Wartburg College is a selective four-year liberal arts college of the Evangelical Lutheran Church in America located in Waverly, Iowa. Wartburg West is in Denver, Colorado.
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Buccal Epithelial Cells with Adherent Bacterial Cells Revealed by Phase Contrast and Fluorescent Staining
Kim Eschweiler, Beth Graham, Andy Koester, Tabitha Peterson
BI 305 Winter Term 2003
Introduction: The purpose of this project was to observe the adherence of bacterial cells to buccal epithelial cells of the mouth. Our hypothesis was that the buccal epithelial cells are viable even with the adherence of bacterial cells.
Methods: The wall of a cheek was scraped with a toothpick and the cells were placed in a 0.9%NaCl solution. Cells were then stained with DAPI and nigrosine and observed by light and fluorescence microscopy. Figure 1a. Fluorescence
Cheek cell viewed at 400X with DAPI stain. Figure 1b. Phase contrast
Cheek cell viewed at 400X . Figure 2. Fluorescence
Another cheek cell viewed at 400X with DAPI. Figure 3. Fluorescence
DAPI stained cheek cell viewed through filter at 400X. Figure 4. Light Microscopy
Cheek cell with bacteria present on and around it at 400X using nigrosine stain. Figure 5a. Fluorescence
Cheek cell show attached bacteria viewed at 400X . Nigrosine stained. Figure 5b. Fluorescence
Same cheek cell viewed at 1000X showing microcolonies of bacteria. Conclusions:
Bacteria do adhere to buccal epithelial cells.
Bacterial cells were not present on all buccal epithelial cells.
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The Effect of Phosphate on Biofilm Algal Numbers
By Riley Lehman, Ryan Martin, Adriana McMullan, Shelly Meyer, Jacob Phillips & Josiah Polito Abstract
After hypothesizing that doubling the phosphate concentration will double the algae numbers, we filled two containers full of water from the Avenue of the Saints pond. For four weeks we sampled the microcosms, centrifuged, and then counted the algae. We also doubled the amount of phosphate in the experimental container each week. Introduction
Phosphate is a very important nutrient that limits the productivity of many algae in freshwater and marine environments. In fact, it is important to have a large supply of phosphorus because the phytoplanktons, kinds of algae, form the base of the food web. These species are responsible for half the photosynthesis on earth, they remove almost as much carbon dioxide from the atmosphere as all land plants, and supply half the oxygen we breathe. We tested our microcosm to see if the added concentration of phosphate was helping the algae grow. Materials and Methods
Our microcosms consisted of two seventeen liter plastic containers each filled with 15 L of water taken from the samples of water in the 45-gallon garbage cans using a one liter graduated cylinder. One container was labeled “Experimental” and the other “Control.” The two containers were stored in the Biology lab’s greenhouse. After placing the two containers in the greenhouse, eight tiles were first cleaned and then...
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Figure 2: Carbon Filter at 40x stained with DAPI. This DAPI test indicates the total count stained and the blue color indicates DAPI stained DNA. This slide shows a possible fungal presence indicated by the long fibrous strands similar to hyphae. Figure 1: Carbon Filter at 40x stained with DAPI. This DAPI test indicates the total count stained and the blue color indicates DAPI stained DNA. It is evident that the amount of microorganisms is smaller compared to the polyfiber filter and shows less diversity. Figure 4: Polyfiber Filter at 40x stained with DAPI. This DAPI test indicates the total count stained and the blue color indicates DAPI stained DNA. This is another example of greater amount and larger diversity of microorganisms found in the polyfiber filter. Figure 3: Polyfiber Filter at 40x stained with DAPI. This DAPI test indicates the total count stained and the blue color indicates DAPI stained DNA. The yellow areas could be from dying or new microorganisms with less DNA. This slide indicates a larger amount and greater diversity of microorganisms were isolated from the polyfiber filter.
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Feb. 17, 2004
Feb. 17, 2004
Exam review
Intro to major teamed writing project
Overview of types of reports & proposals
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A Presentation of the
Ministry on Aging Work Group
Northeastern Iowa Synod
of the ELCA
September 2007
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Thinking Ahead:
“End of Life Considerations”
A Presentation for Senior Adults and their Adult Family Members
A Presentation of the
Ministry on Aging Work Group
Northeastern Iowa Synod
of the ELCA
September 2009
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Lurking Oral Cavity Microbes
Teddy Annang, Lacey Ebert, Hannah Goldammer, Belarmino Hafermann Introduction
There are numerous microbial species lurking in the human mouth, commonly referred to as plaque. Plaque can be either beneficial or detrimental. Dental plaque is a natural flora in the mouth that can lead to oral diseases, tooth loss, discomfort or mortality. Dental plaque build up in the mouth can lead to infections elsewhere in the body. However, by populating the mouth it helps defend the host against the colonization of foreign species.
