Difference between revisions of "Team:UCAS/Hardware"

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                    <a>TEAM</a>
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                    <a>Parts</a>
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                        <li><a href="https://2016.igem.org/Team:UCAS/Part_Collection">Part Collection</a></li>
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                    <a href="https://2016.igem.org/Team:UCAS/Safety">Safety</a>
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                <li><a href="https://2016.igem.org/Team:UCAS/Model">Model</a></li>
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                    <a>HUMAN PRACTICES</a>
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                    <a href="https://2016.igem.org/Team:UCAS/Attributions">Attributions</a>
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                    <h1>Hardware: Bio-pool</h1>
 +
                    <h2 id="js-data-hardware1" class="page-header">Introduction: Activated sludge process</h2>
 +
                    <p style="font-size:18px;">
 +
                        Activated sludge process is now a widely used aerobic biological waste water treatment method. It could remove organic pollutants from water, which can be consumed by microorganism. It was invented by Clark and Gage in the UK in 1912.
 +
                    </p>
 +
                    <p style="font-size:18px;">
 +
                        In the early 20th century, waste water treatment was still a headache for cities, especially for big cities like London. At that time, people happened to discover that some sludge sediment appeared in the stagnant water, and the water above became clear!
 +
                    </p>
 +
                    <p style="font-size:18px;">
 +
                        In fact, the sludge is a biological floc composed of bacteria and protozoa which feed on the organic pollutants, and could oxidize the carbonaceous and nitrogenous matter in the waste water when oxygen presents.
 +
                    </p>
 +
                    <p style="font-size:18px;">
 +
                        Nowadays this principal has developed into various mature industrial processes and bio-reactors. And the basic work-flow is shown in fig 1.
 +
                    </p>
 +
                    <p style="font-size:12px;font-weight:bold;">
 +
                        Reference: <a href="http://www.nesc.wvu.edu/pdf/WW/publications/pipline/PL_SP03.pdf ">Explaining the Activated Sludge Process. Pipeline. Spring 2003</a>
 +
                    </p>
 +
                    <div>
 +
                        <img style="width: 90%" src="https://static.igem.org/mediawiki/2016/d/d9/T--UCAS--hardware_fig_1.png" />
 +
                        <br />
 +
                        <strong style="text-align: center;color: red;">Fig. 1 A generalized, schematic diagram of an activated sludge process</strong>
 +
                        <br />
 +
                    </div>
 +
                    <p style="font-size:12px;width: 80%;font-style:oblique;font-weight:bold;">
 +
                        Originally uploaded by Mbeychok at <a href="https://en.wikipedia.org/wiki/Activated_sludge">here</a>.
 +
                    </p>
 +
                </div>
  
