Using Kanamycin Resistance Bacteria Essay Example for Free

Using Kanamycin Resistance Bacteria Essay Kanamycin is a common antibacterial that interferes with bacterial growth, by inhibiting protein synthesis, and causing the mistranslation of mRNA. Kanamycin is commonly used in chicken feed to keep harmful bacteria from getting into the eggs and producing healthier chickens. Recently reports of severe gastroenteritis have been linked to eating raw or undercooked eggs. This has led to the FDA to look for possible sources of contamination. Scientists have now isolated bacteria from batches of eggs known to cause the illness, and they found that the bacteria are resistant to kanamycin. The contaminated eggs were found to have come from three different chicken farms, Acme, Big AL’s, and Clucky’s chicken farm, that are geographically separate, and are in different states. The scientists also know that there are three different genes responsible for kanamycin resistance, and that these different genes codes for a certain enzyme that alters the kanamycin molecule differently. The enzymes are located between the inner and outer bacterial membranes, and act on the kanamycin after it passes through the outer membrane. The modification of the kanamycin molecule prevents it from being taken up by the inner membrane, preventing it from reaching the ribosomes. Therefore if any bacteria present has one of the three genes for kanamycin resistance, than kanamycin won’t prevent bacterial contamination (Hass C. , Woodward D. , and Ward A. , 2010. ). The purpose of this lab was to determine if there was a shared source of contamination for the three chicken farms, and to make recommendations for steps to prevent further outbreaks. The hypothesis is that all the chicken farms shared the same source of contamination. The guiding questions for the lab are what is the concentration of viable bacteria in the original samples from the three chicken farms? And what is the frequency of resistant bacteria in the original samples? Methods and Materials: This lab is broken up into four different sections. To begin section one of this lab you need to make sure that your lab area is sterile so that there is no contamination of the bacteria. Then each group gets a bacteria sample, and the letter represents which chicken farm the sample came from. Next each group should obtain six plates. Three have kanamycin, and are labeled with a K, and three unlabeled plates. Each group should then put the names of the groups’ member s, date, lab section number, letter of bacteria sample, and label one of each of the three sets of plates, K versus non K, 10-2, 10-4, and 10-6. Then label three, empty, sterile, microtubules with the dilutions, 10-2, 10-4, and 10-6 that will be made. Next using sterile techniques add 990 microliters of water into each microtubule. Afterward mix the bacterial suspension by gently flicking the microtubule, as shown by your TA. Then for each dilution factor, use 10 microliters of the bacterial suspension, and use this as the starting sample to make three-fold serial dilutions. For each dilution factor make sure to keep the bacteria well suspended by flicking the tube before removing each sample, and make sure that a fresh pipette tip is used for each dilution. Then use sterile glass beads to distribute the bacteria evenly on the agar surface of the 10-6 plate by gently swirling the beads in a circular motion. Then using the same set of beads for each plate transfer the beads from 10-6 to 10-4, then 10-2. Each group should then flip the dishes upside down and stack the three dishes together. Lastly tape the stacks together, and label the tape with your group member names, and section number. The plates should be incubated for approximately 24 hours, and then placed in a cold storage room until you are ready to count the colonies (Hass C. , Woodward D. , and Ward A. , 2010. ) For section two of this lab each group will be working as one group with the other groups at your lab bench. To begin you will collect the petri dishes that you prepared before. Remove the tape from the stacks and examine your plates for colonies. Each lab bench will have six tubes containing PCR mix. The orange, blue, and yellow tubes will have primers only, and will have some colonies added to them. The red, green, and pink tubes will have primers with the control plasmid so no colonies will be added to these tubes, as they will be used as positive controls. Second identify and number the antibiotic resistant plates labeled â€Å"K† which have colonies growing on them. Third, use a white pipette tip and dip it into a colony on the plate labeled number one, and dip that into the orange tube, and close the cap. In turn repeat this step using a new