Microbiology Lab Final Study Guide
Microbiology Lab Final Study Guide BIOL 2042
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This 6 page Study Guide was uploaded by kirkmathews on Sunday April 24, 2016. The Study Guide belongs to BIOL 2042 at University of North Texas taught by Dr. Kunz in Spring 2016. Since its upload, it has received 19 views. For similar materials see Microbiology Lab in Biology at University of North Texas.
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Date Created: 04/24/16
1. Rhizopus nigricans & Sabouraud media 2. Aspergillus niger 3. Lacto-Phenol Cotton Blue 4. non-motile, non-photosynthetic, chemoheterotrophic 5. Zygomycota, Basidiomycota, Ascomycota, Imperfect Fungi 6. A mutualistic symbiosis between a fungus and either an alga or cyanobactera 7. The fungus provies structure, while the Cyanobacteria/Alga provide carbohydrates via photosynthesis 8. This is Penicilium, commonly found as fruit mold 9. Penicilium 10. Sacchromyces cerevisiae 11. By color and DNA sequencing 12. Chlamydomonas, in the kingdom protists 13. flagella 14. Spirogyra 15. Volvox, Green alga 16. Diatoms, Gold Alga, has a Glassy cell wall made of Cillica 17. Desmid 18. Chlorophyll a 19. Thylakoid 20. This is anabaena, it produces heterocysts that fix nitrogen 21. Gleocapsa, Oscillatoria, Spirulina 22. Paramecium, pellicle 23. Yes, cilia 24. Vortichella, and before it lands on a target, it is motile by cilia 25. Amoeba- Pseudopods to eat and move, and Euglena, move by flagella 26. Trypanosoma, flagella, and it causes African Sleeping Sickness 27. Plasmodium, it is non motile, the infectious stage is the Sporozoite, and it causes Malaria 28. Trichomonas vaginalis, it moves by use of 5 flagella, and it causes Trichomoniasis 29. Streptomyces griseus, Filamentous gram-positive rod 30. The production of a substance such as an antibiotic by one member of the association, this substance is detrimental to the second member of the association. We used water agar due to it’s semi- selectivity, being that water has no nutrients so other bacteria would not grow. 31. In the soil. 32. Geosmin 33. Streptomyces griseus is responsible for over 50% of the antibiotic production on the market today. 34. Non-cellular, obligate intracellular parasite, can contain DNA or RNA as genetic material 35. The capsid 36. The receptors on the cell’s surface, made up of proteins and glycoproteins 37. Bacteriophage 38. A virus that specifically infects E. coli 39. A lawn, B plaque 40. The lytic phase of viruses consists of the virus hijacking the cells machinery and rapidly producing components for new phages. Once the host cell is depleted of all of its resources, the host cell bursts and phage particles are released to infect other bacterial cells. Plaques are clear. 41. The lysogenic phage consists of the virus’ genetic material being incorporated into the host cell’s DNA and replicating for many generations. During times of cellular stress, the virus can remove itself from the genome and enter the lytic cycle. This incorporation relationship is known as lysogen. Plaques are cloudy. 42. Plaque Forming Units Per Dilution Factor 43. In order to demonstrate a virus infecting an E. coli 44. The viruses were infecting E. Coli 45. In order to create a lawn 46. They are facultative anaerobic, gram-negative, lactose-fermenting rods. 47. Counting bacteria using Dry Weight, or Turbidity 48. 30-300 49. Methylene blue 50. Once the facultative anaerobes use all the oxygen in the solution, Methylene blue acts as the final electron acceptor and turns white in a matter of time proportional to the amount of bacteria per mL. 51. Membrane filtration can be used for large volumes of water, Eosin Methylene Blue or MacConkey + MUG agar is used. For EMB, the inhibitor is the Methylene blue which inhibits the growth of gram positive bacteria, the indicator is the dye responding to a change in pH from the organism producing an acidic byproduct. In the MacConkey agar + MUG the inhibitors are Bile Salts and Crystal Violet. The indicator is the presents of β-glucoronidase from E. coli that fluoresce under UV light at 355 nm. A positive on EMB for E.coli is represented by metallic green sheen colonies. On the MacConkey + MUG, it’s a purple color, that fluoresces under UV light at 355 nanometers. 52. The purpose of the Colisure test is to check if there is any fecal contamination in food/drink. It is qualitative. 53. The enzymes involved in the Colisure test for Coliforms is B-galactosidase, and for E. coli it’s B- glucoronidase 54. Positive for Coliforms – red/magenta result | Positive for E. coli – red/magenta vial result and illuminates under UV light at 355 nanometers. 55. The MPN test is a statistical method to estimate the number of viable microorganisms in a sample. 56. The benefit of this test is that it’s good for measuring very small amounts of bacteria. 57. The indicator in this test is Brilliant Green Bile Lactose Broth. A positive result is a yellow solution. A negative result is a green solution. 58. You streak your highest three dilutions that were positive on an EMB plate. 59. Compact Dry test, and the purpose is to isolate and enumerate coliforms and E. coli 60. You only need 1 milliliter of sample in order to conduct this test. Pink/purple colonies are coliforms, blue colonies are E.coli 61. Magenta Gal & X-Gluc 62. You count each 1 cm by 1 cm square and multiply the average count by 20. 63. We don’t normally want to consume bacteria, especially pathogens. -5 64. 1 x 10 7 65. 