User:Amanda Zimmerman/Notebook/Biology 210 at AU

Name: Amanda Zimmerman, Group 5, The Prairie

January 25th, 2014 Date observed: January 23rd, 2014

Goal/objective:

Procedures:

• -Procedure 1: Hay infusion culture continued
• 1) The culture, without being disturbed, was brought to our group's lab space so observations of the physical appearance could be made
• 2) A sample from the bottom, middle, and top layer were all drawn and made into a wet slide to be observed under a microscope. It is important to note where the organisms are found in order to help identify their nature. Some may be near or far from the plant matter based on how much they need the plant's nutrients
• 3) The organisms found under the microscope were then described in terms of color, mobility, size, etc. and characterized with the help of a Dichotomous key
• -Procedure 2: Preparing and Plating Serial Dilutions
• 1) Four tubes of 10 mL sterile broth were obtained and labeled 10^-2, 10^-4, 10^-6, and 10^-8. A 100 mL micropipeter set and tips were also obtained
• 2) 4 nutrient agar plates were obtained and labeled 10^-3, 10^-5, 10^-7, and 10^-9 with the date and group initials
• 3) 3 agar plus tetracycline plates were obtained (labeled "T") and labeled 10^-3, 10^-5, and 10^-7 with the date and group initials
• 4) The top was then placed on the Hay infusion culture and mixed until the solution was homogenous
• 5) 100 micro liters of the mix were taken from the Hay infusion culture and added to the 10^-2 tube, then inverted several times
• 6) 100 micro liters of the mix were taken from the 10^-2 tube and added into the 10^-4 tube, then inverted several times
• 7) This was then repeated into the 10^-6 then 10^-8 tubes to make the respective dilutions
• 8) 100 micro liters of the mix from the 10^-2 tube were added to the 10^-3 plates of both the nutrient agar and agar plus tetracycline
• 9) The sample was then spread around the plate
• 10) Steps 8 and 9 were then repeated with the 10^-4 tube onto the 10^-5 plates of both the nutrient agar and agar plus tetracycline, and the 10^-6 tube onto the 10^-7 plates of both the nutrient agar and agar plus tetracycline
• 11) Steps 8 and 9 were then repeated with the 10^-8 tube onto the remaining 10^-9 plate of nutrient agar only
• 12) The plates were then incubated at room temperature for 1 week

Observations:

• -Observations of the transect: The transect is covered in several inches of snow from the storm on January 21st-22nd. There are several tracks of foot prints, both human and animal, throughout the transect.
• -Observations of last week's Hay infusion culture:
• Top layer: Brown leaf is floating on top, part of the flower bud is seen in the top layer, the top has turned into a gray-brown opaque film, there are few areas of transparency within the film, no apparent life is on the top unless the film is some sort of fungus
• Middle layer: Composed of an amber-brown transparent liquid, most of the flower bud is in this layer, the piece of grass is submerged in this layer
• Bottom later:Composed of a solid state, soil has all sunk down to the bottom, the wood chip is also on the bottom, dark brown in color
• Other observations: The piece of grass is still green with no apparent loss of pigment, the flower bud has a white spiderweb-like growth hanging off of it, and there is no strong abhorrent smell present.
• If the Hay infusion culture had sat undisturbed for another two months, I would expect the piece of grass to be completely dead, due to the lack of nutrients needed for photosynthesis. Additionally, I would expect the flower bud to have disintegrated, due to the harsh conditions of the culture. I would expect the whole culture to be a musty brown-black concoction, too harsh for anything to survive in.
• The selective pressures of the Hay infusion culture include both the soil's nutrients for growth and survival and also the competition between light and water in overwhelming or saving the plant life. These things affected the composition and livelihood of our organisms.
• -Observations of observed organisms under the microscope of the top layer with possible characterizations:

1) Algae

• Large, green masses
• Multicellular
• Immobile

2) Paramecium Voritcella

• 2.5 ocular spaces on 10x = 25 micrometers
• Clear/colorless
• Very small
• Fast swimmer
• Elongated body
• Moves in a corkscrew fashion

3) Chlamydomonas

• 1 ocular space on 10x= 10 micrometers
• Appears to be a slight green pigment
• Exhibits strange motions
• Appears to be moved by flagella
• Single cell
• Oval shaped

4) Colpidium sp.

• 20 ocular spaces on 40 x= 50 micrometers
• Clear/colorless
• Relatively small
• Oval shaped
• Fast swimmer
• Entirely covered with cilia
• Makes fast/rapid motions

Calculations:

• -Microscope calculations:
• 4x calculations: # ocular spaces x 25 = # of micrometers (not used in this lab)
• 10x calculations: # of ocular spaces x 10 = # of micrometers (ex: Paramecium vorticella= 2.5 ocular spaces on 10x--> 2.5 x 10= 25 micrometers)
• 40x calculations: # of ocular spaces x 2.5 = # of micrometers (ex: Colpidium sp.= 20 ocular spaces on 40x--> 20 x 2.5= 50 micrometers)
• -Serial dilution calculations:
• Started with 100 mLs of Hay infusion in 10 mLs of sterile broth= 10/100= 10^-2

Extra notes:

• -One of the species that demonstrates the needs of life is the paramecium:
• 1) Energy: Paramecium consume bacteria and algae for food, which it then converts into energy to survive
• 2) Cells: Paramecium are composed of membrane-bound cells
• 3) Information: Paramecium have DNA, the information that encodes its life
• 4) Replication: Paramecium reproduce asexually during binary fission, in which they replicate their DNA and pass it into their genetically-idential offspring
• 5) Evolution: Paramecium evolved from their first seen structure in order to feed and protect themselves
• All information from: http://en.wikipedia.org/wiki/Paramecium

Photos: To come of both the jar and diagram of the Hay infusion culture dilutions

January 16th, 2014 Date observed: January 16th, 2014

Goal/objective: Find the designated transect and observe it. List the physical description, biotic, and abiotic factors of the transect. Also, create a Hay infusion culture to be evaluated in one week.

Biotic factors: Grass, leaves, soil, flowers, and bushes Abiotic factors: Fertilizer, concrete bench, metal lining separating the soil from the walkway, "Freidheim Quadrangle" sign, and walkway stones Description of area: This 20' by 20' square is located on the Quad in front of Hurst Hall. It's composed primarily of a spacious grass area. There is an arc of soil, including shrubbery and a portion of the "Freidheim quadrangle" sign. Between the soil area and the walkway is a metal lining. There are very few leaves or flowers left on the shrubbery, due to the extreme wintery conditions. Within that arc is another, smaller arc consisting of a stone walkway and concrete bench.

Methods: Hay infusion culture 1) 10-12 grams of our sample was weighed and placed in a labeled plastic jar with 500 mLs of Deerpark water 2) 0.1 gram of dried milk was added and gently mixed up for about 10 seconds 3) The top of the jar was then removed and the rest of the jar was placed on a lab table with other jar

Conclusions: There isn't anything that I would change for "next time" per se, but I think it would be interesting if we made two Hay infusions and put them in different environments with varying light exposure, temperature, or nutrients and compared later. The abiotic and biotic factors will play more of a role when the weather gets nicer. I expect to see the leaves and flowers budding within a few months. Also, I'm interested in seeing how the nicer weather attracts people to the transect and how they will play a role on the transect's being.

Photos: (To come)