MOD 1

Microbiology: (micro: small ; bio: living organisms ; logia: study off) study of living
organisms that are too small to be seen using naked eye
Six Disciplines: Bacteriology, Parasitology, Mycology, Phycology and Virology
Microbes: nonliving entities and living organisms
Ubiquitous: virtually everywhere
Categories of Microbes
1. Acellular: infectious particles
2. Cellular: microorganisms
Pathogens: infectious agents; microorganisms that cause disease
Non-pathogens: microorganisms that do not cause disease
Microbial allies: helpful microbes
Microbial enemies: harmful microbes
Indigenous microflora: normal flora; microbes that live inside the body
Opportunistic: pathogens which impose risk to immunocompromised individuals
when opportunity is present
Saprophytes: decomposers; organism that lives on dead organic matter
Plankton: microscopic organism in ocean
° Phytoplankton: tiny marine plants and algae
° Zooplankton: tiny marine animals
Biotechnology: use of living organisms to make useful products
Antibiotics: substance used to kill the growth of other microbes
Relevance of Microbiology
1. Medical Microbiology: study of pathogens, its nature, and development of
disease (Epidemiology, Immunology, Clinical Microbiology, and Veterinary
Microbiology)
2. Agricultural Microbiology: study of harmful and beneficial roles of microbes in
plants and crops
3. Sanitary Microbiology: study of risk associated with the production of food and
water
4. Industrial Microbiology: maintenance of essential microorganisms for
commercial enterprises
5. Microbial Ecology: study of relationships between microbes and environment
(Bioremediation: natural pollution fighter; decompose industrial waste in the
environment)
6. Genetic Engineering: gene manipulation; transferring of gene to gene product
(common biochemically engineered microorganisms is E. coli for insulin production)
Microbes Cause Two Categories of Diseases:
Infectious Disease: pathogenic bacteria that colonizes the immunocompromised
persons
Microbial Intoxication: bacteria that produces toxins in vitro
First Microbes on Earth
° Archaea: ancient type of bacteria (Methanogens, Halophiles, Thermophiles)
° Cyanobacteria: found in marine and fresh water
Earliest Known Infectious Disease
° Tuberculosis
° Syphilis
° Parasitic worm infection
1. Schistosomiasis
2. Dracunculiasis
Tape worm infection
Pestilence: first recorded epidemic
Bubonic plague: occurred in 1900 BC
Ebers papyrus: occurred in Egypt
Small pox: occurred in China
Epidemic of Plague: occurred in Rome and Greece
Syphilis: occurred in Europe
 MOD 5

Origin of Virus
1. Virus existed before cells
2. Cells came first
1930s: where electron microscope invented
1940s: where first photographs of viruses obtained
1959: where negative staining procedure developed
Size of Virus: 10-300 nm
Viruses infect humans, animals, plants, fungi, protozoa, algae, and bacterial cells
Virus: smallest infectious agents which contains one type of nucleic acid
Recognition of virus is possible through direct electron microscope or means of
replication
Virus Based on Properties
° Virion morphology
° Virus protein properties
° Genome structure
° Strategies of replication
Virions: complete virus particles
Nucleic Acid: either DNA or RNA
Viral Nucleic Acid
° Single-stranded vs. Double stranded
° Linear vs. Circular
° Segmented vs. Non-segmented
Capsid: protein coat
Nucleocapsid: capsid plus nucleic acid
Capsid Symmetry
° Helical
° Icosahedral
° Complex
Capsomeres: small protein units in capsid
Envelope: membrane that surrounds some virus particles
° Enveloped virus: environmentally labile
° Naked Virus: environmentally stable
Characteristics of Virus
° Presence or absence of envelope
° Antigenic properties
° Number of capsomeres
° Type of genetic material
° Type of host
° Type of diseases
° Target cell
° Shape of capsid
° Size of capsid
Virus Based on Genome they Possess
° Double-stranded DNA or Single-stranded RNA
° Single-stranded DNA or Double-stranded RNA
Shapes of Capsid
° Polyhedral: many sided
° Helical: coiled tubes
° Bullet shaped
° Spherical
Viruses Taxonomic Levels
° Order: virales
° Family: viridae