2.5 x 10 , notice once you move the mL (CFU/ML) from bottom to top, you remove the negative next to the exponent 66. 1 x 10 (Mathematically 10 = 1, so 1 x 1 = 1, which this seems correct as the stock solution hasn’t been diluted at all) 67. A) 1 x 10 -2 B) 1 x 10 -4 C) 1x10 -5 D) 1 x 10 -6 E) 65 Colony Forming Units / 1 x 10 -6 F) 6.5 x 10 Colony Forming Units / mL <Steps to find VCN 68. In 1928, Fred Griffith found out that non-virulent Streptococcus became virulent when injected into mice along with a heat-killed virulent strain of Streptococcus. Only virulent bacteria were recovered from the dead mice. This experiment demonstrates Transformation at it’s finest, as the heat-killed virulent strain of Streptococcus was not virulent alone, but when combined with a non-virulent strain, transformed into a virulent strand. 69. Transformation is the uptake of naked DNA from the environment. 70. It was competent. 71. Acenitobacter ADP 1 and ADP6 72. To demonstrate that the ADP6 that previously could not grow on POB as a sole carbon source, could now take in the naked DNA from lysed ADP1, and grow on POB. 73. Tris buffer – to keep pH at a safe level for DNA EDTA – used as a chelating agent which removed divalent cations that were required by DNA degrading enzymes NaOH – Sodium Hydroxide used to break cell wall Sodium dodecyl sufate (SDS) – used to break cell membrane 74. Acenitobacter and Streptococcus 75. We used Gluclose as a control to demonstrate that both strands could grow on Glucose, and used POB to demonstrate that ADP 6 could not originally grow on POB as a sole carbon source, but the transformed ADP6 can. 76. It is the unidirectional transfer of genetic material that requires a structure called a pilus, and cell to cell contact. 77. Pseudomonas putida PAW15 and 503CA, PAW15 was the donor, and 503CA was the recipient. PAW15 was LEU- TOL+ and 503CA was LEU+ TOL- 78. Pseudomonas putida PAW15 was auxotrophic for leucine, as in it could not produce it’s own leucine 79. 503CA was prototrophic for leucine, as in it could produce it’s own leucine. 80. LEU+ TOL+ 81. The IMVC test is a test that distinguishes E. coli from other enteric bacteria, by use of four key tests. Indol, Methyl Red, Voges Proskauer, and Citrate. E. coli has the Tryptophanase enzyme, and is a mixed acid fermenter, it will show +nMethylRed + Other Enteric bacteria can ferment butanediol, and use citrate as a carbon source, they will show Vogues + Citrat+ The greatest advantage to this test is that you can quickly distinguish two types of enteric bacteria. 82. The indole test for the presence of the enzyme Tryptophanase, the reagent is Kovac’s solution, a positive result is indicated by a red ring. 83. The Methyl Red tests for mixed acid fermenters such as E. Coli. The reagent is Methyl Red, which reacts to a change in the pH of the solution, if the organism produces mixed acids during fermentation, then the media will turn red (at 4-5 pH). 84. The Voges-Proskauer test, tests for the production of Butanediol. α-nepthol and KOH are added to the solution, and if positive, the solution will turn red near the top. 85. The Citrate test, tests for the ability of an organism to use citrate as it’s carbon source. A positive test is indicated by a blue slant. Negative indication is green. 86. This could be either the Glucose or Lactose test. These tests allow the experimenter to see whether the organism uses glucose or lactose as a carbon source, whether or not it ferments, and whether or not it produces gas. 87. This is a SIM test, specifically testing for motility, the right vial is positive for motility. 88. The vial on the left is indole negative, the vial on the right is indole positive. 89. The right vial. 90. The cotton swab on the left is showing a positive oxidase test, we are testing for the cytochrome-c enzyme. 91. Psuedomonas fluorescens 92. This is the Starch test, a positive result shows the presence of the α-amylase enzyme. 93. The reagent is iodine. 94. This is the Urea test, a positive result is a fuchsia color, testing for the enzyme Urease. 95. The solution on the right, due to the presence of ammonia. 96. The mannitol salt agar plate 97. Staphlycoccus saprophyticus 98. 7.5%-10% NaCl (salt) 99. Growth with a yellow surrounding (due to the fermentation of mannitol) 100. Catalase, and the test is the Catalase test also 101. The inhibitor is 5% bile. 102. This is the Bile esculin test. 103. We are testing to see if the organism can 1. Survive on 5% bile, and 2. Can it hydrolyse esculin to form esculetin, which reacts with the ferric ions to turn the media black. 104. The Enteropluri test is used for the identification of Enterobacter. 105. Organisms must be gram-negative rods, and oxidase negative 106. The Enteropluri test conducts 15 biochemical tests. 107. What was the purpose of the Epidemiology experiment? 108. What is Epidemiology? 109. What is an Epidemiologist? 110. What is it called when a disease spread through an entire population in a short time? 111. What is a Pandemic? 112. What are some vectors which spread communicable diseases? 113. What did Serratia marcescens represent in the Epidemiology experiment? 114. What was the “non-pathogenic” organism used in the Epidemiology experiment? 115. What is the function of the CDC? 116. What is a MMWR? 117. What was an experiment we conducted in lab over mutualism? 118. On a P-1000 the dials read 100, what volume is this? P-200 reads 167, P20 reads 025 & Know what basic Microbiology lab equipment is called.
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