Microbiology Notes Summary

OCDEN, Junalyn B.
Microbiology Notes (2015)
LECTURE
NOTES
MICROBIOLOGY
OCDEN, JUNALYN B. (2015)
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OCDEN, Junalyn B.
Microbiology- study of microorganism
Micro- minute, small
Bio-life
Logus- study of
Branches:
1. Bacteriology
- Study of bacteria
- General Branch, involves pathogen/non-pathogen
-Pathogen: disease causing bacteria to Man
-study of medically important bacteria: Pathogens
Example:
Salmonella Typhi
- Eschirichia coli- bacteria that causes Gastroenteritis
- Neisseria gonorrhae
- Treponema pallidum- spirochete; subspecie: pallidum- causes syphilis
- Vibrio cholera- cholera , indigestion of contaminated water
2. Parasitology- study of parasites
Example:
- Entamoeba histolytica- causes Amoebiasis- Protozoa(ingestion of contaminated water)
- Plasmodium falciparum, malaria, vivax, ovale Malaria- Protozoa
Holy 3 because if you have one, you will have all
- Ascaris Lumbricoides- contaminates water and food, 20,000 egss per day
- Trichiuris Trichura
- Hookworms
3. Virology- study of virus
Example:
- Colds- caused by Rhinovirus rhinitis
- AIDS- caused by HIV , retrovirus
- Hepa A- caused by picorna(RNA); Hepa B- Hepadna(DNA)
- SARS(Severe Acute Respiratory Syndrome)
- Ebola
4. Mycology- study of fungi
Example:
- Candida albicans, Aspergillus fumigates, Histoplasma capsulatum
5. Phycology (Alcology)- study of algae
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SCOPE OF MICROBIOLOGY:
1. Agricultural Microbiology- use of microorganism to fertile soil.
*night soil- ex. Urine, feces used as a fertilizer
2.
3.
*Rectal
Food microbiology- example: yakult (Lactus bacillus), yogurt

Sanitary Microbiology
Proper waste disposal
Making water safe for drinking
Water analysis( absence of coli forms)
Making food good for consumer
Swab- no collection of stool
HISTORY:
1. Varro: postulated principle of contagion.
Contagion: - pertained as external forces
- Organism that are bred, minute organisms
- Present in air, water and external forces
2. Roger Bacon: according to him, invisible creatures are present in the environment and it
causes diseases.
3. Girolamo Fracastoro / Fracastorius- he combined the principle of Varro and Bacon- there are
disease causing external forces in the environment that cannot be seen by the naked eye.
(1546)
4. Robert Hooke- He used the microscope to examine a cork and he discovered that there are
little boxes Cell theory (1665)
5. Anton Van LeuwenhoekHave 3 major contributions:
1. First to observe a bacteria called Ahimacules.
2. Descried three basic shapes of a bacteria
Coccibacillispirilli3. Described protozoa
6. Spontaneous Generation/ Abiogenesis
7. Rudolf Virchow
- opposed of the abiogenesis
- life come from non-life
Experiment:
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Putrid meat (exposing it for one week>> formation of maggots
-came up with biogenesis
8. John Needham:
Experiment:
Used chicken brothheatboil sealed in a container CLEAR
9. Francesco Redi:
Experiment:
Putrid Meatplaced in a container and -> NO MAGGOTS
a gauze on top
Conclusion: Maggots came from flies
10.
Edward Jenner (1796)
- Vaccination for small pox (pandemic)
- Attenuated vaccines
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Mathias Schleiden and Theodore Schwann(1838-1839)
- All organisms are made up of cells
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J. Henle- introduced the Germ Theory ut he didnt prove it.
*Louis Pasteur was the one who explained the germ theory further.
13.
Ignas Semmelweiss
- Puerpueral Fever/ Childbirth fever/childbed fever caused by Streptococcus
pyogenes
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John Snow- explained how disease were transmitted especially cholera (transmitted
through water- human sewage)
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Louis Pasteur- father of Microbiology/ bacteriology
Experiment:
Yeastsugar Co2 and ethanol
*discovered fermentation
*air is filled with microorganisms
- Supporter of biogenesis
-Pasteurization- to kill microorganisms specifically those that cause SPOILAGE.
*LTH- Low Temperature Holding/ Bath Method (63-650C for 30mins)
*HTSH- High Temperature Short Time/Dash method of Pasteurization (720C; 15secs)
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17.
Joseph Lister: father of Aseptic Surgery

Robert Koch: father of Bacteriologic Techniques
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-Discovered and came up with pure culture techniques for handling in the lab.
-Described and discovered Bacillus Anthracis- anthray
-Mycobacterium tuberculosis
4 Postulates:
1. Microorganism or causative agent must be present in every case of disease except carriers.>>
an individual that carries the bacteria but doesnt have the signs and symptoms.
Different types of carriers:
a. Active carrier- an individual will acquires the causative agent disease process
recover individual will carry the causative agent indefinitely
b. Passive carrie- an individual acquires the organism and does not experience the
manifestation of the disease but can help in the transmission. Example: Malaria
c. Incubatory- transmit disease during the incubation period. Example: Hepa A

d. Convalescence Carrier- transmits during the recovery period. Example: Chicken pox
2. The microorganism must be isolated and grown and culture outside the body.
Exception: Not all microorganisms grow inside the body.

Example: Mycobacterium leprae- animal armadillo-treponema pallidum-subspecie: pallidumcause
venereal syphilis.
3. Microorganisms must be introduced into a healthy susceptible lab animal. Exception: To those
who are resistant
4. Microorganism must be isolated again and grown in pure culture inside the body. Purpose; for
comparison.
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20.
Walter and Fany Hesse

- Introduced agar-agar
Elie Metchnikoff
- Discovered phagocytes and its functions.
- Cellular mediated immunity or cellular theory of resistance.
Christian Gram: discovered Gram Staining
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*differentiate organism
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Paul Erlich- discovered Salvarsan/ 6C6- treatment for syphilis.

- Introduced HMI( Humoral Mediated Immunity)
*B-cells produced by anto-bodies
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23.
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Alexander Fleming- discovered penicillin from a fungi (penicillum notatum)

Carl Weese- divide organisms into 3 domains: Bacteria, Eukarya, Archaea
Emil Von Behring- Shibasaburo Kitasato
- Discovered dyptheria which is a treatment (anti-toxin)
----------------------January 31,2015--------------------------------Taxonomy
- Classification of all living organism
- Systematic arrangement of organisms into related groups
Classifications:
Related groups; taxon, taxa
I.
Kingdom Classification
a. 2 kingdom classification
-1753 by Carl Von Linne
-divided into 2:
-Static non-motile organisms; example: plantae
-Motile example: Animalia
b. 3 Kingdom Classification
- 1866 by Ernst Haeckl
-divided into 3: animalia, plantae, protista (cover specifically bacteria, fungi,
protozoans, algae)
c. 5 Kingdom Classification
-1969 by Robert Whittaker
-animalia,plantae,protista (protozoa & algae), fungi, Monera( bacteria)
Criteria to come up with 5 kingdom:
a. Cell Type:
Eukaryotes
Animalia, plantae, protista
Cell that have true nucleus
Prokaryotes
Bacteria only
Cells without true nucleus
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b. Cellular Organization:
- Unicellular or multicellular
c. Nutritional type:
1. Food acquisition:
-cell mouth
-Process of ingestion- eukaryotes
-Absorptive- prokaryotes
2. Energy Source:
*phototroph- gain energy from sunlight
*chemotroph- energy from either inorganic/ organic compounds
3. Carbon Source:
* Autotroph self- feeders; get carbon from inorganic substances
*Heterotroh from organic compounds (Carbohydrates, Proteins and Amino Acids)
+ Parasites- rely to a host; feed on living tissue
+Saprophytes-organism that will feed on dead organic matter.
II.
-
System Classification:
1. Phenetic/ Taximetric:
Organism classified according to observable traits or over-all similarities.
Example:
Morphology of a bacteria
Staining Reactions Most bacteria would contain peptidoglycan (can be Gram + or Gram - )
Metabolic Rate of the bacteria
2. Phylogenetic/ Phyletic:
Divided into: - evolutionary development & genetic composition
7 taxonomic levels:
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Kingdom: made up of two or more phylum

Specie: interbreeding organism that can produce fertile off springs; principal natural taxonomic
unit
*Strains- lower than a specie; organism that appear and behave differently from other organism
within the same specie.
Classified as:
o Biovar/Biotype: Biochemical properties
o Morphovar: based on morphology
o Pathovar/ Pathotype: Pollogenecity
o Serovar- antigenenic characteristics
DOMAINS OF LIFE:
- 1978 BY Carl Woese and George Fax
a. Eukarya- different eukaryotes: Animalia, Protista, Plantae, Fungi
b. Bacteria- Prokaryotes- Eubacteria (termed as true bacteria, they have peptidoglycan,
Carbohydrate backbone)
Peptidoglycan 60-100% peptidoglycan;Gram + bacteria
10-20% (Gram bacteria)
*some would have Mycolic Acid instead of peptidoglyca; example: Genus Mycobacterium- would
undergo Acid Fast Stain)
Acid Fast Bacilli- red bacteria; blue background

