User:Wanjiku Wambaa/Notebook/Biology 210 at AU

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Wanjiku Wambaa

Laurie Stepanek

Invertebrate Lab Report

16 February 2015

                                                                Vertebras in Transect

Introduction/Purpose: The objective of this lab is to identify vertebrates to complete the description of life in my transect. Vertebrates are a monophyletic group of animals with backbones. (Freeman, 2014)

Methods: I made the observations around 10am, in the mid 30’s, and very windy making it feel colder than it actually was. This is when I collects the samples from my transect.

Results: In my transect I found some birds in a close by tree. The two birds that were most-likely there were an American sparrow, which is known as Chordata Aves Passeriformes Emberizidae Spizella arborea by classification and a Chirping sparrow, which is known by classification as Chordata Aves Passeriformes Emberizidae Spizella passerine. I know that the grounds keepers built a fence around the transect so that animals would not just come and eat the vegetation in the AU garden. They were avoiding animals such as the white-tailed deer (Animalia Chordata Mammila Artodactyla Cervidae Odocoileninae virginianus), Eastern gray squirrel (Chordata Mammalia Rodentia Sciuridae Sciurus carolinensis), and the European rabbit (Chordata Mammalia Lagomorpha Leporidae Oryctolagus cuniculus).

Discussion: The worms in the well-nourished soil would be a beneficial biotic factor for the birds around my transect so that they could feed their young. This makes the AU farm a particularly good location for the bird’s survival. Deer’s, rabbits, and squirrels would all be attracted to my transect because of its biotic factors that benefit each of these animals exclusively. The factors would be such as the green plants that are there during the Summer season, the acorns, fruits and vegetables that grow in the Fall, and twigs that are there in the Winter time. Temperature, sunlight, and minerals in the transect are all abiotic factors that can allow growth of organisms in the AU farms so that the green plants and vegetation can grow. When these organisms grow it will allow the bigger animals such as birds, deer’s, squirrels and rabbits to be able to habitat. These organisms seen in figure 1 make up the feeding positions in the food web, which are tropic levels. The leaves, grass, fruits, vegetables, flowers, and other photosynthesizing organisms make up the first trophic level. The herbivores such as the squirrels, rabbits and deer make up the second tropic level, while the third and fourth tropic level are primarily carnivores such as a fox. The carrying capacity is the maximum population size a habitat can sustain given the availability of the biotic and abiotic resources in that environment. (Freeman, 2014). The community in my transect include all of the organisms in figure 1. This includes all of the tropic levels. They work together to create my transect.

SW


http://i280.photobucket.com/albums/kk183/shelbybells300/WP_20150218_0011_zps992406d6.jpg






Wanjiku Wambaa

Laurie Stepanek

Invertebrate Lab Report

16 February 2015

                                                           Invertebrates in transect

Invertebrates are animal species that have not developed a vertebral column and they are a diverse paraphletic group of organisms which contain fourteen different phyla. (Freeman, 2014)

Berlese Funnel: We set up the Berlese Funnel to collect Invertebrates that reside in our transect. In order to do this we pored 25ml of the 50:50 ethanol/water solutions into the 50ml conical tube. Then we fit a piece of plastic screening material into the bottom of the funnel. Tape the sides of the screen, so the leaf litter does not fall into the preservative. We carefully put the leaf litter sample in the top of the funnel. Then the funnel was set on a ring stand so that it is held into the tube with the ethanol. I then parafilm the base of the funnel and the tube so the ethanol will not evaporate. A lighted 40 watt lamp was placed above the funnel with the incandescent bulb about 1-2 inches from the top of the leaf litter. We then covered everything with some foil and left the funnel on the bench for a week.

Acoelomate, Pseudocoelomate, and Coelomate are worms with three different internal structures. The differences in these structures affect their movement. Acoelomate worms have a digestive tract, but not a coelom, which is a fluid filled cavity that holds an animal’s organs. (Freeman, 2014) Because of this they are not able to move fast. Psuedocoelomates on the other hand have an incomplete lined body cavity (coelom). That is why it is called “pseudo” meaning false. (Freeman, 2014) These worms can move faster than the acoelomate worms because there is somewhat of a body cavity there that can help with muscle contraction. Coelomate have fully lined fluid coelom where their internal organs stay (Freeman, 2014). Because they can have their muscles against the coelom they are able to move faster than their acoelomate or pseudocoelomate counter parts. An example of these types of worms are earthworms.