° Subfamily: virinae
° Genus: virus
° Species: virus
Bacteriophages: virus that infect bacteria
Bacteriophages Based on Shape
° Icosahedron: spherical shapes with 20 triangular facets
° Filamentous: long tubes assembled into helical structure
° Complex: icosahedral heads attached to helical tails
Steps in Lytic Cycle
1. Attachment of phage to the surface of bacterial cell
2. Penetration: phage injects its DNA into bacterial cell
3. Biosynthesis: phage genes are expressed
4. Assembly: viral pieces are assembled
5. Release: when the host cell burst
Bacteriophages Based on Invasion
° Virulent: phage that lacks the ability to lysogenize
° Temperate: transferred DNA remains into bacterial cell chromosome
Lysogenic Conversion and Transduction: ways in which bacteria acquire new
genetic information
Animal virus: virus that infect humans and animals
Multiplication of Animal Virus
1. Attachment of virus to the cell
2. Penetration: entire virion enter the host cell
3. Uncoating: viral nucleic acid escape the capsid
4. Biosynthesis: viral pieces are produced
5. Assembly: fitting the virus pieces together to produce complete virions
6. Release: virus particles escape from the cells
Interferons: antiviral proteins
Shingles: painful nerve disease
Herpes virus infection: happens when an individual is infected by Herpes viridae
Antiviral agents: inhibits metabolic activities within cellular pathogens
Kaposi sarcoma: cancer common in AIDS patients
Human Immunodeficiency Virus: cause of AIDS
Mimivirus: large double-stranded DNA virus which mimics the bacteria
Plant Viruses: citrus tress, cocoa trees, cauliflower
Viroids: cause of plant disease
Prions: cause of animal and human disease
Kuru: women and children eat human brain
° Poxviridae : largest DNA virus
° Paramyxoviridae : largest RNA virus
° Parvoviridae : smallest animal virus

MOD 2
Pioneers in the Science of Microbiology
1. Anton Van Leeuwenhoek: first person to live bacteria and protozoa (Father of
Microbiology, Bacteriology, and Protozoology)
Contributions
° Simple microscope: tiny glass lenses
° Animalcules: tiny living creatures
° Abiogenesis: theory of spontaneous generation; life arise from non-living materials
Biogenesis: life can only arise from pre-existing life (Rudolf Virchow)
2. Louis Pasteur
Contributions
° Discovered what occurs during alcohol fermentation
° Discovered that life could exist in the absence of oxygen (aerobes: organisms that
require oxygen ; anaerobes: organisms that do not require oxygen)
° Pasteurization: used to kill microbes in many types of liquids
° Germ Theory Disease: specific microbes cause specific infectious disease
° Vaccine development (anthrax, cholera, swine erysipelas, and rabies)
3. Robert Koch
Contributions
° Germ Theory Disease: proved that B. anthracis was truly caused by anthrax using
Koch Postulate
° Characteristics of B. anthracis
° Developed methods for routine bacteriology (fixing, staining, photographing,
cultivating)
- R.J Petri: invent the petri dish
- Frau Hasse: suggest to use agar as solidifying agent
° Tuberculin: protein derided from M. tuberculosis
Careers in Microbiology
° Microbiologist: scientist who studies microbes
- Medical Microbiology: concerned with the transmission of pathogens to protect
people and animals against infectious diseases
- Clinical Microbiology: concerned with the laboratory diagnosis of infectious
diseases of humans
Career Fields within the Microbiology
1. Bacteriologist: scientist who studies the structure, functions, and activities of
bacteria
2. Phycologist: scientist who studies the various types of algae
3. Protozoologist: scientist who studies the activities of protozoa
4. Mycologist: scientist who studies fungi
Microscope: used to observe tiny objects that can’t be seen through naked eye
Types of Microscope
° Simple: has one magnifying lens
° Compound: has more than one magnifying lens (discovered by Hand Jansen and
Zacharias)
- Photomicrographs: photographs taken through the lens system of compound
microscope
° Electron: used electron beam as a source of illumination (Transmission: 2 D and
Scanning: 3D)
Parts of Compound Microscope