Non- Acid Fast Bacilli- blu bacteria; red background
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Cyanobacteria blue-green algae, similar to prokaryotic cells, have similar shape
not totally:
Differences in shape: flat, square shape
*Aquatic photosynthetic- manufacture their own food
c. Archaea- cross between eukaryotes and prokaryotes
Eukaryotes: Genes, metabolic pathways- spend more energy as compared prokaryotes.
TAXONOMY OF BACTERIA
1. Domain- Bacteria
2. Kingdom- Monera
3. Division/Phylum- ends with es
4. Class
5. Order- ends with ales
6. Family- aceae; Nesseriaceae
7. Tribe- aea- Nesseriaea
8. Genus- Neisseria
9. Species- gonorrhea
10.
Strains
Proper naming of bacteria
Binomial System of Nomenclature
-Genus name, Species epithet
Example: Escherichia coli
1. Always italicize or underline
2. Genus name: 1st letter is capitalized and the only name that can be abbreviated.
3. Species epithet: 1st letter is a small letter or capitalize everything
Bergeys manual of Systematic Bacteria
- Adding the common name or trivial name
Example: Escherichia coli Common name: Colons Bacillus
Mycobacterium leprae Common name: Hansens Bacillus
III. BACTERIAL MORPHOLOGY AND CYTOLOGY
A. Morphology: shape, size and arrangement
- Cocci- rounded
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2 cocci- diplococci- produces a horizontal line in 1 plane

Series of cocci /more than 2- streptococci
Neisseria gonorrhea (coffee bean)- red/ Gram
2 planes : tetrad
3 planes: 4 infront ;4 at the back
Multiple Planes: Exhibited by staphylococcus
Prefix Staphyle means grape like cluster
Bacilli- elongated rod shape
Bacilli in pairs- diplobacilli
Bacilli in chains: Strepto bacilli
Bacilli in Chinese letter: example: Corynebacterium diptheriae
Common Name: Kleb-Loeffers- ability to bend /to snap at the middle
Palisade/ Picket Fence:
Example: Mycobacterium leprae
Helical / comma shaped bacteria
Example: Vibrio cholera/ Vibrio comma CN: Commas bacillus
-not a spirilli but looks like one; Gram curved bacilli
Spirochetes: tight rigid coils/ loose irregular coils
a. Treponema
T. pallidum subspecie pallidum
T. pallidum subspecie endemicum
T. pallidum subspecie pertenue
T. carateum
b. Borrelia
B.recurrentis- fever
B. burgdorferi-lime disease
c. Leptospira
L. interrogans serovar ictohemorrhagiae-from rats
L. interrogans serovar canicola- from dogs
B. Cytology
Prokaryotes
1. Cell envelope- stratified (layers)outer structure
3 layers:
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a. Cell membrane- semi-permeable membrane

Components: 40% phospholipids;60% proteins
Fluid- mosaic model glycerol(polar), fatty acids(non-polar)
Functions:
1. Location of transport systems:
Osmosis: increase water,oxygen, Carbon dioxidedecrease conc.
Facilitated Diffusion: from high to low conc.
o Form of transport protein
Active Transport: use of energy- from low to high conc.
Group Translocation: from low to high
Utilizes energy : high energy phosphate compounds
Example: Glucosepyruvic acid
2. Location of Enzyme Systems:
Help in the production of energy
-oxidative phosphorylation
> Biosynthesis: production of macromolecules
> Extracellular Enzymes: produce enzyme from the bacteria released to the environment to
break down any molecule present outside its body.
Example: H2O2 H2O+O2 (bubbles)
3. Chemotaxis: migration of a cell in response to a chemical stimuli.
Example: inside the cell membranesensing proteindetects quality of the chemical
stimuli.
Bacteria to stimuli: positive stimuli
Bacteria away from stimuli: negative stimuli
4. Reproduction:
Prokaryotes have:
Mesosomes- invagination of a cell membrane
b. Cell Wall- rigid structure that encloses the cell membrane

Components:
a. Made up of peptidoglycan: backbone of Carbohydrate
Polymers of disaccharides:
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NAM: N- acetyl muramic
NAG: N-acetyl glucosamine
- Peptide: 4 proteins (tetrapeptides)
Function:
Provide rigidity
protection against osmolysis
Determine difference in Grams reaction:
60% to 100% peptidoglycan (20-80nm); molecules are highly crossed link ;bonded by
interpeptide bridge
Technoic Acid: attached to the cell wall and NAM of a peptidoglycan.
b. Gram-cell wall
10-20% peptidoglycan(8-11nm)
No cross linking, no interpeptide bond
Outer membrane contains phospholipids composed of protein
Outer membrane:
Types:
a. Porins- fixed pore diameter; inside
b. Lipoprotein- smallest attached to a
cell
c. LPS (Lipopolysaccharide)- Function:
bacteria is lased
Responsible for a general signs and
hypovolemic shock)
the porins contains transmembrane channels (antibiotics)

peptidoglycan of a Gram cell wall. Function: stabilize
composed of Lipid A (an endotoxin released if the
symptoms of a disease. (fever hemorrhage, pain, shock
*Polysaccharides:
5 units:
-core polysaccharides
-outer polysaccharides for species identification ; Why? Because of a certain Ag that we
can detect (O Ag/ Ohne Ag/ Somatic Ag)
Gram Staining- differential staining technique through color, shape, arrangement.
Size of bacteria- 0.2-2.0 um by 2.8 um
Inside- intracellular; Outside- extracellular
Reagents or dyees:
1. Crystal Violet: 1 min
2. Grams Iodine- mordant/accentuator; intensifies the color of the primary stain.
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3. Acid Alcohol- (95%); decolorizer (20 secs); removes excess stain around the bacteria
4. Safranin- counter stain or secondary stain
What causes gram positiveness of a bacteria?
-10%-20% peptidoglycan
5. Pathogenecity
6. Stie of Antibiotic Action- penicillin (destroys the cell wall); hydrolysis; cytoserine;
bacitracin; vancomysin
7. Site of Antigen determinant
-Glycocalyx- gelatinous structure outside the cell wall; not all have glycocalyx. Made up
of:
Polysaccharides- examples: Streptococcus pneumoniae CN Preumococcus ,Krebsiella pneumonia
CN: Friedlanders;
Polypeptides: - Polyglutamic Acid example: Bacillus anthracis
Types:
1. Capsule: glycocalyx with a definite shape.
2. No definite shape
Function:
1. Protection against dessication
2. Pathogenecity
3. Anti-Phagocytic
4. Attachment to tissue being infected
5. Antigenic determinant- detect if a bacteria has a capsule ( K Ag)-Kapsel
Periplasm.Periplasmic space: in between of the cell membrane and polysaccharide of the cell
wall.
II. Cellular Apendages:
a. Flagella- for motility, antigenic determinant H Ag(Hauch)
Parts:
1. Filament- whip
2. Basal body- embedded in the cell membrane
3. Hook- between the filament and basal body.
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Eukaryotic: whipping bacteria; counterclockwise:forward; clockwise-tumbling; continuous clockwisestop.
Flagellar arrangement:
1. Atrichous: bacteria has no flagella thus the bacteria is non-motile bu some can release an
enzyme and become motile
2. Monotrichous- one flagella on one side
3. Lophotrichous- group of flagella on one side
4. Amphitrichous- tufts of flagella on both sides
5. Peritrichous- flagella all around the bacteria
b. Endoflagella/ Axial Filament:
Whip-like and for motility; only spirochetes have these
Movement: from left to right
Function: for motility
c. Pili/ Fimbrae
Short hair-like projections
Control protein pilin
Function:
1. Attachment: ordinary/ common pili
2. Pathoggenecity
3. Conjugation- transfer of genes between 2 cells through physical contact (Function of
the sex pili)
III.Cytoplasmic Structures
a. Nucleoid: 1 chromosome rounded/spherical no definite shape and proteins no histones , DNA
chromosomes
b. Plasmids- extrachromosomal DNA
Types:
1. Free plasmids- independent replicate from the DNA itself; can be transferred in the
daughter cells
2. Integrated Plasmids- integrated within the nucleoid; replicate together with DNA.
Example:
F-plasmids- Fertility, produce additional characteristics, responsible for expressing the
function of the sex pili.
R-plasmids- resistance resist antibiotics
3. Bacteriocinogenic Plasmid
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-produce Bacteriocin (protein that kills other bacteria)
4. Degrative Plasmids- withstand the effects of toxins
c. Ribosomes- granular, function for protein synthesis
How to determine ribosomes of:
Prokaryotes(70 s.u.) and Eukaryotes(80s.u.)
-site of anti-bitic action
Selective toxicity (principle)- certain destroy ribosomes inorder to kill the bacteria.
Example: Erythromycin, Tetracycline, Clindamycin, Aminoglycosides
d. Inclusion for food storage of bacteria:
1. Organic compounds: polyhydroxy butyrate
2. Inorganic compounds: Polyphosphates, sulfur
e. Endospores- dormant oval structures from vegetative bacteria- process of sporulation/
sporogenesis
Vegetative bacteria ecposed to extremes and inorder to survive it , it will develop spores.
Process is reversible (Germination)
Cortex: made up of modified peptidoglycan