Table 1:

Organism(Pylum and class) Length in mm # in sample Description

Annelida Oligochaeta (Earthworm) 70mm 1 Has lines across the body but has no clear head or tail

Springtail Primitive insect 100mm 2 Long, has antennas and lines across its body

Arthropoda Archnida (Ground Spider) 90mm 1 All of its legs on one side. Big circular body with mouth fangs

Arthropoda Insecta( Fly) 40mm 1 Very lightly colored wings and dark oval brown body


The range of size of organisms that I measured was between 40mm and 100mm. The Springtail Primitive insect was the largest, while the Ground spider was the smallest. The Springtail Primitive was the most common being that we found two. All the other organisms that we found were only one. SW





Wanjiku Wambaa

Laurie Stepanek

Biology 210 section 7

12 February 2015

Plantae and Fungi

Purpose/Introduction: The objective of this lab was to understand the characteristics and diversity of plants, as well as appreciate the function and importance of Fungi. Land plants evolved from Algae and they are called Plantae. Fungi are not Plantae, but make up their own category where they are simply referred to as fungi. Both plants and fungi share the domain of eukaryotes. (Freeman, 2014)

Material and Method: From my transect, I collected different samples and filled up a plastic bag. From that plastic bag we chose 5 samples to which we were going take to the microscope for further observation. During this time we classified and recorded their vascularization, specialized structures, description, and mechanisms of reproduction. We used a ruler to measure the size and spliced or samples open in order to record their vascular systems. We also observed our fungi that we had left over from the previous week’s lab. Next we set up the Berlese Funnel to collect Invertebrates. We used the funnel to put the rest of our plant matter and taped down a screen so the leaf matter does not fall into the preservative. We then pored 25ml of the 50:50 ethanol/water solutions into the 50ml conical tube. The funnel, tube system was placed under a light for a week.

Data and Observations: There were a variety of plants in my transect. My first sample was a brown leaf that must have fallen from a tree several weeks prior to me collecting it. It was a broad leaf with a network of veins. Being that my transect was a farm, they planted all types of vegetables and Brussels sprouts were one of them. I took a single leaf in order to look at it under the microscope. My third sample was a fungus. It was left over from lab 3. Being that all our samples were vascular, we looked at the fungi to compare non-vascular to that which is vascular. My fourth sample was a fallen twig/branch that I had collected from the AU farm that happened to be in one of the plant boxes. We cut in open to observe in structure. My final sample was a carrot, which is a root vegetable that I pick up from my transect. The data I collected from these 5 samples are shown in Table 1.

Table 1:

Transect Sample Plants Location and # in transect Description Vascularization Specialized Structure Mechanism of reproduction


1 On the ground at AU farm. 4 transect Brown leaf 14 1/2cm. 5 main veins Yes, Phloem and Xylem system. Stomata, a network of veins with a deep vein in the center Dicot

2 In a planting box at AU farm Light green brussel sprout2 1/4cm. Round Yes, Vascular bundles scattered Angiosperm Monocot

3 Fungi # 4 On tet plate 10^-7 White. 1/2cm. circular No Vascular system sporangia Not an angiosperm Neither monocot nor dicot. Asexual reproduction

4 Fallen twig/branch at AU farm 4 transect Light brown 12 1/4cm. about an inch think. long Yes, Vascular bundles in ring parallel veins Dicot

5 Carrot growing in plant box. AU farm 5 1/8cm light orange. Yes, Phloem and Xylem system Network of veins Dicot


Conclusion: In regards to fungi, they grow in Hyphae structure. Some hyphae grow upward and form small, black, globelike structures called sporangia. Inside the sporangia are spore cells. This is their means of reproduction when the sporangia open and the spores are released. Under a dissecting microscope, we classified our own fungi to see if which of the three main groups it fits into. The three groups are Zygomycota, Basidiomycota, and Ascomycota. I assumed that my fungi was a Ascomycota because it looked so similar to mold.

References 1. Bentley, Walters-Conte, & Zeller. 2014. Biology 210 Laboratory: The Diversity of Life Lab Manual. American University: Washington. 2. Freeman, Scott. 2014. Biological Science. Prentice Hall: Boston.

