User:Stephani Wax/Notebook/Biology 210 at AU

23/3/14 Lab 6

The purpose of this lab was to observe the effect of fluoride (in my case) on the development of zebrafish embryos. I set up the experiment by putting 20 live zebrafish eggs in a petri dish with regular water, and 20 eggs in a petri dish that had fluoride water (10 mg/liter). A note to make about this lab is that we did not feed the zebrafish and so they died of starvation before we could take measurements on the final day.

Day 1:

Both petri dishes had 20 live healthy zebrafish eggs

Day 7:

At the end of the first week, in the control dish there were two dead eggs and 18 live, hatched zebrafish. I removed the dead eggs and replaced the water in the petri dish with fresh, control water. The dish with fluoride water had four dead eggs and 16 hatched zebrafish, two of which had died and 14 of which were still alive. I removed the dead eggs and the dead zebrafish, and replaced the water with fresh fluoride water. From both of the petri dishes I took three live fish to preserve, putting them in tricaine solution to knock them unconscious so they could be saved in formaldehyde.

Observing the live fish, I noticed some differences between the control and the test fish. The fish from the fluoride water were slower to respond to movement and light, and I could almost touch them with a pipette before they would swim away. The control fish would react much more quickly even to things like having their petri dish picked up or moved, and they were extremely difficult to collect to preserve, while the test fish were much more lethargic and easy to capture. Casual observation also showed a slight discoloration of the test fish, which were still pale and almost clear, compared to the control fish, which had darkened in color and were more easily visible. Yet there were no observable malformations with the control fish or test fish. Viewing the preserved zebrafish under the microscope revealed some definite differences. The most apparent was the deformed eyes of the test zebrafish that had been in the fluoridated water. Their eyes were lumpy, and in one case almost had a split down the middle, so their vision was probably impeded due to this, which could partly be why they were so unresponsive. They also had different tail/head/body length proportions than the control zebrafish. However, when viewing them under microscope, both had similar heartbeats, viewed at roughly a beat per second.

Control Zebrafish:

Eyeballs: 1 micrometer in diameter, round and regular

Initial fins forming

11 micrometer tail

16 micrometer entire length

No visible yolk sac

Greenish brown in color

Test Zebrafish:

Eyeballs: 0.5 micrometers in diameter, lumpy and irregular.

No initial fins

10 micrometer tail

15 micrometer entire length

No visible yolk sac

Light brown in color

When I returned the next week to check on the zebrafish they had for the most part starved to death. Due to their extreme food deprivation, taking measurements on the remaining live fish would have produced inaccurate results, and so no notes were recorded from that day.

SW

23/3/14 Lab 5

The purpose of this lab was to view invertebrates from our transect and also to learn about the different kinds of invertebrates. First we observed three separate types of worms. The first, the platyhelmenthes, were flatworms. They floated on top of the water, but when the jar was picked up they would fall to the bottom and have to climb up the side of the jar to regain the top. The second worm, the pseudo coelomate, was very small and could only be seen under a microscope. The last worm was an annelida, or an earthworm. It was the most complex of the three.

After this, we looked at the invertebrates that were found in our transect. From the top of the Berlese Funnel, we discovered a type of beetle and also what appeared to be a lice or a mite, but was definitely parasitic. From the bottom, we found the same type of beetle and also was looked like a deer tick. Our transect, a wooded area near a dog park, makes sense to have small parasites, because they probably thrive off the presence of dogs.

Invertebrates from Transect:

Area in test tube, Organism #, Description, Species

Top, 1, 6 legs 2 antenna pincer mouth no wings dark brown, Lice or mite looked parasitic

Top, 2, Small dark teardrop shaped 4 legs, Beetle

Bottom, 3, Oval small legs light brown small head w/ pincers, Deer tick

Bottom, 4, Same as #2, Same as #2

SW

23/3/14 Lab 4

This lab was to teach us about the different kinds of plants and their individual characteristics. We made sure to take our five plant samples from different areas of the transect and also from different species of plants and trees, for the most diversity. All of the specific details are recorded in the table.