Coat: keratin like protein; keratin- rough hard water proof protein always dry. Resists the
effects of chemicals
Crosporium: Carbohydrates, Lipoproteins
-helps the bacteria to gain the ability to form spores.
Function: helps the bacteria to remain alive or viable even in a extreme situation.
Normal Vegetative Bacteria
70%-80%--unfavorable-> 15% water calcium dipicolinate (very resistant to heat)
Common in Clostridium and bacillus:
Gram +
C. Tetani
C.botulinum
C. perfringens
C. welchli
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Spore location:
1. Cental spore: spore at the middle
2. Eccentric: a little bit distant to the middle
3. Sub-terminal: almost at the end
4. Terminal- present at the end
a. Terminal-swollen:
b. Terminal not swollen:
CHAPTER 4: BACTERIAL PHYSIOLOGY
A. Requirements for Bacterial Growth:
1. Temperature Requirement: Bacterial cells will always have an optimum temperature (best at
which a bacteria will multiply rapidly)
Example: 20300C = LL+UL/2= optimum temperature
250C is the usual temperature
Higher than optimum temperature=Slower rate of multiplication
a. Psychrope (-5 150C)- cold-loving bacteria
Example: Legionella pneumophila pneumonia; can be inhaled , found in freezer and air
condition
b. Psychotroph(20450C)- cause food spoilage
Example: Staphylococcus aureus= Staphylococcal Food Poisoning; cream-filled pastries,
frozen products, pasta)
c. Mesophile (25450C)- human pathogens
Example: Staphylococcus aureus
d. Thermopihiles (45-700C)- most are non-pathogenic
e. Hyperthermophiles(701100C)- most heat resistant= Bacillus Stearrothermophilus---used
as a biological indicator in an autoclave
2. Oxygen Requirement:
Oxygen energy. How? Through aerobic respiration
a. Obligare aerobe: Aerobic use oxygen for survival and converted to energy. Example:
pseudomonas can cause UTI- grape like odor.
b. Obligate anaerobe: it doesnt need Oxygen(Bacteroides, Clostridium, Clostridium
prefringens)
c. Facultative anaerobe
-the bacteria is flexible, they can survive without Oxygen.
Example: E. cole, Saccharomytes cerevisiae
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d. Facultative aerobe
-can survive with Oxygen
e. Microaerophile: 2-10% Oxygen
Example: Spirillum volutants- water
Helicobacter pylori- ulcer
f. Heteroaerant anaerobes/ Obligate fermenters
-cannot convert oxygenenergy
Where do they get energy?
Through the process of Anaerobic fermentation
Example: Lactobacillus delbrueckii subspecie bulgarius
Streptococcus pyogenes-sore throat
g. Capnele/Capnophile
5-10%
Ex: Neisseria gonorrhea
3. pH:
a. Neutrophile- 7
b. Acidophile- 1-5.5
c. Alkalinophile- 8
Human pathogens: 6.5-8.5
4. Osmotic pressure
a. Osmophile: survives at Isotonic solution
b. Halophile- can survive with a high salt concentration- salt-loving bacteria
Ex: Staphyl. aureus- can grow in MSA (Mannitol Salt Agar)
Vibrio parahaemolyticus- seafoods (contaminated)
c. Halotolerant- does not need salt for survival but it can grow in it.
5. Nutrition:
a. Non-fastidious- can grow in any envt ,doesnt need special growth factors
b. Fastidious- needs special growth factors
Example: Haemophilus influenza (Type B)
-cause Meningitis in 3-5 years old
Vaccine: HIB (Hemophilus Influenzae type B vaccine)
6. Energy and Carbon Source:
ES
CS
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Photoautotroph
Photoheterotroph
Chemolitotroph
Chemoorganotroph
Sunlight
Sunlight
Inorganic
chemical(H2S, NH3, Fe+
+)
Organic compounds
Carbon dioxide
Organic compounds
Carbon dioxide
Organic compounds
Bacteria Growth Curve:

4 stages:
LAG, LOG, Stationary , Death
BACTERIAL GROWTH CURVE
A. LAG- stage of tooling up
- It will not immediately multiply
- Needs macromolecules for multiplication: enzymes, ribosomes, nucleic acids, ATP
- stage of no growth
B. LOG Phase- Exponential Stage
- Stage of multiplication
- But still considered as an incomplete bacteria
- No cell wall
- Best time to introduce antibiotics
- Produce 1st metabolites
a. Late Log Phase
- It will reach their peak population
- Having a complete cell parts
- Resistant to anti biotics
Start to release peptides and nucleic acids so that living cells would have something to
eat
C. Stationary Phase
- The number of living bacteria is = to the number of dead bacteria
D. Death Phase
- Dead cells are greater in number than the living
E. Period of Prolonged Decline
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-
Purely dead cells
Relationship between the log number cells and time
BACTERIAL GENETICS
- Plasmids, bacterial chromosomes, sex pili
Sexual processes: transformation conjugation transduction
A. Transformation
- The naked DNA from the environment is taken up by the bacterial cells
- Important in recombinant DNA technology
Example: E. colitreat with calcium undergo heat shock DNA extractions placed in a
test tube
B. Conjugation
- Direct transfer of DNA between bacterial cells
Interaction of;
1. F+ with F- cell
F+ :
- Bacteria with a F plasmid
- Contain Bacterial chromosome
- Around it ,sex pili
F- :
- Without a F plasmid
- Contains Bacterial chromosome
- No sex pili
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2. Hfr: High Frequency for recombination
3. F prime cell (F cell)
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2 types:
1. Correct loop out- plasmids
2. Wrong loop out- part of plasmid will dissociate with of bacterial chromosome.
Wrong loop-out: F carry bacterial chromosome with plasmid and plasmids with bacterial
chromosome
C. Transduction:
Transfer of DNA between cells with the use of Bacteriophage (Bacterial virus)
2 parts:
1. Protein coat- outer covering
- Attaches the bacterial surface
2. Small piece of DNA- can also be called as phage DNA
2 basic processes:
1. Generalized Transduction/ General phage life cycle
- 15 mins, produce at least 200 new phage
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VI. HOST MICROBE RELATIONSHIP

Pathology: Pathos: suffering
- Scientific study of diseases
Infection: colonization of microbes
Disease: change in health status of an individual
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Pre-diseased diseased
*Normal Flora:
- 1x1014
- through Microbial Antagonism/ Competitive Exclusion
SYMBIOSIS: relationship where one organism is dependent from the other

a. Commensalism: One would benefit without hurting the other
Example: Corynebacteria- Eyes
Saprophytic mycobacteria- external ears, genitals
b. Mutualism: both would benefit from each other
Example: E. coli- releases vit B and K
c. Parasitism: One organism at the expense of the other
*Probiotics: live microbes that are ingested
*Prebiotics: chemicals mixed with the live microbes to permit their growth and reproduction
*Opportunistic pathogen: Non disease causing in its natural envt but disease causing in its
unnatural envt
*Resorvoir of Infection: animate or inanimate that can serve as shelter to allow their growth
multiplication and possible transmission.
1. Human Reservoir: principal reservoir
Example: Carriers
2. Animal Reservoir: zoonotic diseases
Example: Bacillus anthracis
3. Non-living reservoir:
- Soil: night soil
- Water: can be focally contaminated
*Transmission:
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1. CONTACT TRANSMISSION:
a. Direct Contact: no intermediate object between 2 inviduals
b. Indirect contact: presence of intermediate odject
Infected Infected Object(animate/inanimate=called fomites)N-II.
c.
-
Droplet nuclei:
Modified respiratory movements
Through droplet spread and travel less than 1m and doesnt stay long in air
Sneezing ,coughing
Example: Pneumonia, Pertusis/whooping cough (c.a.: Bordetella petussis)
2. Vehicle Transmission:
a. Water Borne Transmission:
- Ingestion of contaminated water
- Example: Cholera, Amoebiasis, Gastroenteritis, Typhoid fever
b. Food Borne Transmission:
- SFP
c. Air borne Transmission:
- Can travel in more than 1 meter (through dust particles)
- Stay long in air
- Example: TB, measles
3. Vector Transmission:
-Anthropods:
a. Mechanical: Passive process
b. Biological: Active process
MicrobesAnthropods multiply
Portals of Entry
1. Mucous membrane:
Respiratory System- through inhalation
GIT- ingestion
GUT-sexual
2. Skin:
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3.
4.
-
Through the openings- pores, follicles

Parental Route:
Direct inoculation of the microorganism underneath the skin
Puncture (needle stick injuries), bites
Transplacental Route:
Mother fetus
DEVELOPMENT OF DISEASE:
1. Incubation Period:
- Definite: 10-14 days

- Variable: rabies (example)
Factors affecting period:
1. Virulence
2. Number of infecting microorganisms
3. Host resistance
2. Prodromal Period:
Short: early mild signs and symptoms
3. Period :
Overt/ Severe Signs and symptoms
4. Period of decline
Starting to recover; Prone to infection
5. Convalescence Recovery
MICROBIAL PATHOGENECITY AND VIRULENCE
Virulence Factors:
1. Adhesion/Adhesiveness:
- Substances containing adhesion (aka ligand)
In the form of glycoprotein/lipoprotein
Location: in the glycocalyx, fimbriae, flagella, endoflagella
In the body:
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There are surface receptors
Bacteria containing adhesin:

- Streptococcus mutans tooth decay
Location of adhesion: glycocalyx
- Escherichia coli
Location of adhesion: fimbriae
- Neisseria gonorrhea
Attach to the GUT, pharynx, eyes
Extra genital gonorrhea

