Abby PapariellaSection 7April 13, 2015Unknown #19Unknown A: Family: Micrococcaceae. Genus: Micrococcus. Species: Micrococcus Luteus.Unknown B: Family: Enterobacteriaceae. Genus: Escherichia. Species: Escherichia Coli.Papariella2I. Abstract In this microbiology lab each student was told to choose a random tube containing a bacterium that was unknown to the student. In order to correctly determine the family, genus and species of the bacterium there was a protocol and flowchart that the student must follow. The objective of this lab is for the student to explain, through extended tests and observations, the identity of the unknown bacterium. The experiment began with streaking for isolation, which separated the mixed media into large and small colonies. The colonies where then first tested by Gram staining, which told about their cell wall properties. Unknown A was gram positive and showed as purple, so it was then tested on its ability to grow with or without oxygen by the Fluid Thioglycollate test. The organism was strict aerobe, which means it requires Oxygen to survive. Because of these results, it was then tested for the presence of two enzymes amylase and oligo 1,6 glucosidase using a starch hydrolysis test. Because there was no clearing zone on the test it came back negative. This proves that Unknown #19 A is in the Family Micrococcaceae, the Genus Micrococcus, and the Species Micrococcus Luteus. After the Gram test was performed on Unknown #4B, it showed up as red, so it is Gram negative. Because of this it was then tested for its ability to grow in the presence or without the presence of oxygen, it proved as a facultative anaerobe, which means it can grow with or without oxygen. Next it was tested for its ability to ferment glucose and preform lysine decarboxylation, it was positive for both. Lastly, the SIM testwas performed which tests for sulfur reduction, indole production and motility. It was positive for these properties resulting in Unknown #19 B being in the Family Enterobacteriaceae, the Genus Escherichia and the Species Escherichia Coli. Unknown #19 A is Micrococcus Luteus and Unknown #19 B is Escherichia Coli.Papariella3II. Introduction In the 1660s, an Englishman, Robert Hooke first observed “little structures” that he believed werealive. He observed these through a microscope and led to even further observation by a Dutch linen merchant, Antonie van Leeuwenhoek. Leeuwenhoek, also known as the Father of microbiology, observed many different particles, including linen drapes; to discover these movingthings he called animalcules. Many new ideas were introduced to the world around this time period, such as the introduction of aseptic techniques, by an English surgeon, Joseph Lister. Although these aseptic techniques were very much more limited than what we have today, as theywere only things such as disinfecting your hands, they were a big step in the history of microbiology. Later, in the 1800s many infectious diseases were being identified, due to this antibiotics were being developed. In this same time period, due to the much advancement in microbiology, Hans Christian Gram, discovered Gram staining. This remains the universal basis for, as used in this class, bacterial classification and identification (Cowen, 2012).The recognition and identification make it possible to discover what may be going on with someone by being able to quickly and sufficiently treat them. By knowing some properties about different bacterium one may know what antibiotic or treatment a patient must receive. This may mean saving a life. Which makes the proper identification of organisms be very important. For example, if a patient were infected with something, that if not treated quickly, would cause death. One must know what bacterium it may be and the properties of it in order to know how to effectively treat and save the patient. To identify and unknown bacterium there are many tests, procedures, protocols and observations one must follow. Many microorganisms test are incubated with the bacterium and then quickly show the results (positive or negative) but some must be incubated for some time period, and then be observed. After it is determined whether the unknown bacterium is a Gram positive or negative, there are many different pathways and protocols one should follow.Papariella4The objective of this unknown report is to recall all the tests and protocols learned throughout the course in order to determine a bacterium unknown to the student preforming the tests. The learned process of introducing bacteria into some media and then observing the results and interpreting them are very important in this report. The identification of properties for certainbacterium is very important so that one may know what antibiotic to prescribe or what treatment to take in order to prevent widespread disease or an epidemic from occurring.Papariella5III. Materials and Methods Aseptic technique Aseptic technique refers to being sterile so that it decreases the possibility of those handling the bacteria to be infected and the contamination of culture and surrounding surfaces and objects. When entering the classroom, wipe off bench top and wash hands. When exiting theclassroom, do the same procedure. Gloves must be worn at all times, and when one is finished with the gloves they are disposed in the biohazard can. Closed toed, shoes must always be worn, and hair must be tied back. When placing bacteria on a slide, the inoculating loop must be incinerated before and after, to prevent the spread of the bacteria. Streak for isolationThis method was used in order to get the mixed culture to separate into individual cell colonies on the plate agar. The cells that are isolated will grow into colonies, which will only consist of the original cell type. These isolated colonies will be used in order to proceed with the tests. When preforming this test you will streak the bacteria in a zigzag pattern between the quadrants.Papariella6Gram stainingGram staining is used to determine the cell membrane properties of an organism. This testis very specific and one must pay close attention to the amount of time required for each reagent to stay on before being rinsed. After following the aseptic technique, add a small drop of water toa clean slide. Using a sterilized loop, smear the organism across the slide. The slide must be dry before the next step, which can be done using heat fixing. Apply the primary stain,