pipette tip each time for colonies two and three, in the blue tube, and the yellow tube respectively. Finally load all six tubes into the PCR machine, and you TA will help you run them. While the PCR machine is running each group can begin working on section three of the lab. To begin with each group will look at the bacteria plates, and count the number of colonies. If the colonies are distributed evenly in the plate then you can divide the plate into four quadrants and just count one quadrant and multiply that number by four. However if they are not, you must count all of the colonies. If there is more than 800 colonies on a plate record the number as lawn growth. Finally record the number of colonies for each plate and use these numbers to calculate the concentration of viable bacteria in the original sample, and the frequency of antibiotic resistant bacteria in the sample. In the last section for the lab each group will be using gel electrophoresis to run their bacteria DNA. Each lab bench will make, and run one gel electrophoresis per table. Once the gel is ready to be loaded, load five microliters of PCR DNA ladder into the first well, as a standard. This should be found in a tube in and ice bucket. Next add two microliters of 6x loading dye into the six sample tubes. The dye should be mixed in thoroughly by gently pipetting up and down after adding the dye. Following that you should load fifteen microliters of each sample into the following six wells. Since lane one will have the DNA ladder lane two starts the samples using the orange tube, then the blue, yellow, red, green, and pink tubes go into lanes three, four, five, six, and seven respectively. Once all the samples are loaded turn on the electrophoresis machine, and wait until the bromophenol blue tracking dye has migrated at least half the length of the gel. Lastly using gloves carefully remove the gel and carry it to the UV light box to view, and photograph the gel (Hass C. , Woodward D. , and Ward A. , 2010. ). Results: The results of this experiment show that the farms do not share the same plasmid that carries the antibiotic resistance gene. Table one shows the individual group data for the concentration and frequency of the antibiotic resistant bacteria. Table two shows the overall frequency of antibiotic resistant bacteria for code A which was taken from Acme Farm, for the section. Table three shows the section data for the overall frequency of antibiotic resistant bacteria, for all three farms, and which plasmid corresponds to that bacteria code. The results showed that for code A which was Acme farm, their resistant bacteria carried plasmid A. For code B, Big Al’s, and code C Clucky’s chicken farm, their resistant bacteria carried plasmids B, and C respectively. Figure A shows the gel electrophoresis picture for the bacteria code A. This figure shows that code A does in fact carry the plasmid A. Discussion:  Based on our data we can conclude that the three farms had different sources of contamination because the three farms all had different strands of resistant bacteria, as shown by the gel electrophoresis pictures from each farm. Figure one shows the plasmid that correlates to bacteria code A which came from Acme Farm. Based on the results shown in table 3 we learn that our hypothesis that all three farms shared a contamination source was wrong. The three farms each carry a different plasmid that is resistant to the antibiotic so their contamination sources must be different. The overall trends from this data are that there was an overwhelming amount of bacteria in almost every case for the 10-2 dilution factor, and the frequencies of viable resistant bacteria were low so that means there was not a lot of resistant bacteria. Some possible sources of error were the DNA samples were not placed properly in the gel so the electrophoresis was not as reliable, or a fresh pipette tip was not used for each dilution which would have messed up the dilutions. Additional experiments that can be done are use three different farms from the previous experiment and see if the same results are obtained. Our research was significant because it showed that there was not a common source of bacteria for the farms, and that bacteria can have multiple strands of DNA that could be resistant to an antibiotic. The significance of the guiding questions was to give practice calculating the concentrations and frequencies of bacteria. Doing these calculations also gave us an indication of how reliable or data could be based on the amount of viable specimen. Recommendations for the farms would be to figure out where the bacteria is coming from and find a way to keep it from the chickens, or to use a different antibiotic that has less resistant strands.

