microbiology
the study of organisms too small to be seen by unaided eye
living members of the microbial world
bacteria, archaea, alga, fungi, protozoan
nonliving members of the microbial world
viruses, viroids, prions
6 applications of microbiology/microbes to our lives
1. food production
2. biodegradation
3. biotechnology
4. genetic engineering
5. synthesis of commercially valuable products
6. medical microbiology
when was microbiology developed?
in 1674 with the discovery of microscopes and microbes
Antony van Leeuwenhoek
invented microscope. observed animalcules (today known as microorganism and microbes)
Robert Hooke
also credited with the discovery of microbes and microscopes. describes common bread mold in 1665
Francesco Redi
disproved theory of spontaneous generation by demonstrating that worms on rotting meat came from eggs of flies landing on meat (1668)
Louis Pasteur
disproved theory of spontaneous generation by demonstrating that air is filled with microorganisms and contact of broth with microorganisms is required for the growth of microorganisms in the broth. also developed swan-necked flasks
John Tyndall
provided by further experiments that Pasteur was correct
Ferdinand Cohn
discovered endospores
Robert Koch
demonstrated that anthrax is caused by bacteria. developed pure culture techniques
Joseph Lister
started the use of antiseptics in surgery
Hans Christian Gram
developed gram staining technique
Alexander Fleming
discovered the first antibiotic, penicillin
golden age of microbiology
1875-1903
How do we study microorganisms?
1. by observing them using microscopes
2. by making it easier to see cell structures using dyes or stains
3. by growing them in the lab using artificial culture media
immersion oil
used with the 100 X objective lens to maximize resolution
contrast
the ability of a microscope to make an image clear
resolving power
measurement of how clear the image is
magnification
measurement of how big an image can be
light microscopes
light passes through a specimen and then through a series of magnifying lenses, magnifying the image up to 1000X. has 6 different types
electron microscopes
beams of electrons are passed through specimen and then a series of electromagnetic lenses, magnifying the image more than 100000X. it has 2 types
atomic force microscopes
a very sharp probe moves across the surface of the specimen, feeling the bumps and valleys of the atoms. it can produce detailed images of individual atoms on a surface. it has a resolving power much greater than that of EM
basic dyes
most commonly used dyes
ex. methylene blue, crystal violet, safranin
acidic dyes
sometimes used dyes. used in negative staining
ex. india ink or nigrosine
simple staining
a single dye is used to visualize bacteria
differential staining
two dyes are used to distinguish between different groups of bacterial. the two most frequently used techniques are gram staining and acid-fast staining
special staining
one, two or more dyes are used to stain specific structures inside or outside the cells
ex. capsule staining, flagellar staining, endospore staining
fluorescent labeling
fluorescent dyes are used to stain total cells, a subset of cells, or certain proteins on cell surface
gram staining
type of differential staining. most commonly used staining technique in microbiology. it is used to identify bacteria belonging to two major groups: gram positive and gram negative. it is based on fundamental difference in cell wall structure of bacteria belonging to these groups
acid-fast staining
type of differential staining. it is used to detect bacteria of the genus Mycobacterium. it differentiates acid-fast bacteria from non-acid fast bacteria. acid-fast have high concentrations of mycolic acid in their cell wall
coccus/cocci
spherical
bacillus/bacilli
rod-shaped
coccobacillus/coccobacilli
short rods
vibrio
short curved rods
spirillum/spirilla
curved rods forming spirals
spirochetes
long helical cells with flexible cell walls
pleomorphic
many shapes
strepto-
chain
diplo-
pair
tetrads
groups of four
sarcinae
cubical packets
What structures are prokaryotic cells made up of?
1. cell envelope- cytoplasmic membrane, cell wall, capsule and slime layer
2. other structures outside the cell- flagella and pili
3. cytoplasm
structures inside the cell- nucleoid, plasmids, endospores, and others
cytoplasmic membrane
STRUCTURE: a thin delicate structure that surrounds the cytoplasm. it is made up of two layers of lipid molecule (phospholipid bilayer) embedded with proteins.
