MCB 11 - LE 1

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Microbiology

Study of organisms and agents too small to be seen by the naked eye (

can be single celled or acellular
has nucleic acid genome
capable of replicating the genome

in new definition of microbiology, organisms are defined there as...

Thiomargarita magnifica (9000 micrometer)

largest bacterium

understanding the living world of microscopic organisms (basic bio science)
applying this understanding to humankind and planet earth (applied bio science)

two themes of microbiology

basic microbiology

Interest in specific group of microorganisms and their biology.

Virology

viruses

Mycology

fungi

Phycology

algae

Protozoology

protozoa

Bacteriology

bacteria

applied microbiology

Applying our understanding of microbial life processes for the benefit of humankind and planet Earth.

Saccharomyces cerevisiae

yeast responsible for making bread rise, it metabolize carbohydrates and produce CO2

miasmatic odors (mal'aria)

Ancient Greeks attributed disease to bad air

aqueducts

in rome they built this to bring fresh water into the city

Cloaca Maxims

giant sewer by Rome that carried waste away and into river Tibet

Hippocrates

Father of Western Medicine

Thucydides

Father of Scientific History

Zacharias Janssen

invented the first compound microscope (3

Athanasius Kirche

suggested to be the first to observe microorganisms using compound microscope

Antonie van Leeuwenhoek

coined the term wee animalcules to describe microscopic organisms that he observed using his handcrafted single

Robert Hooke

He published the book, "Micrographia". He used simple compound microscope (30x) then discovered "cells" in cork and elongated stalks in fungi

Spontaneous Generation Theory

Life emerges from non

Biogenesis Theory

Life begets life. Living organisms can only arise from other living organisms, not from non

Francesco Redi

founder of Experimental Biology. He tested spontaneous generation by placing meat in open and sealed jars. Maggots appeared only in the open jars, where flies could lay eggs.

John Needham

He boiled broth (but not long enough) to kill microorganisms and then sealed it in a jar. broth becomes cloudy then he observed the growth of microorganisms in the broth.

Lazzaro Spallanzani

He repeated Needham's experiment but boiled the broth longer and sealed the jars tightly. No microorganisms grew in the sealed jars.

Franz Schulse

exposed broth to air that had passed through strong acids. No growth of microorganisms in the broth.

Theodor Schwann

boiled nutrient solutions and exposed them to air that had passed through a heated tube. no microorganisms grew.

Georg Friedrich Schröder & Theodor von Dusch

used flasks with long necks to prevent airborne particles from reaching the broth. also filtered air through cotton wool (plugging). no microorganisms grew.

Louis Pasteur

Father of Modern Microbiology. used swan

John Tyndall

gave the final blow to theory of spontaneous generation. demonstrated that dust carries microorganisms. also provided evidence for the existence of exceptionally heat

germ theory of disease

in this theory, diseases are believed to be caused by specific agents called germa

Girolamo Fracastoro

disease was caused by invisible living creatures

Agostino Bassi

showed that a disease (muscardine) of silkworms was caused by a fungus (Beauveria bassiana)

Miles Joseph Berkeley

demonstrated that the Great Potato Blight of Ireland was caused by a fungus

Heinrich Anton de Bary

Founding father of Plant Pathology and founder of Modern Mycology. showed that smut and rust fungi caused cereal crop diseases

Ignaz Philipp Semmelweis

savior of mothers. introduced asepsis (absence of pathogens) in obstetrical wards to prevent transmission of fever. he introduced the policy of hand

chlorinated lime solutions (calcium hypochlorite)

what did Semmelweis advised to wash with?

Joseph Lister

father of antiseptic surgery. provided indirect evidence that
microorganisms were the causal agents of disease. He prevented postoperative infections.

phenol or carbolic acid

what did Lister used in surgical dressings and heat

Louis Pasteur

showed that the pébrine disease of silkworms was caused by a protozoan (Nosema bombycis)

Robert Koch

established the relationship between Bacillus anthracis and
anthrax (the disease).

Koch's Postulates

set of four criteria established by the German physician Robert Koch in the late 19th century to determine whether a specific microorganism is the cause of a disease.

1. microorganism must be absent from healthy organisms but present in diseased organisms

2. microorganism must be isolated from the diseased organism and grown in pure culture

3. cultured microorganism cause the same disease to healthy organisms

4. the microorganism must be reisolated from experimented infected host and identified as original pathogen

Koch's four postulates

Variolation

An early method of trying to protect people from smallpox. In this process, material (usually pus) from a person infected with smallpox would be deliberately introduced into the skin of a healthy person. This was done in hopes that the person would develop a mild form of the disease and, as a result, become immune to more severe infections of smallpox.