SW

Wanjiku Wambaa Laurie Stepanek Section 007 5 February 2015

Introduction/Purpose: The objective of lab 3 is to understand the characteristics of bacteria from examining its growth and both the tetracycline plates and the plates with nutrients that we plated last week. Viewing the tetracycline plates allowed us to see how a bacterium responds to antibiotics. Since tetracycline is an antibiotic there will most likely be little to no growth on those plates. I predict that plates without it will have growth, being that there was a lot of life in our Hay Infusion Culture. The nutrients on the plates will only encourage growth in general, whether it is of bacteria or archaea. In the lab we also did Gram staining of some colonies, where we observed there size, shape, arrangement, and motility. We also did PCR in the lab to help us understand how DNA sequences are used to identify species.

Materials and Methods: We counted the number of colonies on each of the plates we had, then we selected four plates to look at more in depth. I selected Agar 10^-3 and 10^-7 along with Tetracycline 10^-9 and 10^-3. We observed the wet mounts using with a microscope using 10x and 40x. After that, we did a gram stain procedure with the four plates, where we observed different aspects of the colony and depending on whether it stain pink or purple we decide whether it was positive or negative. We also set up a PCR for 16S sequencing. Then we transferred a single colony of bacteria into 100micro liters of water in a sterile tube. This incubated at 100 degrees C for ten minutes in a heat block. We centrifuged it for 5minutes at 13,4000rpm. We then added 20 micro liters of a primer and water mixture to our labeled PCR tube. The final step was transferring the 5micro liter of supernatant liquid from centrifuged samples to the 16S PCR reaction and placing the tube in the PCR machine.

Data and Observations: The first thing I did was observe my Hay Culture. It was the same color. The smell was less strong, but still noticeable. There were also half as much water in our container then there was the previous week.

Table 1: 100-fold Serial Dilutions Results Dilution Agar Type Colonies Counted Conversion Factor Colonies/mL

10^-3 Nutrient 350 X10^3 350,000

10^-5 Nutrient 250 X10^5 250,000

10^-7 Nutrient 120 X10^7 120,000

10^-9 Nutrient 300 X10^9 300,000

10^-3 Nutrient+ tet 8bacteria 1fungi X10^3 9,000

10^-5 Nutrient+ tet 12bacteria 4fungi X10^5 12,000

10^-7 Nutrient+ tet 11bacteria 4fungi X10^7 11,000

10^-9 Nutrient + tet 6bacteria 2fungi X10^9 60,000


Table 2: Bacteria Characterization

Colony Label Plate Type Colony Description Cell Description Gram

1 Agar 10^-3 Round purple/grayish color 4cm diameter Clear, long, clustered with no motility Positive

2 Agar 10^-7 Round, looks like 2 colonies have not yet separated. Yellow. 2cm No motility, colored, long and clustered Negative

3 Tet 10^-9 7cm Round, shiny light yellow No motility, tannish/brown Negative

4 Tet 10^-3 6cm Round orange/yellow Clear, almost sphere, but vary in shape, small clusters, no motility, Negative


Conclusions and Future Directions: While observing the different plates I saw many things that made ones with and without tetracycline stand apart from each other. The first thing that stood out to me was how there were many more colonies on the plates without tetracycline than the one with. This was expected. There was also more diversity in the colonies that were in the plates with only nutrients. One thing that I found interesting is that fungi had grown on all of my plates with tetracycline, but on none of the plates with the nutrients. I would have guessed that if it could grow on the plates with an antibiotic, it would be able to grow on a plate that has more nutrients to encourage bacteria growth.

Image 1 of agar plates http://i280.photobucket.com/albums/kk183/shelbybells300/WP_20150129_0101_zps6a58c2c4.jpg

Image 2 of tet plates http://i280.photobucket.com/albums/kk183/shelbybells300/WP_20150129_0111_zps7084cf3b.jpg

Image 3 of tet pltaes http://i280.photobucket.com/albums/kk183/shelbybells300/WP_20150129_0121_zpsf3daae4f.jpg

Image 4 of tet plates http://i280.photobucket.com/albums/kk183/shelbybells300/WP_20150129_0131_zpsf29b5f5d.jpg

Image 5 of stained plate http://i280.photobucket.com/albums/kk183/shelbybells300/V__5A8F1_zps9c7ffa5b.jpg

Image 6 of stained plate http://i280.photobucket.com/albums/kk183/shelbybells300/V__AED31_zpsbd565ab0.jpg

