Community Public Health Prepared by: Christian M.

Sanuco,RMT

FUNDAMENTALS OF BACTERIOLOGY

BACTERIAL MORPHOLOGY
A. Shape. Along with the other properties, shape is used to identify bacteria. It is determined by the
mechanism of cell wall assembly.
1. Bacilli – Rods
2. Cocci - Spheres
3. Spirochete - Spiral
a. Vibrio – Straight rod or with single rigid curve
b. Spirillum – Rigid helical cord
c. Spirochete – Flexuous helical cord
4. Pleomorphic organism – vary in size and shape
5. Altered shape – Due to antibiotics that affect cell wall biosynthesis
B. Arrangement
1. Pairs – Diplococci and Diplobacilli
2. Chains – Streptococci and Streptobacilli
3. Grape-like clusters or Bunch of grapes – Staphylococci
4. Group of four or Tetrads – Peptoccous
5. Packed of eight or Cuboidal – Sarcinae
6. Palisades or Chinese letter – Corynebacterium
C. Size: The unit of measurement is micrometers (0.001mm = to 1micrometer)
1. Smallest pathogenic bacillus – Haemophilus (0.2 x 0.5µm)
2. Largest pathogenic bacillus – Bacillus anthracis (1 x 3-10µm)
D. Capsule: The mucilaginous envelope that surrounds some bacteria
1. Polysaccharide – Structure surrounding a bacterial cell and is external to the cell wall
2. Poly-D-glutamic acid – The only capsule of Bacillus anthracis
3. Slime layer – loosely arranged extracellular material that surrounds bacterial cells; found in biofilm
(ex. S. epidermidis uses heart valve catheters)
Function: May increase the virulence by preventing phagocytosis (immunoevasion)
Encapsulated microorganisms
Some Streptococcus pneumoniae
Nasty Neisseria meningitidis
Killers Klebsiella pneumoniae
Have Haemophilus influenzae
Some Salmonella typhi
Capsule Cryptococcus neoformans
Protection Pseudomonas aeruginosa
Community Public Health Prepared by: Christian M. Sanuco,RMT

E. Cell wall: Also known s the peptidoglycan layer or murein layer.

Functions:
a. Protects bacteria against osmotic pressure
b. Gives shape to the bacteria
c. Confer the gram’s reaction of the bacteria
d. Usual target of antimicrobial drugs

1. Gram-positive cell wall

 Composed of very thick protective murein layer
 Consist of glycan chains of alternating N-acetyl-D-glucosamine (NAG) and N-acetyl-D-muramic
acid (NAG)
 Teichoic acid – negatively charged and contributes to the negativity of the cell wall; it may
also bind and regulate the movement cations into and out of the cell
2. Gram-negative cell wall
 Composed of thin layer of peptidoglycan.
 Periplasmic space – between outer membrane and gram-negative cell wall. It is here that
secreted proteins (mannoproteins), which gives the bacteria an ability to resist permeability
against foreign matters.
3. Acid fast cell wall
 Has a gram-positive reaction
 Concurs resistant to drying
 Contain waxy layer of glycolipids (mycolic acid) bound to the exterior of the cell wall.
Acid-fast organism
NO Nocardia
M Mycobacterium
I Isospora
L Legionella
C Cryptosporidium
Others: Rhodococcus, Gordonia, Tsukamurella, Corynebacterium

4. Bacteria without cell wall BUT contains sterols(3 layer of cell membrane) in their cell membrane.
- Mycoplasma
- Ureaplasma
Community Public Health Prepared by: Christian M. Sanuco,RMT