 +
                <div class="bs-docs-section">
 +
                    <h2 id="js-data-hardware2" class="page-header">Our Device</h2>
 +
                    <h3 id="js-data-hardware2-1" style="color:#777777;">Overview</h3>
 +
                    <p style="font-size:18px;">
 +
                        We designed and assembled a hardware(shown in fig 2&3 and the Hardware Operation Demonstration Video) imitating the biochemical pools in waste water treatment plants (WWTPs) at the ratio of 1:50, including handmade microporous tube aerators and flux control and monitor equipment for air and water, which could be used for the experiments of variable aerobic biological treatment methods in laboratory, and the working capacity tests for new types of engineered microorganism under the real conditions in WWTPs.
 +
                    </p>
 +
                    <p style="font-size:18px;">
 +
                        On top of the traditional biochemical pool construction, we adopted pitched V-shaped bottom and vertical water inlet, and added partitions with holes in the pool, which could improved the practical performance of the working organism.</p>
 +
                    <div>
 +
                        <img style="width: 90%" src="https://static.igem.org/mediawiki/2016/9/9e/T--UCAS--hardware_fig_2.jpg" />
 +
                        <br />
 +
                        <strong style="text-align: center;color: red;">Fig. 2 Our whole set of device</strong>
 +
                        <br />
 +
                    </div>
 +
                    <div>
 +
                        <img style="width: 90%" src="https://static.igem.org/mediawiki/2016/6/6a/T--UCAS--hardware_fig_3.jpg" />
 +
                        <br />
 +
                        <strong style="text-align: center;color: red;">Fig. 3 Bio-pool blueprint</strong>
 +
                        <br />
 +
                    </div>
 +
                    <video width="800" height="600" controls="controls">
 +
                        <source src="https://static.igem.org/mediawiki/2016/0/05/T--UCAS--Hardware_Operation_Demonstration_Video.mp4" type="video/mp4" />
 +
                    </video>
 +
                    <h3 id="js-data-hardware2-2" style="color:#777777;">Parts</h3>
 +
                    <b style="font-size:18px;">
 +
                        Micro-porous tube aerators
 +
                    </b>
 +
                    <p style="font-size:18px;">
 +
                        To ensure effective aeration, the aerator is required to provide small and homogeneous bulbs. So we made holes on steel and PVC pipe, and assemble them together. Thus we made micro-porous tube aerators by hand.(shown in fig 4&5)
 +
                    </p>
 +
                    <div>
 +
                        <img style="width: 90%" src="https://static.igem.org/mediawiki/2016/2/20/T--UCAS--hardware_fig_4.jpg" />
 +
                        <br />
 +
                        <strong style="text-align: center;color: red;">Fig. 4 Micro-porous tube aerator</strong>
 +
                        <br />
 +
                    </div>
 +
                    <div>
 +
                        <img style="width: 90%" src="https://static.igem.org/mediawiki/2016/0/02/T--UCAS--hardware_fig_5.JPG" />
 +
                        <br />
 +
                        <strong style="text-align: center;color: red;">Fig. 5 Holes on the steel and PVC pipe</strong>
 +
                        <br />
 +
                    </div>
 +
                    <p style="font-size:18px;">
 +
                        There are two types of tube aerators: the long ones are placed at the V-shaped grooves, while the short ones are placed at the sludge collecting grooves near the front and behind walls.(shown in fig 6&7)
 +
                    </p>
 +
                    <div>
 +
                        <img style="width: 90%" src="https://static.igem.org/mediawiki/2016/5/5d/T--UCAS--hardware_fig_6.JPG" />
 +
                        <br />
 +
                        <strong style="text-align: center;color: red;">Fig. 6 Two types of tube aerators</strong>
 +
                        <br />
 +
                    </div>
 +
                    <div>
 +
                        <img style="width: 90%" src="https://static.igem.org/mediawiki/2016/4/45/T--UCAS--hardware_fig_7.JPG" />
 +
                        <br />
 +
                        <strong style="text-align: center;color: red;">Fig. 7 The placement of aerators</strong>
 +
                        <br />
 +
                    </div>
 +
                    <b style="font-size:18px;">
 +
                        Pitched V-shaped bottom
 +
                    </b>
 +
                    <p style="font-size:18px;">
 +
                        To make sludge collection and removal more convenient and efficient, we constructed V-shaped and pitched bottoms with a groove to collect the sludge produced in the purifying process.(shown in fig 8)
 +
                    </p>
 +
                    <div>
 +
                        <img style="width: 90%" src="https://static.igem.org/mediawiki/2016/9/92/T--UCAS--hardware_fig_8.png" />
 +
                        <br />
 +
                        <strong style="text-align: center;color: red;">Fig. 8 Pitched V-shaped bottom</strong>
 +
                        <br />
 +
                    </div>
 +
                    <b style="font-size:18px;">
 +
                        Vertical water inlet
 +
                    </b>
 +
                    <p style="font-size:18px;">
 +
                        To realize equivalent water-in, we came up with the vertical water inlet, making from PVC pipe with small holes on both sides.(shown in fig 9)
 +
                    </p>
 +
                    <div>
 +
                        <img style="width: 90%" src="https://static.igem.org/mediawiki/2016/a/ac/T--UCAS--hardware_fig_9.jpg" />
 +
                        <br />
 +
                        <strong style="text-align: center;color: red;">Fig. 9 Vertical water inlet</strong>
 +
                        <br />
 +
                    </div>
 +
                    <h3 id="js-data-hardware2-2-3" style="color:#777777;">Blueprints of the Bio-pool</h3>
 +
                    <div>
 +
                        <img style="width: 90%" src="https://static.igem.org/mediawiki/2016/b/bc/T--UCAS--hardware_fig_10.png" />
 +
                        <br />
 +
                        <strong style="text-align: center;color: red;">Fig. 10 Three-view diagram of the bio-pool</strong>
 +
                        <br />
 +
                    </div>
 +
                    <div>
 +
                        <img style="width: 90%" src="https://static.igem.org/mediawiki/2016/0/0e/T--UCAS--hardware_fig_11.png" />
 +
                        <br />
 +
                        <strong style="text-align: center;color: red;">Fig. 11 Size of the bio-pool</strong>
 +
                        <br />
 +
                    </div>
 +
                    <h3 id="js-data-hardware2-2-4" style="color:#777777;">Test</h3>
 +
                    <p style="font-size:18px;">
 +
                        At first, we plan to make the engineered bacteria attach to some substrate materials like plastics. However, the model organism we choose to use, E. coli, cannot express bio-film protein naturally. We tested the attach property of the bacteria using materials provided by Nanjing-China, and the result is negative.
 +
                    </p>
 +
                    <p style="font-size:18px;">
 +
                        Thus, we chose Activated Sludge Method instead of Contact Oxidation Method which uses plastic substrate with working organism attached in the form of bio-film.
 +
                    </p>
 +
                    <p style="font-size:18px;">
 +
                        To test whether the engineered bacteria could grow and work in the bio-pool, we prepared 30L 1:10 diluted LB with 15g ≥10%tetracycline hydrochloride(for livestock and poultry raising) and 200mL saturated E. coli, added them into one bio-pool and started aeration as is shown in fig 12.
 +
                    </p>
 +
                    <div>
 +
                        <img style="width: 90%" src="https://static.igem.org/mediawiki/2016/2/2e/T--UCAS--hardware_fig_12.jpg" />
 +
                        <br />
 +
                        <strong style="text-align: center;color: red;">Fig. 12 Test (initial)</strong>
 +
                        <br />
 +
                    </div>
 +
                    <p style="font-size:18px;">
 +
                        After 40 hours, it could be seen observably that the liquid in the pool became unclear with plenty of bacteria suspended in the water.(shown in fig 13)
 +
                    </p>
 +
                    <div>
 +
                        <img style="width: 90%" src="https://static.igem.org/mediawiki/2016/3/3e/T--UCAS--hardware_fig_13.jpg" />
 +
                        <br />
 +
                        <strong style="text-align: center;color: red;">Fig. 13 Test (40h later)</strong>
 +
                        <br />
 +
                    </div>
 +
                    <p style="font-size:18px;">
 +
                        When it come to the simulating waste water used in the experiment, we received the recommendation from Nanjing-China. The standard formula of simulating waste water is as following:
 +
                    </p>
 +
                    <p style="font-size:18px;">
 +
                        Glucose 0.3 g, tryptone 0.1 g, yeast extract 0.01 g, CH3COONa 0.15 g, NaCl 0.05 g, MgSO4·7H2O 0.236 g, K2HPO4·3H2O 0.147 g, NH4Cl 0.18 g, DI water 1 L, 115℃ 30 min sterilization.
 +
                    </p>
 +
                    <p style="font-size:18px;">
 +
                        The total phosphorus(TP) of the simulating waste water is 20 mg/L since all of the phosphorus in the water comes from the K2HPO4·3H2O added. By changing the  amount of K2HPO4·3H2O, we can adjust the TP in the simulating waste water according to experimental demand.
 +
                    </p>
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                </div>
  
