Microbiology Laboratory SARCINAE

BACTERIAL MORPHOLOGY *Forming a “cube like” structure, group of 8

*Classifying bacteria based on shape. Ex. Micrococcus luteus

 Clinically significant bacteria (0.25nm to STAPHYLOCOCCUS

1nm x 1nm to 3 nm)
*Divides in multiple planes and forms “Grape
 Bacteria in general (0.2nm to 2nm x
like” / “Broad sheet” appearance
2.8nm)
Ex. Staphylococcus aureus
COCCI
BACILLI
*Meaning berries.
*Meaning small staffs
*Round, Oval, Elongated or Flattened on one
side. *Rod shaped, rectangular, slender, club shaped.

*0.5nm to 1.0nm. *0.5nm to 2nm

CELL ARRANGEMENT CELL ARRANGEMENT

DIPLOCOCCI SINGLE BACILLUS
*Remain in pairs after dividing. *most common

*Moves sideways during division. *usually found in singles.

Ex. Neisseria gonorrhoeae DIPLOBACILLI

STREPTOCOCCUS In pairs after division

“Chain like” STREPTOBACILLI
*Resulted from continuous division Found in long chains.
Ex. Streptococcus pyogenes PALISADE

Ex. Streptococcus mutans *Resemble a picket fence or cigar pocket.

TETRADS *Brought about by slipping of cells during cell

division
“Square”
CHINESE LETTERS
*Divides in two pairs & remains in group of
four. *Referred to as “cuneiform arrangement”

*X, Y, L, and V arrangement, caused by snapping

of cells during cell division.
COCOBACILLI 3 CARDINAL TEMPERATURE

*Intermediate between cocci and bacilli MINIMUM TEMPERATURE

*Short stamped bacilli *Minimum temperature at which the organism

can grow.
SPIRALS
OPTIMUM TEMPERATURE
*Have one or more twist
*Temperature at which organisms grow best
*Never straight
MAXIMUM TEMPERATURE
CELL ARRANGEMENT
*Highest temperature at which an organism can
VIBRIO grow.
*Curvy rods GROUPS OF BACTERIA ACCDG. TO
*Comma shaped TEMPERATURE

Ex. Vibrio cholerae THERMOPHILE

SPIRILLUM  Min: 45 degrees Celsius

 Optimum: 50 to 0 deg. Celsius
*Thick, rigid cell wall  Maximum: 250 deg. Celsius

*Has Flagella PSYCHROPHILE

*Helical and coiling (curvy)  Min: 0 degrees Celsius

 Optimum: 15 deg. Celsius
“Cork screw” resemblance
 Maximum: 20 deg. Celsius
*Shorter than spirochete

SPIROCHETE MESOPHILE

*Flexible  Min: 15 to 20 degrees Celsius

 Optimum: 20 to 40 deg. Celsius
*Helical or coiling
 Maximum: 45 deg. Celsius
*Axial Filaments

Ex. Treponema palidum

PH
BACTERIAL PHYSIOLOGY
ACIDITY OR ALKALINITY
PHYSICAL REQUIREMENTS FOR BACTERIAL
3 GROUPS
GROWTH

TEMPERATURE
ACIDOPHILE “Facultative halophiles”

Grows in pH below four Ex. Staphylococcus aureus

ALAKINOPHILE CHEMICAL REQUIREMENTS: NUTRIENTS

Grows in pH greater than eight MAJOR ELEMENTS

NEUTROPHILE CARBON

Grows in pH between .5 to 7.5 *Structural backbone of living matter

IN THE LABORATORY *Needed for all organic compounds

*Bacteria often produce acids that could inhibit *Half of the dry weight of bacteria is made up
their own growth. of carbon.

*To maintain the proper pH, buffers are NITROGEN AND SULFUR
included in the medium.
*Used in protein synthesis
OSMOTIC PRESSURE
NITROGEN AND PHOSPHOROUS
3 GROUPS
*Used in DNA and RNA synthesis
OSMOPHILE
NITROGEN
*Requires higher osmotic pressure
*Nitrogen fixation: use of gaseous nitrogen
*Not pathogenic directly from the atmosphere

“Extreme Halophile” SULFUR

HALOPHILE *Synthesize sulphur containing amino acids and

vitamins such as THIAMINE and BIOTIN
*Requires higher concentration of salts (30%)
PHOSPHORUS
*Lives in sea water
*Synthesis of nucleic acids
“Obligate Halophiles”
*Phospholipids of cell membrane
Ex. *requires higher concentration of salts
(30%) *ATP synthesis

