User:Adriana N. Hooks/Notebook/Biology 210 at AU

Zebrafish

Drug: Ethanol Dosages: Dosage 1 → 2 mL 1% ethanol (pre-made) *24 wells total

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

OBSERVATIONS

2/22 ZebraFish Observations

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

A3 (generally observed from dissect 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 micrometer

Treatment: initially didn’t react, then moved once around dish.

A3 (from dissect scope): no egg inside well

A4 (from dissect scope): found to be unresponsive to stimuli dead

C6 (from dissect scope: no egg inside well

dead fish preserved. Labeled AH DG BB 003 (1)

B2 observed (depression slide) eye movement: pectoral fins developed: stable mouth: N/A heart: N/A pigment: yellow with dark green/purple spots yolk size: about 20 micrometers

(all fish were fed)

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.

A1 observed yolk sac: 14 eye movement - eyes are dark yellow/green tint with dark spots and purple. fins constantly moving

B6 observed yolk - 30 mouth visible

Treatment: all alive

D6 (dissect scope): didn’t respond to stimuli initially → closer tip = response. appear more transparent with strip of yellow on back. not as responsive.

B2 observed: yolk: 15 eye movement: stable fin movement: constant

A5 observed: mouth: visible fin movement: rapid (abnormal) yolk: 28 eye movement: stable

2/24 Zebrafish Observation

a3 [control] - removed and preserved, assumed dead bc still not hatched.

control - cleaned

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

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

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

B5 - nothing in the well

A2 (dissect 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 (dissect scope): Head: deformed, head too big? swim patterns: in spiral facing bottom of well

C6 (dissect 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 nm (x2.5) fin movement mouth visible light yellow-green, (brown-green spine), spots are black/dark brown head size (mouth-fin): 30 nm

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

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 eye: 10

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

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 (picture) death by fungus/bacteria?, D1 dead by fungus/bacteria (picture at 6:05)

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

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 eye:14

Treatment

Still alive: Preserved:

B2, (previously observed) dead

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

3/1 Zebrafish Observations

Control:

• all fish were fed, wells were cleaned

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 eye:14

B6 observations:

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 head: 33 (25) eye: 14 (10)

3/2 Zebrafish Observation

Control:

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 head size: 23 total length: 125

Treatment:

→ Ethanol provides environment for bacteria and fungus = more death than control → control decomposed and disintegrated while ethanol grew fungus. → minimal growth changes.

Tet Plates

Procedure I: Quantifying and Observing Microorganisms File:TetTable

Plate Observations Tet (+) [for slides chose: 10^-5] >shape: convex >white bacteria appeared “fuzzy”. Shape described as: umbonate >10^-3 and 10^-9 shape appear to be circular >10^-5 appears to be filamentous

Tet (-) [for slides chose: 10^-9] >shape: raised - some were convex >orange, yellow/white bacteria. (white bacteria not similar to white bacteria found in Tet (+)) >10^-3 = punctiform >other plates = circular / irregular

Hay Infusion and Protist Identification

Observation

Previous findings: During the first Hay Infusion observation, in week two, there was a noticeable "swampy" odor along with a thing layer of film. Small amounts of mold were visible along the film and fungi, already on the bench before hay infusion mixture, increased.

Recent findings: There was still a swamp-like odor similar to fertilizer. The film found from the previous week increased (in thickness) along with white mold growing. Some mold was found growing under the layer of film. The berries that were in the jar starting as red berries and were orange during week 3 observation. The leaves and branches under the water appeared to be the same in terms of color a decay/vibrancy while the ones above water were discolored and limp. Water line receded.

Protist Identification

(a) taken from the mold on first (film) layer. [total of 2 found in this section] lead to identification: colored(green/brown), single, motile cells, one flagella =Euglena

colored(green), colony of many, flat, disc-shaped, 16+ cells =Gonium

(b) taken from center of jar, open water [total of 1 found in this section] colored(green), single, motile cells, one flagella =Euglena

(c) taken from under the leaf [total of 1 found in this section] colored(green), single, motile cells, two observed locomotor flagella, cell elongated with narrow posterior =Chilomonas

(d) taken from area under the bench - closest to mold & fungi piece [total of 1 found in this section] colorless single, motile cells, creeping (sliding), -no visible rays, but assumed =

Transect section 3

Aerial Diagram File:HooksTransectDrawing.pdf

Abiotic and Biotic factors

Biotic: (1)Gnat/Spider (2) Berries (3) Squirrels (4) Birds (5) Trees

Abiotic: (1) Rock (2) Cigarette (3) Candy wrapper (4) Bench (5) Lamp post

Description of Transect The transect section given was section 3. Within the post in the area, many habitats were discovered, one being a birds nest. On the same tree, there was a sign attached reminding people to respect the area. In a near row of trees and shrubs was a row of fallen berries that had an unpleasant scent. Along the first turn of the sidewalk was a bench made out of wood that seemed to be rotting and around the whole transect was plenty of trash of candy wrappers and cigarettes.