Antibiotic Resistance Lab

In the first part of the experiment, we plated donor bacteria, which was chloramphenicol resistant, on a Nal plate. Because no donor bacteria would grow on a Nal plate, this was a way of ensuring that the sample of donor E. Coli bacteria was pure.

A constant/controlled variable is the yeast that is used. This would affect the results because different yeasts could affect the rate of fermentation. This will be controlled by using the same bottle of yeast for all tests.

The focus of this lab was to identify which plasmid (PFG or Pglo) was inserted into the E. coli culture. In order for the E. coli to be transformed, the E.coli must first be made competent. The cells were made competent by incubating the cells in calcium (2+) and then in cold temperatures. A quick heat shot opens pores in the cell membrane and allowed plasmid DNA to enter the cells. The plasmid that was inserted into the E. coli culture had antibiotic-reistance gene and a reporter gene. A reporter gene is a gene that is easy to detect, and for this lab it could be Pglo or PFG. The cells were placed in plates that contained antibiotics to kill any bacteria that did not uptake the plasmid, this is necciary because about 1% of bacteria will be transformed. This allowed only cells that up took the DNA to

There was another area of testing, in which we tested the effectiveness of common household spices on bacteria and yeast. My group tested Paprika, which showed to have no effect on the Serratia. There was a lab-malfunction, in which we tested Serratia twice, which inhibit the results of Paprika on yeast. However, other groups tested different spices/food which showed to be very effective; vinegar and honey (on Serratia), and garlic for both bacteria and yeast.

The first task we had to perform in this lab was diluting an Escherichia coli culture and compare it to the

The two most important sequences on the plasmid that will be used are the ORI bacteria and the antibiotic resistance. The ORI is the origin of the bacteria and is an important part of the bacteria because it does the job of replication. The antibiotic resistance is equally important because it is used to see which bacteria take in the plasmid. In the lab there will be two micro test tubes used. One tube will be labeled +plasmid and the second tube will be labeled -plasmid. Both test tubes will contain the E. coli bacteria and 0.25mL of calcium chloride (CaCl2), but only the +plasmid tube will receive 10pL of pGLO plasmid. There will also be four agar dishes used in this experiment. They will be labeled as follows: LB/Amp+, LB/Amp-, LB+, and LB-. The LB/Amp+ dish will have luria broth, ampicillin, and the pGLO plasmid. The LB/Amp- dish will contain luria broth, ampicillin, and will not contain any plasmid. LB+ also contains luria broth, will not contain ampicillin and will contain the plasmid. The last dish, LB-, will have the luria broth, and will not have neither ampicillin nor the plasmid inserted. The LB/Amp- and LB- dishes act as the control group because they will be used to observe changes that occur without the plasmid. The LB/Amp+ and LB+ will act as the experimental dishes because the plasmid will

To determine how E. coli develops in different environments, the researcher will be investigating how E. coli inhibits different materials in an aqueous environment. During the experiment, the researcher will then calculate the colonies in a 1 centimeter radius in the most dense spot of the petri dishes of each material. Then he will find the averages of the 3 petri dishes of each material and compare with the results of each

Still holding the vial at a 45 degree angle, the swab was inserted into the vial to soak up the S. marcescens for 10 seconds. Partly lifting the lid of one petri dish, shielding it over top like an umbrella, we inoculated S. marcescens onto the agar growth medium in the petri dish. We repeated these steps 2 more times to inoculate all 3 petri dishes. Once all three plates were inoculated with the S. marcescens we removed our forceps from the alcohol. We air dried them, and then passed them through the flam to sterilize them. Using the sterile forceps we placed the c disc in the center of a petri dish and press down lightly to ensure that it stayed. It is then labeled it C, for the control group, 5 for the replicate number, along with the date and time of the class. Again we sterilize the forceps and place the T disc into the next petri plate. It is labeled the same way but with a T for tetracycline. The T disc contained 30ug of the antibiotic. We repeat the sterilizing of the forceps one more time and place the Chl disc on the remaining petri plate and labeled it Chl for chloramphenicol. The Chl disc contained 30ug of the antibiotic. Once all 3 petri dishes are labeled they were placed upside down to prevent water droplets from falling onto the nutrient agar substrate. The last step to the experiment was to clean up. We placed our swabs in a waste beaker, cleaned up our work stations, and then washed our hands with soap and