Ex. Vibrio parahaemolyticus *Cofactor for enzymes

HALO TOLERANT OXYGEN

*Can withstand an environment with high salt *Poisonous to organisms

concentration (2%)
*Dependent on enzyme systems
*inability to convert oxygen can cause problems OBLIGATE AEROBES
*Requires constant exposure to
TOXIC FORMS oxygen.
*Singlet oxygen
FACULTATIVE AEROBES
 Normal molecular oxygen in a higher *Primarily anaerobes that can
energy state tolerate the presence of oxygen.
 High reactive
ANAEROBES
*Superoxide radicals *Lives in the absence of oxygen
 “Superoxide anion” *Lacks the enzymes SOD and CAT
*Accumulation of toxic radicals may
 Formed in small amounts during
inhibit growth.
respiration
 Highly unstable
OBLIGATE ANAEROBES
*Peroxides Requires an environment completely
free of oxygen
 From hydrogen peroxide produced
from the neutralization of superoxide FACULTATIVE ANAEROBES
radicals Primarily aerobes that can tolerate
the absence of oxygen
*Hydroxyl radicals

 Most reactive form MICROAEROPHILES

 Formed in the cytoplasm by ionizing *Refers to bacteria that require low
radiation concentration of oxygen (2 to 7%)
*Cannot grow in the presence of 20
OXYGEN: GROUPS OF BACTERIA to 21 % oxygen
*Cannot grow in the absence of
AEROBES
oxygen
*Able to live in the presence of oxygen
CAPNOPHILES
*Has enzymes that convert toxic oxygen *Requires higher concentration of
derivatives. Carbon dioxide (3 to 5 %)
*Normal concentration of carbon
 Superoxide dismutase
dioxide (1%)
Converts molecular oxygen to
hydrogen peroxide
O2 + O2 +2H H2O2 +O2
 Catalase
Hydrogen peroxide is still toxic so its
converted to water and oxygen.
2H202 2 H20 +O2
BACTERIAL GROWTH AND REPRODUCTION
DEFINITION OF TERMS:
*Refers to an increase in bacterial NUMBER,
NOT an increase in the SIZE
*Mode of reproduction: BINARY FISSION
INSERT PIC
For example, if 5 cells were allowed to divide 9
times, this would result in
5 x 2^9 = 2560 cells
To calculate the number of generations a
culture has undergone, cell numbers must be
converted to logarithms. Standard logarithm
values are based on 10. The log of 2(0.301) is
used because one cell divides into two.
Number of generations=

Log number of cells (end) – log number of cells (beginning)

*The interval of time between successive binary
0.31
fission of a cell or population.
To calculate the generation of time for a
*Time required for a cell to divide and its population:
population to double.
Minutes/generation=
GENERATION TIME
60 min/hr x hours
*Varies in bacterial species
Number of generations
 Staphylococcus aureus: 15 minutes
As an example, we will calculate the generation
 Mycobacterium tuberculosis: 15
time if 100 bacterial cells growing for 5 hours
hours
produced 1,720,320 cells:
 Treponema paallidum: 33 hours
14 generations=
*Depends on the condition the bacteria is in
Log 1,720,320 – log 100
 Optimum condition: faster GT
0.301
LOGARITHMIC REPRESENTATION OF
BACTERIAL POPULATIONS 21 minutes/generation=

60 min/hr x 5 hours
As a cell divides, the population increases
exponentially. Numerically this is equal to 2 14 generations
(because one cell divides into two) raised to the
number of times the cell divided (generations) ;
GENERATION TIME: IMPORTANCE
2^number of generations
*Useful determining the amount of time that
To calculate the final concentrations of cells:
passes before disease symptoms appear
Initial number of cells x 2^number of generations =
Number of cells
*Determine the effect of newly developed BATCH CULTURE SYSTEM
preservatives on the culture.
 Single batch of medium only
PHASES OF GROWTH  No additional nutrients are added
 Waste products are not removed
LAG PHASE
CONTINOUS CULTURE SYSTEM
 Adapting to new environment
 No growth rate  “Open culture system”
 Phase of physiologic youth  Additional of nutrients
 Intense metabolic activity  Removal of waste
 Synthesis of enzymes and various  Chemostat
molecules.  Turbidostat

LOG PHASE

 Undergoes binary fission at the fastest

rate
 Has the shortest and constant
generation time
 “Exponential phase”
 Phase of balanced growth

EVENTS:

 Symptoms of infection
 Appearance of colonies

STATIONARY PHASE

 Has the highest number of cells

 Reproduction= death
 “Plateau Phase”

FACTORS OF DEATH

 Nutrient depletion
 pH change
 Waste accumulation

DEATH PHASE

 Dead cells > Viable cells

 Negative exponential growth

IN VITRO ENVIRONMENT