User:Jennifer G. Desimone/Notebook/Biology 210 at AU

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Lab 6 Embryology & Zebrafish Development 3/26/15 Purpose: The purpose of this lab was to study how environmental conditions affect embryonic development in zebrafish. For our experiment, we tested the effects of alcohol on embryonic development. This experiment with the zebrafish could also be used as a model for study in other organisms, like humans who have fetal alcohol syndrome.

Methods and Material: We used 1.5% ethanol as the environmental condition in our experiment. 20 zebrafish were placed in a petri dish with 20 mL distilled water for a control group and 20 zebrafish were placed in a separate petri dish filled with 20 mL of 1.5% ethanol. These zebrafish were observed over a course of 14 days, with observations on day 1, 4, 7, and 14. The observations made were body size and length, eye diameter and movement, heart rate, pectoral fin size, mouth development, and general movement. Also the stage the fish were in was also observed and how many had survived. At day 4, 10 mL of water was removed and 10 mL added and dead embryos were removed. At day 7 and day 11, 10 mL of water was added and 10 mL of ethanol was added and dead embryos were removed. One drop of food was added on each observation day.

Data and Observations: On day 1, all 20 zebrafish embryos were alive and growing. They were in the stage 20-somites. On day 4, 18 fish were alive in the control group and only 12 in the alcohol group. The control group was in the protruding mouth stage and the experimental group was in the pec-fin stage. The control group was slightly bigger at 0.4 cm and the experimental group was only 0.3 cm. The fish exposed to ethanol were more jittery and swam very fast in bursts. On day 7, only 2 fish were alive in the control but 8 were still alive in the experimental. The 2 fish that were alive were still further along in development than the experimental. The fish in ethanol also had an increased body cavity and had obvious physical abnormalities. On day 12, all 40 fish were dead.

Conclusions: The result of the experiment support my hypothesis that if the fish were exposed to 1.5% ethanol, then they would have physical abnormalities. The control group did develop more fully and the fish exposed to ethanol had different eyes, with jittery movement, and a very large body cavity. Although the experimental group died by day 14, so did the control group. By day 7, there were more fish in ethanol than there were in water. This was not what I had expected. There may have been disruption from another lab section. When we did observations on day 7, our petri dish had been cracked for the control group. Both petri dishes were also not found in the spot where we had put them after observations on day 4. Human error could have been a huge factor in this study.


4.8.15 A data table and images would improve this entry. Missing 16S sequencing data for Transect study. SK

2.24.15 Good start. Need more detail in data and methods. Missing vertebrate results, discussion and food web. SK

Lab 5 Invertebrates 2/19/15 Purpose: The purpose of this lab was the understand the importance of invertebrates and to learn how simple systems evolved into more complex systems.

Methods and Materials: First, 3 different invertebrates were observed; acoelomates, pseudocoelomates, and coelomates. After this, the invertebrates were collected from our Berlese Funnel, which contained a 50:50 ethanol/water solution that was set under a light with a flask to collect water. The funnel was broken down and liquid and organisms were poured onto a petri dish and observed under a dissecting microscope.

Data and Observations: The Planaria (acoelomate) was tiny and curled and extended alone the side of the jar. It had a head and moved like a snake. The nematode was in a elongated S shape and extended and crawled slowly the move. The annelida was accordian like, it was flexible and curled up the stretched out to move. We identified 4 different kinds of invertebrates. The first was a springtail. It was a smaller than 1 mm and had clearish body with legs and antennas. We found 2 of these in our sample. We also found a pillbug, soilmite, and proturan. The biggest and most visible was the proturan. It was long and black with antennas. It was about 2 mm long. The soilmite was by far the smallest and the hardest to spot. It was black as well.

Conclusions: We found our transect is home to multiple different invertebrates in our transect and there are probably much more not observed. All of these organisms and plants come together and form an ecosystem. These organisms must have competition for food since some of them have similar diets which results in survival of the fittest.


