MICROBIOLOGY

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• Microorganisms (microbes) - all organisms that are too

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small to be seen by the unaided eye. Beneficial agents in

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the fields of biotechnology and medicine.

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Pathogens - disease-causing microbes

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• Microbiology - scientific discipline that carefully studies

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identifies

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and characterizes microorganisms.

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Classification of Microorganisms

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Acellular

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Not made up of

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Viruses - acellular

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infectious

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true cells

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agents

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• Cannot

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incapable of replicating

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outside host cells

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reproduce

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Structure

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independently

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Core - nucleic acid

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Considered

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can be RNA or DNA

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non-living

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(never both)

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Viruses and

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prions

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ii.

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Capsid -

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(misfolded

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material protecting the

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proteins)

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sure made up of

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ili.

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Cellular

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microbes

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Prokaryotic Cells

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Do not have

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true nucleus

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nuclear

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membranes

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membrane-

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bound

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organelles.

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without envelope

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Envelope viruses -

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with envelope

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Sons to looks tour

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Banana acterom Monera

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unicalla

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(rokaryotic

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bacteria) and

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cyanobacteria (formerly

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-

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blui walen algae)

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peptidoglycan

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Unicellular

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prokaryotic

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in extreme conditions

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Extremophiles

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Eukaryotic Cells

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Have

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Protozoa/Protozoan

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true nucleus

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(Kingdom Protista

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Domain

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Eukarya)

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nuclear

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membranes

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unicellular

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eukaryotic

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non-photosynthetic

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membrane-

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bound

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organelles.

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Fungi (Kingdom Fungi

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Domain Eukarya)

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non-photosynthetic

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eukaryotic

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Microscopic fungi

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include the unicellular

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yeasts and the

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multicellular molds

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capable of carrying out

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both sexual and

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asexual reproduction

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Yeast cells via

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budding

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Molds via apical

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extension

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-

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Cell wall is made up of

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chitin

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Algae (Kingdom Protista

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Domain Eukarya for

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microscopic

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unicellular

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algae)

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Photosynthetic

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eukaryotic organisms

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Some are unicellular

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while others are

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multicellular

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Cell wall is

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predominantly made up

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of cellulose

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Disciplines in Context with Microbiology

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Study

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Microorganism

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Bacteriology

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Bacteria

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Clinical Bacteriology

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Pathogenic bacteria

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Virology

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Virus and virus-like agents

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Mycology

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Microscopic yeasts and

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molds

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Phycology (Algology)

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Protozoology

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Microscopic algae

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Eukaryotic and unicellular

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protozoans

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Parasitology

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Parasites (some protozoans

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are parasitic)

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Development of Microbiology

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Early Beginnings: Discovery of Cells and Very Small

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Organisms

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• Zacharias Janssen

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-

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One of several individuals who discovered two convex

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lenses (two-lens instrument) make small objects

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appear larger

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• Giovanni Faber

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• Coined the term “microscopio” or microscope for the

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two-lens system

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• Robert Hooke

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Used a 25x microscope

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• 1665: Published the book Micrographia which

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detailed his studies using the 25x microscope

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• Studied a piece of cork using his microscope

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used the

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term “cella” to describe “great many little boxes” he

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observed.

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• First person to describe and draw a microorganism

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(mold)

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• Anton van Leeuwenhoek

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Came up with a 200x microscope

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• Coined the term animalcules for hundreds of tiny

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living animals (probably protozoa and algae) he

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observed

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Understanding Disease Transmission: Uprise of

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Epidemiology

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Epidemiology - study of the distribution and determinants of a

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disease in a specified population.

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Naturalists

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Believed that miasma (bad air) is origin of epidemics

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emanating from rotten organic matter

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• Ignaz Semmelweis

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Hungarian obstetrician

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• Observed the mortality of pregnant women due to

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puerperal fever (childbed fever)

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Disease was more prevalent in the ward

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handled by medical students than in the ward

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run by midwifery students (29% death vs 3%

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death)

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Concluded that the source of contagion must be

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from cadavers on which the medical students

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previously had been performing autopsies before

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entering the wards. On the other hand

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midwifery

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students did not work on cadavers

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Directed his staff to wash their hands using

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chlorine water before entering the maternity

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ward leading to fewer fatalities due to childbed

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fever

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• John Snow

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Father of Epidemiology

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• Investigated 1854 London Cholera Epidemic

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concluded that cholera was waterborne and not

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spread by miasma

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Spot-mapping - interviewing the sick and healthy

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Londoners and the location of cholera cases were

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plotted in a map. It was found that most cholera

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cases were clustered and were getting their water

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source from the street pump in Broad Street

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Snow requested the parish Board of Guardians to

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remove the street pump handle

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effectively

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breaking the spread of the disease

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Classical Golden Age of Microbiology

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• 1854: Microbiology blossomed and continued until the

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advent of World War I.

