User:Dominic L Genuario/Notebook/Biology 210 at AU

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Transect Description Group 3's transect is located near Bender Arena and the amphitheater of American University. It is an environment which seems to be heavily impacted by humans since it is within a garden and includes sidewalks and benches. It is relatively flat, but has small mounds of dirt and mulch which rise above the sidewalks. Upon initial investigation, possible biotic components are birds, squirrels, spiders, trees, bushes, ferns, some grasses, fungi, and most likely protists and other bacteria. The birds are possible since a nest was discovered in a tree within the transect. The trees are also conducive to squirrels. A spider was viewed directly, as were the trees, bushes, ferns and grasses. The fungi were present on the benches. Obvious abiotic components are the sidewalks, benches, rocks, dirt, cigarettes, candy wrappers, and street lamps which were present.

Image:GenuarioTransect.png

Image courtesy of Google Maps. Image is oriented due north. Transect roughly outlined in red. --DG

Identifying Algae & Protists.

Purpose The purpose of this lab is to observe microscopic eukaryotic organisms within the lab and transect and to classify them using a dichotomous key.

Data & Observations The Hay Infusion has a swamp-like smell and is yellow overall in appearance. There is no apparent life on top of the liquid. The organisms observed are varied with some very motile protozoa and other less-motile photosynthesizing algae. The motile protozoa don't seem to photosynthesize and instead rely on other sources of energy. One observed algae appeared to be from the genus chlamydomonas. Organisms within this genus are generally phototrophic, and have a single flagellum; the observed organism was roughly 25 micrometers in diameter.

Conclusions If the Hay Infusion grew for two more months there would be an increase in bacteria, fungi, and microscopic organisms until the food supply was diminished. Since it is such a closed ecosystem there would be a point at which much of the life would die off and an equilibrium relying on photosynthesis would be reached.

Microbiology & Observing Bacteria

Purpose The purpose of this lab is to learn to characterize bacteria based on motility, gram stains, and colony morphology.

Data & Observations The Hay Infusion has an increased film, a lower water line, the same putrid smell, more growth on the wood, more dirt on the bottom, water which is still yellow, and mold growth under the film.

Image:GenuarioTable1.png

The Tet positive plates have orange yellow colonies that are fewer in number and the colonies are convex and umbonate from the agar plate surface. The Tet negative plates have smaller white colonies that are much greater in number, and less uniform in formation. These white colonies also seem to be raised, and these plates also have condensation in them unlike their Tet positive counterparts. From each sample, from the Tet positive and negative plates, and the yellow and white colonies, the most recognizable bacteria were small coccus shaped formations. Because the bacteria cells found were so small and all coccus in shape the overall motility was determined to be immobile.

Image:GenuarioSerialDilution.png

Conclusions It is hypothesized that the appearance and smell change from week to week due to bacterial growth and mold growth. It is unlikely that Archaea will have grown on the agar plates made from the Hay Infusion since Archaea thrive in extreme environments, environments not found on AU's campus. The difference between the Tet positive and Tet negative plates demonstrates the difference in the type of bacteria growth. It's clear that the bacteria which survived on the Tet positive plates are antibiotic resistant, a trait which is rarer. On the other hand, the bacteria on the Tet negative plates had no factors inhibiting their growth which meant they could multiply extensively. As mentioned, the tetracycline reduced the total number of bacteria and fungi. It seems that only two species were unaffected by the tetracycline. Tetracyclines are a wide range of antibiotics which used to affect almost all bacteria, both gram positive and gram negative, through the inhibition of protein synthesis. Now, however, many bacteria have acquired resistance, which reduces the effectiveness of the drug. (Source: http://www.toku-e.com/Assets/MIC/Tetracycline%20hydrochloride.pdf)

Plantae & Fungi

Purpose The purpose of this lab is to investigate the importance and diversity of Plantae and Fungi.

