User:Kira G. Flaherty/Notebook/Biology 210 at AU

Lab 2: Introduction
Observing the Hay Infusion Culture and Using a Dichotomous Key to Identify Organisms
Due 1/29/15
Kira Flaherty
This experiment included three parts. The first part was using a dichotomous key to identify one known organism. The second part of the experiment was to observe the Hay Infusion Culture that was made using the sample from the transect from last week. Different organisms of two different niches were identified from the Hay Infusion Culture. The third part of this experiment was to create serial dilutions in preparation for a future lab that focuses on the microbiology of the Hay Infusion Culture. The purpose of this experiment was to observe and identify the organisms that are living in the Hay Infusion Culture from a certain transect and to successfully identify organisms using a dichotomous key, as well as preparing for observing bacteria from the culture in next week's lab. (Bentley, Walters-Conte, Zeller 2015)
Methods and Materials
Part 1: Getting Familiar with Using a Dichotomous Key
For the first part of the experiment, a wet mount of known organisms was retrieved. This was done by using a glass slide, Protoslo and a cover slip to prepare a slide that was viewed under a microscope. When it was placed on the microscope, it was viewed at 4X objective and the 10X objective. One organism that was found was the focus of study and its characteristics, including the shape, color and colony size, were described. It was measured using the ocular micrometer within the lens of the microscope. A dichotomous key was used to identify the organism that was the focus of study. (Bentley, Walters-Conte, Zeller 2015)
Part 2: Identifying Organisms of Different Niches from the Hay Infusion Culture
The Hay Infusion Culture was observed. Two different niches within the culture were observed. One niche was from the bottom surface of the culture and the other niche was from the surface of the water. A wet mount of each was prepared on a slide with Protoslo and a cover slip that was viewed under a microscope. Three different organisms from each niche's wet mount were identified and described, making a total of six organisms. Each organism was measured using the ocular micrometer. Other characteristics of each organism were noted including the general look/shape, a picture of the organism and the organism's method of motility. (Bentley, Walters-Conte, Zeller 2015)
Part 3: Preparing to Study Bacteria from the Hay Infusion Culture through Serial Dilutions
Three tubes of 10 mL sterile broth were retrieved. One was labeled 10^-4, one was 10^-6 and the last was 10^-8. Six agar petri dishes were retrieved that included three that had tetracyline and three that did not. The plates that had tetracyline were labeled so that they didn't get mixed up with the other three plates. For the tetracycline plates, one was labeled as 10^-5, one was 10^-7 and one was 10^-9. The same labeling process was done for the plates without tetracycline. The Hay Infusion Culture was then gently mixed by swirling the jar with the lid on. A micropippetor was used to transfer 10 μL of the culture to the 10^-4 broth tube. The tube was swirled to mix the contents. Using the micropippetor again, 100 μL from the mixed 10^-4 tube was then added to the 10^-6 broth tube, which was also mixed by swirling. Then, 100 μL from the 10^-6 tube was added to the 10^-8 tube, which was mixed by swirling. 100 μL from the 10^-4 broth tube was then added to each of the 10^-5 petri dishes (one with tetracycline and one without). The same was done for the 10^-7 petri dishes, but using the 10^-6 broth. The same process was done for the 10^-9 petri dishes, but using the 10^-8 broth. For each petri dish, the broth was spread across the agar by using a sterilized glass spreader. The lids were placed on the petri dishes. The petri dishes with the tetracycline were wrapped in tin foil. All of the petri dishes were set aside to be incubated for a week in room temperature. (Bentley, Walters-Conte, Zeller 2015)
Observations and Results
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Figure 1: Microscopic View for Looking at a Known Organism in Part 1 of Procedure
From the dichotomous key, the above organism was identified as Gonium. It is a disc-shaped organism, with a green coloring. The colony size for this organism was about 16 cells. It was measured as about 10 μM using the ocular micrometer.
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Figures 2 & 3: Images of the Hay Infusion Culture from Transect 2
Above is two different views of the Hay Infusion Culture that was performed with the sample from transect 2.
Observations
The smell of the culture was not a good smell-- It was a smell that was comparable to sewage-like smell. There was some growth on the surface, looks like molds. The two niches that were observed were the bottom of the jar and the surface of the water.
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Figure 4: Oberved and Identified Organisms from Two Different Niches of the Hay Infusion Culture
Above are pictures and descriptions for each of the six organisms from the Hay Infusion Culture. Three different organisms for each niche was identified using a dichotomous key, as shown in the image. They were measured with the ocular micrometer. Plant matter was included in the observed wet mounts so there would be more of a variety of organisms. Some organisms may live off or use plants in some way to enhance survival while some organisms don't rely on plant matter at all. All of the organisms were motile, and either protozoa or algae.
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Figure 5: Serial Dilutions This is a diagram of the process of creating the serial dilutions. These petri dishes will allow the microbiology of the Hay Infusion Culture to be observed after a week for incubation.
Conclusion and Discussion
There was no need for a hypothesis and prediction for this experiment because it focused on successfully using a dichotomous key and to observe the organisms that were in the Hay Infusion Culture. In Part 1 of the procedure, the known organism was identified as Gonium. It was a green, disc-shaped organism and followed the description and diagram of the Gonium organism given by the dichotomous key. The dichotomous key also was beneficial in identifying the organisms of the two niches from the Hay Infusion Culture. The protozoa seemed to be more present on the surface niche, while the algae was more present on the bottom niche. If the Hay Infusion Culture grew for another two months, then I would predict more organisms of different forms and species to be present, as well as more growth of mold. The water would also continue to evaporate, which may affect the organisms that are able to survive and the carrying capacity. There would probably be an organism or several that could survive the best, so it or they may take over the culture and be the most present organism.

