The algae arrived today! It came in to glass vials filled with agar, and it looked like streaks of green smeared on top. In the morning, I finished sterilizing about 20 L of tap water by boiling it in a kettle. I poured 400 mL into each of the 21 half-liter bottles.
For the experimental set-up, I covered a tank with foil on all sides to provide the algae with ambient lighting. Matt spent much of the day helping me build a stand for the UV lights and wiring the lights. Thanks to Matt, I simply plug in a cord, and the algae will have the bright light they need for optimum growth.
I scraped some algae off from the agar and transferred it to two starter culture bottles with some growth medium in each.
I am also much indebted to Julie, who talked me through every step and walked over to Parsons to grind and weigh out the caffeine and ibuprofen pills. I used pipettes to measure out the needed volumes of DEET and antibacterial soap. All the chemicals were deposited into stock solutions, which will be transferred to the other 15 bottles with the algae once the algae have time to grow over the weekend.
Friday, August 6, 2010
Thursday, August 5, 2010
Day 19 - Got it!
For the past three days, I have been puzzling over finding consistent data to use for the concentrations of each PPCP. I've scoured Web of Science, visited sources cited in related papers, and searched for USGS and EPA reports, but I have not managed to find consistent numbers to average for each PPCP.
Today I decided to try emailing the Massachusetts Water Resources Authority and their research director to request data on PPCPs if they have it. I did not expect him to respond. To my surprise, no more than two hours after I sent the email, he sent me a whole spreadsheet with concentrations of PPCPs, including the four that I need. I was beyond relieved - I no longer needed to worry about being unable to find concentrations of PPCPs to use. I can now focus my experiment on data from Massachusetts wastewater, thereby making it better controlled.
I tried sterilizing tap water today. The kettle at Sea Grant takes about 20 minutes to boil water. And the water took even longer to cool enough to pour into algae bottles. I poured the water into a beaker first to measure out 400 mL, then transferred the warm water to the algae photobioreactors.
Algae will be here tomorrow!
Today I decided to try emailing the Massachusetts Water Resources Authority and their research director to request data on PPCPs if they have it. I did not expect him to respond. To my surprise, no more than two hours after I sent the email, he sent me a whole spreadsheet with concentrations of PPCPs, including the four that I need. I was beyond relieved - I no longer needed to worry about being unable to find concentrations of PPCPs to use. I can now focus my experiment on data from Massachusetts wastewater, thereby making it better controlled.
I tried sterilizing tap water today. The kettle at Sea Grant takes about 20 minutes to boil water. And the water took even longer to cool enough to pour into algae bottles. I poured the water into a beaker first to measure out 400 mL, then transferred the warm water to the algae photobioreactors.
Algae will be here tomorrow!
Day 17 - Lights, camera, action!
We finally finished the new control box for the water sampler video today. It's hard to believe that only a couple of weeks ago, I was fascinated by my own first control box, which I had gone through many struggles to set up. This time, the control box took only a day and a half, and the wiring and soldering seemed so much more straight-forward for me.
I stood in front of the camera, showing how to set up the solenoid valve and collecting canister. Under the pressure of the camera, I felt clumsier than usual, especially with little things such as ripping off electrical tape. We filmed mostly in one take, with a couple of pauses in the middle as I tried to attach the zip ties, but the canister kept slipping away! We'll have to do some minor editing to connect the various parts of the video before it is released for the whole world to see.
Thankfully, Matt helped me to clean out a back room for my research project. I have half of a countertop to myself in this cramped space, but I think it will work. Today I took the subway to Central Station and bought ibuprofen and antibacterial soap (triclosan) in as high concentrations as possible for my project. In the evening, I bought more gang valves and an extra air pump at Petco, 98.25% DEET insect repellent, and pure caffeine pills. I will have to figure out the dilutions for the ibuprofen, triclosan, DEET, and caffeine tomorrow.
I stood in front of the camera, showing how to set up the solenoid valve and collecting canister. Under the pressure of the camera, I felt clumsier than usual, especially with little things such as ripping off electrical tape. We filmed mostly in one take, with a couple of pauses in the middle as I tried to attach the zip ties, but the canister kept slipping away! We'll have to do some minor editing to connect the various parts of the video before it is released for the whole world to see.
Thankfully, Matt helped me to clean out a back room for my research project. I have half of a countertop to myself in this cramped space, but I think it will work. Today I took the subway to Central Station and bought ibuprofen and antibacterial soap (triclosan) in as high concentrations as possible for my project. In the evening, I bought more gang valves and an extra air pump at Petco, 98.25% DEET insect repellent, and pure caffeine pills. I will have to figure out the dilutions for the ibuprofen, triclosan, DEET, and caffeine tomorrow.