Plants from Transect:

Sample #, Location, Description, Vascularization, Leaves/special characteristics, Seeds flowers other reproductive parts

1, Under pine tree, Limber pine – bundle of needles, Seeded, Bundles of 5 needles, Cone

2, Under pine tree, Limber pine cone scaled, Seeded, Scales thick at tip, Cone

3, Vine w/ big leaves top half of transect, Big leaves – five curves on each, Stems non-seeded vascular, 5 curves on each leaf one leaf per stem, No flowers but stems

4, Long vine bottom half of transect, Small Leaves, Stems, Lots of leaves pointy 6 leaves per stem, No flowers stems

5, Near the center, Small stick brown, Vascular seeded, 8 protrusions off of stick, From a tree (not the pine)

We also observed fungi under the microscope. When looking through a microscope, the sporangia are visible. Sporangia stick off of the fungi, and are a form of asexual reproduction, making them very important.

We then set up Berlese funnels to collect invertebrates, so we can observe them next week. 

SW

23/3/14 Lab 3

This lab was meant to observe the bacteria that grew in our transects, and whether it was antibiotic resistant or not. We started with another check on the hay infusion culture, which revealed that the bad smell had gotten even stronger since the week before.

There were definite differences between the plates treated with tetracycline and those that had not been treated. The tet+ plates had significantly more of the round, orange bacteria, which had grown with much less frequency on the non treated plates, where other bacteria apparently had more ability for growth and expansion. But it was clearly much more antibiotic resistant, and had taken over all three of the tet+ plates. A few other strains of bacteria had grown on the tet+ plate that was the least diluted, but the orange bacteria was clearly the most hearty antibiotic resistant bacteria from our transect.

We then took three samples to observe under a microscope and to gram stain. We took bacteria from three different plates that all had significantly different appearances. We looked at the orange antibiotic resistant bacteria from the tet+ 10^-5 dilution, and then took one from the 10^-7 and one from the 10^-9 dilutions that had no tetracycline. After observing characteristics we made gram stains of the three. The 10^-9 dilution failed to gram stain, most likely because of some issue during the staining process, but the other two bacteria showed up to be gram negative.

Bacteria from Transect:

Colony Label, Tet+ or NA, Colony Description, # of colonies, Cell description, Gram + or Gram –

10^-7, NA, Wrinkled shape undulate margin umbonate elevation cream/white color, 1, Brownian Bacillum Diplobacilli, Pink – gram negative

10^-9, NA, Circular shape entire margin convex elevation cream/yellow color ,4, Brownian Cocci, Unable to tell

10^-5, Tet+, Irregular shape raised elevation light yellow/orange color, 16, Brownian Spirilum, Pink – gram negative

At the end of this lab, we set up all three bacteria to start PCR preparation for DNA sequence identification next week.

SW

23/3/14 Lab 2

This lab was to view the protists from our transect found in the hay culture infusion. The hay infusion culture smelled strong and terrible. A layer of mold was forming on the top of the liquid, which was brownish. The dirt had settled on the bottom, and on top of the dirt were the leaves and twigs.

My group took samples from two different niches – a plant niche and a dirt niche. We observed four organisms, two from the plant niche and two from the dirt niche. We found chlamydomonas in both of the niches. The chlamydomonas were five micrometers in diameter, and round in shape. They had a darker outline and a slightly see through middle part. In the dirt niche we also found a paramecium, that moved by stretching out and then pulling itself forward, shrinking again. This meant it varied from 1.5 to 7.5 micrometers in length. In the plant niche we also found what looked like a euglena. It was about 30 micrometers, and was more tear drop shaped.

Protists Found in Transect: Niche, Size, Type

Dirt Niche: 1.5 – 7.5 micrometers, (Paramecium?)

5 micrometers, Chlamydomonas

Plant Niche: 5 micrometers, Chlamydomonas

30 micrometers, Euglena

If the hay infusion had been left to sit for another two months, the leaves and twig would most likely have decomposed and mixed in with the dirt on the bottom of the culture. The mold that had started forming would probably have gotten much more plentiful and taken over more of the culture.

Then we set up the serial dilutions for the next week. We made dilutions at concentrations of 10^-3, 10^-5, 10^-7 and 10^-9 for the regular agar plates and concentrations of 10^-3, 10^-5 and 10^-7 for the agar plates treated with tetracycline. We then left these to sit until next class.

SW