3.3.3 Termites Experiments
Overview
We ZJU-CHINA 2015 are pursuing an effective and environmental-friendly way to manage termites. It will be unwarranted if we only engage in idle theorizing about termites behavior and only do the genetic engineering experiments. Thus we carry out a series of termites experiments in order to achieve our goals, which are:
- Our baits mixed with bacteria can attract the termites
- Worker termites carry the baits in their mouth and return to the nest, being alive
- Worker termites feed other termites in the nest through the process called trophallaxis
- The termites that receive food die because of the toxin in the baits
Bait attractiveness
To kill the termites, first of all, we must make sure that termites will eat our baits. In any case, we cannot pry open termites'mouth and stuff in our food. Thanks to our advisor Professor Mo, who is an authority on termites control, we are provided with fantastic baits for termites. In order to confirm its attractiveness, we put a pinch of the bait in the petri dish together with a pinch of normal pine wood powder and see how the termites are attracted and distributed in the dish.
Figure 1 Examination of infected D.saccharalis midgut by scanning electron microscopy.
Chosen compound toxicity verification
Figure 2 top and side view of the tcdA1 prepore (left) and pore (right) prntamer.
Before the gene manipulation and capsulation experiment begin, it is important to make sure that the toxins we chose are toxic enough to termites. Avermectin is naturally produced in Streptomyces Avermitilis. Toxic protein TcdA1, TcdB1, Plu0840 and Plu1537 are natural product of Photorhabdus luminescens TT01. Accordingly, we feed the termites with Streptomyces Avermitilis and Photorhabdus luminescens TT01 and note down the mortality rate and speed. Besides, other insects are tested under the same condition to detect the toxicity to these insects.
Cellulase and lysozyme activity measurement
It is vital for termites to keep alive when they are carrying the food back to their nest. If they die on the way, the toxin cannot be brought and what's more, the toxin may spread into the environment. Thus we capsulate the bacteria with cellulose. To ensure the cellulose can be carried by termites on the way and can be digested when ate, we measure the cellulose activity in different parts of termite's gut. Besides, lysozyme activity is measured to ensure that bacteria can be break in termite's gut and release the toxin.
Figure 2 top and side view of the tcdA1 prepore (left) and pore (right) prntamer.
Verification of the release delay effect due to the CNC capsulation
Figure 2 top and side view of the tcdA1 prepore (left) and pore (right) prntamer.
To ensure the termites will not die during the carrying stage, we design the CNC capsulation and expect a release delay effect. Thus, we feed the termites with capsulated Streptomyces Avermitilis and expect that the dying speed will be lower, which illustrate that the capsulation have the release delay effect.
Trophallaxis verification
Trophallaxis has been well proved existed in the Coptotermes formosanus Shiraki. However, we want a concise demonstration so that everyone can have a clear understanding of the trophallaxis process. Therefore, we conduct an experiment. Termites are fed with stained food. A group are dyed in blue and the other group are dyed in red. We mix termites from two groups and observe the color change. Besides, soldiers termites who can only get food by trophallaxis were also added in each group.
Figure 2 top and side view of the tcdA1 prepore (left) and pore (right) prntamer.