User:Lauren S. Lit/Notebook/Biology 210 at AU

3/23/14: Lab 6: Zebra Fish

Introduction:

The objective of this lab is to determine the effect of retinoids on zebra fish embryos and the development from juveniles into adult zebrafish. It was hypothesized that too much retinotropic acid in an environment will kill the zebrafish embryos and have impairments on their development. Rentinoids are known as vitamin A

Materials and Methods:

1. 3 Petri dishes were set up. 1 with 20 mls of H2O, one with 20 mls of retinoic acid and one with 2 mls retinoic acid and 18 mls of H2O. 2. 20 zebrafish embryos were placed in each petri dish. 3. The fish embryos were observed at 4,7,11, and 14 days.

Observations:

Day 7:

 - 100% Acid: smaller, lighter in color, less developed organs, they are having difficulty swimming and twitch instead. small swim bladder, lighter in pigment
 - Control (Just Water): faster moving, 1 unhatched, 16 alive, 2 developed but dead, 1 unhatched dead, quick healthy fins, large black eyes
 - 2ml acid/18ml water: Bigger than the control, dark developed organs, 18 alive, 2 developed dead, fast heartbeat, large swim bladder

Day 12:

 - 100% Acid: All Dead, organs are blurring and decaying
 - Control (Just Water): loss of control of the front fins, 6 alive, 10 dead, twitching tail, definite dark organs
 - 2ml acid/18ml water: lighter organs, fins are not moving, some have started decaying, 7 alive and 11 dead.

Results/Conclusions:

The Datas shows that too much retinoid acid is hazardous to the development stages of zebra fish. it is possible that there is a threshold of how much acid should be in the environment and a toxicity amount. The organisms survived well without the acid but the diluted acid fish developed quicker and stronger but did not live as long. While vitamin a is important it is also imperative to ensure we are not receiving too much vitamin A. LL

1/23/14: Lab 5: Invertebrates

Introduction:

The objective of the lab is to observe the characteristics of invertebrates and to understand how simple systems evolved into more complext systems. It is predicted that there will be very few animals in the transect and those that around in the transect will most likely be worms.

Materials and Methods:

1. Acoelomate, psuedocoelomates and coelmates were observed under the disecting scope 2. The burlese funnel was taken down and the preservative solution was placed in a petri dish 3. Organisms from the dish were observed and measured using a disecting microscope

Observations:

Acoelomates - Planaria: They move by gliding their bodies

Pseudocoelomates - nematodes: whip like movement

Coelmates - Annelida: The body contracts and expands to move

All organisms were taken from the West Virgina Leaf Little because no animals were found in the leaf litter.

  - Organism 1: beetle - 2.5 cm long - Large, brown, 6 legs, antenae, pinchers
  - Organism 2: Scorpian - 4mm - small, brown, pinchers, 4 small legs
  - Organism 3: Flea - 2mm - small, brownish black, 6 legs, suckling mouth
  - Organism 4: Arachnid - 75 micrometers - small, black, 8 legs
  - Organism 5: Lice - 1mm - Small, black, chewing mouth parts

Conclusion

Since our transect leaf litter had no animals we viewed the West Virginia Leaf Litter Samples. The most common organism was the arachnids. The beetle was the largest organism at 2.5cm and the lice was the smallest at 1mm.

2/23/14 - Lab 4: Plantae and Fungi

Introduction:

The objective of the lab is to observe the characteristics and diversity of fungae and plants. Plants evolved from green aquatic algae, which closely resemble bryophytes which are known as moss. Byrophytes are non-vascular. As the plants began to specialize more and grow on land the tracheophytes started to develop with a vascular system.

Materials and Methods:

 -A plastic bag was filled with leaf litter to ensure it contains dirt and dead leaf sediment.
 -5 plant samples were taken from the transect.
 -The samples were observed to characterize the plants vascularization and reproductive qualities.

Observations and Data:

1. Horsetail - located in the middle of the transect. It is a bush like cyclinder plant. It is vasularized but has no leaves. The plant is seedless.

2. Located in the outer edge of the transect. The plant has long thin leaves and is vascularized however it is low to the ground. The plant is seedless.

3. Red Tree with vascularization through its roots. The tree has small green leaves with red stems. It is seedless.

4. Long thin plant. Located in the middle of the transect. It is vasularized. It has fuzzy flowers which are part of the plants reproduction system.

5. Small green plant that looks like moss. It is not vascularized and it contains small green leaves. The plant has no seeds.

6.Lilly - The lilly is vascularized. It is a momcot

7. Moss is a bryophye and is not vascularized because it is low to the ground.

Conclusions:

Plants are a very diverse species that have adapated to fit a variety of different environments. LL.

2/16/14 - Lab 3:Microbiology and Identifying Bacteria with DNA

Introduction:

Materials and Methods:

1.Using the swirled up Hay infusion from Lab 1 and 2, 100 ml of the hay infusion culture was added to 10mls of nutrient broth into a tube. This made a 1:100 dilution or 10^-2. Next 100microliters from that tube was placed into 10mls of nutrient broth to make a dilution of 1:10,000 or 10^-4. Next 100microliters from tube 4 was placed into 10mls of nutrient broth to make a dilution of 1:1,000,000 or 10^-6. Next 100microliters from tube 6 was placed into 10mls of nutrient broth to make a dilution of 1:10,000,000 or 10^-8.