Therefore: transmission: oro-genital
Treponema pallidum
Location: endoflagella
2. Capsule:
Function: anti-phagocytic
- Body can produce Antibody for a bacteria with a capsule
Examples:
- Streptococcus pneumoniae
- Klebsiella pneumoniae
- Bacillus anthracis
- Haemophilus influenzae
- Yersinia pestis
3. Cell wall components:
a. M-protein:
- Streptococcus pyogenes (heat and acid resistant)
Function: anti-phagocytic
b. Fimbriae
- Neisseria gonorrheae
- Attachment
c. Opa-outer membrane protein
- Function: attachment
26
OCDEN, Junalyn B.
d. Mycolic acid/ Hydroxymethoxy acid
- Aka waxy lipid
4. Enzymes:
- Especially Exoenzymes
a. Condigest materials between cells
b. Promote blood clot
c. Promote digestion of blood clot
- Staphyococcus aureus:
5. Coagulase: promotes the formation of blood clot
o Converts fibrinogen to fibrin
6. Kinase: specific: Staphylokinase
For streptococcus: Streptokinase
No WBC bacteria will release kinase bacteria will spread
7. Hyaluronidase:
- Hydrolysis hyaluronic acid
In connective tissues
Function: bind the cells and maintain the shape of the eyeball
- Help the bacteria spread
- Spreading factor
8. Beta lactamase: Staph. aureus
- Aka penicillinase
9. Collagenase: help spread Gas gangrene
- Destroys collagen
Function: strength and support
Present in: Clostridium perfringens
Cause Gas gangrene/ Anaerobic Myonecrosis
10.
IgA Protease
Present in: Neisseria gonorrheae, Neisseria meningiditis
- Produce IgA (body fluids)
27
OCDEN, Junalyn B.
- Mucosal membrane---- prevent attachment of bacteria to IgA
5. Antigenic Variation:
Pathogens will alter the shape to resist destruction by anti bodies

6. Invasins
- proteins that will rearrange Actin
Present in: cell membrane
Function: part of the cytoskeleton
Proteins invasion
Attack actin
Rearrange
Salmonella typhimurium
- Ingestion of contaminated seafoods
Releases invasin
Attack the actin in cell
Result: water splash appearance or membrane ruffling
7. Toxins- poisonous substances produced by microbes
-- Toxigenicity
* Toxemia- presence of toxin in blood
*Toxoid- weakened exotoxin
Process to weaken exotoxin:
- Heat
- Formaldehyde treatment
- Iodide treatment
28
OCDEN, Junalyn B.
Function: provoke immune system to produce anti toxin.
2 types:
1. Exotoxin- produced together with growth and metabolism
Characteristics:
- Proteins in form of enzymes
- Lethal substances: small amout
- Soluble in body fluids
- Readily spread
Considered to be disease specific
Types:
a. A-B toxin
- Most common type of toxin
- Contains 2 sub-units (A &B)
A- Active enzyme components
B- Binding component (released first)
b. Membrane Disrupting Toxin
Function: disrupts the cell membrane
How will it destroy the membrane?
1. Through protein channels
Staphylococcus aureus- lyses phospholipid membrane , destroys the phospholipid portion
Clostridium perfringens
Function: kills phagocytes, help bacteria escape within phagocytes
c. Super antigens
- Produce intense immune response
29
OCDEN, Junalyn B.
Provoke Immune system

To release T cells
For CMI
-it will release a lot of

T cells
- Increase cytokines
(effect of releasing increase cytokines: onset of
fever, chills, vomiting, diarrhea, shock, death)
Examples:
1. Leukocidin:
Under MDT
Function: destroy WBC
2. Hemolysin:
- Under MDT
- Destroys RBC
3. Streptolysin:
- MDT
- Destroy RBC, WBC & other cells in the body
Type:
SLC- Streptococcus lysin O Oxygen labile ASTO/ASCT (Anti Streptococcus Lysin O test)
SLS- Streptococcus Lysin S Oxygen Stable
4. Diptheria Toxin:
- Under A-B toxin
- Function: inhibit protein synthesis
- Corynebacterium diptheriae: can produce toxin if infected with lysogenic phage
5. Erythrogenic toxin:
- Under super antigen
- Produce 3 toxins: A, B, C
- Produced by Streptococcus pyogenes
- Function: destroys wall of blood vessels
6. Botulinum toxin:
Classification: A-B neurotoxin
30
OCDEN, Junalyn B.
Function: inhibit the release of acetylcholine
Neurotransmitter
- Relaxed/ lack tone
Botulism: burring vision/ cause of Flaccid Paralysis
7. Tetanus toxin:
- An A-B toxin neurotoxin
It inhibits the release of Glycine
Blocks the relaxation of muscles
- Muscles will contract Spasmodic contraction contract
- > Spastic Paralysis
8. Vibrio enterotoxin:
- Can also be termed as
- A-B toxin
: Cholera toxin
9. Staphylococcal Enterotoxin:
- Super antigen
- Fumction: similar with vibrio enterotoxin
2. Endotoxins:
- Produced in the outer membrane (Lipid A)
- Common in G- except : Listeria monocytogenes
When?
- During bacterial lysis
- During bacterial multiplication
Released:
- Have some effects: fever, nausea, body aches, weakness
- Typhoid fever, Meningococcemia
All cocci are G+ except: Neisseria, Branhamella, Veilonella

All bacilli are G- except:
31
OCDEN, Junalyn B.
Non spore forming: Corynebacterium, Listeria, Erysiphelothrix

Spore forming: Clostridium and Bacillus
Other: Lactobacillus
I.
Gram positive cocci:
- Micrococcus: resemble staphylococcal species
- Peptococcus: only anaerobic G+ cocci
GENUS STAPHYLOCOCCUS
STAPHYLOCOCCUS
Characteristics:
Gram + cocci in clusters
Aerobic to Facultative Anaerobic

Grow readily in any ordinary media (BAM)
Colonies: round, smooth, raised, glistening
Composed of 33 species and 17 are pathogenic
Medically Important species:
Staph. aureus- can be gray to deep golden yellow colonies
Staph. epidermidis- gray to white
Staph. saprophyticus
Others:
Staph. lugdenensis
Staph. hominis
Staph. warneri
Resistant to : effects of heat, drying, 9% NaCl
Anti- microbial: 3% Hexachlorophene
Reactions: catalase + , Non-motile (no cell appendages) , non-spore forming
STAPHYLOCOCCUS AUREUS
Method used: Slide Method
How long? 5-10 seconds
32
OCDEN, Junalyn B.
CHARACTERISTICS
Gram + cocci in
clusters
Aerobic to
facultative
anaerobic
Most pathogenic G+
cocci in clusters
Why? Presence of 15
virulence factors
Only Staphylococcus
that is coagulase +
1. Bound Coagulase
Clumping factor
Attached to the
cell wall
Directly interacts
with fibrogen
2. Free coagulase
Detected
through:
- Tube method
- Extracellular
enzyme
- Contain (CRF)
Coagulase Reacting
Factor
Mannitol Fermenter
DNAse +
NO3 reduction test
+
VIRULENCE FACTORS
Catalase
- Break down
H2o2O2+H2O
- To protect
bacteria
against
toxicity
Coagulase
Staphylokinase/
fibrinolysin
Hyaluronidase
Betalactamase/
penicillinase
DNAse:
Function:
- Liquefies
purulent
discharges
- Help in the
formation of
Pus
Hemolysin
Panton
Valentine
Leukocidin
(PVL)- destroy
WBC
TSST (Toxic
Shock Syndrome
Toxin)- cause
by TSST 1
Exfoliation/
Exfollative
toxin- peeling/
desquamation
DISEASES
1. CUTANEOUS DISEASES:
- Isolated on skin
- Can cause:
Folliculitis aka
Stye
Carbuncledifference: extent
of
infection;from:epide
rmis to
dermis;manifestfluid to pus,
Furuncle- deeper
infection; from
epidermis to
subcutaneous tissue,
can infect epidermis
to fibrous tissue
Acne
Impetigo- small
vesicles with pus
2. DEEP TISSUE
INFECTIONS:
Pneumonia
- Inflammation of
the wall of the
lungs
Osteomyelitisinfection of
bone and bone
marrow
Meningitis
Endocarditisinflammation of
the inner layer
33
LAB TESTS
1. Gram Staining:
collection
through: skin
swabbing, G+
cocci in
clusters
2. Catalase +
3. Coagulase +
4. DNAse +
5. Mannitol + can ferment
salt
6. BAM: Medium to
large
colonies,
smooth,
entire,
slightly
raised,
translucent,
creamy yellow,
beta-hemolytic
7. NO3 reduction
test +
OCDEN, Junalyn B.
SSSSStaphylococcal
Scalded Skin
Syndrome
Alpha Beta
Delta Gamma
Toxinsfunction:
cytotoxinsdestroy
different cells
in the body
Protein Asimilar to the
fxn of the
coagulase,
resist the
action of
phagocytes and
resist the
process
opsonization
Capsule
of the heart
3. TOXIGENIC DISEASES:
1. SSSS- due to
exfoliation;
common to
neonates
2. TSS- common to
those who use
tampons; causes a
drop in blood
pressure= drop in
blood volume
resulting to:
Hypovolemic
shock, fever,
rashes
3. Staphylococcal
Food Poisoningincubation: 18-24
hours; Signs and
symptoms:
abdominal pain,
vomiting, head
ache, nausea,
diarrhea
STAPHYLOCOCCUS EPIDERMIDIS
CHARACTERISTICS
Coagulase
Oppurtunistic
pathogen if
immunocompromised
and Normal flora
in the skin and
VIRULENCE FACTOR
Slime layer- help
DISEASES
Sepsisconsidered as
devise related,
related to
catheter and
prosthetic valves
bacteria attach
to surfaces
34
LABORATORY TESTS
1. Grams stain
2. Catalase +
3. Coagulase
AntibioticNovobiocinsusceptible
4. BAM- small to
OCDEN, Junalyn B.
mucus membrane if
immunocompetent
Endocarditis
STAPHYLOCOCCUS SAPROPHYTICUS
CHARACTERISTICS
VIRULENCE FACTOR
Gram + in
clusters
Catalase +
Coagulase
Opportunistic
pathogen
Normal flora in
the GenitoUrinary Tract
DISEASES
LABORATORY TESTS
1. BAM: large
colonies,
entire,color
white to yellow
to orange
colonies,
glistening,
smooth, opaque,
butyrous, convex
No specific
Enzyme ureaseresponsible for
UTI (signs and
symptoms:
dysuria, fever,
flank pain)common to
pantyliner use on
a daily basis
B. STREPTOCOCCUS:
- Streptococcaceae
Characeristics:
-
medium, grayish
to white, nonhemolytic,
translucent
colonies
Streptococci in chains; in pairs (S. pneumoniae)