A. Define the term selectively toxic. Why is it an important feature of antimicrobial agents? Selective toxicity is the ability of a chemical or drug to kill a microorganism without harming its host. Selective toxicity is important to microbial agents because it enables these agents to inhibit or kill a microorganism by interacting with microbial functions or structures different from those of the host thereby showing little or no effect to the host.

The purpose of the investigation is to determine what amount of antibiotic is best against bacterial resistance. The independent variable in the investigation is the amount of the antibiotic. The dependent variable is the bacterial resistance. The control group is the petri dish that receives the lowest amount of the antibiotic solution and the constants are the bacteria and the amount of bacteria used in the investigation. There is an estimate of nearly 600,000 people who die each year around the world because they contracted an infection that was no longer able to be treated because the bacteria became immune to the drugs that were supposed to suppress the infection.

The goal of antimicrobial susceptibility testing is to predict the success or failure of antibiotic therapy. Tests are performed and measure the growth response of an isolated organism to a selected drug or medicine. The results of antimicrobial susceptibility testing ought to be combined with clinical information and skill once choosing the foremost appropriate antibiotic. There are many factors to think about in selecting an appropriate antibiotic for treatment of a bacterial infection. The foremost basic of those is whether the causative organism is prone to a selected antibiotic, and the way much of that antibiotic are necessary to inhibit or kill the organism. Whereas we tend to do

According to the results of the experiment located on Image 1 in the results section, the phenotype of the E Coli demonstrated a change when exposed to the plasmid pGlo. This conclusion disproved the hypothesis. All four agar plates were exposed to LB broth (Luria broth). This broth is the source of food and nutrients E Coli used to grow and divide. As demonstrated by Image 1, there was growth in the in the agar plates -LB, +LB/amp, and +LB/amp/arab. The substantial growth that was demonstrated in the -LB agar plate (estimated at 75-80% growth) was due to this was the standard E Coli without any ampicillin or pGlo present. Moreover, the agar plate for +LB/amp showed a margin of growth. The percentage of growth was estimated at a 45-50% growth and signified this in individual colonies. This plate contained the plasmid pGlo. This plasmid is coded with GFP protein, antibiotic resistance gene, and the araC gene (www.bio-rad.com). Even though the agar plate contained the antibiotic ampicillin which kills the E Coli bacteria, the E Coli still grew because the plasmid pGLO was present. This growth demonstrated that the E Coli bacteria absorbed the pGLO plasmid and revealed a change in the E Coli’s phenotype. There was a margin of growth because the E Coli was subjected to the transformation solution (CaCl2) and the heat shock. A

Regular low doses of antibiotics that aren’t strong enough to destroy or kill bacteria provides a suitable environment for bacteria to develop mechanism of survival or to become resistant. The bacteria acquires ways to fight off antibiotics through hindering antibiotics from reaching their target cells .For example changing the permeability of cell walls or distorting the structure of cells or completely replacing them . So, in brief antibiotic resistance happens whenever an antibiotic losses its ability to effectively kill or control the growth of bacteria.

get sick, antibiotics are the answer. The more times you use a drug, the more

There seems to be an ongoing battle between the evolution of bacteria and modern medicine. This has become a serious problem in hospitals as bacterial resistant infections continue to be on the rise despite the practice of standard precautions that treat all patients as if they have an infectious disease. One of the leading public health threats today is meticillin-resistant Staphylococcus aureus or MRSA. With all of the advancements in medicine, it is hard to believe that we do not have the ability to manufacture new antibiotics that would target these resistive organisms. The problem is not only confined to the United States but affects people globally. According to Bartlett, Gilbert, & Spellberg (2013) this is being considered a