What a busy but incredible week this has been. I received a phone call from a fellow intern Sunday night. His project revolves around a specific type of algae that can be grown, undergo an oils extraction which can be used for bio-fuel. As fascinating as this sounds I know very little about this concept. He stated that the algae he was harvesting "died" and that the project was a failure and that he was going to trash the water and start over. I explained that there is no such thing as "failure" in science and asked him to meet me in the lab Monday morning to discuss what else this could mean. He agreed to meet and talk with me before trashing the water. As I stood in front of his tanks that morning I saw cloudy water with a light green hue, sediment at the bottom of the tank, and a lot of hard work that had been done. The brainstorming began. I started asking myself a few basic questions:
1. Why did the algae die? Something killed it or the growing conditions were not optimal
2. What is the sediment on the bottom of the tank? Put it on a slide and look under a microscope.
3. What would kill the algae? Most likely a microbe, collect it, stain it, identify it.
4. Was the original algal sample tested for microbial growth when it arrived? No
5. If a microbe attacked the algae where did it come from? The lab or where it was shipped from
6. What can we do to prevent this from happening again? Identify the cause and implement a procedure to restore the tank.
7. Can we restore the algal growth? If we can identify the cause of death and eliminate it, it is possible to restore the algal growth.
8. What can we learn from this? Without a proper protocol and constant monitoring, algae will die. Also there could be a benefit of a microbe that kills algae.
9. What are possible benefits of a microbe that eliminates algae? A natural algaecide for fish tanks.
Now I was ready to meet my friend. When he arrived my excitement at all the new possibilities was contagious. A new project had begun, but there was a lot of work to be done and only two of us. Lucky for us, a perfect storm entered (Jeremy) and the electron cloud was formed. Identifying the contaminate of the water was the first order of business. I collected water from the tank, the pure algal culture, and the bucket with the rest of the algal water that Tony brought back from the algal lab in Carlsbad, Ca. I placed each specimen in a centrifuge for 10 minutes at 16 RPM. I extracted the supernatant added more sample and centrifuged again. I repeated this two more times until I has a sufficient pellet at the bottom of the PCR tube. I placed a sample of the pellet on a microscope slide and viewed each sample under the microscope. I visualized algae and other microbes. I decided to do a quick gram stain which ensued in an argument between the three of us about the color of our new friend. I sad the color was purple but the boys were adamant that the color was pink (I was wrong). Because there was indecision, Jeremy streaked a TSA plate (to isolate a colony), and a macconkey plate (to see if it was indeed gram negative or gram positive). After 24 hours in the incubator, there was no growth. Jeremy asked if it were possible that the microbe was anaerobic. We decided to light a candle to see. Tony was busy testing the water sources for salinity, pH, and buying fish (to see if the bacterium would negatively impact them). He also learned how to successfully perform a gram stain (perhaps it is possible to teach an old dog new tricks). A specimen of contaminated water was obtained (100mL) and 1 disc of clindamycin and 1 disc of penicillin was added to this water. 24 hours later there was zero microbial growth. Two feeder goldfish were obtained and placed in a bowl with a mixture of 30 mL of contaminated water with 1 liter of DI water. Salinity was 896 PPM. After the fish were added, the salinity dropped to 696 PPM. the following afternoon an additional 30 mL of contaminated water was added to the fish bowl. 48 hours later, salinity was 1620 PPM, 30 mL contaminated water was added today and salinity is currently 2090 PPM. When the fish were first added the salinity decreased. That was the only time a decrease in salinity was observed. MSA plate was streaked today with contaminated water to see if the bacteria is sensitive to salt.
This is what we have learned:
1. The bacterium are anaerobic, gram negative, cocci, that start as round spheres with spike-like projections and mature into what appears to be filaments that grow in a bunch and break away from one another and infiltrate water. Many other tests need to be done before identification can be made.
2. The algal water arrived and had a salinity of around 5 PPM. After contamination the salinity was so high that the meter read "error". Either the bacteria or the algal death increased the salinity in the water.
3. A combination of clindamycin and penicillin killed the bacteria in an isolated study.
4. The bacteria does not appear to harm fish.
This has been an amazing week. I have truly loved this experience with this electron cloud. You boys are the best! I look forward to working with you both when we return from holiday.
No comments:
Post a Comment