° Ocular lens: X10 magnifying lens
° Revolving nosepiece: holds the objective lens
° Objective lens: used to magnify objects
° Stage: where specimen is placed
° Stage adjustment knob: used to move stage
° Iris diaphragm: used to adjust the amount of light passing through condenser
° Condenser: used to focused light onto specimen
° Collector lens: controls the amount of light entering the condenser
° Rheostat control knob: controls the amount of light emitted from the light source
° Field diaphragm: used to adjust the amount of light passing through the collector
lens
° On/off switch: used to turn the light source
° Base: contains the light source
° Condenser control knob:
° Fine and coarse adjustment knob: used to focus the objective lens
° Arm: supports the binocular body and revolving nosepiece
° Binocular body: holds the ocular lens in their proper locations

MOD 6
Domain Bacteria contains 5 subclasses, 23 phyla, 32 classes, , 77 orders, 14
suborders, 182 families, 871 genera, and 5,007 species and divided into three
phenotypic categories:
a. Gram-negative
b. Gram-positive
c. Lack a cell wall
• A bacterium’s gram reaction, cell shape, and morphological arrangement are
important in organism’s identification
• Bacteria varies in size (0.2 um to 10), shape or arrangement
Shapes of Bacteria
• Spherical (Cocci)
° singly, pairs (diplococci)
° chains (streptococci)
° clusters (staphylococci)
° packets of four (tetrads)
° packets of eight (octads)
Important Medical Cocci
1. Neisseria spp.
2. Enterococcus spp.
3. Staphylococcus spp.
4. Streptococcus spp.
• Rod-shaped (bacilli)
° singly, pairs (diplobacilli)
° chains (streptobacilli)
° elongated cocci (coccobacilli)
Important Medical Bacilli
1. Enterobacteriaceae
2. Pseudomonas aeruginosa
3. Bacillus spp.
4. Clostridium spp.
• Spiral-shaped (spirilla)
° Vibrio spp.
° Campylobacter spp.
° Treponema spp.
Pleomorphic: ability to exist in variety of size and shapes
Micoplasma: ability to resist antibiotics
Arrangement of Size
1. Prions
2. Virus
3. Bacteria
4. Parasites
Preparation for staining: bacteria are smeared onto a glass microscope slide, air-
dried, then fixed
Common Methods of Fixation
° Heat fixation: smear is passed through a bunsen burner flame
° Methanol fixation: flood with absolute methanol for 30s
Purpose of Fixation
1. Kills organisms
2. Preserves morphology
3. Anchors the smear to the slide
Types of Stains
1. Simple Stain: used to determine bacterial shape and morphologic arrangement
(Structural staining procedures: used to observe bacterial capsules, spores, and
flagella)
2. Differential Stain: the gram and acid-fast staining: done to enable differentiation
between one group of bacteria to another
a. Gram staining procedure: developed by Hans Christian Gram; serve as a clue
when identifying particular bacterium
(a). Gram-positive bacteria: if bacteria were not decolorized, they will be blue to
purple
(b). Gram-negative bacteria: crystal violet was removed from cells and it will turn
from pink to red
(b). Acid-fast staining: developed by Paul Ehrlich; used for identification of
mycobacterium
3. Structural Stain
a. Capsule stains: determine whether organisms encapsulated
b. Flagella stains: determine whether organism posses a flagella
c. Endospore stains: determine where organism is spore-former
• Motility can be demonstrated by stabbing the bacteria into a tube of semisolid agar
or by using the hanging drop technique
Motile: if bacterium can swim
Non-motile: if bacterium can’t swim
• Non-motile organisms grow only along the stab line while motile will spread away
• Bacterial motility is most often associated with the presence of flagella
Bacterial colony: pile of bacteria; it is characterized based on:
a. margin
b. elevation
c. color
d. overall shape
e. size
Atmospheric Requirements
a. Obligate aerobes: needs 20-21% atmospheric oxygen
b. Microaerophiles: needs at least 5% atmospheric oxygen
c. Obligate anaerobe: can only grow in an anaerobic environment
d. Facultative anaerobe: can survive in either the presence or absence of oxygen
e. Aerotolerant anaerobe: does not require oxygen but can survive in atmospheric
O2