Facultative AnaerobicCapnophilic
Fastidous
Non- encapsulatedAvirulent; Encapsulated Virulent
Catalase
Classification of streptococci in chains:

1. ACADEMIC CLASSIFICATION/ BERGEYS :
- Baced on optimum temperature
35
OCDEN, Junalyn B.
a. PYOGENIC:
- 370C
Streptococcus pyogenes
b. VIRIDANS:
- Thermophiles; 450C
Streptococcus mitis: causes endocarditis, sinusitis, Abscess (painful inflammation with
pus)
Streptococcus salivarius: causes dental caries
Streptococcus thermophilus: used to enhance the flavor of cheese; non- pathogenic
c. ENTEROCOCCUS/ FAECALIS:
- 100C AND 450C
Streptococcus faecium
Streptococcus faecalis
Streptococcus bovis
Streptococcus equines
o All causes UTI and endocarditis
d. LACTIC:
- 100C
Streptococcus lactis
Streptococcus cremoris
o Both enhances the flavor of cheese
2. SMITH BROWNS:
- Based on hemolysis
-
a. Alpha-hemolytic aka Green Streptococcus/ Viridans

Partial destruction of RBC
Gives of greenish color
b. Beta-hemolytic
Complete destruction of RBC produced by pyogenes
36
OCDEN, Junalyn B.
Streptococcus pyogenes, Streptococcus haemolyticus, Streptococcus agalactiae
-
c. Gamma hemolytic:
No destruction of RBC
Streptococcus lactis, Streptococcus cremoris, Streptococcus faecalis
d. Alpha Prime:
2 zone:
Inner> Alpha; Outer>Beta
3. REBECCA LANCEFIELD
- Classified based on the presence and absence of
C-polysaccharide (function: attachment to tissues)
a. GROUP A
Type of hemolytic pattern: Beta-hemolytic
Streptococcus pyogenes- encapsulated, copy paste
(found in oral cavity)
VIRULENCE FACTOR:
1. Protein F:
Function: attachment to epithelial cells
2. M protein:
Function: anti-phagocytic, acid and heat resistant
3. Streptokinase
4. Hyaluronidase
5. Streptodornase- like the function of DNAse
6. Streptolysin: Types O(labile) and S(stable)
7. Capsule
8. Erythrogenic Toxin/ Pyrogenic Toxin
~red- rashes, scarlet fever
9. C-polysaccharide- for attachment to tissues
10.
Lipotechoic acid- for attachment to tissues
DISEASE:
1. Impetigo/Pyoderma
2. Erysipelas- presence of small red painful rashes
3. Streptococcus pharyngitis/ Streptococcus sorethroat
37
OCDEN, Junalyn B.
-dysphagia
-if left untreated for 1-2 weeks, the bacteria release erythrogenic toxin
causing:
4. Scarlet Fever/ Scarlatina: Fever, rashes, strawberry tongue
5. Rheumatic Fever:
Suffer from: carditis = inflammation of all the walls of the heart; nodules; polyathritis
6. AGN: Acute Glomerulonephritis
Experience: Edema, Hypertensive, hematuria, proteinuria (increase in albumin)
7. Puerpueral Fever
8. Pneumonia
9. Sequelae- condition that arises from previous complication
TESTS:
Grams Stain
BAM- grayish white, transparent to translucent, MAT appearance, large zone of beta hemolysis
CAN (Columbia Agar with Colistin & Nalidixic Acid)
PEA (PhenylEthyl Alcohol Agar)
b. GROUP B
- beta hemolytic
Streptococcus agalactiae
Animals: Bovine mastitis
Humans: causes: neonatal minigitis (neonates); septicemia (adults)
Lab Tests:
BAM: small, translucent, flat, glossy, narrow zone of beta hemolysis
c. GROUP C
- beta hemolytic
- Animal pathogens
Streptococcus equi, Streptococcus equisimilis
38
OCDEN, Junalyn B.
d. GROUP D
ENTEROCOCCUS
- Alpha/gamma
- Strep. faecalis & faecium
- Wound, subacute bacterial
endocarditis, UTI, septisimia
NON-ENTEROCOCCUS
gamma
- Strep. mitis &bovis
- subacute bacterial
endocarditis, UTI, septisimia
e. GROUP F & G
- beta hemolytic
Difference in BAM:
F
Gray, MAT, narrow zone of beta
hemolysis
G
Gray,MAT, wide zone of beta
hemolysis
STREPTOCOCCUS PNEUMONIAE/ DIPLOCOCCUS PNEUMONIAE/STREPTOCOCCUS LANCOELATUS

CHARACTERISTICS
G+ cocci in pairs/ short chains
Lancet shape bacteria
80% cause of pneumonia
Non-motile encapsulate
Facultative
anaerobicCapnophilic
Fastidious
Non spore forming
VIRULENCE FACTORS
1. Capsule
2. Pneumolysin Ocause
destruction of
cells;
dermatoxic;
cytotoxic
3. Neuramidasespreading
factor; destroys
Neuraminic
acid~attached to
the mucous
membrane in the
respiratory
system
4. Amidaseautolytic
enzyme; for the
39
DISEASE
1. Lobar
pneumoniainvade
bilobe;
abrupt onset
of signs and
symptoms
such as
fever,
chills,
dysphea,
cough(sputum
: rusty
colored
sputumcontain
blood
linged)
2. Bacteremia
LAB TESTS
1. Grams
Stain
2. BAM:
small,
glistening
, mucoid,
shiny,
alphahemolytic
3. PEA
4. CAN
5. NQ
OCDEN, Junalyn B.
bacteria to
multiply/cell
division
5. PPP: Purpura
Producing
Principleautolysis;
caused by
amidase; effect:
dermal
hemorrhage
3. Meningitis
4. Sinusitis
5. Otitis media
BIOCHEMICAL TESTS:
1. BACITRACIN SUSCEPTIBILITY TEST:
Purpose: determine the effect of small amounts of Bacitracin
Streptococcus pyogenes- easily inhibited by Small amounts og Bacitracin
+ result: zone of inhibition
2. Bile Esculin Agar:
- Agar slant
Streptococcus and Enterococcus
o Grow in 40% Bile
o Hydrolyze esculin (producing: esculetin and ferric ions)
Causes: color change in Ferric ammonium citrate (ph indicator)
+ result: dark brown color(formation of phenolic iron compound) indicative of Enterococcus
faecalis
3. Bile Solubility Test:
- Uses Bile/Bile Salt Solution /Sodium desoxycholate
Readily lyses Strep. pneumoniae because of the presence of Intracellular
Autolytic Enzyme
+ result: no bacterial growth indicative of Strep. pneumoniae
4. CAMP: Christie Atkins Munch Petersen Agar
- Differentiate group A from group B Strep.
40
OCDEN, Junalyn B.
Group B Strep: contains a diffusible extracellular protein a CAMP FACTOR which
synergistically acts with betalysin of another bacteria( Staph. aureus) to
produce an enhanced zone of hemolysis
+ result: arrow head zoneof hemolysis
-result: Strep. pyogenes
5. HIPPURATE HYDROLYSIS TEST:

- DETECT : HIPPURICASE
In a tube:
--Hippuric acidGlycine+Benzoic acid = deaminationNinhydrin
+ result: red color indicating Streptococcus agalactiae

Result: yellow to pink
6. LAP: LEUCINE AMINO PEPTIDASE

- IDENTIFY CATALASE cocci
Leucine-B-napthylamineLap from bacteria-> B-napthylamine + cinnamaldehyde
+ result: red
-result: yellow
7. OPTOCHIN TEST:
- Check the action of optochin (ethylcupreine hydrochloride)
Result: > 14mm= Streptococcus pneumoniae
< 14mm= other Strep
= 14= other streptococcus; equivocal/intermediate
8.
-
PYR TEST:
Almost the same with Lap
Used to identify G+ cocci
Utilize reagent L-pyrrolidonyl-B-napthylamineidentify presence of L-pyrrogluramyl-aminopeptidase produce: B-napthylamine + NN methyl amino cinnamaldehyde
+ red; - orange
41
OCDEN, Junalyn B.
9. PYRUVATE BROTH:
- DETERMINE ability of microorganism if it can utilize pyruvate differenciate Enterococcus
faecalis & Enterococcus faecium.
Green to yellow: E. faecalis
Yellow Green: E. faecium
10.
SALT TOLERANCE TEST:
BHI(media)+ 6.5% NaCl+ Glucose+ (pH indicator) bromcresol purple
- Differentiate Enteroccocus from non-enterococcus
Result: turbidity= bacterial growth; no color change negative
If turbid; purple to yellow Enterococcus
No turbidity;no color change Non-enterococcus
NVS OR NUTRITIONALLY VARIANT STREPTOCOCCI
-
Fastidious: need substance such as pyridoxal(add Vitamin B6), thiol