FUNCTION: it is selectively permeable allowing the movement of gases water and small molecules across the PL Bilayer through simple diffusion and osmosis, the proteins embedded serve as gates for the transport of larger molecules into the cell, helps to produce energy from food through the ETC
electron transport chain
1. electrons are transferred from one protein to the other in the chain
2. in the process, proteins release protons to the outside of the membrane
3. the collection of protons immediately outside the membrane forms a force called proton motive force
4.the PMT is used by the cells as a source of energy
cell wall
STRUCTURE: a strong and slightly rigid structure outside the cytoplasmic membrane
FUNCTION: gives shape to the cell, peptidoglycan provides strength to the cell wall, the outer membrane blocks many toxic compounds from entering the cell
peptidoglycan
alternating series of subunits NAG and NAM joined to form glycan chains
gram-positive cell wall
thick peptidoglycan layer, teichoic acid present, no outer membrane, no lipopolysaccharide, no porin proteins
gram-negative cell wall
thin peptidoglycan layer, teichoic acid absent, outer membrane present, lipopolysaccharide present, porin proteins present
capsules and slime layers
STRUCTURE: a distinct and regular gel-like layer outside the cell wall; an irregular gel like layer outside the cell wall. both composed of either polysaccharides or polypeptides
FUNCTION: help the bacteria attach to different surfaces, avoid the natural defense system of the host
flagella
STRUCTURE: long protein structures that originate from the cytoplasmic membrane and protrude out from the cell surface
FUNCTION: help the bacteria move from one place to another. carries out the movement of chemotaxis
pili
STRUCTURE: shorter and thinner than flagella. also move much slower
FUNCTION: some help bacteria cells attach to cell surfaces, some help bacterial cells move on solid surfaces, the sex pilus helps function in reproduction by joining two bacterial cells for transfer of DNA
chromosome
known as the nucleoid. a single, circular, double-stranded DNA molecule. tightly packed in the cell. contains all the genetic information required by the cell
plasmids
circular, double-stranded DNA molecules that can multiply independent of the chromosome. much smaller than the chromosome. most carry genes for antibiotic resistance
endospores
a unique type of cells produced by some bacteria. can remain dormant for a long time. they are highly resistant to high temperature, desiccation, and toxic chemicals
taxonomy of prokaryotes
the grouping of microorganisms based on similar properties: classification, identification, and nomenclature
how are prokaryotes classified?
1. phenotypic classification- based on morphological features and physiological characteristics (shape, size, arrangement, ect.)
2. molecular or genotypic classification- based on similarity in the nucleotide sequences (DNA and RNA)
3 domains of living organisms
1. Bacteria ( P)
2. archaea ( P)
3. eucarya (E)
species
a basic unit: group of closely related isolates or strains
genus
a collection of similar species
how are prokaryotes identified?
1. phenotypic characteristics: microscopic morphology, culture characteristics, metabolic properties, unique proteins and carbohydrates, types and amounts of fatty acids
2. molecular or genotypic characteristics: the similarity in unique nucleotide sequences, the similarity in rRNA and rDNA sequences
microscopic morphology
determines the size, shape, and staining characteristics. gives quick information, sometimes enough to make a presumptive diagnosis
culture characteristics
looks at the morphology of bacterial colonies growing on culture media
metabolic characteristics
performs biochemical tests on bacterial samples
dichotomous key
a flowchart of tests that give either a positive or negative result. a number of biochemical tests are performed on the bacteria to identify them to the species level
how to bacteria multiply?
prokaryotic cells divide by binary fission
exponential growth
when cells are actively dividing, the population doubles after each division
generation time
the time it takes for a bacterial cell to double. it depends on the bacterial species and growth conditions
how do bacteria grow?
1. singly
2. in mixed groups--by forming biofilms
biofilms
communities of different bacterial species encases in polysaccharides known as EPS
the importance of biofilms on human health
1. dental plaque leads to tooth decay, gum disease
2. many infections are complicated biofilms (ear infections)
3. more resistant to treatment with antimicrobial agents
pure culture
a population of bacteria that developed from a single bacterial cell. it is very useful in identifying and studying particular species. needs aseptic techniques (sterile)
batch cultures
considered closed systems as nutrients are not renewed and wastes are not removed from the culture
what are the phases of the bacterial growth curve?