F. Outer membrane
1. Only in gram-negative bacteria
2. Composed of Proteins, phospholipids and lipopolysaccharide.
a. Lipopolysaccharide is vital to evade the host defenses, it also contributes negative charge of the
bacterial surface and is comprised of:
 Lipid A: major constituent; toxic
 Core polysaccharide
 O polysaccharide
 Endotoxin
G. Cell membrane
1. Essential component of both gram (+) and gram (-) bacteria.
2. It is the site for bacterial energy metabolism.
H. Endospores
1. Function: survival response to certain adverse nutritional condition, such as depletion of certain
resource
2. Highly resistant to desiccation, heat and various chemicals.
3. Structure poses a core that contains many cell components, a spore wall, a cortex, a coat and a
exosporium.
 Core contains calcium dipicolinate, which aids in heat resistance within the core.
4. Spore forming microorganism.
 Bacillus and Clostridium
Clostridium tetani Terminal spore
Clostridium botulinum Subterminal spore
Bacillus anthracis Central spore
5. Endospore stains:
- Schaeffer-fulton stain = stains endospore green
- Doemer’s stain
- Writz-conklin
I. Flagella: bacteria possess a flagellum for locomotion
a. Atrichous: no flagella
b. Monotrichous: single flagellum at one pole
c. Amphitrichous: single flagellum at each pole
d. Lophotrichous: tuff of flagellum at one pole
e. Peritrichous: Flagellum all over the organism
 Common in pathogenic bacteria ( monotrichous and peritrichous )
1. Motility can be seen best at 25 ° C at growth of 18 hours (young culture) and for nonmotile (35-37 °C)
Community Public Health Prepared by: Christian M. Sanuco,RMT
2. Flagellar stains: contains tannic acid that coats flagella and precipitates during stains.
a. Leifson
b. Fisher and conn
c. Gray
3. Endoflagella: also known as the axial filament, this is found commonly in spirochetes
Spirochete organism:
 Borella
 Leptospira
 Treponema
4. Types of motilities.
 Tumbling motility: Listeria
 Darting motility: campylobacter
 Gliding motility: Capnocytophaga
 Swarming motility: Proteus
J. Pili/Fimbriae: hair like extensions that extends to the environment
TYPES of Pili
Common pili: For attachment
Sex/ conjugate pili: for conjugation
Community Public Health Prepared by: Christian M. Sanuco,RMT

FUNDAMENTALS OF BACTERIOLOGY

BACTERIAL GROWTH AND REPRODUCTION

A. Reproduction
- Bacteria reproduce via binary fission
B. Growth cycle
a. Lag phase: little or no multiplication but enzymes are active
b. Logarithmic/ Exponential phase: organism grows at maximum rate (most sensitive to antimicrobials).
c. Stationary phase: Growth ceases because nutrients are exhausted or toxic metabolic product are
accumulated
d. Decline/Death phase: Bacteria slowly decreases viably but count remain.
C. Mechanism of Bacterial Gene Transfer
1. Transformation
 Direct uptake DNA from surrounding environment
 Allows for evolution of DNA over time.
 Very useful techinique in micro labs
 Introduce genes to bacteria for replication
2. Conjugation
 Transfer gene from one cell to another via pili: requires physical contact of two organisms
 Transferred via plasmid
 PLASMIDS: small DNA molecule within cell
3. Transduction
 Transferred via bacteriophage
 Virus picks up DNA, transfer to another bacteria
 Cycle of bacteriophage
a. Lytic cycle
b. Lysogenic cycle

