The two bacteria I have are (B.cereus) (E.aerogenes). Goals:1.To isolate and identify your bacteria.2.To use the scientific method to reason through a series of questions (tests) and answers (results) to arrive at your conclusions.3.To write your dichotomous chart for your unknowns. 4. Write a short summary of the variety of known pathogen activity for both of these organisms. Please, seek two references for the current outbreak(within 10 years), beyond your book to support your finding. How bad are your bugs? Set up: 1. Write Unknowns on the first line. Section One: Your steps to identifying your unknown. This section needs to include step, what was done? Why done (purpose)? And observations, descriptions, and results of test, conclusion-or what were the results that lead to the next step? Why? Then write up the next step, etc…Section Two: Summarize the entire contents of section one in paragraph form for both bacteria.Section Three: Summarize the results in a dichotomous chart similar to the ones in your lab book appendix.Section Four: Write several paragraphs introducing your bacteria as a pathogen as observed elsewhere.EXAMPLE: (using factious organisms)Unknown report name: Tatiana JonesBacteria: Staphylococcus too harder and Klebsiella nowayoutaMy steps:One: Gram stain my unknown both purpose: to observe fresh culture at 24 hr growth, hoping to see both organize observations/results: Gram + cocci seem to be in chains, Gram –cocci or rods?Two: Made a streak isolation platePurpose: to start isolation, hoping to get separate distinct coloniesObservations/results: Where growing, no separate colony formation, but some halo around and beyond streaks and dots within streaks.Thought/conclusions: that one is nonmotile and the other is motile. Three: Did a Gram stain on the halo and where dots were found? Purpose: to see if the growth was-different organism.Observation/result: More Gram –in halo area way from inoculation site and more mixed Gram + and –from dot area on inoculum. Thoughts/conclusion: Gram –must be motile and Gram+ must be nonmotile. Four (+): Followed up with Gram +, by inoculating an MSA plate.Purpose: Mannitol Salt Agar has a high salt and mannitol sugar-more selective forGram + bacteria. Observations/results: Increased growth was seen on MSA, media turned yellow.Thoughts/conclusion: probably Gram +, can tolerate high salt and use mannitol as a carbohydrate source. Fifth (+): Gram stain a couple of growth spots from MSA. Purpose: to confirm or disprove the isolation of Gram +.Observation: mainly Gram + cocci, now in staph groups.Thought: mostly isolated organisms, probably in the Staph family.Sixth(+): Inoculated a PEA-blood agar plate or another MSA. Purpose: to isolate Staph completely, PEA-blood agar inhibits Gram –growth. Observation: Growth on PEA-blood, did not hemolyze blood (gamma hemolytic).Thought: Definitely not Strep, most likely Staph.-Gram stain.Seventh(+): Ran a catalase test on culture in MSA. Purpose: to see if this Staph? Can detoxify hydrogen peroxide to water and oxygen?Observation: yes –positive.Thought: yes this Staph has the catalase enzyme.Most likely: Due to MSA results and catalase, did not hemolyze blood, Gram + staph may have either Gram + Staphylococcus aureus Staphylococcus too harder.Eighth(+): Ran an oxidase test on the organism on the MSA plate. Purpose: to see if this organism has cytochrome oxidase to distinguish between aureus or too harder. (found test results on internet at firstname.lastname@example.orgObservation: negative for oxidase, no black on Q tipConclusion: The organism lacks cytochrome c, so does fermentation or oxidative phosphorylation without cytochrome c. I must have staphylococcus too harder.STEPFour Gr-: etc…-repeat steps relevant to Gr-bacteria if your Gram staining showed negative results. Summary: I was given culture with both a Gram + cocci and a Gram –rod. After several tests and attempts at isolating these into two separate cultures, I believe that I did isolate them both. The Gram + species (strain) atPEA-Blood selection media step, all could see is Gram-positive cocci. I believe that the Gram-positive cocci are Staphylococci too harder because it is definitely staph, by the morphology of gram stains and gram + coccidia gram staining. This staphylococcus can tolerate high salts, utilize mannitol as a carbohydrate source, does not hemolyze RBC, has the catalase enzyme but not the oxidase enzyme. For the Gram -…. That is all gram –rods. Unknown Project Requirements:1.You are given names of two bacteria: one Gram+, and one Gram-microorganism. You have to describe for each microorganism what steps must be taken for identification. First, you need to describe Gram staining for each. 2. Starting with the first Gram staining (see above) you need to describe the possible results of every test and every staining that you would perform during the identification of this particular microorganism. The description can be in a form of drawing, photographic pictures, tables, or simply a description of your expected results. 