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                    <h2 id="js-data-hardware3" class="page-header">Acknowledgment</h2>
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                        We appreciate the help provided by Nanjing-China in our hardware test.
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                        More details at:<a href="https://2016.igem.org/Team:UCAS/Collaborations">UCAS: Collaborations</a> & <a href="https://2016.igem.org/Team:Nanjing-China/Collaborations">Nanjing-China: Collaborations</a>
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                            <a href="#js-data-description1">The mass production and abuse of antibiotics</a>
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                            <a href="#js-data-description2">The emerging problem of antibiotic residues</a>
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<p>iGEM is about making teams of students making synthetic biology projects. We encourage teams to work with parts and build biological devices in the lab. But we are inclusive and want all teams to work on many other types of problems in synbio. Robotic assembly, microfluidics, low cost equipment and measurement hardware are all areas ripe for innovation in synbio. </p>
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Teams who are interested in working with hardware as a side project are encouraged to apply for the hardware award.
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<h5>Inspiration</h5>
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<p>You can look at what other teams did to get some inspiration! <br />
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Here are a few examples:</p>
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<li><a href="https://2015.igem.org/Team:TU_Delft">2015 TU Delft  </a></li>
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<li><a href="https://2015.igem.org/Team:TU_Darmstadt">2015 TU Darmstadt</a></li>
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<li><a href="https://2015.igem.org/Team:Cambridge-JIC">2015 Cambridge JIC</a></li>
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Revision as of 20:56, 19 October 2016

Hardware

Hardware: Bio-pool

Introduction: Activated sludge process

Activated sludge process is now a widely used aerobic biological waste water treatment method. It could remove organic pollutants from water, which can be consumed by microorganism. It was invented by Clark and Gage in the UK in 1912.