2. Four nutrient agar plates were labeled. 10^-3, 10^-5,10^-7,10^-9. Another set of Agar plates with tetracycline were labeled with the same labels.

3. 100 microliters of the 10^-2 solution was placed on the surface of the 10^-3 agar plate and the 10^-3 agar plate with tetracycline. 100 microliters of the 10^-4 solution was placed on the surface of the 10^-5 agar plate and the 10^-5 agar plate with tetracycline. 100 microliters of the 10^-6 solution was placed on the surface of the 10^-7 agar plate and the 10^-7 agar plate with tetracycline. 100 microliters of the 10^-8 solution was placed on the surface of the 10^-9 agar plate and the 10^-9 agar plate with tetracycline.

4. The plates were inoculated at room temperature for 1 week.

5. The plates were observed

Observations and Data:

Table 1: 100-Fold Serial Dilution Results

 - 10^-3 --> 120 Colonies --> 1,200,000 colonies/ml
 - 10^-5 --> 30 Colonies --> 30,000,000 colonies/ml
 - 10^-7 --> 4 Colonies --> 400,000,000 colonies/ml
 - 10^-9 --> 21 Colonies --> 1.7*10^-10 Colonies/ml
 - 10^-3 + Tetracycline --> 23 Colonies -->230,000 Colonies/ml
 - 10^-5 + Tetracycline --> 1 Colonies -->1,000,000 Colonies/ml
 - 10^-3 + Tetracycline --> 1 Colonies -->100,000,000 Colonies/ml\

Table 2: Colony Observations:

 -10^-5 Nutrient Agar --> Dark Purple, Circular, Entire edge, Flat --> 1 colony --> Quick motility with cilia --> Gram negative
 -10^-5 Tetracycline --> Cream, circular edge -->1 colony --> brownian movement (drifting) -->Gram negative
 -10^-3 Tetracycline --> yellow, small, convex, entire edge --> 230,000 colonies --> no movement --> Gram positive

Image:IMG 2107.jpg

Results and Conclusions:

There is some antibiotic resistance on the 10^-3 tetracyline plate. This is seen when comparing the number of colonies on the Nutrient agar plate and the tetracline plate. There are less colonies on the nutrient plus tetracycline plates compared to the just nutrient plates. This indicates the antibiotics are working to kill the bacteria. If the bacteria grew anyway, which is known as antibiotic resistance. There is clear evidence that the presence of the tetracycline is enough to really prevent and limit bacteria growth. there is roughly 25species of the bacteria which were unaffected by the tetracycline. 

The gram stain performed showed us that the bacteria from plate 10^-3 with tetracycline is gram positive and is antibiotic resistant to tetracyline. while the other 2 samples from 10^-5 and 10^-5 with tet. were both gram negative. Both gram positive and gram negative are known to have species that are resistent to tetracyline. LL

[edit] 2/9/14 - Lab 2: Identifying Algae and Protists Introduction: Understanding and Identifying Unicellular Organisms, specifically Algae and Protists. Algae and Protists have very specialized characteristics. While they are different organisms they both come from a common ancestor and are both unicellular eukaryote. Materials and Methods: - A wet mount was created using 8 different known organisms --> A drop of the specimen solution containing various organisms were placed on the slide and covered with a coverslip -The wet mount and specimens were viewed under tthe microscope -Once an organism was located it was characterized using a Dichotomous Key Observations and Data: 1. Euglena - 20-25 micrometers - multicellular 2. Chlamydomonas - 10 micrometers - multicellular 3. Paramecium Caudatum - 180 micrometers

Hay Infusion -Smalls musty -mold is growing on the sides -Leaves and plants are on the top layer -Algae is growing on the very bottom of the jar -algae coating on the top. -Top of Jar: Brown Algae

-Immobile
-10 micrometers in length

-Bottom of Jar:

-Spirostimum
-brown algae
-Colpidium - 62 micrometers

Conclusions: The hay infusion is expected to grow more mold agae and other organisms. The smell is expected to get progressively worse. The niches are expected to become more specialized and more organisms with more distinct roles will develop. LL

[edit] 1/31/14 - Lab 1: Biological Life at AU The objective of Lab 1 Proceedure 1 is to understand Natural Selection. To get an understanding of natural selection 3 members of the volvacine line were observed under the microscope. They included Chlamydomonas, gonium, and volvox. Chlamydomonas Number of cells: 1 Colony Size: 23 Functional specialization Cells: Flagellum for movement and chloroplast Gonium Number of cells: 75-100 Colony Size: 3 Functional specialization Cells: Flagellum Vovox Number of cells: about 500 Colony Size: 1 Picture to be added here. The objective of part 2 is to understand the biotic and abiotic aspects of a niche at American University outside the Katzen Arts Center. Abiotic Factors Pebbles, large rocks, little rocks dead leaves wood chips bricks soil Biotic Factors Cotton Tails Red Bushs Grass Weeds Picture to be added here. In the future I will be going back to the niche to observe any changes that may have occured to the factors, individually and how they interact together. LL. Good start. Could be improved with more substance. Address red text from protocol include detailed descriptions, analysis and conclusions. For more instructions see TA notebook. SK

1/26/14 - Lauren Lit's Lab Notebook - Bio210. LL