D. Nutritional Requirements
1. Heterotrophs/organotrophs: require preformed organic compound (sugar, amino acids) for growth.
2. Autotrophs/Lithotrophs: do not require preformed organic compound for growth because they can
synthesize them from inorganic compounds and carbon dioxide.
Community Public Health Prepared by: Christian M. Sanuco,RMT
E. Energy requirements
1. Phototrophs: uses light as energy
2. Chemotrophs: uses redox potential generated by various chemical reaction
F. Oxygen requirements
1. Aerobe: grows well in the presence of atmospheric free oxygen
a. Obligate aerobe- cannot grow in absence of free oxygen. Contain superoxide dismutase, which
protects from the toxic oxygen.
b. Microaerophile – requires reduced amount of free oxygen
2. Anaerobe- grows well in the absence of free oxygen.
a. Obligate aerobe- cannot grow in the presence of atmospheric free oxygen, lack of superoxide
dismutase.
b. Facultative anaerobe- Fundamentally an aerobe but can grow in the absence of free oxygen
c. Aerotolerant – does not grow well, but survives in the presence of atmospheric free oxygen
Obligate aerobes Obligate Aerotolerant Microaerophiles Facultative
anaerobes anaerobes
P.aeruginosa Actinomyces Lactobacillus Campylobacter Most
Nocardia Bacteriodes Propionibacterium Helicobacter pathogenic
Bacillus Clostridium bacteria are
Mycobacterium Fusobacterium facultative
CorynebacteriumEubacterium anaerobe
Provotella
G. Capnophile- growth enhanced by the increased CO2
Streptococcus
Campylobacter
Haemophilus
Helicobacter
Moraxella catarrhalis
Mycobacterium
Pathogenic neisseria
H. Thermal Requirements
1. Psychrophilic: Grows below 10°C-------Listeria moncytogenes (blood bank contaniminants)
2. Mesophilic: Grows at 20-40 °C----------Most pathogens
3. Thermophilic: Grows at 50-55°C--------Effective as biologic indicators.
I. Incubation conditions

Aeorobe Ambient air w/c contains 21%O2+ 0.03%CO2

Anaerobe 0% O2, 5-10% CO2, 5-10%H2 + 80-90N2
Capnophiles 5-10% CO2+ 15% 02
Microaerophiles 5-10% 02+ 8-10% CO2
Community Public Health Prepared by: Christian M. Sanuco,RMT

Fundamentals of Bacteriology
STERILIZATION AND DISINFECTION
A. Terminology
1. Sterility – total absence of viable microorganism as assessed by no growth on any medium
2. Bacteriocidal – kills bacteria
3. Baceriostatic- inhibits growth of bacteria
4. Sterilization – Removal/killing of all microorganism
5. Disinfection- Removal/killing of disease-causing microorganism in inanimate object
6. Antisepsis- any procedure that inhibits the growth or multiplication of bacteria either complete
destruction or not

B. Methods of Sterilezation and Disinfection

1. Moist heat – Coagulates protein
a. Boiling – 100°C for 15-30 mins kills all vegetative organisms but not all spores and viruses.
b. Fractionation – alternate heating
i. Tyndalization – flowing steam for 30 minutes on 3 successive days
ii. Inspissation – 75-80°C for 2 hours on 3 successive days
c. Autoclaving- uses steam under pressure, 121°C at 15psi for 15-30mins
i. Most effective method for sterilization
ii. Can kill spores and viruses
iii. Uses pressure to raise the temperature
iv. Biologic indicator (Bacillus stearrothermophilus)
d. Pasteurization: recommended for milk and dairy products
i. Batch: low temperature holding (63°C for 30mins
ii. Flash: Hight temperature at 72°C for 15 seconds
2. Dry heat- Kills by oxidation
a. Oven 160-180°C for 1-2 hours
b. Flame – to sterilize inoculating loop.
3. Filtration
a. Asbestos filter- 98% effective
b. Membrane filter- 100% effective
4. Radiation
a. UV light
b. Ionizing Radiation (gamma rays. X-rays)
5. Ethylene oxide gas (COLD STERILIZATION)
 Effective for materials that cannot be autoclave ex. Rubber, plastic tubes
Community Public Health Prepared by: Christian M. Sanuco,RMT
6. Disinfectants and antiseptics
a. Quaternary ammoniums -inactivate bacteria by binding their hydrophobic and lipophilic groups
and interacting with their cell membrane altering metabolic processes.
b. Phenolics- denature proteins
c. Chlorine- oxidizes organic matter
d. 2% aqueous glutaraldehyde - for lensed instrument
e. Chlorhexidine- is a diphenyl cationic analog that is a useful topical disinfectant.

BIOLOGIC INDICATORS
Autoclave Bacillus stearrothermophilus
Ionizing radiation Bacillus pumilis
Dry heat oven Bacillus subtillis var niger
Ethylene oxide gas Bacillus subtillis var globigii