By not brushing the teeth regularly an exponential increase of bacteria can occur. Using biofilms, the objective would be to observe this population increase. Materials and Methods
To conduct the experiment, plaque samples were scraped from the molars of a volunteer’s oral cavity, obtained four times at different intervals. The first sample was obtained immediately after brushing and flossing of the teeth to serve as the control. The second sample was taken after fasting for six hours. Another sample was taken an hour after one meal, followed by a final sample taken 24 hours after the first sample.
With each of the samples we inoculated 2 wells of a 12 well plate containing 5mls of TSB with 10% sucrose and a coverslip. Each sample plate was then incubated at 35ºC for 24 hours.
A LIVE/DEAD staining technique was used, as seen in the Biofilm Preparation for Staining handout, to compare live and dead bacteria from the biofilm.
Co...
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400x 100x 200x A B C Epithelial cell Epithelial cell Attached bacteria Figure 1 E. coli attachment to human bladder epithelial cells. (A) After two hours, 29214 has begun to stick to the tissue cells. (B) 700417 floods the field as the bladder cells have deteriorated 18 hours after inoculation. (C) After 29 hours, stock E. coli congregate and attach on a bladder cell; this strain does not appear as pathogenic because the tissue cells seem healthier at 29 hours when compared to those in Fig 1(B). Biofilm Formation and Adherence of
Uropathogenic E. coli Jeffrey Voreis, Steven Nus & Adam Ostendorf
Winter 2004, BI 412 Introduction
Biofilms are a group of sessile bacteria attached to a surface. They secrete a slime matrix which surrounds and protects them. Chemical signals accumulate and trigger gene expression which may initialize biofilm production. Growing these films is difficult in the laboratory because they form very slowly.
Sessile bacteria secrete antigens and stimulate antibody production. Biofilms facilitate survival of bacteria because they protect against antimicrobial attacks due to the thick layer of slime and its slow diffusion rate. Furthermore, limited nutrition leads to a metabolic rate reduction that also protects the bacteria encased in a biofilm by reducing their susceptibility to metabolic attacks. By using a biofilm for protection, bacteria may infect surrounding tissues while avoiding attack.
Uropathenogenic Escherichia coli...
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What You’re Swimming With Matt Fox and Erin Wright
What You’re Swimming With Matt Fox and Erin Wright
Materials and Methods: Two- 20 ml samples were taken from the filter and wall of a hot tub. 2 ml of each sample were mixed with 10 ml of TSB media with 5% glucose and 5% sucrose and placed into a Petri dishes containing coverslips to provide a surface for biofilm growth. The dishes were then incubated for 24 hours at 35C on a Big Bill Thermolyne® rotary shaker. The two samples were also inoculated onto TSA plates in order to grow colonies for the following day.
The colonies grown from the filter sample and from the wall sample were used to inoculate plates of mannitol salt agar, blood agar, starch, pseudomonas isolate agar, eosine methylene blue, and spirit blue lipid hydrolysis. Dilutions of 10-2 to 10-9 were made by scraping coverslips of each sample to determine the number of organisms on the 4.4 cm2 coverslip.
Coverslips were then stained with DAPI and fluorescent gram stain was completed (Molecular Probes, Biofilm Preparation for Staining, 2004). Results: Table 1: Agar plate results from Isolate # 1 and 2 from bacterial growth from the filter sample. The biochemical tests indicate that Isolate #1 is likely a Pseudomonas. It grew very well on PIA and Blood agar showing that it could be harmful. Isolate #2 does not appear to be harmful from the biochemical tests run. Introduction:
A report released from the Centers for Disease Control proposed that hot tubs are not as harmless as they appear. The article talks about outbreaks of pseudomon...
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Biofilm Formation and Adherence of Uropathogenic E. coli Steven Nus, Adam Ostendorf & Jeffrey Voreis Winter 2004 Figure 2 After 8 hours appeared like this picture of strain 700417. 400x 100x 200x A B C Epithelial cell Epithelial cell Attached bacteria Figure 4 E. coli attachment to human bladder epithelial cells. (A) After two hours, 29214 has begun to stick to the tissue cells. (B) 700417 floods the field as the bladder cells have deteriorated 18 hours after inoculation. (C) After 29 hours, stock E. coli congregate and attach on a bladder cell; this strain does not appear as pathogenic because the tissue cells seem healthier at 29 hours when compared to those in Fig 1(B).
Over the course of a 48 hour trial, the three strains of E. coli displayed varying degrees of adherence and pathogenicity. After examining the fields on all slides, we could determine little to no difference in adherence to epithelial cells. The three strains attached to the tissue cells, but they also stuck to the glass slide despite rinsing multiple times with PBS while rocking. The adherence to the cells may have been due to the expression of type 1-pili. An extracellular layer of proteins and biofilms may have facilitated adhesion to the slide.
The tissue cells began degrading five hours after inoculation, yet those with the stock strain did not seem as severely damaged. The control cells appeared normal at the end of the trial, so we believe the uropathogenic strains exh...
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