Allen Ginsberg Essay -- Biography Biographies Essays

Allen Ginsberg Allen Ginsberg "saw the best minds of his generation destroyed by madness" ("Howl"). He struggled through family conflicts and homosexuality throughout his adolescence, and then he went on to become one of the most read poets of his time. Allen was a strong man who never allowed anything get the best of him, including fear. He made a list of all his fears, large and small, and then worked his way through them, ridding himself of one fear after another (Mitchell 30). His influence on everyone he came in contact with carries on even after his death, and many writers dedicate their time to documenting his life as it affected them. Readers of his poetry say he has "a delicate lyrical style reminiscent of certain seventeenth century poets" (Brinnin 49). Allen Ginsberg, father of the beat generation, was the embodiment of the ideals of personal freedom, nonconformity, and the search for enlightenment. Irwin Allen Ginsberg was born on June 3, 1926 in Newark, New Jersey, and soon after moved to Paterson, New Jersey ("Modern American Poetry"). He was his parent's second child, preceded by one brother, Eugene, who was named after a speaker his father was impressed with as a young child (Miles 30). His father, Louis Ginsberg, was a high school teacher and a moderate Jew Socialist, and Naomi, his mother, was a "radical communist and irrepressible nudist who went tragically insane during early adulthood" ("Literary Kicks"). Naomi grew up speaking Yiddish and learned to play the mandolin when she was young. She went to Barringer high school, which is where she met Louis Ginsberg in 1912, when they were both only seventeen (Miles 12). Often Naomi, who also suffered through recurrent epileptic seizures and a severe form of... ...shes it. Works Cited "Allen Ginsberg." Literary Kicks. Feb 2002. http://www.charm.net/~brooklyn/people/allenginsberg.html. Brinnin, John Malcolm and Bill Read ed. Twentieth Century Poet: American and British (1900-1970). St. Louis: McGraw-Hill Book Co., 1970. Charters, Ann. "Allen Ginsberg's Life." Modern American Poets. Feb. 2002. http://www.english.uiuc.edu/maps/poets/g_1/ginsberg/life.htm. Ginsberg, Allen. "Howl." March 2000. http://www.charm.net/~brooklyn/poems/howl.html. Kramer, Jane. Allen Ginsberg in America. New York: Fromm International Pub., 1997. Miles, Barry. Ginsberg: A Biography. New York: Simon and Schuster, 1989. Mitchell, Adrian. "The Man Who Set Me on Fire." New Statesman April 1997: 30(2) Mitgang, Herbert. Dangerous Dossiers: Exposing the Secret War Against America's Greatest Authors. New York: D.I. Fine, 1988.

Leadership and Employee Morale

Assignment Leadership and Employee Morale Dorothy â€Å"Micki† Gould Kaplan University Organizational Behavior MT302 Professor Rhonda Shannon May 09, 2012 Unit 8 Assignment Leadership and Employee Morale The ten truths, just learning them is not enough. It is crucial to good leadership to apply them. (Kouzes & Posner, 2010) * You make a difference. Believe in yourself. Believe you can make a difference. If you do not believe it, neither will anyone else. * Credibility is the foundation of leadership. Mean what you say and say what you mean.Do not be misleading; be honest and upfront about everything. * Values drive commitment. If you do not know what you stand for, find out. You need to know what you value. * Focusing on the future sets leaders apart. How can you lead if you do not know where you are going? What is the end goal; do not think you are the only person who sees it or that can get you there. * You cannot do it alone. Remember to lead you have to have followers; you cannot lead if you do not include other people in your plans and have a team. Trust rules. Trust and respect, you cannot get either one without giving it. * Challenge is the crucible of greatness. Do not be afraid of change. If it is not working, ask for suggestions and /or look for different ways to implement changes. Make the changes necessary to grow. * Either you lead by example, or you do not lead at all. Do not expect someone or anyone to do something you are not willing to do yourself. This also goes back to credibility. Do as I do not just, as I say. * The best leaders are the best learners.Strive to learn. Going back to challenges and changes, if it is not working, find a new way to do it. Also, remember you can learn from your followers and team members. * Leadership is an affair of the heart. Have passion in what you are doing, or you cannot lead your team members to want what is best. This is my favorite. My job has a saying: Have a Heart H = Help everyone you can E = Enjoy your job and your customers A = Always â€Å"be there† for your co-workers and customers R = Respect everyone, especially the difficult peopleT = Truth is always the answer (Management, 2012) â€Å"All aspects of transformational leadership—are leaders able to motivate followers to perform above expectations and transcend their self-interest for the sake of the organization. Individualized consideration, intellectual stimulation, inspirational motivation, and idealized influence all result in extra effort from workers, higher productivity, higher morale and satisfaction, higher organizational effectiveness, lower turnover, lower absenteeism, and greater organizational adaptability. (Judge & Robbins, 2007, p. 391) References Judge, T. A. , & Robbins, S. T. (2007). Organizational Behavior (14th ed. ). Upper Saddle River, NJ: Pearson Education, Inc. Retrieved March 22, 2012 Kouzes, J. , & Posner, B. (2010, August). Leadership Truths. Leadership Excel lence, 27(8), 15. Retrieved May 11, 2012 Management, B. (2012). Employee Handbook. BMG Employee Handbook. Brundage Management. Retrieved May 12, 2012

Big Brother in 1984 by George Orwell - 1423 Words

Big Brother Big Brother is a term used in the book 1984 by George Orwell. This term is used to describe a person or an organization that gains total control over people’s lives, it exercises complete control by doing things such as, creating a new language, destroying history, taking away rights so the people become powerless and all sources of communication are recorded and stored by â€Å"Big Brother†. George Orwell predicted this would happen in his book 1984 he predicted that we would live in a society in which everything is controlled and monitored. 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