Data & Observations The first sample was taken from the branch of a tree which was >25ft tall. The sample was 4.5cm tall and 7.5cm wide, was determined to be a dicot, and had netlike veins and spiky leaves. The second sample from the branch of a 3ft tall bush was determined to be a dicot and was 4.5 cm tall and 10.5 cm wide. The third sample from a stem of yellow grass about 2.5ft in height had fibrous seeds from a singular stem and was 24cm tall and 5.5cm wide. The sample from a branch of the low green plants covering the ground was a dicot, with many leaves per branched which were tiered and inconsistent in size. The sample was 8cm tall and 7.5cm wide. The final sample was taken from the stem of a yellow grass growing alone next to a bush and had many stems from the origin with many seeds growing on each stem.

Conclusions Sporangia are the enclosures in which spores are formed, and many organisms, plants and fungi included, form sporangia at some point in their life cycle.

Invertebrates & Vertebrates

Purpose The purpose of this lab is to observe the invertebrates and vertebrates range of body size, body plan, mechanism of motility, mechanisms of reproduction, sensory systems and nutritional requirements

Data & Observations

Image:GenuarioInvertebrates.png

5 observed organisms: Arthropoda Arachnida Acarina, Big Brown Mite Arthropoda Insecta Psocoptera Arthropoda Arachnida Acarina, Mite C Arthropoda Arachnida Acarina, Mite C Unknown ant-like organism

Conclusions All organisms found were under 20 micrometers in length. The big brown mite was the largest and it seems that mites are the most common organism in leaf litter from the transect. The two samples are different in what organisms they contain, however, they do both contain mites, the specified common organism. Two bird species which might appear in this transect are Passerine Passeridae, sparrows, and Passerine Passeri, or songbirds. The seeds, worms, and other small invertebrates would assist the birds as well as the trees which provide shelter. The food web of this transect most likely has bacteria at the bottom which are preyed upon by protists and mites. These mites may also suck juices from plants and feed on fungi. Above these mites in the web, of organisms that were observed, would be squirrels although the squirrels likely don’t eat the mites and instead eat nuts, fungus, and insects. On a similar level to the squirrels would be the birds who consume seeds, worms, and other small invertebrates. The insects likely present in the transect would fall on a level similar to the mites. The carrying capacity of this ecosystem wouldn't be extremely large for larger organisms such as birds, however it does provide plentiful nutrients for microscopic life.

Zebra Fish Development

Purpose The purpose of this experiment is to compare and contrast the development of zebrafish in a standard water solution versus a 1% ethanol solution in order to model possible effects of mild fetal alcohol syndrome.

Data & Observations Zebrafish Experiment Drug: Ethanol Dosage 1 → 2 mL 1% ethanol (pre-made) *24 wells total

Control → 2 mL of control water only *24 wells total

2/22 ZebraFish Observations

Control:

A1: alive A2: alive A3: unhatched A4: alive A5: alive A6: alive B1: alive B2: alive B3: alive B4: alive B5: alive B6: alive C1: alive C2: alive C3: alive C4: alive C5: alive C6: alive D1: alive D2: alive D3: alive D4: alive D5: alive D6: alive

Control: Fish circled dish multiple times and continued to squirm to other side.

A3 (Observed from dissecting scope): unhatched → still at about 48 hpf

A1 observed (depression slide) Eye movement: stable Fin development: developed and stable movement Color: yellow tint with dark green dots and purple Yolk size: 15 micrometers

Treatment:

A1: alive A2: alive A3: NO EGG A4: dead (2/22) - preserved A5: alive A6: alive B1: alive B2: alive B3: alive B4: alive B5: alive B6: alive C1: alive C2: alive C3: alive C4: alive C5: alive C6: NO EGG D1: alive D2: alive D3: alive D4: alive D5: alive D6: alive

Dosage 1: Initially didn’t react, then moved once around dish.

A3 (Dissecting scope): No egg inside well

A4 (Dissecting scope): Found to be unresponsive to stimuli, presumed dead

C6 (Dissecting scope): No egg inside well

Dead eggs preserved. Labeled AH DG BB 003 (1)

B2 observed (depression slide) Pectoral fins developed: stable Mouth: N/A Heart: N/A Pigment: yellow with dark green/purple spots Yolk size: about 20 micrometers

Poked with pipette tip for movement

2/23 Zebrafish Observations

A3 [ control ] - still hasn’t hatched.

Control - all are alive: Movement: immediate reaction when pipette tips was placed in cell. circled and squirmed around dish multiple times.