Wednesday, August 4, 2010
Day 18 - Math, math, math
This morning we finished filming instructions for how to wire the water sampler. We had to stop in the middle, because the camera battery kept running out, so we waited for it to charge. I felt like I was on a DIY or cooking show, giving instructions on how to make a certain dish or decorate the house in a certain way. The biggest difference is that the Sea Perch Channel is much more useful.
Sarah let me borrow fifteen 500 mL bottles to use as photobioreactors for the algae. I labeled and washed them, then set them up in rows in a large plastic bin. There will be 3 trials for each chemical, as well as for the control.
I am trying to find more data on concentrations of PPCPs in U.S. wastewater, but I'm not having much luck. I've seen data from Germany, China, Spain, and other countries, but only one or two reports from U.S. wastewater treatment plants. Many articles also show graphs of the removal rates of certain compounds after treatment, but the exact concentrations in micrograms/L are unclear. Julie says I may have to directly contact a few of the authors, asking them for the raw data. My goal is to make the experiment as relevant and true to real life situations as possible, so I need to have data from a variety of wastewater sites across the U.S. Ideally, once I average the concentrations, I can calculate how much of each compound to put in the algae.
Julie helped me to think through how to dilute the compounds I have into the desired concentrations of migrograms/L. It is quite confusing, because we have to make stock solutions to add to the bottles and therefore calculate how much of each compound to add to the stock solutions. Also, we took a long time to research the density of the antibacterial soap so that we could convert a certain volume into mass. Luckily, the density was almost equal to that of water. I hope we have scales precise enough to measure out fractions of milligrams!
The algae is supposed to arrive on Friday. To-do for Thursday:
Sarah let me borrow fifteen 500 mL bottles to use as photobioreactors for the algae. I labeled and washed them, then set them up in rows in a large plastic bin. There will be 3 trials for each chemical, as well as for the control.
I am trying to find more data on concentrations of PPCPs in U.S. wastewater, but I'm not having much luck. I've seen data from Germany, China, Spain, and other countries, but only one or two reports from U.S. wastewater treatment plants. Many articles also show graphs of the removal rates of certain compounds after treatment, but the exact concentrations in micrograms/L are unclear. Julie says I may have to directly contact a few of the authors, asking them for the raw data. My goal is to make the experiment as relevant and true to real life situations as possible, so I need to have data from a variety of wastewater sites across the U.S. Ideally, once I average the concentrations, I can calculate how much of each compound to put in the algae.
Julie helped me to think through how to dilute the compounds I have into the desired concentrations of migrograms/L. It is quite confusing, because we have to make stock solutions to add to the bottles and therefore calculate how much of each compound to add to the stock solutions. Also, we took a long time to research the density of the antibacterial soap so that we could convert a certain volume into mass. Luckily, the density was almost equal to that of water. I hope we have scales precise enough to measure out fractions of milligrams!
The algae is supposed to arrive on Friday. To-do for Thursday:
- Find more data, especially for DEET, so that I can make accurate averages
- Sterilize water
- Set up grow lights
- Make dilutions
Monday, August 2, 2010
Day 16 - Videos and Project Work
Over the weekend, I bought supplies for my project: an air pump, gang valves, airstones, and tubing. I will set up photobioreactors (bottles) with algae growing in each, then add amounts of four PPCPs - caffeine, DEET, triclosan, and estrogen (or ibuprofen) - in each bottle. I am using Chlorella vulgaris, a hardy strain of algae commonly used for wastewater nutrient removal. The ultimate long-term goal of the project is to find out the extent to which wastewater needs to be purified before it can be used to grow algae for biodiesel. The short-term goal is to discover the effects of these four PPCPs on algae growth.
Last week I found journal articles with data from all over the world, but I could not find data targeted to the United States. Today Julie helped me to locate relevant data on PPCP levels in the United States, and I will consolidate the data into a database to determine the amount of each chemical I will choose to expose the algae to. By conducting journal searches systematically and patiently, we uncovered much data that I was not able to uncover last week by myself. I've discovered that organization is absolutely key to attaining any results in a scientific procedure, from planning to final conclusions.
Cole and I have already filmed a video on constructing the frame for the Sea Perch to go on the Sea Perch website (the old video was unclear). Jordan wants us to make another set of videos to go along with the online instructions for making a water sampler. This entails constructing a new control box to use in the video and attach to the water sampler. We've begun working on the new control box and will hopefully be finished by tomorrow, in time for filming.
Last week I found journal articles with data from all over the world, but I could not find data targeted to the United States. Today Julie helped me to locate relevant data on PPCP levels in the United States, and I will consolidate the data into a database to determine the amount of each chemical I will choose to expose the algae to. By conducting journal searches systematically and patiently, we uncovered much data that I was not able to uncover last week by myself. I've discovered that organization is absolutely key to attaining any results in a scientific procedure, from planning to final conclusions.