WW

Wanjiku Wambaa Laurie Stepanek 210-007 Section 7 January 29, 2015

Protist vs Algae

Purpose: They are three Domains: Archaea, Bacteria, and Eukarya. Protists and algae are two of six kingdoms and they are a part of the Eukarya Domain. Protists get nutrients from consuming other organisms, while algae make their own food through photosynthesis (Bentley et. al, 2014). Last week, we collected samples from our transect and made a Hay Infusion Culture. In Lab 2, we are able to examine it. The objective of this lab is to see the differences as well as the similarities between algae and protists. To do this, we also practiced using a Dichotomous Key, which was designed to help identify any group of organisms that was in our transects sample. The Hay culture is an ecosystem with two distinct niches, the surface and the bottom. I hypothesize that organisms that need light for photosynthesizing will be on the surface of the Hay Infusion Culture leaving the organism that do not photosynthesize primarily at the bottom.

Data and Observations: I brought the Hay Infusion Culture back to my table carefully so that I would not disturb the organisms. As I was bringing it, it smelled awful, but I could tell that there was life present. The more I moved the culture the strong the stench became. The water in the culture was dark brown in color and it looked cloudy. There were sticks, dirt, and several other components in it that had both sunk to the bottom and some floating at the top as well. We took looked at our culture under the microscope for a better observation. We looked in both the surface and at the bottom. The areas are all in the same ecosystem, but are at different niches because they are competing for different resources and there is a carrying capacity at each niche (Bentley et. al, 2014). As shown and described in figure 1, I saw an Arcella, Pandorina , Gonium, and Euglena on the surface. Three out of the four organisms that I looked at of the top layer of my culture; Pandorina , Gonium, and Euglena are all algae and photosynthesize ,which makes sense why they would want to be on the surface of the culture to get more sunlight for them to build up more energy. As shown and described in figure 2, I saw another Arcella, an Actinosphaerium, a Gonium, and a Stentor at the bottom. Being that Acella, Actinosphaerium ,and Stentor are all non- photosynthesizing organisms it supports my hypothesis that those organism do not need much sunlight, such as the majority of the organism that were on the surface.

Materials and Methods: I observed the size, shape, movement, and color of the organisms in my culture by preparing a wet slide with a drop from my Hay Infusion Culture. After we observed them we identified them using the dichotomous key. To look at prokaryotic organisms in our Transect we had to inoculate nutrient agar petri dishes. I first shook the Hay Infusion Culture. I had eight labeled tubes of 10mLs sterile broth and I took a micropipettor that was set to 100microliters and added it to the sterile broth. This was a 1:100 dilution. I then swirled the inoculated tube to mix it. To plate serial dilutions, we pipetted 100uL from the tubes and put it onto its corresponding plate. We had four plats of the agar petri dishes with tetracycline and another four without it. We left the dishes at room temperature for the bacteria to divide and multiply.

Conclusions:

It did turn out that the majority of the organisms that I found on the surface were photosynthesizing organism and the organisms at the bottom were not. This supported my hypothesis that the organisms that photosynthesize would be primarily on the top of my culture. 

Figure1: http://i280.photobucket.com/albums/kk183/shelbybells300/WP_20150129_0051_zpsf4b82cb7.jpg

Figure 2: http://i280.photobucket.com/albums/kk183/shelbybells300/WP_20150129_0061_zps34bce487.jpg

References 1. Bentley, Walters-Conte, & Zeller. 2014. Biology 210 Laboratory: The Diversity of Life Lab Manual. American University: Washington. 2. Freeman, Scott. 2014. Biological Science. Prentice Hall: Boston. WW






Wanjiku Wambaa Laurie Stepanek 210-007 Section 7 January 26, 2015

Observing a Transect on American University Campus

Purpose: An ecosystem has both biotic and abiotic components in it and for this lab we aimed to observe the biodiversity in the transect and prepare a Hay Infusion Culture. We do this so that we can characterize the protest, bacteria, plants, and animals in an ecosystem.

Data and Observations: I was in transect number four. The transect was the university garden where we observed and took samples from a 20 by 20 meter dimension of our designated area. The garden was next to the university’s tennis counts at the bottom of a hill. As shown in figure one, the space we focused in on had four boxes all with different things growing in them. Box one had broccoli. Box two and three did not have anything growing in them due to the ice inside of it that had stopped plants from sprouting. Despite the cold weather and ice box four had brussel sprouts. The biotic components that were in our transect were the broccoli, the Brussel sprouts, the grass, the leaves, and the branches. Some abiotic components that I saw was the ice that were in boxes two and three, the soil that was both in and out of the boxes, the planting boxes themselves, some rocks ,and a fence that surrounded the perimeter.