1. lag phase
2. exponential (log) phase
3. stationary phase
4. death phase
5. decline phase
lag phase
phase in which the number of cells does not increase. the cells being synthesizing enzymes required for growth
exponential (log) phase
phase in which the cells divide at a constant rate. generation time is measured in this phase. bacteria are most sensitive to antibiotics in this phase
stationary phase
phase in which nutrient levels in the medium becomes too low to sustain growth. total numbers of bacteria remain constant. some bacteria die, others grow
death phase
phase in which the total number of viable cells decrease. bacterial cells die at a constant rate
decline phase
phase in which some fraction of bacteria may survive and become adapted to tolerate worsened conditions
continuous cultures
also known as open systems. bacterial growth is maintained at exponential phase by constantly adding nutrients to the culture medium and removing wastes from the culture medium
what are the environmental factors that affect the growth of bacteria?
1. temperature requirements
2. oxygen requirements
3. pH requirements
4. water availability
psychrophile
temperature range: -5 C to 15 C
ex. pseudomonas
psychrotroph
temperature range: 20 C to 30 C
optimal growth temperature: above 25 C
ex. listeria
mesophile
temperature range: 25 C to 45 C
optimal growth temperature: 37 C
ex. staphylococcus aurous; most disease causing bacteria belong to this group
thermophile
temperature range: 45 C to 70 C
ex. bacilius thermophius
hyperthermophile
temperature range: 70 C to higher
ex. bacilius thermophilus stereothermophilius; belong to archaea
obligate aerobe
grows only when oxygen is available. requires oxygen for respiration. produces superoxide dismutase and catalase
facultative anaerobe
grows best when oxygen is available, but also grows without it. uses oxygen for respiration if available. produces superoxide dismutase and catalase
obligate anaerobe
cannot grow when oxygen is present. does not use oxygen in the energy harvesting process. does not produce superoxide dismutase and catalase
microaerophile
grows only if small amounts of oxygen are available. requires oxygen for respiration. produces some superoxide dismutase and catalase
aerotolerant anaerobe
grows equally well with or without oxygen. it does not use oxygen in the energy harvesting process. produces superoxide dismutase but not catalase
neutrophiles
grow optimally at pH 5 to 8, optimum pH is near 7. most microbes belong to this group
acidophiles
grow optimally at pH below 5.5
alkaiphiles
grow optimally at pH above 8.5
halotolerant
withstand up to 10% salt concentrations
ex. staphylococcus
halophiles
require high salt concentrations (up to 3%)
extreme halophiles
require extremely high salt concentrations (up to or greater than 9%)
what are the nutritional factors that affect the growth of bacteria?
1. required elements
2. growth factors
3. energy sources
required elements
major elements that make up cell components
heterotrophs
bacteria that use organic carbon
autotrophs
bacteria that use inorganic carbon as CO2 (by carbon fixation)
growth factors
certain molecules that some bacteria cannot synthesize and need to obtain from external sources
amino acids, vitamins, purines, pyrimidines
non-fastidious bacteria
bacteria that can synthesize their own growth factors, and therefore, have wide metabolic capabilities
ex. E. coli
fastidious bacteria
bacteria that have complicated nutritional systems
ex. Meisseria gonorrheae
phototrophs
bacteria that obtain energy from sunlight
chemotrophs
bacteria that extract energy from chemicals
photoautotrophs
energy source: sunlight
carbon source: CO2
photoheterotroph
energy source: sunlight
carbon source: organic compounds
chemolithoautotroph
energy source: inorganic chemicals
carbon source: CO2
chemoorganohetertroph
energy source: organic compounds (sugars, amino acids, ect.)
carbon source: organic compounds
general purpose media
culture media that support the growth of all bacteria that do not have specific growth requirements
complex media
culture media that contain a variety of ingredients
chemically defined media
culture media that are composed of exact amounts of pure chemicals
selective media
special type of culture media that inhibits growth of certain species
differential media
special type of culture media that contains substance that microbes change in identifiable way
Joseph Lister
british physician who started the use of antiseptics in surgery
sterilization
the removal of all microorganisms--including endospores and viruses
disinfection
the elimination of most pathogens. some viable microbes remain
disinfectants
used on inanimate objects- may be called biocides, germicides, bactericides
antiseptics
used on living tissues
pasteurization
the brief heating to reduce the number of spoilage organisms, and to destroy pathogens. used on foods and inanimate objects
what are methods of control of microbial growth?