2.19.15 Good entry, contained all most of the relevant information. A data table like the one from the manual would have improved the clarity of the data. SK

Lab 4 Plantae and Fungi 2/12/15

Purpose: The purpose of this lab was to understand the characteristics and diversity of plants and to appreciate the function and importance of fungi.

Methods and Materials: We walked to our transect near Cassell and Hughes Hall and collected five plant samples from our transect and a leaf litter in a separate ziploc bag. We collected berries, 2 different kinds of leaves, tree bark, and moss. We examined the five samples and described their size and shape, vascularization, specialized structures, and mechanisms of reproduction. After observing the plants, we set up a Berlese funnel using the leaf litter we collected from our transect and 50:50 ethanol/ water solution, and set it under a light fixture with a flask under it to collect the funneled water.

Data and Observations:

Berries: The berries were round and red and there were a bunch of them connected to one stick. We collected them off of a small tree. They were about .6 cm and fall under gymnosperms seed plants. We identified the vascularization of the berries to be xylem and pholem, which transport water and nutrients through a stem. The berries were determined to be dicot because it had two cotyledon when split apart.

Moss: The moss was found on a rock and was green and grassy on top of soil in large patches. The sample was about 1.2 cm in height. It falls under non-vascular plants, more specifically bryophyta. The vascularization was determined to be rhizoid in order to obtain water from their environment. The moss was also monocot.

Tree bark: The tree bark was tan and very stiff in a cylinder shape. It would found on the ground under a tree, most likely the one it was from. It was about 15.5 cm long. The vascularization was determined to be xylem and phloem and it was determined to be dicot.

Leaf: The leaf was brown and very dry and crunchy. It was about 5 cm long and was found on the ground in a pile of other leaves of the same type. The vascularization was xylem and phloem. The leaf was dicot because it has a network of veins.

Long leaf: The long leaf was found on the ground as well but in a different area as the other leaf. It was long, thing, and green. It curled a little but it the majority was straight, it was about 35.5 cm. The vascularization was xylem and phloem and it was monocot because it has parallel veins.

The fungi sporangia are important because they contain the spores. The spores lead to alternation of generations and allows the plant to reproduce with haploid cells. The fungi observed under the microscope was a mushroom.

Conclusions: Our transect is home to many diverse plants and fungus. The different features of each individual plant gives variation for their functions and the way they obtain nutrients and water.


Lab 3 Microbiology and Identifying Bacteria with DNA Sequences 2/11/15

Purpose: The purposes of this lab were to understand the characteristics of bacteria, observe antibiotic resistance, and understand how DNA sequences are used to identify species.

Methods and Materials: The growth on the agar plates were observed to see if any bacteria or possibly fungi had grown on the surface of the agar and the colonies were counted. A wet mount was made of three types of bacteria from our agar plates without tetracycline because no bacteria grew in the presence of tetracycline on our plates. The slides were observed under 10X and then 40X to get a description of the cell. After this gram stain slides were made in order to see if the bacteria was gram positive or gram negative. Over a staining tray each slide was smeared with crystal violet for one minute then washed off with a wash bottle filled with water. Then the three slides were smeared with Gram's Iodine mordant for one minute and rinsed with water as well. Then the slides were decolorized by pouring 95% alcohol for 10-20 minutes and rinsed with water. The smear was finally covered in safranin stain for 20-30 seconds and then washed with water. The slide was then observed under a microscope without a coverslip at low magnification at first, then under 40X and the 100X oil immersion objectives.

