Professional Documents
Culture Documents
Introduction To Bacteriology Part I
Introduction To Bacteriology Part I
HISTORY
Important Events in the Development of
Microbiology
1590-1595 Hans Jansen developed the first compound microscope.
1676 Antonie van Leeuwenhoek discovered animalcules.
1786 Otto Friedrich Muller produced the first classification of
bacteria.
1798 Edward Jenner introduced the smallpox immunization through
cowpox inoculation.
Important Events in the Development of
Microbiology
1838-1839 Theodor Schwann and Matthias Jakob Schleiden
introduced the cell theory.
1858 Rudolf Virchow proposed the theory of biogenesis.
1861 Louis Pasteur used the aerobic and anaerobic
terminologies in describing the metabolism of yeast.
1880 -described the weak strain of organisms as attenuated in
developing vaccines
1881- developed the anthrax vaccine.
1885 -produced the rabies vaccine.
1867 Joseph Lister published his work on antiseptic surgery.
Ignaz Semmelweis (1816-1865)
He demonstrated that routine handwashing can prevent the
spread of diseases.
1875 Ferdinand Colm used the genus name Bacillus in
classifying bacteria.
HETEROTROPHS
-require complex organic sources such as glucose
FASTIDOUS
-require additional nutrients such as vitamins, hemoglobin etc.
ENVIRONMENTAL FACTORS INFLUENCING
GROWTH
A. PH
-pathogenic bacteria grow best at neutral pH
-culture media area adjusted to pH 7.0-7.5
B. TEMPERATURE
1. Psychophiles-(10-20 degrees celcius)
2. Mesophiles-(20-40 degrees celcius)
3. Thermophiles -(50-60 degrees celcius)
ENVIRONMENTAL FACTORS INFLUENCING
GROWTH
C. GASEOUS COMPOSITION
1. Obligate aerobes
2. Microaerophilic
3. Facultative anaerobes
4. Obligate anaerobes
5. Capnophilic- requires increased CO2
6. Microphilic- requires decreased CO2
BACTERIAL GROWTH
• Binary fission:one cell divides into 2 cells
• Generation time/ doubling time- time required for once cell to divide
into two cells
BACTERIAL GROWTH (PHASES)
1. Lag phase
2. Log phase
3. Stationary phase
4. Death or decline phase
Bacterial Genetics:
1. Genotype
2. Phenotype
3. Bacterial genome/bacterial
chromosome
4. Plasmid
TRANSPOSONS
Bacterial Genetics:
5. 6. Insertion sequences-small mobile nucleic acid strands
capable of activating or inactivation genes
7. mutations-changes that occur in the DNA code
8. Point mutation- change in one nucleotide base; leads to a
change in a single amino acid within a protein; single base is
replaced with different one
9. Frameshift mutation- one or few nucleotide pairs are deleted
or inserted in the DNA
GENETIC RECOMBINATION
• MECHANISMS:
1. Transformation- uptake and incorporated of naked DNA into
bacterial cell
2. Transduction- transfer of bacterial genes by bacteriophage
3. Conjugation- transfer of genetic material from a donor to
recipients strain which requires close contact between 2 cells
-mediated by the formation of a sex plus
-plasmids and chromosomal genes can be replicated and transferred
by this method.
Bacteriology:
CONTROL OF
MICROORGANISMS &
SPECIMEN COLLECTION AND
HANDLING
STERILIZATION
-removal/killing of all forms of life
DISINFECTION
-removal/killing of pathogenic organism but not
necessarily bacterial or other spores
*disinfectants:applied to inanimate objects
*antiseptics: applied to living tissues
FACTORS THAT INFLUENCE THE
DEGREE OF KILLING
1. Types of organisms
2. Number of organisms
3. Concentration of disinfecting agent
4. Amount of organic soil present
5. Nature of the surface to be disinfected
6. Temperature of pH of process
7. Length of contact time
8. Type of water available (hard vs soft)
9. Biofilms(community of bacteria)
10. Compatibility of disinfectants
Types of organisms according to resistance
to killing
1. Bacterial spores most resistant
2. Mycobacteria
3. Non-lipid viruses
4. Fungi
5. Bacteria (vegetative cells)
6. Lipid viruses (enveloped) least resistant
Device Classification
1. Critical materials- invade sterile tissues; require
sterilization
2. Semicritical materials-come in contact with
mucuos membranes; require high level
3. Noncritical materials- come in contact with
intact skin; require intermediate-level to low-
level disinfection.
Device Classification
I. Physical Methods
A. Heat
4. Boiling
o 100 °C in 10mins
o Only disinfection is achieved (tuberculocidal)
5. Pasteurization
o 63 °C for 30mins or 72°C for 15 secs (tuberculocidal)
o Usually employed in the food industry
II. Filtration
Mechanism of action:
1. Reaction with components of the cytoplasmic membrane
2. Denaturation of cellular proteins
3. Reaction with thiol(-SH) groups of enzymes
4. Damage to RNA and DNA
III. Chemical Methods
A. Alcohols
o Most common: Isopropyl and ethyl (70%)
o Non-sporicidal, non-tuberculocidal
o Denatures proteins
B. Aldehyde
1.Formaldehyde
2.Glutaraldehyde
o germicidal and sporicidal
o Kills through inactivation of DNA and RNA
o Sterilizer for medical equipment
III. Chemical Methods
C. Halogens
1.Iodine(mainly used as antiseptics)
o Non-sporicidal,non-tuberculocidal
F. Phenolics
oDisrupts cell wall
oPrecipitate proteins
oSome may have tuberculocidal acitivity
III. Chemical Methods
G. Gases
Use of Anticoagulants:
Sodium polyanetholsulfonate (SPS);most common
Sodium amylsulfate (SAS)
Heparin : viral culture
Citrate and EDTA should not be used
Use of Holding and Transport Media
• *keeps the organism viable while inhibiting
replication
Examples:
Carey-Blair
Modified-Stuart transport media
Amies
Storage of Specimens:
Refrigeration (4 °C): urine, viral
blood spn, catheters, swabs
Incubator (37 °C) : CSF, bacterial
blood spn, cultures on agar plates
Room temperature (25°C): spns
for fungus isolation
Freezing (-70 °C) : if processing
will be delayed more than 4 days
Labelling and Rejection of Specimen
I. Requisitions
II. Unacceptable/Sub-optimal spns
1.non-matching requisition and specimen
2. Leaking specimen
3. Syringes with needles attached
4. Stools contaminated with urine or barium
5. Anaerobes from inappropriate sources/containers
6.Unpreserve specimen older than 2 hours
7. Refrigerated blood cultures for bacteriology
8. dried-up specimen
9. Specimen in formalin
10. Specimen in non-sterile container
11. QNS Specimen
Bacteriology:
DETECTION METHODS
Universal Precautions
1. Barrier protection-gloves, masks, lab gown, puncture resistant
biohazard container(color)
2. Engineering controls – protects while working(BSC) from aerosols
a. Heat
b. UV
c. HEPA(High efficiency particulate air) – removes 0.3um particle
BSC classes
CLASS DESCRIPTION