1. physical
2. chemical
3. other
physical control methods
1. using heat to destroy microorganisms and viruses: incineration, autoclaving, boiling, pasteurization
2. using filtration to remove microbes from liquid or air: membrane filters, HEPA filters
3. using radiation to destroy microbes: ionizing radiation and UV radiation
chemical control methods
using germicidal chemicals
alcohols
chemical control method which is antiseptics or disinfectants. organic solvents. destroy cells by coagulating essential proteins and damaging lipid membranes. kill vegetative cells of bacteria and fungi, but do not destroy endospores and some naked viruses
aldehydes
chemical control method which is a disinfectant. it destroys microbes by inactivating proteins and nucleic acids.
ex. formaldehyde, glutaraldehyde, and orthophthalaldehyde
ethylene oxide gas
chemical control method which is a disinfectant. used to sterilize heat or moisture sensitive items. it destroys microbes by reacting with proteins
halogens
chemical control method which is a disinfectant. it destroys microbes by damaging proteins and cell components.
ex. chlorine and iodine
chemical preservatives
food preservatives that are non-toxic and safe
ex. weak organic acids, nitrate and nitrite
low-temperature storage
refrigeration and freezing--hold bacteria at non growing phase
reducing available water
salt and drying
how do we choose the appropriate control method?
1. the type of microbe
2. the population size of microbes
3. the type of object
4. the level of toxicity, activity, and effectiveness of the germicidal chemical
5. the cost of the treatment process
mycology
the study of fungi
mycoses
diseases caused by fungi
superficial- pertaining to the skin (external)
systemic- of the organs (internal)
yeasts
single-cellular fungi
molds
multi-cellular fungi
hyphae
threadlike filaments that make up fungi
mycelium
a visible mass of hyphae
spores
the reproductive organ of fungi. they germinate into hyphae
sporangia
a sac-like structure that the spores are produced in
ex. aspergillus and rhizopus
conidia
asexual spores borne on hyphae; produced by both fungi and bacteria of the genus streptomyces
what are two ways that fungi are useful?
1. some produce antimicrobial medicines-- penicillin
2. some can be used to produce important molecules such as human insulin and hepatitis B vaccine
what are three ways that fungi are harmful?
1. some can grow on or in the body and cause diseases
2. some cells or spores may cause allergic reactions
3. some produce toxins that are harmful if ingested-- aspergillus can cause cancer
candidial skin infection
fungal disease caused by candida albicans (yeast) on the skin
vulvovaginal candidiasis
fungal disease caused by candida albicans (yeast) of the vagina
malaria
disease caused by the protozoan plasmodium
giardia
disease caused by the protozoan giardiasis
african sleeping sickness
disease caused by the protozoan trypanosoma
amebiasis (diarrhea)
diseased caused by the protozoan entamoeba
hookworm disease
infection from a hookworm (necator americanus and ancylostoma duodenale)
characteristics: anemia, weakness, fatigue, physical and intellectual disability in children
tapeworm disease
infection from either a fish tapeworm or a beef tapeworm (diphyllobothrium latum or taenia saginata)
characteristics: few or no symptoms, sometimes anemia
cysticercosis
infection from a pork tapeworm (taenia solium)
characteristics: variable symptoms depending on location and number of eggs that form larval cysts in the body
african sleeping sickness
infection of trypanosomes from the tsetse fly (arthropod)
characteristics: sleepiness, headache, coma
malaria
infection of plasmodium species from a mosquito (arthropod)
characteristics: chills, bouts of recurring fever
lyme disease
infection of borrelia burgdorferi from a tick (arthropod). known as the number one vector dsease
characteristics: fever, rash, joint pain, nervous system impairment