Paramecium Bursaria was one of the organisms found in the Hay Infusion Culture, from the surface of the water niche. It is a protozoa. The Freeman texts describes something alive by being able to use energy, having cells, processing information through genes and transferring/receiving information in the environment, replicating itself and continuing to go through evolution. Because this Paramecium Bursaria is alive, it does all of these things. It is unicellular, which means the entire organism is a cell, so it follows that requirement. It is able to obtain energy by eating other microorganisms and storing food in its central vacuole. It then gets energy from that stored food to move and other things to survive. Paramecium communicates with its environment by being able to sense different chemicals and using cilia to recognize its surroundings. Paramecium most often reproduce asexually, but can also reproduce through sexual reproduction (Parker 1982). There are different forms of Paramecium, some more advanced, which can show how evolution is continuing to happen.

For the next time observing organisms under a microscope, better diagrams and observations can be made. This would make identifying and making inferences about the organism easier. It would also make the data more valuable because different parts of the organisms could be identified and the information would be more credible.
References
Bentley, M., Walters-Conte, K. & Zeller N. K. 2015. A Laboratory Manual to Accompany: General Biology II. Department of Biology: American University, Washington, D.C. 18-24.
Parker, Sybil P. 1982. Paramecium. Synopsis and Classification of Living Organisms, 1. Retrieved from http://101science.com/paramecium.htm.

1.27.15 Excellent first entry. Well structured, clear image and thorough description of procedures. Keep up the good work. SK