In the early 20th century, waste water treatment was still a headache for cities, especially for big cities like London. At that time, people happened to discover that some sludge sediment appeared in the stagnant water, and the water above became clear!

In fact, the sludge is a biological floc composed of bacteria and protozoa which feed on the organic pollutants, and could oxidize the carbonaceous and nitrogenous matter in the waste water when oxygen presents.

Nowadays this principal has developed into various mature industrial processes and bio-reactors. And the basic work-flow is shown in fig 1.

Reference: Explaining the Activated Sludge Process. Pipeline. Spring 2003


Fig. 1 A generalized, schematic diagram of an activated sludge process

Originally uploaded by Mbeychok at here.

Our Device

Overview

We designed and assembled a hardware(shown in fig 2&3 and the Hardware Operation Demonstration Video) imitating the biochemical pools in waste water treatment plants (WWTPs) at the ratio of 1:50, including handmade microporous tube aerators and flux control and monitor equipment for air and water, which could be used for the experiments of variable aerobic biological treatment methods in laboratory, and the working capacity tests for new types of engineered microorganism under the real conditions in WWTPs.

On top of the traditional biochemical pool construction, we adopted pitched V-shaped bottom and vertical water inlet, and added partitions with holes in the pool, which could improved the practical performance of the working organism.


Fig. 2 Our whole set of device

Fig. 3 Bio-pool blueprint

Parts

Micro-porous tube aerators

To ensure effective aeration, the aerator is required to provide small and homogeneous bulbs. So we made holes on steel and PVC pipe, and assemble them together. Thus we made micro-porous tube aerators by hand.(shown in fig 4&5)


Fig. 4 Micro-porous tube aerator

Fig. 5 Holes on the steel and PVC pipe

There are two types of tube aerators: the long ones are placed at the V-shaped grooves, while the short ones are placed at the sludge collecting grooves near the front and behind walls.(shown in fig 6&7)


Fig. 6 Two types of tube aerators

Fig. 7 The placement of aerators
Pitched V-shaped bottom

To make sludge collection and removal more convenient and efficient, we constructed V-shaped and pitched bottoms with a groove to collect the sludge produced in the purifying process.(shown in fig 8)


Fig. 8 Pitched V-shaped bottom
Vertical water inlet

To realize equivalent water-in, we came up with the vertical water inlet, making from PVC pipe with small holes on both sides.(shown in fig 9)


Fig. 9 Vertical water inlet

Blueprints of the Bio-pool


Fig. 10 Three-view diagram of the bio-pool

Fig. 11 Size of the bio-pool

Test

At first, we plan to make the engineered bacteria attach to some substrate materials like plastics. However, the model organism we choose to use, E. coli, cannot express bio-film protein naturally. We tested the attach property of the bacteria using materials provided by Nanjing-China, and the result is negative.

Thus, we chose Activated Sludge Method instead of Contact Oxidation Method which uses plastic substrate with working organism attached in the form of bio-film.

To test whether the engineered bacteria could grow and work in the bio-pool, we prepared 30L 1:10 diluted LB with 15g ≥10%tetracycline hydrochloride(for livestock and poultry raising) and 200mL saturated E. coli, added them into one bio-pool and started aeration as is shown in fig 12.


Fig. 12 Test (initial)

After 40 hours, it could be seen observably that the liquid in the pool became unclear with plenty of bacteria suspended in the water.(shown in fig 13)


Fig. 13 Test (40h later)

When it come to the simulating waste water used in the experiment, we received the recommendation from Nanjing-China. The standard formula of simulating waste water is as following:

Glucose 0.3 g, tryptone 0.1 g, yeast extract 0.01 g, CH3COONa 0.15 g, NaCl 0.05 g, MgSO4·7H2O 0.236 g, K2HPO4·3H2O 0.147 g, NH4Cl 0.18 g, DI water 1 L, 115℃ 30 min sterilization.

The total phosphorus(TP) of the simulating waste water is 20 mg/L since all of the phosphorus in the water comes from the K2HPO4·3H2O added. By changing the amount of K2HPO4·3H2O, we can adjust the TP in the simulating waste water according to experimental demand.

Acknowledgment

We appreciate the help provided by Nanjing-China in our hardware test.

More details at:UCAS: Collaborations & Nanjing-China: Collaborations