Materials and Methods: We observed the 20 by 20 meter dimensions of the transect that was marked with four Popsicle sticks. Here, I took a sterile 50mL conical tube to collect samples of the ground vegetation. I also collected anything else around that would help me get a good representation of what was in my transect and put it in the tube. Other than soil, in my sample were twigs, leaves, ice, grass, pieces of broccoli and pieces of brussel sprouts. Back in the lab we made a Hay Infusion Culture. To do this we placed 10 to 20 grams of our soil/ground vegetation sample in a plastic jar with 500 mLs of Deerpark water. I added .1 gm of dried milk to the jar and put the lid on so that I could mix the items in the jar gently. We did this for 10 seconds. After mixing I took the lid off of the jar and felt the jar open.

Conclusions and Future Directions: As we continue to monitor the transect, I predict that there will be more biotic organisms. Wanjiku Wambaa Laurie Stepanek 210-007 Section 7 January 29, 2015 Protist vs Algae Purpose: They are three Domains: Archaea, Bacteria, and Eukarya. Protists and algae are two of six kingdoms and they are a part of the Eukarya Domain. Protists get nutrients from consuming other organisms, while algae make their own food through photosynthesis (Bentley et. al, 2014). Last week, we collected samples from our transect and made a Hay Infusion Culture. In Lab 2, we are able to examine it. The objective of this lab is to see the differences as well as the similarities between algae and protists. To do this, we also practiced using a Dichotomous Key, which was designed to help identify any group of organisms that was in our transects sample. The Hay culture is an ecosystem with two distinct niches, the surface and the bottom. I hypothesize that organisms that need light for photosynthesizing will be on the surface of the Hay Infusion Culture leaving the organism that do not photosynthesize primarily at the bottom.

Data and Observations: I brought the Hay Infusion Culture back to my table carefully so that I would not disturb the organisms. As I was bringing it, it smelled awful, but I could tell that there was life present. The more I moved the culture the strong the stench became. The water in the culture was dark brown in color and it looked cloudy. There were sticks, dirt, and several other components in it that had both sunk to the bottom and some floating at the top as well. We took looked at our culture under the microscope for a better observation. We looked in both the surface and at the bottom. The areas are all in the same ecosystem, but are at different niches because they are competing for different resources and there is a carrying capacity at each niche (Bentley et. al, 2014). As shown and described in figure 1, I saw an Arcella, Pandorina , Gonium, and Euglena on the surface. Three out of the four organisms that I looked at of the top layer of my culture; Pandorina , Gonium, and Euglena are all algae and photosynthesize ,which makes sense why they would want to be on the surface of the culture to get more sunlight for them to build up more energy. As shown and described in figure 2, I saw another Arcella, an Actinosphaerium, a Gonium, and a Stentor at the bottom. Being that Acella, Actinosphaerium ,and Stentor are all non- photosynthesizing organisms it supports my hypothesis that those organism do not need much sunlight, such as the majority of the organism that were on the surface. Materials and Methods: I observed the size, shape, movement, and color of the organisms in my culture by preparing a wet slide with a drop from my Hay Infusion Culture. After we observed them we identified them using the dichotomous key. To look at prokaryotic organisms in our Transect we had to inoculate nutrient agar petri dishes. I first shook the Hay Infusion Culture. I had eight labeled tubes of 10mLs sterile broth and I took a micropipettor that was set to 100microliters and added it to the sterile broth. This was a 1:100 dilution. I then swirled the inoculated tube to mix it. To plate serial dilutions, we pipetted 100uL from the tubes and put it onto its corresponding plate. We had four plats of the agar petri dishes with tetracycline and another four without it. We left the dishes at room temperature for the bacteria to divide and multiply. Conclusions:

It did turn out that the majority of the organisms that I found on the surface were photosynthesizing organism and the organisms at the bottom were not. This supported my hypothesis that the organisms that photosynthesize would be primarily on the top of my culture. 

References 1. Bentley, Walters-Conte, & Zeller. 2014. Biology 210 Laboratory: The Diversity of Life Lab Manual. American University: Washington. 2. Freeman, Scott. 2014. Biological Science. Prentice Hall: Boston.

WW


Figure 1: http://openwetware.org/images/c/c2/WP_20150126_002-1-.jpg