Pleomorphic
Satellite cells- bacteria can grow around another type of bacteria (such as Staphylococcus
aureus can grow in Enterobacteria)
LAP, have enzyme Arylamidase
ABRIOTROPHIA
2 species:
Normal flora in the oropharynx
1. A. defectives
2. A. adjacens
Both causes:
Bacteremia, Endocarditis, conjunctivitis, endophthalmitis: intraocular cavities inflammation,
Keratitis: inflammation of the cornea of the eyes, ICK: Infectious Crystalline Keratopathyneedle/fern shape deposits in cornea, Otitis media
Treatment: penicillin: less susceptible
Susceptible to: Clindamycin, Erythromycin, Chloramphenicol, Vancomycin, Rifampin
42
OCDEN, Junalyn B.
Tests:
1. Pyridoxal Requirement Test
2. Satellite test
3. LAP
FAMILY: NEISSERIACEAE
CHARACTERISTICS:
- Gram diplococcic
- Kidney bean or coffee bean shape
- Flatten adjacent sides
- Fastidious
- Capnophilic (5-10% CO2)
- Non-motile
- Non-encapsulated, NSF
- Catalase +
- Oxidase +; deep purple
- CHO fermenters
CTA (Cystin Trypticase Agar)- consist of Tryptone, L-cystine (source of nutrients);
sodium chloride and sodium sulfate(source for ions); ph indicator: phenol red (red to
yellow)
Tube media: agar butt
4 tubes: Same CTA content: difference: carbs which are Glu, Mal, Suc, Lac
NEISSERIA GONORRHEAE
Characteristics:
- Pathogenic
- Glucose fermenter
- Easily inhibited by: drying, heating and action of chemical disinfectants
Habitat: anorectal area, GUT, inflammation of the oropharynx and laryngophanrynx, conjunctiva
VF:
1. Pili/fimbrae: for attachment
2. Outer membrane:
a. POR aka Protein 1: for entrance of nutrients/ serves as a passageway
b. OPA aka Protein 2: attachment
c. RMP aka Protein 3 or Reduction Modifiable protein
3. LOS (Lipo Oligo Saccharide)- scarring of tissues
43
OCDEN, Junalyn B.
4.
5.
6.
7.
8.
LPS (Lipo Poly Saccharide)- destroy epithelial cells, responsible for toxicity
LiP/H8: for attachment
FBP or Iron binding protein: if Iron is low; FBP will look for Iron
IgA protease 1- splits and deactivates IgA
Beta- lactamase
Disease:
Subdivided into 2 according to organs of infected:
A. GENITAL GONORRHEA
MOT: sexual intercourse
Usually form infected females to non-infected males
Divided into 2:
1. MALE G.G:
-symptomatic type
- incubation: 2-7 days
-S/S: urethritis:
- dysuria: painful burning sensation during urination with a presence of
yellowish purulent discharge
- prostatitis
- Epididymitis:20 days for a sperm cell to mature
- Steriltity/infertility
-
2. FEMALE G.G:
Asymptomatic
Incubation: 8-10 days
S/S: vaginitis, cervixitis, bloody discharge, dysuria related to UTI, PID (Pelvic
Inflammatory Disease/ Salpingitis), Sterility/Infertility
B. EXTRA GENITAL GONORRHEA

1. Opthalmia Neonatorum
2. Pharyngitis
3. Proctitis: inflam. Of the anus
44
OCDEN, Junalyn B.
TREATMENT:
- Doxycycline
- Ceftriaxone
- Quinolones: Ciprofloxacin, Ofloxacin, Levofloxacin
LAB DIAGNOSIS:
1. Grams Stain: Gram diplococcic intracellular, extracellular
2. Culture:
a. Thayer-Martin (TM)
-
Type
o
o
o
o
of CAM with enrichment supplements:

Isovitalex
Colistin- inhibit growth of G- bacilli
Nystatin- inhibit growth of yeast
Vancomycin- inhibit growth of G+ bacteria
b. Modified Thayer Martin:

Addition of trimethoprim
c. Martin-Lewis (ML)
Difference:
Addition of anisomycin (anti-fungal)
Increased concentration of vancomycin
d. NYC:
Appearance: transparent medium
Composition: lysed horse blood, horse plasma, yeast dialysate, colistin, vancomycin,
nystatin, trimethoprim
Advantage: permit growth of Mycoplasma hominis, ureaplasma urealyticum
e. CAM:
Small, grayish white, convex, translucent, shiny, smooth, irregular margins
f. JEMBEC (John E. Martin Biological Environmental Chamber)

Contains CO2 generating system (citric acid, Na2CO3)
45
OCDEN, Junalyn B.
Other tests:
1. RIA (radio Immuno Assay)

2. ELISA (Enzyme Link Immunosorbent Assay)
3. Molecular Assay :
Non-amplified probe assay (chemiluminescent test) detect: Gonococcal/Ribosomal RNA
Amplified Assay: for large screening
4. Particle Agglutination Method:

Phadebact
Gonogen II test
NEISSERIA MENINGITIDIS
CN: Meningococcus
G-diplococci, fastidious, capnophilic, Non-motile, NSF
Diff.: encapsulated , opportunistic
Ferments: glucose, maltose
Habitat: anorectal; NF: nasopharynx
VF:
1.
2.
3.
4.
5.
Capsule
Outer membrane: similar to POR formation of outer membrane
LPS: site of endotoxic activity
IgA protease
Pili
DISEASE:
1. Meningitidis
- Through inhalation, droplet spread
- Kernigs ; Budzinskis
- Fever, stiff neck, vomiting, headache
2. Meningococcemia
- Bacteria in blood
- Fuminant; explosive; death : 6-8 hours
46
OCDEN, Junalyn B.
-
Fever, stiff neck, vomiting , head ache and formation of petechiae
*Waterhouse Friedrichsen Syndrome (WFS):

-
dessiminated intravascular coagulation

Hemorrhagic adrenalitis
Low blood pressure, coma, death
Polysaccharide Capsule Antigen:
**13 serogroups: 5 medically important: A,B,C,Y, W135

Prophylaxis: giving treatment before a patient suffers from complication
- Rifampin, Ceftriaxone, Chloramphenicol
LAB TESTS:
1.
2.
3.
4.
5.
-
Grams Stain: G- dc
Culture: TM,MTM,ML
CAM: mucoid, moist, grayish white, medium, rough
JEMBEC
Moraxella Catarrhalis/Branhamella
CAM: lare,non-pigmented, gray, opaque, smooth, give off Hockey Puck consistency
DISEASES:
Otitis media, Pneumonia, sinusitis, cause Respiratory Tract Infection in the elderly and in
patients with COPD (Chronic Obstructive Pulmonary Disease)
OTHER SPECIES:
N. cinerea
N. mucosa
N. flavescens
N. sicca
N. subflava
**can grow in MAC
GRAM + BACILLI:
47
OCDEN, Junalyn B.
1. Non spore formers:

Corynebacterium , Listeria,; both are catalase + and non branching; Erysipelothrix (Catalase
-)
2. Spore Formers:
Clostridium and bacillus
CORYNEBACTERIACEAE
Genus: Corynebacterium
1. Corynebacterium diptheriae
CN: Kleb Loefflers Bacillus
Characteristics:
- Pleomorphic
- Exhibit Chinese Letter Arrangement
- Palisade/picket fence
- Club shape
- Non-motile, Non-encapsulated, fastidious
- Aerobic Facultative anaerobic
- Contains:
o Metachromatic Granules/Volutin/ Babes-Ernst Granules
Intracellular polyphosphate granule
Have high affinity to nuclear stains
o LAMB
Granule: RED- used to absorb pollutants esp. in waters
HABITAT:
- Nasopharynx, oropharynx and skin
VF:
1. Diptheria Toxin:
AB toxin
A- Inactivates E-F2(Elongation Factor 2)/EEF2 (Eukaryotic Elongation Factor 2)
Function: responsible for the formation of protein
Causes: cell necrosis(cell death) & neurotoxic effects
48
OCDEN, Junalyn B.
** .1ug/kg body weight
Disease:
1. Respiratory Diptheria
MOT:
- Respiratory Droplets: coughing, sneezing, laughing
- Indirect contact: with the use of fomites
IP: 2-5 days
S/S:
1. Fever
2. Sorethroat/Pharyngitis
3. Bluish skin
4. Loud barking sound (during cough)
5. Hissing sound (during inhaling)
6. Occurrence of pseudomembrane of the throat: thick grayish white adherent membrane on the
throat and over the tonsils
PREVENTIVE MEASURE:
1. DPT: tatenus given at 2 months or 6 months around 1-4 years old
Problem: lasts for 10 years thus another injection during the 12th year; for adults: booster
is given
TREANTMENT:
1. Diptheria and anti-toxin:
Oral: erythromycin and penicillin
LAB DIAGNOSIS:
1. Grams Stain
Cbd contains arabinose, galactose, short chains of mycolic acid
2. LAMB: identify metachromatic granules
CMN (N=Nocardia)
3. Culture:
a. CTBA (Cystine Tellurite Blood Agar) black
b. MT (Modified Tinsdale) gray to black colonies with a dark brown halo.
49
OCDEN, Junalyn B.
c. Loefflers medium with serum and egg: from beef extract (pulp: causes coagulation)
conclusion: exhibit proteolytic activity
4. PCR (Polymerase Chain Reaction)
5. Tissue Culture Test
6. Elekss Test:
-
Immunodiffusion test to verify if a certain strain of corynebacterium is toxigenic
Serogroups: gravis, mitis, belfanti, intermedius