Nutrional Requirements Fastidious Organisms: organism that requires nutrition
to grow in the laboratory
• Many pathogens cause disease because they possess capsules, pili, endotoxins
(components of cell wall of Gram-negative bacteria), or because they secrete
exotoxins and exoenzymes
Pathogenicity: ability to cause disease
Molecular Diagnostic Procedures: test that analyzes the organism’s DNA or RNA
Many archaea are extremophiles (lived in extremely hot, dry or salty environment)
° Thermophiles: extremely hot
° Halophiles: extremely salty
° Acidophiles: extremely acidic
° Psychrophiles: extremely cold
° Alkaliphiles: extremely alkaline
° Piezophiles: extremely high pressure
 MOD 3

Cell: fundamental unit of any living organism

Metabolism: chemical reactions that occur within a cell
Prokaryotes or prokaryotic cells: cells which includes bacteria and archaea
Eucaryotes or eukaryotic cells: cells which contains true nucleus and many
membrane-bound organelles
Cytology: study of the structure and function of cells
Eucaryotes (caryo refers to nut or nucleus)
• Cell membrane: skin around the cell composed proteins and phospholipids
• Nucleus: controls the functions of the entire cells
a. Nucleoplasm: base material of the nucleus
b. Chromosomes: embedded in the nucleoplasm (Genome: complete collection of
genes)
c. Nuclear membrane: skin around the nucleus
• Cytoplasm: gelatinous, nutrient matrix (Cytosol: semifluid portion of the
cytoplasm)
• Endoplasmic reticulum: highly convoluted system of membranes
a. Rough endoplasmic reticulum: many ribosomes are attached
b. Smooth endoplasmic reticulum: no ribosomes are attached
• Ribosomes: consist of rRNA and protein (Polysomes: cluster of ribosomes)
• Golgi complex: communicates with ER
• Lysosomes: contains lysozyme and digestive enzymes that break down foreign
material (Autolysis: breaking of entire cells)
• Peroxisomes: membrane-bound vesicles where hydrogen peroxide is both
generated and broken down
• Mitochondria: energy necessary for cellular function provided by ATP molecules
(major energy-carrying molecules within the cells)
• Plastids: sites for photosynthesis (Chloroplast: type of plastid which contains
photosynthetic pigment called chlorophyll)
• Cytoskeleton: present throughout the cytoplasm is a system of fibers
(Microtubules: tubules composed of spherical protein subunits called tubulins)
Three Types of Cytoskeletal Fibers: microtubules and microfilaments (essential for
cell division), and intermediate filaments
• Cells wall: external structures that provide rigidity, shape and protection
a. The cell walls of algae contains polysaccharide—cellulose—found in cell wall of
plants
b. The cell walls of fungi contains polysaccharide —chitin—found in exoskeletons of
beetles and crabs
c. Mesosomes: where cellular respiration takes place
• Chromosome: control center of bacterial cell wall (Bacterial nucleoid: occupied
space within a bacterial cell
• Plasmids: circular molecules of double-stranded DNA
• Cytoplasm: mixture of all materials required by the cell for its metabolic functions
• Polysomes: site of protein synthesis
• Cytoplasmic granules: occur in certain species of bacteria
• Bacterial Cell Wall: defines the shape of bacterial cells, consist of peptidoglycan
(macromolecular polymer)
• Glycocalyx: gelatinous material produced by the cell membrane
a. Slime layer: not highly organized and is not firmly attached to the cell wall
b. Capsule: highly organized and firmly attached to the cell wall
• Flagella: protein that enable bacteria to move
Types of Flagellar Arrangement
a. Peritrichous: flagella all over the surface
b. Lophotrichous: tuft of flagella at one end
c. Amphitrichous: one flagellum
• Cilia: organelles of locomotion (Ciliates: cilia found in some species of protozoa)
Prokaryotic Cell Structure: ten times smaller than eukaryotic cells
• Cell membrane: similar to eukaryotic cell in structure and function
• Pili: hairlike structures composed of polymerized protein called pillin
Types of Pili
a. Type that enables bacteria to attach surfaces
b. Sex pilus: enables the transfer of genetic material from one bacterial cell to
another