Edward Jenner

He used a vaccination procedure to protect individuals from smallpox. He used pus from cowpox blisters to inoculate to healthy human, which stimulates immune response and will later protect individuals from smallpox exposure in the future. Cowpox virus is closely related to variola, the causative agent of smallpox

Pasteur and his co

workers

Attenuation

Incubation of cultures for long intervals between transfers caused pathogens to lose their ability to cause disease. Growing the pathogen in an unnatural host weakens it. These two processes is called?

Attenuation of rabies

Rabies vaccine was developed through?

porcelain bacterial filter

device designed to separate bacteria from liquids or gases.

Charles Chamberland

developed porcelain bacterial filter

Martinus Beijerinck

One of the founders of virology. He published results on the filtration experiments demonstrating that tobacco mosaic disease is caused by an infectious agent smaller than a bacterium, which he called "virus".

contagious living fluid

contagium vivum fluidum

Paul Ehrlich

developed 606th compound SALVARSAN (organic arsenical to treat syphilis)

Alexander Fleming

discovered the "miracle drug" penicillin from Penicillium

Louis Pasteur

Demonstrated the role of yeasts in fermentation. developed the process of pasteurization to preserve wine.

Eduard Buchner

fermentation could occur in yeast extracts free of cells. cell

Ferdinand Cohn

discovered the existence of heat

Beijerink

discovered nitrogen fixation (bacteria convert nitrogen gas into a usable form for plants)

Winogradsky

discovered lithotrophy (using inorganic compounds for energy) and chemoautotrophy (using inorganic compounds for both energy and carbon)

10^

6 m

10^

9 m

adequate magnifying power, high resolving power, provide good contrast, serves your purpose

Good microscope

Light microscopes

use light waves and mirrors

simple microscope

short focal length, only has 1 lens, has magnification of 300x

Compound or complex microscope

has 2 sets of lenses, has magnification of 1000x

electron microscope

use electron beams as energy source, has higher magnification and resolving power, for objects smaller than 0.2 mm in diameter, in vacuum

bright field light microscopy

In this microscope, objects under study are darker, while the microscopic field is brightly lit. Used to observe gross morphology (basic shapes and structure) of microorganisms.

Dark Field Light Microscopy

in this microscope, the background of the microscopic field is dark, and the objects being studied appear luminous or bright. Used for invisible specimens in bright field microscope that cannot be stained as it can be distorted. Also useful for visualizing structures like flagella or ciliary motion.

Phase Contrast Light Microscopy

enhances contrast in specimens that are transparent, making it easier to view fine details. It is based on variations in refractive indices. It's often used to study internal structures of cells. It doesnt need staining or fixing.

Fluorescence Microscopy

type of microscope that uses fluorescent dyes (called fluorochromes) to make parts of a specimen glow under certain lighting conditions. visualizes specimens that
fluoresce. It can detect immunological reactions because it uses fluorescently labeled antibodies to bind to specific antigens, which helps visualize and localize proteins

Confocal Microscopy

Uses laser that can scan multiple z

Transmission Electron Microscope (TEM)

used to examine the internal ultrastructure of cells and other specimens. It provides highly detailed images of thin sections of samples, revealing structures at the microscopic and even atomic level. TEM is widely used to examine viruses at very high magnifications because viruses are often too small to be seen.

Scanning Electron Microscope (SEM)

used to study the surface features of specimens in 3D. It provides detailed, high

1. Smear Preparation

2. Fixation

3. Staining

Three basic steps in staining microorganisms

Smear

a thin, dry film of microorganisms

Fixation

refers to the process of preserving biological samples (cells, tissues, or microorganisms) in a state as close to their natural state as possible

kills cells, makes cells sticky, increases their diameter

purposes of fixation

heat fixation

type of fixation that is through direct flame or steam

chemical fixation

type of fixation that uses alcohol

staining

application of biological dyes

Dyes (stains)

organic compounds carrying chromophoric ions. make cell's internal and external structures more visible with the increased contrast with the background

basic or positively charged dye

These dyes have a positive charge and thus bind to negatively charged cellular components.

blue or purple

expected color in positively charged dye

Acidic or Negatively Charged Dyes

These dyes have a negative charge and usually bind to positively charged components, such as background areas

background is pink or red, cells are colorless or transparent

expected color on negatively charged dye

neutral stains

combination of both positively charged and negatively charged components.

simple staining

involves using only one dye to color the cells.

Positive/Direct Staining

The dye stains the cells directly because the positively charged dye binds to the negatively charged components of the cells.

same color as the color of the dye used, typically violet or green

expected color in positively/direct staining

negative/indirect staining

This method uses an acidic dye that stains the background and leaves the cells colorless or luminous.

background is pink or red, while cells are colorless or transparent

expected color in negative/indirect staining

differential staining

This method uses two or more dyes or reagents to differentiate between different types of cells or structures.

gram

staining

purple

expected color on gram positive bacteria

pink

expected color on gram

Acid

Fast Staining

Ziehl

Neelsen technique

Kinyoun technique

A cold method that uses a stronger dye and detergent to penetrate the cell wall.

red or pink

expected color on acid