Introduction
Bacteria and Protists of an AU Transect and the Volvocine Line
Due 1/26/15
Kira Flaherty
There were two parts of this experiment. The first was to study organisms within the Volvocine Line to observe evolution . These organisms were forms of green algae. The second part of the experiment was to observe and discuss the abiotic and biotic components that make up a transect acting as an ecosystem. Transect #2 was the focus of study. With the transect, we will be able to study the protists and bacteria that are within our transect through a Hay Infusion Culture process. Each transect has abiotic and biotic parts that make up the transect that is acting as an ecosystem. We can make conclusions and discuss our transect by observing the abiotic parts and the living organisms. The purpose of this experiment is to study and observe the organisms that are living in our assigned transect, and also observe evolution by looking at organisms of the Volvocine Line. (Bentley, Walters-Conte, Zeller 2015)
Methods and Materials
Part 1: Organisms of the Volvocine Line
In Part 1, 3 different green algae organisms of the Volvocine Line were observed. The organisms were the unicellular Chlamydomonas, multicellular Gonium and the most advanced Volvox. A table was created that included each organism in its own column. Each column was broken up into rows for various characteristics including number of cells, colony size, specialization of cells, motility, isogamous or oogamous and the picture of the organism. A microscope was used to study and identify these characteristics of these organisms. Slides for each organism were created by using a pipette, Protoslo, to slow the organism down, and a cover slip. The colony size of the cells were measured using the ocular micrometer within the lens of the microscope. (Bentley, Walters-Conte, Zeller 2015)
Part 2: Transect
The transect was found. An aerial view of the transect was sketched. All abiotic and biotic features of the transect were included and labeled. With a 50mL conical tube, a sample of the soil, vegetation and other things that made up the transect was retrieved. A Hay Infusion Culture was made to observe in future labs. To make the Culture, 10-12 grams of the sample of the transect was placed in a clear jar. 500 mL of bottled water was then added to the jar. 0.1 grams of dried milk was put into the jar and the contents of the jar were mixed. The jar was placed somewhere safe (somewhere it can stay for a week without being harmed) without the lid. After a week went by, the protists and bacteria from the Hay Infusion Culture were observed. (Bentley, Walters-Conte, Zeller 2015)
Observations and Data
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Figure 1: Observed Transect
This is an image of the transect. The location of the transcript was on AU's campus, right near Hughes Hall. There were several abiotic and biotic components of the transect, as listed and labeled in the image. The top of the picture is north, the right is east, the left is west and the bottom is south.
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Table 1: Observing Various Organisms of the Volvocine Line to Observe Evolution
This is a table that was used to compare Chlamydomonas, Gonium and Volvox. Volvox is the most advanced. Evolution can be observed by seeing the advancements in Volvox when compared to Chlamydomonas and Gonium.
Conclusion and Discussion
This particular experiment did not require a hypothesis or prediction because the purpose was just to observe evolution by looking at green algae of the Volvocine Line and to observe a transect, then set up a Hay Infusion Culture for future observations. Looking at the table of the Chlamydomonas, Gonium and Volvox, evolution can be observed. The most simplistic of the organisms is Chlamydomonas: unicellular, small colonies, no specialization of cells, isogamous and motile through the flagella. Then it is Gonium: multicellular, small colonies as well, no specialization of cells, motile (in a matrix) and isogamous. The Volvox is the most advanced and evolved of the three with about 1500 for number of cells, much bigger colonies than Gonium and Chlamydomonas, specialization of reproductive cells, nonmotile and oogamous.
The transect included many abiotic and biotic components. The biotic components included grass, trees, moss, bushes, insects, bacteria and other living organisms. The abiotic components included snow, dead leaves, fallen twigs, rocks, signs, a bench and soil. Many different species and classes of organisms were living within this same transect, which makes it a biodiverse ecosystem. The organisms within the ecosystem all have different niches and interact in ways that enhance survival (Bentley, Walters-Conte, Zeller 2015). A Hay Infusion Culture was made to be able to observe other organisms that can't be seen with the naked eye within the transect.
There is always room for improvement. For next time, something like a transect can be looked at even closer and the observations can be more precise. This will take more time and effort, but will be more beneficial to the outcome of the experiment.
References Bentley, M., Walters-Conte, K. & Zeller N. K. A Laboratory Manual to Accompany: General Biology II. Department of Biology: American University, Washington, D.C. 12-17.