Spores: capable of forming thick-walled spores
Robert Hooke
- English physicist who published the Micrographia
- First person to used the term cells
German botanist Matthias Scheleiden and German zoologist Theodor Schwann
concluded that all plant and animal tissues were composed of cells—known as cell
theory
Rudolf Virchow: German pathologist who proposed the theory of biogenesis — that
life can only arise from pre-existing life
 MOD 4

Prokaryotic Cell Reproduction: simple compared with eukaryotic cell division

Binary fission: cell splits in half to become two daughter cells
DNA replication: before prokaryotic cell divide, chromosome must be duplicated
Generation time: time it takes for binary fission to occur
Bacterial generation time: 10 mins to 24 hrs
According to Bergey’s Manual of Systematic Bacteriology, taxonomy
(science of classification of living organisms) consist of three areas:
1. Classification: arrangement of organisms into taxonomic groups (known as taxa)
2. Nomenclature: assignment of names according to international rules
3. Identification: process of determining whether an isolate belongs to one of the
established
Carolus Linnaeus: Swedish scientists who established the science of taxonomy
Binomial System
• First name: Genus
• Second name: specific epithet
“First and second names together are referred as species”
sp: used to designate a singles species
spp. : used to designate more than one species
Robert H. Whittaker: the one who proposed the Five-Kingdom System of
Classification
• Bacteria and Archaea: Kingdom Prokaryote (Monera)
• Algae and Protozoa: Kingdom Protista (Protist)
• Fungi: Kingdom Fungi
• Plants: Kingdom Plantae
• Animals: Kingdom Animalia
Carl R. Worse: scientist who devised the Three-Domain System Classification: in this
system, there are two domains of prokaryotes (Archaea and Bacteria) and one domain
(Eukarya)
Comparison between Eukaryotic and Prokaryotic Cells
EUKARYOTIC CELLS
PROKARYOTIC CELLS
Plant Type Animal Type
Biologic All plants, fungi, All animal and All bacteria
distribution and algae protozoa
Nuclear Present Present Absent
membrane
Membranous Present Present Absent
structure
Microtubules Present Present Absent
Cytoplasmic 80S 80S 70S
ribosomes
Chromosomes Composed of DNA Composed of DNA Composed of DNA
and proteins and proteins only
Flagella or cilia Present Present Present
Cell wall Present Absent
Photosynthesis Present Absent Present
 MOD 7

Protozoology: study of protozoa

Protozoologist: scientist who study protozoa
Examples of Free-living Pond Protozoa: Naegleria fowleri and Acanthamoeba
Protozoan’s Life Cycle
° Motile Trophozoite: dividing stage
° Non-motile Cyst: survival stage
Parasitic protozoa: absorb nutrients from the body of host
Classification of Protozoa based on Locomotion
° Flagellated protozoa: moved by means of flagella
° Amebae: moved by means of pseudopodia
° Non-motile protozoa: protozoa that lacks pseudopodia
° Ciliates: moved by means of cilia
Fungi: garbage disposers of nature
Mycology: study of fungi
Mycologist: scientist who study fungi
Saprophytes fungi: fungi that lives on organic matter in water and soil
Parasitic fungi: fungi that lives within animals and plants
Parts: Hyphae, Mycelium, Thallus
Spore Reproduction
° Spore reproduction: fusion of two gametes
° Asexual reproduction: form other than fusion of gametes
Fungi based in Phylum
1. Lower fungi
° Zygomycotina: bread moulds
° Chytriodimycotina: not true fungi
2. High fungi
° Ascomycotina: yeast and fungi that cause plant disease
° Basidiomycotina: fleshy fungi that cause plant disease
3. Deuteromycotina: 5fth phylum; no sexual reproduction
Types of Fungi
1. Yeast: eukaryotic cell which lacks mycelia
° Saccharomyces cerevisiae: baker’s yeast
° Candida albicans: most commonly isolated in humans
° Cryptococcus neoformans
2. Moulds: fungi which are often seen in water, soil, and food
Two Types
° Aerial hyphae: above the surface of mould
° Vegetative hyphae: beneath the surface
3. Fleshy fungi: encountered in forest
Fungal Infections of Human
° Superficial mycoses: fungal infection om outermost layer of human body
° Cutaneous mycoses: fungal infection on dermis