OTHER SPECIES:
- C.jeikeium: opportunistic
- C. urealyticum: urease (UTI)
- C. pseudotuberculosis: sub-acute relapsing lymphadenitis
- C. xerosis: pharyngitis
- C. ulcerans: diphtheria like disease
- C. pseudodipthericum : low resistance to infection and endocarditis
2. Listeria monocytogenes
Characteristics:
- Non-encapsulated
- Catalase + and esculin
- Voges Proskauer
- Non-motile at 350C and motile at 250C (tumbing motility)
- Chains/pairs
- Habitat: GIT; females: GUT(pregnant and growing fetus)
VF:
-
1. Listeriolysin:
Hemolytic ,cytotoxic, help bacteria in phagocytes
DISEASE:
Listeriosis: ingestion of contaminated milk, direct contact with animals, transplacental
50
OCDEN, Junalyn B.
INCUBATION: 2 months
S/S: (lasts for 2-3 days)
Fever, sore throat, diarrhea
a. IN PREGNANCY:
- Fever, sore throat, diarrhea
b. INTRAUTERINE LISTERIA:
The fetus will experience granuloma formation and if persists it will lead to Granulomatosis
infantiseptica which can cause: premature abortion and still birth
c. NEONATAL LISTERIOSIS:
S/S: fever, sore throat, diarrhea, meningitis, septicemia and Granulomatosis infantiseptica
is common
d. LISTERIOSIS IN IMMUNOCOMPROMISED PATIENTS
Experience: meningitis and septicemia
LAB DIAGNOSIS:
*SPECIMEN: placenta
COLD ENRICHMENT METHOD:
Nutrient brothincubate at 40C for wks and mossub cultureCAM: small, white, smooth,
translucent, beta-hemolytic
*motility test:
N.Bincubate: 1-2 hrs at room temperature; result: umbrella-like patternincubate
over night and check again for an umbrella-like pattern
3. ERYSIPELOTHRIX RHUSIOPATHIAE/E. INSIDOSA
- pleomorphic
Long filaments=rough colonies
Short filaments= smooth colonies
ZOONOTIC: pigs and cattles (fishes??)
VF:
1. Neuroamidase: cause: arteritis (inflammation of the wall of the arteries) and
thrombocytopenia
51
OCDEN, Junalyn B.
2. Hyaluronidase
DISEASE:
ANIMALS:
Swine Erysipelas- rashes, dermatitis
- Inflammation of the skin and joints
- Transmission: direct contactMAN:
MAN:
Erysipeloid:
- Inflammation of the hands and fingers
- Itchy burning sensation (skin)
- Complication: Septicemia and endocarditis
LAB DIAGNOSIS:
CAM, CAM, CNA, HBT: Human Blood Bilayer Tween
GRAM + BACILLI SPORE FORMERS

Genus: Bacillus
BACILLUS ANTHRACIS
CN: Anthrax Bacilli
Habitat: Soil
MOT: direct contact, into wounds, inhalation, ingestion
TYPES OF ANTHRAX:
1. Cutaneous Anthrax (95%):
MOT: skin-skin contact
Experience: Malignant pustule black necrotic lesion
Areas: arm, neck, face
2. Pulmonary Anthrax aka Woolsorters disease (5%):
- Through inhalation
S/S: pneumonia, flu-like symptoms, Resp. Tract Infection, there is Hemorrhagic Mediastinitis (detected
through chest X-ray)
3. Bacteremic Anthrax:
- Via wounds
52
OCDEN, Junalyn B.
- Rare; causes sepsis
4. GIT anthrax aka violent enteritis
- Ingestion of dairy products
- Severe form
S/S: nausea, fever, vomiting, abdominal pain, bloody diarrhea
Absence of PXO2= Sterne Strain (3F2 Strain); specimen: pus from skin lesion or blood or sputum
CHARACTERISTICS
-
Aerobic
Spore: sub-terminal
non swollen
Resistant to:
disinfectants, UV
light, heat, drying
Encapsulated
Size: large (3-4 um)
Square/ovoid(circula
r)
In chains; bamboo
fishing rod
appearance
Non-motile
Gamma
hemolytic/Nonhemolytic
Zoonotic bacteria ;
in herbivores
VIRULENCE FACTORS
- encoded in plasmids; pXo1 & pXo2
a. capsule: produced by pXo2
b. Polysaccharide somatic antigen:
destroys somatic cells
c. Protein Somatic antigen: destroy

somatic cells
d. Anthrax toxin: produced by pXo1
3 PROTEIN FACTORS:
1. Protective Antigen:
Function: Binding Component;
help in the entry of EF
Combine with EF to form=Edema
toxin
Combine with LF to form= Lethal
toxin (Major virulence factor of
Bacillus anthracis)
2. Edema Factor:
Aka Adenylyl cyclase
Cause the increase production of
CAMP (Cyclic Adenosine Mono
Phosphate); CAMP fxn: for effects of
hormones in cells
Effect of Edema Factor: Loss of
function of Neutrophils
Result: tissue necrosis and massive
53
DISEASE
LAB DIAGNOSIS
Anthrax
Biochemical Test:
1. Gel liquefaction/Gelatin
Swab Culture:
+ result: inverted fir
tree
2. VP +
Serological Test:
1. ELISA: detect anti
lethal toxin and anti
edema toxin
Direct Detection Methods:
- Use of TSI, urea agar,
Nutrient agar(+ 5 mg
Manganese
Sulfate/liter)
- Spore Stain: use of
Malachite green; green
spores-red cells
2. Culture:
BAM: medium-large,
flat, gray, irregular,
gamma/nonhemolytic, swirling
projections (termed
specifically as Medusa
head aka curled hair
OCDEN, Junalyn B.
edema
3. Lethal Factor:
Specific: zinc mettaloprotease
Function: Inactivates protein kinase
(kinase is for the regulation of
normal function of cells)
54
lock appearance)
CAM
PEA
PLET(Polymyxin
Lysozyme EDTA
Thallous Acetate) and
Bicarbonate Agar:
induces capsule
formation
For both PLET and BCA,
it specifically isolate
and identify
B.anthracis;
capnophilic incubation
for 2 hours ar 370C
Egg yolk agar:
+ result: wide zone of
lecithinase
OTHER TESTS:
1. Red line alert test:
- Presumptive test; after
BAM
- Colonies are from BAM
- Immunochromatograph
ic test
+ result: red line on
test and control
If there is no line in
control and present in
test = invalid
2. Ascoli (from: Alberto
Ascoli)
- Thermoprecipitation
test
- Use tissue extract
- Anthrax anti serum
OCDEN, Junalyn B.
3. String of Pearls:
- Differentiate B.
anthracis from B.
cereus
- Susceptibility test to
penicillin (PCN)
Young colonies( 12-18
old)
-source of colonies:
BAM
Use another agar:
55
OCDEN, Junalyn B.
BACILLUS CEREUS
CN: Fried Rice Bacillus
MOT: ingestion; contaminated food(fried rice) & soil
CHARACTERISTICS
Similar with Bacillus
anthracis
Difference:
- Grows best at 450C
- Beta hemolytic
- Semi solid agar;
motility: swarming
- Beta lactamase
VIRULENCE FACTORS
DISEASE
1. Enterotoxin: cause of
food poisoning
2. Pyrogenic toxin
1. Food poisoning:
a. Diarrheal Type:
S/S: diarrhea, profuse
diarrhea (watery; no fecal
debris)
- Easily inactivated
with heat/ Heat
labile
- 18 hours of
incubation
b. Emetic type:
- Heat stabile
- Incubation: 2-6
hours
S/S: abdominal pain,
profuse vomiting, nausea,
fever
- Self-limiting disease;
recovery: 24 hours
2. Keratitis
3. Panophthalmia: all
layers of conjunctiva
is infected
4. Post traumatic eye
infection
5. Endocarditis
6. Bacteremia
7. Meningitis
8. Osteomyelitis
9. Pneumonia
56
LAB DIAGNOSIS
1. VP +
2. BAM: large, feathery,
spreading beta
hemolytic
OCDEN, Junalyn B.
BACILLUS SUBTILIS
- Common laboratory contaminant
BAM: large, gray, dull, flat, ground glass appearance, pigmented (colors: brown, pink, yellow, and orange)
OTHER SPECIES:
C. thurigiensis, B. sphaericus, B. popillae/ paenibacillus, B. larvae, B. lentimorbus : insect pathogens
1. Early/Fernandez
- If the wheal appear at 24-48 hours
2. Late aka Mitsuda
- 3-4 weeks
GRAM BACILLI
Family: Enterobacteriaceae
Habitat: GIT
MOT: Ingestion; fecal-oral route
Non spore forming
Anaerobic to Facultative anaerobic
Generally motile: except Shiegella, Klebsiella
Catalase +; except: Shiegella and dysenteriae type I
Oxidase
Carbohydrate fermenter ; specifically: Glucose
Media:
1. BAM: large,gray, smooth
2. MAC
3. MAC with sorbitol agar
- Differential media: E coli 0157:H7 from other E. coli
4. EMB: lactose/sucrose
5. HEA: Hektoen Enteric Agar
6. XLD: Xylose Lysine Desoxycholate
7. BHI
8. FT
9. SSA
57
OCDEN, Junalyn B.
10.CIN: Cepsulodin Irgasan Novobiocin
- Grow Yersinia enterolitica
ESCHERICHIA COLI
CN: Colons Bacillus