Cole and I have already filmed a video on constructing the frame for the Sea Perch to go on the Sea Perch website (the old video was unclear). Jordan wants us to make another set of videos to go along with the online instructions for making a water sampler. This entails constructing a new control box to use in the video and attach to the water sampler. We've begun working on the new control box and will hopefully be finished by tomorrow, in time for filming.
Day 15 - Parsons Tour
Sometimes opportunities don't fall into place to you unless you go looking for them. I told Brandy a couple of days ago that I am interested in environmental engineering, and she immediately tried to arrange a tour of Parsons Lab for me and Cole. I am often astounded by Brandy's efficiency, organization, and persistence. Within a day's notice, Sheila of Parsons Lab agreed to meet us at 11:30 a.m. on Friday.
Sheila was very friendly and eager to acquaint us with the lab. She entered nearly every lab and bothered the researchers and grad students there to give us a brief description of their research. We got to witness cutting-edge research, from hydrodamics on a microscopic scale to extremophiles in a laboratory environment. Environmental engineering is a smaller department; as Sheila says, the environmental engineering department is like a family. The few undergrads therefore get research opportunities and positions that larger departments may not offer.
Several of the labs in Parsons have gas chromatographs, complicated pieces of equipment that I was hoping to use for measuring algae lipid profiles. However, without EHS training and a professor to supervise, I am unable to access the equipment over the summer. I wish I could learn to use all of the machines in Parsons, from the gas chromatographs to the fluorocytometers, which were made by Sheila herself. For now, I will have to conduct a simpler version of the project - measuring cell density and cell count instead of lipid profiles - without forgetting my dream of extending the scope of the project in the future.
Sheila was very friendly and eager to acquaint us with the lab. She entered nearly every lab and bothered the researchers and grad students there to give us a brief description of their research. We got to witness cutting-edge research, from hydrodamics on a microscopic scale to extremophiles in a laboratory environment. Environmental engineering is a smaller department; as Sheila says, the environmental engineering department is like a family. The few undergrads therefore get research opportunities and positions that larger departments may not offer.
Several of the labs in Parsons have gas chromatographs, complicated pieces of equipment that I was hoping to use for measuring algae lipid profiles. However, without EHS training and a professor to supervise, I am unable to access the equipment over the summer. I wish I could learn to use all of the machines in Parsons, from the gas chromatographs to the fluorocytometers, which were made by Sheila herself. For now, I will have to conduct a simpler version of the project - measuring cell density and cell count instead of lipid profiles - without forgetting my dream of extending the scope of the project in the future.
Day 14 - Research & Testing Water Samplers
Today I spent the morning in the Hayden Humanities Library doing research for my project, trying to find relevant articles on algae for biodiesel and PPCPs to read. The Hayden Library is one of my favorite spots on campus that I've seen so far. The 2nd floor reading room is unbelievably spacious with windows that stretch from the ground to the ceilings. Sunlight streams in and makes one eager to study or complete homework on one of the long wooden tables.
There have been few studies done on the effects of PPCPs on algae. I was trying to find average national concentrations of the four PPCPs I would like to test - triclosan, DEET, estrogen, and caffeine - in wastewater. I was able to access reports from various countries, but was having trouble finding data targeted towards the U.S.
In the afternoon, we took our Sea Perches over to the OETL and tested the water samplers. The OEX camp was in the process of finishing their wind turbine; the room was filled with the smell of burning styrofoam and the noises of drills and saws. For some reason, Kraken kept spinning in circles, heading helplessly towards the right. I was dreading the inevitable - having to completely redo that motor. Jordan took a look at the problem and immediately advised me to simply re-epoxy the propellor to the propellor shaft. It turns out that the propellor had only come loose; there was no need to worry about completely re-waxing another motor!
There have been few studies done on the effects of PPCPs on algae. I was trying to find average national concentrations of the four PPCPs I would like to test - triclosan, DEET, estrogen, and caffeine - in wastewater. I was able to access reports from various countries, but was having trouble finding data targeted towards the U.S.
In the afternoon, we took our Sea Perches over to the OETL and tested the water samplers. The OEX camp was in the process of finishing their wind turbine; the room was filled with the smell of burning styrofoam and the noises of drills and saws. For some reason, Kraken kept spinning in circles, heading helplessly towards the right. I was dreading the inevitable - having to completely redo that motor. Jordan took a look at the problem and immediately advised me to simply re-epoxy the propellor to the propellor shaft. It turns out that the propellor had only come loose; there was no need to worry about completely re-waxing another motor!
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