° Subcutaneous mycoses: fungal infections on dermis and underlying tissue
° Systemic mycoses: fungal infections on internal organs
Antifungal Agents: Nystatin, Amphotericin, or 5-fluorocytosine
Lichens: combination of algae and fungus (this process is known as symbiotic
relationship)
Mutualism: wherein both parties benefit
Photobiont: algae in lichen
Mycobiont: fungus in lichen
Slime moulds: found in rotting clogs
Cellular slime moulds: lowest level of cellular differentiation
Plasmodial slime moulds: haploid cells fused to become diploid cells
Algae: photosynthetic eukaryotic organism
Cellulose: polysaccharide
Diatoms: algae that live in both fresh and seawater
Diatomaceous: used in filtration system, insulation, and abrasives
Dinoflagellates: microscopic photosynthetic algae
Cytostome: primitive mouth
Prototheca: one genus of algae which cause protothecosis
- Algae are protist
- Dinoflagellates are responsible for red tides
- Most protozoa are single-celled free-living microorganisms
- Asexual Spores are called conidia
- Single yeast cells are blastospores
- Yeast are produce through binary fission
- Moulds and fungus produce mycotoxins known as microbial intoxications
° Dermatophytes: cutaneous that lives on dermis
° Mycoses: fungal infections
° Slug: phagocytosis
Algae cells have:
• Pellicle: thicked cell membrane
• Stigma: light sensing organelle
• Flagella
° Algae are classified as green, golden brown, or red
Components of Green Algae
• Desmids: unicellular algae
• Volvox: multicellular algae
• Euglena: possessed by both algae and protozoa
• Chlamydomonas: biflagellate algae
• Spirogyra: filamentous algae
Important Medical Moulds: Aspergillus and Penicillium
Protothecosis: subcutaneous lesion that looks crusty
Dimorphic fungi: fungi that live either as yeast or moulds
Micro organism’s Group
° Bacteria
° Fungi
° Protozoa
° Algae
° Parasites
- Bacteria move by means of flagella whereas protozoa use either flagella, cillia, or
pseudopods
- Motile organisms found in pond water can be observed using went mount method or
hanging drop preparation
Wet mount methods: placing specimen on a glass slide
Hanging drop preparation: specimen is inverted over a slide
Brownian movement: random movement produced by currents in liquids
DEFINITION OF TERMS
a. Aseptic technique: used to eliminate pathogens to prevent contamination
Importance of Aseptic Technique: prevent microbiologist from infection, from
contamination of their work environment, clinical specimens, cultures, and subcultures
b. Antiseptic technique: developed by Joseph Lister (common antiseptic agent: )
c. Sepsis: presence of pathogens in blood or tissue (asepsis: absence of pathogens)
d. Antisepsis: prevention of infection
e. Sterile technique: use to exclude all pathogens from particular area
BACTERIAL GROWTH
Binary fission: multiplication of bacteria
Generation time: time it takes to undergo binary fission
Example:
• Pseudomonas and Clostridium spp. (10 minutes)
• Staphylococcus and Streptococcus spp. (20 minutes)
• Mycobacterium tuberculosis (18-24 hours)
Short generation time: bacteria are rapid growers
Long generation time: bacteria are slow growers
Difficult to grow: fastidious organisms
Phases of Bacterial Growth
1. Lag phase: no increase in bacterial cells
2. Log phase: exponential increase in bacterial cells
3. Stationary phase: plateau in number of bacterial cells
4. Death phase: exponential decrease in bacterial cells
Factors that Affect Bacterial Growth
• Availability of Nutrients: nutrients serve as energy source such as carbon,
hydrogen, oxygen, nitrogen, sulfur, and phosphorus
• Moisture: cells are composed of 70-95% of water making water essential for life
(desiccation: drying process)
• Temperature
1. Thermophiles: loves high temperature (ex: Acinetobacter)
2. Mesophiles: grows at moderate temperature (mesophilic: indigenous microflora
that grew at body temperature)
3. Psychrophiles: loves cold temperature
4. Psychroduric organisms: loves warm temperature (fecal matter left by Artic
explorers contains E. coli)
• pH
1. Acidophiles (2-5): prefer environment containing sulfurous gases (E. coli and
Lactobacillus spp.)
