1. UNIT 2: HISTORY OF MICROBIOLOGY & BACTERIAL GENETICS

2. Unit 2 Overview: 1. History of Microbiology Video Koch’s postulates 2. Bacterial Growth and Metabolism Growth Requirements for Bacteria Bacterial Metabolism Culturing Bacteria and Sterile Technique 3. Bacterial Genetics Central Dogma in Bacteria Bacterial Transformation Bacterial Conjugation

3. HOW DID MICROBIO BECOME A FIELD?

4. 1665: Robert Hooke reported that living things were composed of little boxes, or cells 1858: Rudolf Virchow said cells arise from preexisting cells Cell theory: All living things are composed of cells and come from preexisting cells

5. Figure 1.2a The First Observations 1673-1723: Antonie van Leeuwenhoek described live microorganisms http://www.ucmp.berkeley.edu/history/lee uwenhoek.htmlhttp://www.ucmp.berkeley.edu/history/lee uwenhoek.html

6. Leeuwenhoek’s microscope

7. The Debate over Spontaneous Generation ( a very long lasting debate….) Spontaneous generation: The hypothesis that living organisms arise from nonliving matter; a “vital force” forms life Biogenesis: The hypothesis that the living organisms arise from preexisting life

8. ConditionsResults Three jars covered with fine net No maggots Three open jarsMaggots appeared From where did the maggots come? What was the purpose of the sealed jars? Spontaneous generation or biogenesis? Evidence Pro and Con 1668: Francesco Redi filled 6 jars with decaying meat

9. ConditionsResults Nutrient broth heated, then placed in sealed flask Microbial growth From where did the microbes come? Spontaneous generation or biogenesis? Evidence Pro and Con 1745: John Needham put boiled nutrient broth into covered flasks

10. ConditionsResults Nutrient broth placed in flask, heated, then sealed No microbial growth Spontaneous generation or biogenesis? Evidence Pro and Con 1765: Lazzaro Spallanzani boiled nutrient solutions in flasks

11. ConditionsResults Nutrient broth placed in flask, heated, not sealed Microbial growth Nutrient broth placed in flask, heated, then sealed No microbial growth Spontaneous generation or biogenesis? Evidence Pro and Con 1861: Louis Pasteur demonstrated that microorganisms are present in the air

12. Figure 1.3 The Theory of Biogenesis Pasteur’s S-shaped flask kept microbes out but let air in

13. The Golden Age of Microbiology 1857–1914 Beginning with Pasteur’s work, discoveries included the relationship between microbes and disease, immunity, and antimicrobial drugs

14. The Germ Theory of Disease 1835: Agostino Bassi showed that a silkworm disease was caused by a fungus 1840s: Ignaz Semmelweis advocated hand washing to prevent transmission of puerperal fever from one OB patient to another 1860s: Applying Pasteur’s work showing that microbes are in the air, can spoil food, and cause animal diseases, Joseph Lister used a chemical disinfectant to prevent surgical wound infections

15. Thomas Eakins The Gross Clinic 1875The Agnew Clinic 1889

16. Lister and Semmelweis

17. Fermentation and Pasteurization Pasteur showed that microbes are responsible for fermentation Fermentation is the conversion of sugar to alcohol used to make beer and wine Microbial growth is also responsible for spoilage of food Bacteria that use alcohol and produce acetic acid spoil wine by turning it to vinegar (acetic acid) and bacteria that utilize lactose to produce lactic acid transform milk into yogurt

18. Figure 1.4 Fermentation and Pasteurization Pasteur demonstrated that these spoilage bacteria could be killed by heat that was not hot enough to evaporate the alcohol in wine Pasteurization is the application of a high heat for a short time

19. Pasteurization Reduces spoilage organisms and pathogens Equivalent treatments 63°C for 30 min High-temperature short-time: 72°C for 15 sec Ultra-high-temperature: 140°C for <1 sec Thermoduric organisms survive

20. The Germ Theory of Disease 1865: Pasteur believed that another silkworm disease was caused by a protozoan 1876: Robert Koch proved that a bacterium causes anthrax and provided the experimental steps, Koch’s postulates, to prove that a specific microbe causes a specific disease

21. Robert Koch (pronounced “coke”) - German physician and bacteriologist -Lived 1843-1910 -Developed a criteria for determining whether a given bacteria is the cause of a given disease: -Known as Koch’s Postulates The Germ Theory of Disease

22. Koch’s Postulates 1.The microorganism must be found in all organisms suffering from the disease, but not in healthy organisms. 2. The microorganism must be isolated from a diseased organism and grown in pure culture. 3. The cultured microorganism should cause disease when introduced into a healthy organism. 4. The microorganism must be again isolated from the inoculated, diseased experimental host and identified as identical to the original specific causative agent. The Germ Theory of Disease

23. Koch’s Postulates Figure 14.3

24. Koch’s Postulates Figure 14.3

25. Procedures Overview

26. Postulate 1 The microorganism must be found in all organisms suffering from the disease, but not in healthy organisms. 1.Compare yogurt and milk and define the symptoms of “yogurtness”: - microscopic observations - textures, consistency - smell - pH Milk simulates a “healthy” sample Yogurt simulates a “diseased” sample

27. The microorganism must be isolated from a diseased organism and grown in pure culture. Postulate 2 2. Observe the cultures using a microscope and compare the different types of colonies. 3. Inoculate 3 separate petri dishes: Heathy individual- milk Diseased individual- yogurt Control bacteria- E.coli (control) 4. Grow cultures overnight at 37 0 C

28. Postulate 3 The cultured microorganism should cause disease when introduced into a healthy organism. 5. Inoculate fresh milk with bacteria colonies from the petri dishes 6. Incubate overnight 37 0 C 7. Assess symptoms of the subject (pH, smell, texture). Are these the same symptoms of “yogurtness”?

29. The microorganism must be again isolated from the inoculated, diseased experimental host and identified as being identical to the original specific causative agent Postulate 4 8. Observe yogurt and milk under the microscope: Can the bacteria be matched to the original culture? Got Yogurt?