3. Every test that you describe must have a logical explanation as to WHY you need to do it, WHAT you would see as a result of the test.4.After outlining the results of your tests, you need to conduct Flow Chat (dichotomous chart)for your unknown bacteria clearly showing what tests you performed and why narrowing down the list of possible microorganisms in each group.5.After completing Flow Chat for yourGram+ and Gram-microorganisms, you need to write a report based on the proposed tests for both, Gram+ and Gram-bacteria, confirming your conclusions by literature references (textbook and lab manual are included). 6. You need to conclude Unknown Project Report by brief Summary describing specific characteristics of both, Gram+ and Gram-, microorganisms that you identified, highlighting potential pathogenic activity for both of these organisms, unique characteristics, and other specific features. 7. As you write your Unknown Report, you need to use at least three appropriate sources of references from the scientific literature database. Do not use your textbook to support your findings. You can use the list of Journals that are posted for Photo Quiz, or other relevant peer-reviewed Journals. Copying and pasting Journal’s URL is not an appropriate reference format. You need to arrange your references using EXACTLY the same format as you find in one of the Journals listed, or in the Journal that you choose to use. Unknown ProjectEvaluation Criteria (100% total Project’s grade):1.Ability to follow logical steps proposing tests for bacterial identification –20%2.Logical and conclusive Flow Chat –20%3.Supporting data presentation –20%4.Ability to follow the rubrics –15%5.Logical and science-based explanations of every step and tests performed–15%6.Support of the statements and conclusions by cited literature –5%7.Literature references made inappropriate format –5%_________________________________________TOTAL 100%. Guidelines for Students’ PapersThe goal of analyzing research papers and writing your own paper based on that analysis, is:To attain the ability to comprehend and analyze research literature;To use literature search tools to generate a pool of reliable references to support your point of view;To conduct research papers and to appreciate differences between the structure of research papers that are based on authors’ own experimental results and review papers that are based on up-to-date literature analysis. Sections of Students’ Paper.1.Abstract.Write no more than 4 –5 sentences:highlight the importance of the topic of your paper;very briefly describe what type of study or experimental approaches have been used to test a proposed hypothesis (include a very limited description of hypothesis or authors points of view on the topic);make a brief statement about the results that authors acquired and whether or not you agree with author’s conclusions.2.Introduction.Write 3 –5 sentences for each paragraph listed below:what so far is known about the microorganism of your paper’s topic;why authors decided to find new approaches in diagnosis or treatment of given microorganism;what was authors’ hypothesis (detailed explanation of proposed hypothesis and its possible outcome as to the improvement of treatment, understanding of the disease pathway, or development new diagnostic techniques); why authors chose the methods that they used for experiments;what have authors shown by their experiments (has been hypothesis proven? were results of experiments applicable to practical needs of biomedical science?).3. Materials and Methods. Describe study subjects or objects (patients, animal models, cells), list all groups of the subjects or objects used in experiments. Describe experimental design (treatment, challenges, techniques that have been used to assess the changes during experimental procedures). Practically you will be copying this part from the original paper, however, it is important to describe everything in your own words applying your own understanding of the methods used.4.Results.Describe what results in authors derived from performed experiments. This part you will also copy from the original paper, however, describe the results in your own words and make your comments (1-2 sentences) at the end of the result for every performed experiment. 5.Discussion.Write 3 –5 sentences for each paragraph below:did the authors prove their hypothesis?what other experts in the field think/write about the subject of study?state your opinion about the subject of the study, proposed hypothesis, methods of testing the hypothesis, and interpretation of the results.make realistic suggestions about experimental settings and methods. Support every statement that you make in Introduction, Results or Discussionsectionsby cited literature. Use 5 to 10 reliable references (scientific journals, NIH and CDC websites, and textbooks). The references MUST be arranged in the format that was used in the original paper, which you are using to write your paper (the list of references you can find at the end of the paper). To site reference in your text, use the number corresponding to the number of references in your references list. DO NOT “re-use” references! These means do not use the references that authors of the original paper used! Find your own.
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