Control:

A1: alive A2: alive A3: unhatched A4: alive A5: alive A6: alive B1: alive B2: alive B3: alive B4: alive B5: alive B6: alive C1: alive C2: alive C3: alive C4: alive C5: alive C6: alive D1: alive D2: alive D3: alive D4: alive D5: alive D6: alive

A1 observed Yolk sac: 14 micrometers Eye movement - Normal, eyes are dark Color: Yellow/green tint with dark spots and purple. Movement: Fins constantly moving

B6 observed Yolk: 30 micrometers Mouth visible

Treatment:

A1: alive A2: alive A3: NO EGG A4: dead (2/22) - presv. A5: alive A6: alive B1: alive B2: alive B3: alive B4: alive B5: alive B6: alive C1: alive C2: alive C3: alive C4: alive C5: alive C6: NO EGG D1: alive D2: alive D3: alive D4: alive D5: alive D6: alive

D6 (Dissecting scope): Didn’t respond to stimuli initially → closer tip resulted in response. Appears more transparent with strip of yellow on back. Not as responsive.

B2 observed: Yolk: 15 micrometers Eye movement: stable Fin movement: constant

A5 observed: Mouth: visible Fin movement: rapid (abnormal) Yolk: 28 micrometers Eye movement: stable

2/24 Zebrafish Observations

A3 [control] - removed and preserved, assumed dead because still unhatched.


Control:

A1: alive A2: alive A3: presv. (2/24) A4: alive A5: alive A6: alive B1: alive B2: alive B3: alive B4: alive B5: alive B6: alive C1: alive C2: alive C3: alive C4: alive C5: alive C6: alive D1: alive D2: alive D3: alive D4: alive D5: alive D6: alive


A1 observed: HR → 90 bpm yolk: about 20 micrometers mouth: visible fin movement: stable eye size: 10 micrometers eye movement: constant

B6 observed: yolk sac: NONE HR → 120 bpm eye size: 11 micrometers eye movement: stable fin movement: N/A mouth: visible

treatment: all are alive, 1 missing movement: more than control, constant. Treatment:

A1: alive A2: alive A3: NO EGG A4: dead (2/22) - presv. A5: alive A6: alive B1: alive B2: alive B3: alive B4: alive B5: empty B6: alive C1: alive C2: alive C3: alive C4: alive C5: alive C6: NO EGG D1: alive D2: alive D3: alive D4: alive D5: alive D6: alive

B5 - nothing in the well

A2 (dissecting scope): responded initially to pipette movement then didn’t.

B2 observed (depression slide): Unable to observe, kept swimming into wall. Assumed to be because it is avoiding the light source from the microscope Lowered light: was able to view for short amount of time after searching. Unable to see clearly to observe so turned light up again: immediately swam away. - continued to swim into wall.

A5 observed - mouth: visible, can see mouth eating shrimp brine that came with observation fin movement: rapid/abnormal dark appearance (darker than previous observation) eye movement: minimal HR → N/A

2/25 Zebrafish Observations

control:

C4 (dissecting scope): Head: deformed, head too big? swim patterns: in spiral facing bottom of well

C6 (dissecting scope): Head: deformed, head/back appears large and abnormal Swimming patterns: on side Response: on side very late reaction to stimuli

Preserved A2, B2, C2.

A1 observed HR → 105 bpm eye movement, size: 10 micrometers fin movement mouth visible light yellow-green, (brown-green spine), spots are black/dark brown head size (mouth-fin): 30 micrometers

B6 observed HR → 128 bpm very active fin movement head size: 32 micrometers eye size: 12 micrometers

Control:

A1: alive A2: presv. (2/25) A3: presv. (2/24) A4: alive A5: alive A6: alive B1: alive B2: presv. (2/25) B3: alive B4: alive B5: alive B6: alive C1: alive C2: presv. (2/25) C3: alive C4: alive C5: alive C6: alive D1: alive D2: alive D3: alive D4: alive D5: alive D6: alive

treatment: all alive

Preserved A1, B1, C1

B2 observed HR → 52 bpm ? eye movement: rapid, left to right/no pattern hard to swim, darts forward in short distances. fin movement: sporadic lots of spots, darker yellow/green tint. more noticeable head (appears bigger than body): 27 micrometers eye: 10 micrometers