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• Louis Pasteur |

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Considered as Father of Bacteriology

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• First to correctly explain that fermentation involves

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converting sugar into alcohol (wine) by yeasts in the

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absence of oxygen

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• Concluded bacteria causes souring and spoilage of

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wine and dairy products

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Developed pasteurization technique as practical

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solution for wine disease (souring). Original method

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was heating the wine to 55°C after fermentation but

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before aging

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• Proposed Germ Theory of Disease (Specific

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pathogens are responsible for specific infectious

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disease)

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• Unable to isolate any causative agents to completely

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validate the germ theory

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• Robert Koch

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• Verified the germ theory using Koch’s Postulates

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(formalized standards when relating a specific organism to

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a specific disease)

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• Koch’s postulates

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a.

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The same microorganisms are present in every

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case of the disease

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b.

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The microorganisms are isolated from the tissues

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of a dead animal and a pure culture is prepared.

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Microorganisms from the pure culture are

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inoculated into a healthy

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susceptible animal. The

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disease (same disease in postulate 1) is

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reproduced.

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d.

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The suspected pathogen must be re-isolated from

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the tissue

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e.

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specimens of the experimental animal and shown

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to be the same as the original

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Possutatu 2

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The microorganisni

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Allows microscopic observation and classification of

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bacteria according to shape

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Bacterial Classification

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Based on Cell Wall Structure

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Gram Positive

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Gram negative

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Thick

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Thick

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Spherical

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peptidoglycan

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peptidoglycan

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Based on Shape

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Bacilli

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Rod-

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shaped

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wall

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no outer

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wall

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no outer

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lipid

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membrane

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lipid

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membrane

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Exhibit purple

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Exhibit purple

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color

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color

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Gram positive bacteria

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Gram negative bacteria

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botulinum

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Staphylococcus

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Postulate 3

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pure cunture are inoculated

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Competition Between the Koch Lab and The Pasteur Lab

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Pasteur Lab - focused on mechanism of infection & immunity.

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Louis Pasteur with Charles

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Chamberland

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Attenuated (weakened) the

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bacterial cells of

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chicken cholera

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Louis Pasteur

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Applied attenuation to anthrax

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and used it to sheep

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Louis Pasteur with Emile Roux

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Roux: tested rabies vaccine to

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dogs

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Pasteur: tested the rabies

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vaccine to Joseph Meister

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Identified the diphtheria toxin

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Alexander Yersin & Emile

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Roux

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Eli Metchnikoff

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Discovered phagocytosis

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Koch Lab - procedural methods for isolation

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cultivation

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identification of pathogens.

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Robert Koch

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Friedrich Loeffler

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Georg Gaffiky

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Paul Ehrlich

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Shibasaburo Kitasato

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Emil von Behring

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Identified

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cultured

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discovered the following

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Mycobacterium tuberculosis

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Bacillus anthracis

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Vibrio cholerae

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Isolated Corynebacterium

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diphtheriae

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Cultivated typhoid bacillus

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Suggested dyes might control

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bacterial infection

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Isolated Clostridium tetani

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Developed antidiphtheria toxin

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Independent Scientists in Classical Golden Age of Microbiology

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Gerhard Hansen

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Observed bacterial cells in

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leprosy patients

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Albert Neisser

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(Mycobacterium leprae)

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Discovered Neisseria

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gonorrhoeae

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Charles Laveran

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Discovered that malaria is

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caused by a protozoan

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(Plasmodium falciparum)

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Theodore Escherich

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Discovered the bacterium

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responsible for infant diarrhea

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Richard Pfeiffer

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Identified a bacterial causative

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agent of meningitis

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(Haemophilus influenzae)

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Shibasaburo Kitasato and

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Alexander Yersin

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Independently discovered the

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bacterium causing plague

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(Yersinia pestis)

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Jules Bordet and Octave

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Gengou

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Cultivated Bordetella pertussis

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(pertussis bacillus)

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Common Techniques in Microbiology Section

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1

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Gram Staining

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Invented by Hans Christian Gram

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Classifies bacteria according to cell wall structure

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(thickness of peptidoglycan and presence of outer lipid

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membrane)

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monildormis

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Streptococcus

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cholerac

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Corynebacteriun

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Streptococcur

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2. Acid Fast Stain

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Used mainly for the differentiation of acid-fast

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organisms from non-acid fast organisms.

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Most useful in the detection of Mycobacterium

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tuberculosis.

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3

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KOH (Potassium Hydroxide) Preparation

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utilizes 10% potassium hydroxide which dissolves

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the keratin of skin scrapings

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nails

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detect fungal elements such as spores and hyphae

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4. Culture

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Isolation of the bacterium from the actual site of

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infection

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Allowing the organism to grow in vitro

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Allows microbiologists to come up with a sufficient

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population of the pathogen which eventually be used

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for identification of the pathogen.

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5. Sensitivity Testing/ Antimicrobial Susceptibility

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Testing (AST)

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• Done after culture to determine the most appropriate

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antibiotics in treating bacterial infections.

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• Most routinely used technique is the Kirby-Bauer

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technique (disk diffusion susceptibility test).