2. Alkaliphiles (8-10): found in intestine and soil laden (V. cholerae)
• Osmotic pressure and salinity: pressure exerted on cell membrane
1. Isotonic solution: equal concentration
2. Hypotonic solution: less concentration (RBC: hemolysis Bacteria:
plasmoptysis)
3. Hypertonic solution: greater concentration (RBC: crenation Bacteria:
plasmolysis)
Halophilic: loves salt
Haloduric: halotolerant
• Barometric pressure: Piezophiles: thrive deep in ocean (Archaea: capable of
living in the deepest part of ocean)
• Gaseous atmosphere
1. Obligate aerobes: 20-21% oxygen and 78-79% nitrogen
2. Microaerophiles: 5% oxygen
3. Obligate anaerobes: killed by the presence of oxygen
4. Capnophiles: 5%-10% carbon dioxide
INOCULATION OF CULTURE MEDIA
Inoculation of a liquid medium involves adding specimen to medium while inoculation
of a solid medium involves the use of sterile inoculating loop (streaking)
CULTURING BACTERIA IN LABORATORY
° Successful attempts of culturing: Joseph Schroeter, Ferdinand Cohn, Oscar
Brefeld
° Culture techniques: • Gelatin: first solidifying agent in Koch media • Agar:
replaced gelatin
Petri dish: container for solid and bacterial culture (Robert Koch, Fanny Hesse,
Richard Petri)
° Joseph Lister: first person to obtain a pure culture of Streptococcus lactis (Louis
Pasteur and Robert Koch: contribute to Germ Theory of Disease
CULTURE MEDIA
Artificial media: used to culture media
Based on Composition
• Chemically-defined or synthetic: all ingredients are known
• Complex or non-synthetic: ingredients are not known
Based on Consistency
• Liquid media: tubed media (ex: TSB)
• Semi-solid media: contains 0.5-1% solidifying agent (ex: sulfide)
• Solid media: poured in petri dish (Agar: polysaccharide used as solidifying agent)
INCUBATION
Incubation: incubating in appropriate atmosphere, moisture level, and temperature
(35-37°C)
Types of Incubators
1. CO2 incubators
2. Non-CO2 incubators
3. Anaerobic incubators
INHIBITING GROWTH OF MICROBES
Sterilization: elimination of all microbes including spores
Physical Method
1. Heat: inexpensive method of sterilization (temp and time)
• Thermal death point: lowest temperature that kill all organisms
• Thermal death time: length of time necessary to sterile a pure culture
2. Dry heat: provides effective sterilization of metals (incineration: burning of
contaminated materials)
3. Moist heat: more effective than dry heat
4. Autoclave: pressure cooker that destroys all microbes
5. Cold: microorganism’s metabolic activities are slowed (C. botulinum and C.
perfringens causes food poisoning)
6. Desiccation
7. Radiation: these are x-rays and gamma and beta rays
8. Filtration: used to filter cells from liquids
9. Gaseous atmosphere:
• Aerobes and microaerophiles: can be killed by removing oxygen from
environment
• Obligate anaerobes: can be killed by adding oxygen to environment
• Gas gangrene: caused by Clostridium
Disinfection: elimination of all microbes except spores
1. Pasteurization: disinfect liquids
2. Disinfectant: disinfect inanimate objects
3. Antiseptics: disinfect skin and living tissues
4. Sanitization: reduce microbial populations
Microbicidal agents: antiseptics that kill microbe
1. Bactericidal agent: kills bacteria
2. Sporicidal agent: kills bacterial endospores
3. Fungicidal agent: kills fungi
4. Algicidal agent: kills algae
5. Viricidal agent: kills virus
6. Pseudomonicidal agent: kills pseudomonas agents
7. Tuberculocidal agent: kills M. tuberculosis
Microbistatic agents: chemical that inhibits the reproduction of micro