A5 observed HR → 148 bpm swimming sideways rapid fin movement minimal eye movement transparent with slight yellow/green tint. not many spots head: 40 micrometers eye: 11 micrometers

Treatment:

A1: presv. (2/25) A2: alive A3: NO EGG A4: dead (2/22) - presv. A5: alive A6: alive B1: presv. (2/25) B2: alive B3: alive B4: alive B5: empty B6: alive C1: presv. (2/25) C2: alive C3: alive C4: alive C5: alive C6: NO EGG D1: alive D2: alive D3: alive D4: alive D5: alive D6: alive

2/26 Zebrafish Observations

Control:

A1: alive A2: presv. (2/25) A3: presv. (2/24) A4: alive A5: alive A6: alive B1: alive B2: presv. (2/25) B3: alive B4: alive B5: alive B6: alive C1: alive C2: presv. (2/25) C3: alive C4: alive C5: alive C6: alive D1: alive D2: alive D3: alive D4: alive D5: alive D6: alive

A1 observations: HR → around 100 bpm Normal eye movement, eye size: 10 micrometers Active fin movement Same color as yesterday [light yellow-green, (brown-green spine), spots are black/dark brown] head size: 30 micrometers

B6 observed HR → around 120 bpm Very active eyes and fins Head size: 32 micrometers Eye size: 12 micrometers

Treatment:

A1: presv. (2/25) A2: alive A3: NO EGG A4: dead (2/22) - presv. A5: alive A6: alive B1: presv. (2/25) B2: alive B3: alive B4: alive B5: empty B6: alive C1: presv. (2/25) C2: alive C3: alive C4: alive C5: alive C6: NO EGG D1: alive D2: alive D3: alive D4: alive D5: alive D6: alive

B2 observations: HR → Not determined Sporadic swimming, eye movement, and fin movement Noticeable dark green spots Head size: 27 micrometers Eye size: 10 micrometers

A5 observations: HR → Around 150 bpm Abnormal swimming Normal fin movement, less eye movement Some light green spots Head size: 40 micrometers Eye size: 11 micrometers

2/29 Zebrafish Observations

Control: A1 dead death by fungus/bacteria?, B4 alive but no movement or reaction to stimuli. heart visibly beating, C4 dead by fungus/bacteria?, D1 dead by fungus/bacteria

Still alive: A4, A5, A6, B3, B5, B6, C1, C3, C5, D2, D3, D4, D5, D6 Preserved: A2, B2, C2


Control:

A1: dead 2/29 A2: Pres. 2/25 A3: unhatched A4: alive A5: alive A6: alive B1: alive B2: Pres. 2/25 B3: alive B4: alive B5: appears empty B6: alive C1: alive C2: Pres. 2/25 C3: alive C4: dead (2/29) C5: alive C6: dead D1: dead (2/29) D2: alive D3: alive D4: alive D5: alive D6: alive

A1 (previously observed) dead. translucent, in process of decomposition, dark colored bacteria and fungi swimming around within and outside of fish carcass

B6 observed HR → N/A swimming patterns: normal (healthy, not lacking in energy) darts to various positions spontaneously fin movement: normal head: 33 micrometers eye:14 micrometers

Treatment:

A1: Preserved (2/26) A2: dead (2/24) A3: no egg A4: no egg A5: alive A6: alive B1: Preserved (2/26) B2: dead (2/29) B3: alive B4: dead (2/29) B5: alive B6: alive C1: Preserved (2/26) C2: alive C3: alive C4: alive C5: alive C6: dead (beginning) D1: alive D2: dead (2/29) D3: alive D4: alive D5: alive D6: alive

B2, (previously observed) dead

A5 HR → N/A swimming patterns: away from light fine movement: golts around eye size: 10 micrometers head size: 22 micrometers

3/1 Zebrafish Observations

Control: 11 Alive: A4, A5, A6, B3, B5, B6, C1, C3, D2, D3, D5

B6 Observations:

  • complication: observed fish from B5 well by mistake

HR → N/A swimming patterns: normal (healthy, not lacking in energy) darts to various positions spontaneously fin movement: normal head: 33 micrometers eye:14 micrometers

