and centrifuge tubes with a piece of cotton placed at the opening to prevent them from escaping while still allowing air exchange.

    Feeding on sucrose-peptone agar (sited by ATCC) when you choose the centrifuge tubes as their living environment; a ratio of composition below is the way to feed the fruit flies in a larger container. The larval need the other formula to grow up whether living in the centrifuge tubes or the larger transparent container. The adult feeding formula without peptone can also alive the oriental fruit flies but not breeding.

    Larva formula Adult formula ratio Yeast extraction 35g Sucrose: yeast extraction: peptone Sucrose 60g 3:1:1 Oat 120g Sodium benzoate 1g Hydrochloric acid 4.5ml, 37% Water 450ml

    The advantage of cultivating in large container is avoiding the tight space that oriental fruit flies can almost only walk and then causing the growth medium spoil easily. On the other hand, cultivating the fruit flies in the centrifuge tubes can easily distinguish the situation of the oriental fruit flies, for example, gender, health, quantity to take the correct actions.

    How to prevent COMTAMINATION?

  • Adding preservatives, for instance, sodium benzoate, and decreasing environmental humidity can decease the growth of the mold on agar.

  • Sterilize cotton according common lab procedure or just buy pre-sterilized one.

    • How do you KEEP the oriental fruit flies in place during experiments?

  • Using carbon dioxide gas to induce comatose and try your best to contain the gas within the tube and the oriental fruit flies should be unconscious after 10-20 seconds.

  • Induce coma under low temperature.

    • How about the methyl eugenol? Are there any suggestions on attracting the male oriental fruit flies or the actual situation that farmer using it to prevent the oriental fruit fly pest?

  • It is best to complete experiments using methyl eugenol before 2:00 PM.

  • Several time experiments may cause the male oriental fruit flies became desensitized to methyl eugenol, so it's important to change the sample you test on the experiments routinely to avoid the factor to interfere the result.

  • The attraction traps putting in the orchard need to add pure and undiluted methyl eugenol to capture the oriental fruit flies. It also needs some cottons or gauzes to separate the oriental fruit flies and methyl eugenol to prevent the chain of fruit flies and eventually causing the death.

  • It's not a good idea to set the trap when (a) it's a rainy day (b) it's not the harvest season of the guava or tangerine, or the number of the fruit flies captured will not high.


    • Is the common characteristic among the hemolymph of Bactrocera dorsalis and other insects sufficiently confirmed the accuracy of the experiment result?


    Can the designed M. anisopliae actually invade B. dorsalis?


    Acknowledgements

    Beitou orchard S. J. Chen Grower

    Beitou orchard Z. L. Liao Grower

    Bioresource Collection and Research Center G. Y. Liou Fellow

    Chiayi Agricultural Experiment Branch P. H. Chen Research assistant

    Kaohsiung District Agricultural Research and Extension Station, COA, EY M. N. Tseng Research director

    National Chung Hsing University, the Department of Entomology S. Y. Hwang Professor

    National Chung Hsing University, the Department of Entomology Y. Y. Tseng Research assistant

    Taiwan Agricultural Research Institute, Applied Zoology Division M. Y. Chiang Research assistant

    Taiwan Agricultural Research Institute, Applied Zoology Division Y. B. Huang Professor /p>