User:Philippa J. Hazlewood/Notebook/Biology 210 at AU

Embryology and Zebrafish Development February 25th, 2015

Purpose: This aim of this lab was to demonstrate the different stages of embryonic development across different organisms. In addition, an experiment was set up to explore how environmental conditions affect embryonic development.

Methods: The zygotes of the starfish, the frog and the yolk of an egg were all examined and their embryological features were compared with a human zygote. Then an experiment to determine how constant light or dark affected the development of the zebrafish was prepared. First the zebrafish embryos were observed and their developmental stage was determined. One control group and two test groups were prepared by filling three petri dishes with 20 mL of Deerpark water. 20 healthy translucent embryos were transferred into each dish using a dropper pipet. One dish was kept in a drawer in total darkness, another under a lamp in total light, and a third acted as a control. Every two or three days the eggs were observed and recorded. Empty or dead egg cases and 10 mL of water were removed from the dishes and 25 mL of fresh water was added.

Data and Observations:

Plantae and Fungi February 11th, 2015

Purpose: The purpose of this lab was the gain and understanding of diversity amongst plants and of the role and functionality of fungi.

'Methods: 'The first part of the lab was conducted outdoors at the transect site. First a leaf litter sample was created from the biotic materials in the transect. To do so, about 500g of dead leaves, plant matter, and a little of the top layer of the soil were transferred into a plastic bag. In a separate bag a representative sample was taken by nondestructively collecting small pieces from five different plants. Once back inside the lab the plant matter that was collected was inspected and identified as either monocot or dicot. Elsewhere in the lab fungi was observed under the microscope. Three different types of fungi were observed and classified as of the zygomycota, basidiomycota, or the ascomycota division. The prepare for the following week, a Berlese Funnel was set up using the 500g of dead leaves and plant matter collected at the beginning of the lab.

Data and Observations: It was found that there were both dicot and monocot organisms residing in our transect. The specific observations made are outlined on the chart below.

It was also found through observation that the mushroom is a member of the basidiomycota division, that black bread mold is a member of the ascomycota division, and rhizopus mold is a member of the zygomycota division.

Mushroom:

Rhizopus Mold:

Microbiology and Identifying Bacteria with DNA Sequences February 3rd, 2015

Purpose: The purpose of this experiments was to gain an understanding of the characteristics of bacteria through observing antibiotic resistance. We also began investigating how DNA sequences are used to identify species. It was predicted that the plates that contained tetracycline would host less bacterial growth, or at least fewer varieties of bacteria, because only antibiotic resistant strains of bacteria would be able to thrive in this environment.

Methods: To begin this investigation the agar plates that were prepared the week before were retrieved and observed. A rough total of the number of colonies on each plate was recorded as well as the differences between plates with and without tetracycline. Met mounts were then prepared from bacteria in two colonies from the nutrient agar plate and two from the tetracycline plate. The wet mounts were observed using the 10x and then with a 40x. 4 gram stained slides were then prepared and observed using bacteria from the same colonies. Finally, one bacteria from each plate was selected for future PCR analysis to amplify the 16S rRNA gene in order to accurately identify the bacteria.

Data and Observations: There were clear differences between the tetracycline and the nutrient agar plates. The nutrient agar plate had significantly more distinct bacterial growth than the tetracycline plate.

However, there were difficulties with the lab equipment so the bacteria collected weren't observed past the 10x objective lens so they could not be accurately identified.

Conclusions: It was concluded that the tetracycline definitely impacted and hindered the bacterial growth, however certain strains continued to thrive.

Great job! Make sure you include everything that is on the rubric I posted on Blackboard- the dilution table will be helpful for you when you're writing up the results for your final paper. ML

Identifying Algae and Protists January 27th, 2015

Great job! Same as before, photos would definitely help your notebook. I really like the way you break your notebook into sections, keep it up! ML

Purpose: The purpose of this experiment was to gain experience identifying protists and algae under a microscope by using a dichotomous key and use this skill to identify and organisms that inhabit the hay infusion that was created the week prior. It was predicted that a variety of protists would be found in the hay infusion sample, with the most sophisticated and variety of specimens found near plant matter by the top of the jar because it has the most access to nutrients and oxygen.