Control:

A1: dead 2/29 A2: Pres. 2/25 A3: unhatched A4: alive A5: alive A6: alive B1: dead B2: Pres. 2/25 B3: alive B4: dead 2/29 B5: alive B6: alive C1: alive C2: Pres. 2/25 C3: alive C4: dead (2/29) C5: dead (3/1) C6: dead D1: dead (2/29) D2: alive D3: alive D4: dead (3/1) D5: alive D6: dead (3/1)

Treatment: 9 alive: A5, B3, B6, C3, C4, D1, D3, D4, D6

B2 → dead

A5 Observations: HR → N/A fin movement: consistent swim pattern: fish actively moves away from light and other stimuli (pipette tip) total length: 140 micrometers head: 33 micrometers eye: 14 micrometers

Treatment:

A1: Preserved (2/26) A2: dead (2/24) A3: no egg A4: no egg A5: alive A6: dead (3/1) only body outline visible B1: Preserved (2/26) B2: dead (2/29) B3: alive B4: dead (2/29) B5:Appears empty B6: Alive C1: Preserved (2/26) C2: dead (3/1) C3: alive C4: alive C5: dead (3/1) C6: dead (beginning) D1: alive D2: dead (2/29) D3: alive D4: alive D5: dead (3/1) D6: alive

3/2 Zebrafish Observation

Control:

A1: dead 2/29 A2: Pres. 2/25 A3: unhatched A4: alive A5: dead (3/2) A6: alive B1: dead B2: Pres. 2/25 B3: alive B4: dead 2/29 B5: alive B6: alive C1: alive C2: Pres. 2/25 C3: alive C4: dead (2/29) C5: dead (3/1) C6: D1: dead (2/29) D2: alive D3: alive D4: dead (3/1) D5: alive D6: dead (3/1)

B5 Observed HR → N/A Swim patterns: stable, stays in light, minimal golting, smoother swimming. Reacts to stimuli (pipette tip) fin movement: stable eye size: 8 micrometers head size: 23 micrometers total length: 125 micrometers

Treatment:

A1: Preserved (2/26) A2: dead (2/24) A3: no egg A4: no egg A5: dead (3/2) A6: dead (3/1) only body outline visible B1: Preserved (2/26) B2: dead (2/29) B3: dead (3/2) disintegrated B4: dead (2/29) B5:Appears empty B6: alive C1: Preserved (2/26) C2: dead (3/1) C3: alive C4: dead (3/1) not disintegrated C5: dead (3/1) C6: dead (beginning) D1: dead (3/2) D2: dead (2/29) D3: dead (3/2) D4: dead (3/2) D5: dead (3/1) D6: dead (3/2)

3/3 Zebrafish Observations

Control:

A1: dead 2/29 A2: Pres. 2/25 A3: unhatched A4: alive A5: dead (3/2) A6: alive B1: dead B2: Pres. 2/25 B3: alive B4: dead 2/29 B5: alive B6: alive C1: alive C2: Pres. 2/25 C3: dead (3/3) C4: dead (2/29) C5: dead (3/1) C6: D1: dead (2/29) D2: dead (3/3) D3: alive not responding to stimuli D4: dead (3/1) D5: dead (3/3) D6: dead (3/1)

Surviving #: 7

Treatment:

A1: Preserved (2/26) A2: dead (2/24) A3: no egg A4: no egg A5: dead (3/2) A6: dead (3/1) only body outline visible B1: Preserved (2/26) B2: dead (2/29) B3: dead (3/2) disintegrated B4: dead (2/29) B5:Appears empty B6: dead (3/3) body still visible C1: Preserved (2/26) C2: dead (3/1) C3: dead (3/3) disintegrated C4: dead (3/1) not disintegrated C5: dead (3/1) C6: dead (beginning) D1: dead (3/2) D2: dead (2/29) D3: dead (3/2) D4: dead (3/2) D5: dead (3/1) D6: dead (3/2)

Conclusions Not only did the ethanol lead to mutations and impaired growth, it also seemed to be conducive to the growth of fungi. For unknown reasons, when the zebrafish in the ethanol died their bodies quickly disintegrated leaving only an outline whereas the fish that passed away in the control group didn't immediately decompose.

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