Data and Observations: In our Hay Infusion Culture the water was very dark and did not smell as bad as the previous week. Our agar plates only had bacteria on the plates without tetracycline indicating no different strands survived the antibiotics. There were three different types of bacteria. Our table for the amount of colonies counted is shown below:

Bacteria Colony Data: If the bacteria was gram positive, then it would be purple and if it was gram negative it would be pink. The gram stain sticks to a thick cell wall, so if the bacteria has a thick cell wall then it is gram positive. The bacteria from colony 1 was determined to be Coccobacillus. They were a very pale yellow or peach color and round. They were also gram negative. The bacteria from colony 2 were smaller but still a pale yellow. These were slightly rod shaped, gram positive, and determined to be Fusobacterium. The bacteria from colony 3 were more orange and round. We did not draw a conclusion as to what type of bacteria they were or if they were gram positive or gram negative.

Conclusions: Antibiotic resistance was observed in several other groups and three different types of bacteria from the same culture were observed from our own transect. The three different types of bacteria shows that bacteria are extremely diverse and come from multiple different niches within a similar environment.


Lab 2 Identifying Algae and Protists

Purpose: The purposes of this lab were to practice using a dichotomous key, understand characteristics of algae and protists, and examine algae and protists from my transect.

Methods and Materials: After obtaining our Hay Infusion Cultures that had been sitting for a week, we took samples from two different niches in the jar, one from the bottom in the soil, and from the top in the foggy film. A wet mount was made for both niches and three organisms were observed using a microscope and dichotomous key from each niche.

Data and Observations: Our Hay Infusion Culture smelled extremely awful, similar to a sewer. It was very dirty and the dirt and twigs floated to the bottom while the leaves floated to the top accompanied by a foamy film on the top of the water. It was also very foggy. We observed 6 different types of protists. Three were from the bottom of the Hay Infusion Culture and three were from the top of the culture. The three protists observed at the surface were concluded t o be paramecium aurella, colpidium, and paramecium bursaria. The paramecium aurella was about 150 μm and was long and skinny. The colpidium was about 70 μm and was almost crescent shaped. This protist would move slowly, but then have a burst of energy and move very fast. The last one on the surface, paramecium bursaria, was about 70 μm as well and was very fast. This protist moved by using cilia. On the bottom, the three protists we observed were ameoba, chlamydomonas, and arcella. The ameoba was very small, only 12.5 μm and was not a consistent shape. The chlamydomonas was about 5 μm and was circular and colorless. It was found swimming around. The last was arcella, it was the largest at about 40 μm and did not move very fast. Serial dilutions of the bacteria samples were prepared for the next lab.

Conclusions: The organisms observe differed because of the environment from which they lived in. If the Hay Infusion were to grow for another two months then amount of organisms would decrease significantly. As the amount of resources diminish so will the organisms, Just as natural selection would support, the organisms that are more fit, or bigger and faster, would survive longer and the organisms that are not as fit would die off and the amount of different organisms in the culture would decrease.

-JD 2/5/15

Lab 1 Biological Life at AU 1/15/15

Purpose: The objectives for this lab were to understand how natural selection drives the evolution of different species and to make observations about the characteristics of a niche.

Data and Observations: Our transect was located on north campus near Hughes and Cassell Hall. It was a 20 by 20 square consisting of abiotic and biotic factors. Some of the abiotic factors were a bench, a sign, rocks, dead leaves and metal; biotic factors included insects, trees, fungus, moss, and grass. In our 20 by 20 square there was a rock path leading to a bench and 1 large tree accompanied by little trees and bushes. A metal sign stood at the front of the transect and dead leaves covered the majority of the ground along with moss.

Methods and Materials: Before the lab we were given a 50 mL conical tube. At our transect we collected samples of dirt, leaves, grass, and twigs in the tube to observe later for research. Back in the lab we used the tube to make a Hay Infusion Culture. We placed 10 grams of the soil and plants of our sample in the tube in a plastic jar filled with 500 mL of deer park water and added 0.1 gm of dried milk, then shook it for 10 seconds. The Hay Infusion Culture were left to sit for further observation for future labs.

Conclusions: The Hay Infusion Culture contains multiple different niches and looks like it would contain many types of bacteria. -JD 2/5/15