Methods: To establish an understanding of the dichotomous key wet mounts of known organisms were prepared and observed with a microscope. After this, two more wet slides were created, one using a sample from the top of the hay infusion culture and one using a sample from the bottom. Three different organisms were located and attempted to be identified from each sample. To prepare for the next weeks lap serial dilutions of the hay infusion culture were prepared and plated. 100 uL of the culture added to a tube with 10mL of sterile broth. 100uL were then taken from that tube and added to another also containing 10 mL of sterile broth. This process was repeated two more times. A 100 uL sample from each tube was plated onto separate nutrient agar plates and placed aside for next weeks lab.

Data and Observations The specimen gave off an extremely rancid swamp like odor. The top of the hay infusion was sealed with a green layer of thick mud like matter and the water had turned brown. The darkest sediment had accumulated on the bottom of the jar in mud like clumps. In the specimen that was derived from the top of the jar contained a variety of organisms, some of which were difficult to identify. The first organism spotted was peranema, a single celled organism 30 um. After that a colony of volvox cells were identified at 400 um. The last organisms spotted in this sample was unidentifiable using the dichotomous key. It changed shape but was generally round and oval containing many organelles and 60 um. It contained cilia and moved around in circles. The sampled the was derived from the bottom of the jar yielded a different variety of organisms. The first to be identified was colpidium, an organisms 50 um large. The second was chlamydomonas a single celled organisms that is 5um. The last final identified organism was gonium at 20 um. Sketches of the identified organisms are pictured below.

Conclusions: It was concluded the the initial predictions were fairly accurate because of the organisms identified on the bottom half of the jar chlamydomonas was found, a protist that is known to be of the most primitive in the volvocine line.

Biological Life at AU January 27th, 2015

Great job on your notebook so far! I really like that you date each entry and clearly differentiate through headings, keep it up! In the future though, including tables and photos are really helpful and will help get your point across not only to me, but to you as well. When you have to pull all this information together for the transect assignment, having coherent photos and tables will help you remember what you did and found. Great job! ML

Purpose: This lab was conducted in multiple parts. The purpose of the first half was to gain further understanding of the mechanisms behind evolution, namely natural selection, by investigating the subtle differences between different organisms in the Volvocine line through a microscope. During the second half, we investigated the biotic and abiotic components in a niche on AU's campus.

Methods: The first half of the experiment was conducted by creating and examining slides of live Chlamydomonas, Gonium, and Volvox cells under the microscope at 10x magnification. For each specimen, the number of cells that were present in the field of view, the size of the colony, whether or not there was specialization of cells happening in the organism, the mechanisms of motility, and if the organisms was Isogamous or Oogamous were all recorded. Crude sketches of the organisms were also taken. For the second part of the experiment I was assigned transect 5 to observe. Pictures and detailed notes of this niche were taken to record the biodiversity of the community living within this ecosystem. Biotic and abiotic components to this niche were recorded and a sample of the soil/ground vegetation was collected in a sterile 50 mL conical tube. To create a hay infusion, 10 grams of the sample were placed in a plastic jar with 500 mL of deer park water and 0.1 gm of dried milk. The jar was mixed gently for 10 seconds and then placed aside in the lab without a lid, for the following weeks experiment.

Data and Observations

Transect 5 was mostly comprised of grassland with a small planted area that spanned from corner D to corner C. Within this section a variety of biotic components were found, including a rose bush, a different variety of bush, ferns, clovers, and grass. Throughout the transect abiotic components were found such as soil, stone, a small corner of a cement wall, snow, wood chips, and fallen leaves.

Conclusions: A variety of living and nonliving components make up the ecosystem of transect 5.