Characteristics: Motile, Anaerobic, Non-fastidious, short/plump bacilli
VF:
1. Capsule/Microcapsule/ B Antigen
2. Endotoxin- responsible for fever
3. Pili/Fimbrae
DISEASES:
1. UTI (80%) Community Acquired
- In the urinary bladder(cystitis)
- In the kidneys (pyelonephritis)
- Start experiencing: painful urination, high frequency in urination, increase in WBC (pus cells)
- Experience: Fever, Flank pain (low back pain)
2. Neonatal meningitis
3. Septicemia
4. Gastroenteritis
ETEC (ENTERO TOXIGENIC ESCHERICHIA COLI)

VF:
a. Pili/Fimbrae
b. LT: heat labile toxin; function: regulate secretion of water and electrolytes
c. ST: stable toxin; function: regulate secretion of water and electrolytes
-
Disease: Travellers diarrhea/ Childhood diarrhea

Watery diarrhea, abdominal pain, vomiting, nausea, low grade fever
EIEC (ENTERO INVASIVE ESCHERICHIA COLI)

-
Colonize large intestine species ; electrolytes of the large intestine

Severe type: watery to bloody diarrhea dysentery (blood in stool), severe abdominal pain
58
OCDEN, Junalyn B.
EPEC (ENTERO PATHOGENIC ESCHERICHIA COLI)

VF:
a. Pili
b. Intimin cause destruction of microvilli of the small intestine
Disease:
1. Infantile diarrhea
- Soft stool, fever, nausea, vomiting, abdominal cramps
EHEC (ENTERO HEMORRHAGIC ESCHERICHIA COLI)

-
VTEC: VEROTOXIN PRODUCING ESCHERICHIA COLI AKA CYTOTOXIN

Attacks/destroys vascular endothelial cells
Diseases:
Start at:
1. Hemorrhagic colitis
- Bleeding inside the large intestine
S/S: watery bloody diarrhea, little occurrence/no fever
2. HUS (HEMOLYTIC UREMIC SYNDROME)
- Destruction of RBC
- Experience: Microangiopathic hemolytic anemia
RBCs are microcytic hypochromic
Can lead to Acute Renal Failure; platelets will go down and will lead to: thrombocytopenia
EAEC (ENTERO AGGREGATIVE ESCHERICHIA COLI)

-
Contains pili that causes watery diarrhea
LAB DIAGNOSIS:
1.
2.
3.
4.
5.
BAM: beta-hemolytic, opaque, cream, smooth, 2-4 mm

EMB: green metallic sheet; fermented lactose and sucrose
HEA: yellow colonies
XLD: yellow colonies
MAC: pink, flat, dry ; lactose fermenter
BIOCHEMICAL TEST:
59
OCDEN, Junalyn B.
1.
2.
3.
4.
5.
TSI: A/A g
LIA: purple slant/purple butt
IMVIC: ++- Starch Agar test
MUG test (4-methyl-umbeliferyl-B-d- glucoronide)
6. Acetate Utilization Test

- Acetatecarbonundergo Alkalinization + result: green to blue
7. Spot Indole Test +
8. PYR
9. PAD
EDWARDSHIELLA TARDA
Gram bacilli, motile, non-lactose fermenter, produces H 2S, opportunistic
Disease: Wound Infection, Liver Abscess
SHIEGELLA
-
Non-motile, non-encapsulated, non-lactose fermenter (generally)

Anaerogenic (does not produce gas) , non-fastidious
NON- MANNITOL FERMENTER

60
OCDEN, Junalyn B.
Shiegella dysenteriae CN: Flexners Bacillus
Disease: Philippine Dysentery
MANNITOL FERMENTERS
NON-LACTOSE FERMENTER:
Shiegella flexneri CN: Flexners Bacillus
Shiegella boydii CN: Boyds Bacillus
Disease: British Dysentery
LATE LACTOSE FERMENTER
Shiegella Sonnei CN: Sonne duvals
Disease: U.S. dysentery
EWINGS CLASSIFICATION:
Group
Group
Group
Group
A: Shiegella dysenteriae
B: Shiegella flexneri
C: Shiegella boydii
D: Shiegella Sonnei
VF:
1. Enterotoxin
DISEASE:
1. Shiegellosis/Bacillary dysentery
MOT: fecal-oral route
IP: 1-4 days
S/S:
1. Abdominal pain (severe)
2. Waterybloody diarrhea
3. Tenesmus- painful straining during defecation and incomplete feeling of defecation
4. Myalgia- muscle weakness
LAB DIAGNOSIS:
Culture:
EMB, MAC, SSA colorless colonies
XLD red colonies
61
OCDEN, Junalyn B.
BIOCHEMICAL TEST:
1. TSI: K/A
2. IMVC (A,B,C): V + - D: - + - 3. LIA
SALMONELLA
-
Most are motile, aerogenic (except : Salmonella typhi)

2200 serotypes
Differentiate based on:
o O antigen- present on the cell wall aka as cell wall/Somatic Ag
o H antigen- Flagellar Antigen- heat labile
o E antigen- Envelope Antigen/Surface Antigen; example: Vi Antigen
COMMON SEROTYPES:
1. Salmonella cholerasuis
2. Salmonella typhi/typhosa CN: Eberths Bacillus
- Anaerogenic;
3. Salmonella enteritidis; CN: Gardners Bacillus
a. Salmonella paratyphi A
b. Salmonella paratyphi B/ Salmonella Schotmuelleri
c. Salmonella paratyphi/ Salmonella hirschfeldii
d. Salmonella typhimurium
e. Salmonella sendaii
f. Salmonella pullorum non-motile
g. Salmonella gallinarum- non-motile
h. Salmonella derby
DISEASES:
1. Salmonellosis
a. Enterocolitis
- Caused by Salmonella typhimurium (ingestion of contaminated oysters)
I.P.: 6-48 hours
-- Vomiting, watery to bloody diarrhea, abdominal pain, nausea
b. Enteric Fever:
2 types:
B1. Typhoid fever- caused by Salmonella typhi
B2. Paratyphoid fever
62
OCDEN, Junalyn B.
Mot: FecalOral route
Ip: 10-14 days
DIVIDED INTO 4:
1st week: sustained fever, constipation, diarrhea; bacteria is present in the intestines
2nd week: sustained fever , rose spots: adults-10-12 in the abdomen while for children 10-12 in the face; bacteria is
present in the intestines
3rd week: Bacteria is capable of Intestinal perforation, Hemorrhage- gain access to gallbladder and spleen
4th week: period of convalescence
Salmonella cholerasuis, Salmonella typhi, Salmonella paratyphi
2. Bacteremia: s/s high _____ fever
LAB DIAGNOSIS:
Culture:
EMB and MAC- colorless
SSA- colorless black
XLD- black
BSA-Bismuth Sulfite Agar
BGA-Brilliant green Agar; all Salmonella sps except S. typhi appears to be opaque and colorless
Serology:
a. Widal Test: non-specific
- An agglutination test for Enteric
1st week: blood
2nd week: feces/urine
3rd week: blood
b. Typhidot/ Typhoid ELISA

Rapid immunochromatographic test
63
OCDEN, Junalyn B.
CITROBACTER SPECIES
- Motile
- Normal flora of the colon
Citrobacter freundeii and Citrobacter diversus: contain 3 virulence factors
1. Endotoxin
2. Capsule
3. Adhesion proteins
- Cause nosocomial infection
Citrobacter freundeii
Citrobacter diversus
Indole
+
H2S
+
KCN
+
TSI
K/Ag H2S
A/Ag
IMVC
-+-+
++--
KLEBSIELLA PNEUMONIAE/BACILLUS MUCOSUS CAPSULATUS

CN: FRIEDLANDERS BACILLUS
-
Non-motile
Encapsulated ; NF test +
Aerobic
VF: Endotoxin, Capsule, Adhesion proteins
DISEASE:
1. Pneumonia
- Ulceration and necrosis of the respiratory tract
- Thick mucoid bloody sputum (currant jelly sputum)
2. UTI
3. Arthritis
4. Meningitis
5. Septicimia
LAB DIAGNOSIS:
CULTURE:
(on any culture especially Mac and EMB)
- Large, mucoid
64
OCDEN, Junalyn B.
EMB: purple
MAC: red/pink (lactose fermenter)
KLEBSIELLA GRANULOMATIS
OLD NAME: CALYMMATOBACTERIUM GRANULOMATIS

- Found with mononuclear endothelial cells
Donovan body: surrounded by WBC
Wrights / Giemsa Stain
+ result: blue with granules
safety pin appearance
DISEASE:
1. Donovanosis/Granuloma inguinate
- Small red beefy bumps
- Either genital/anus
KLEBSIELLA OXYTOCA
-
Non encapsulated
Non-motile
Non-lactose fermenter
ENTEROBACTER AEROGENES
OLD NAME: AEROBACTER AEROGENES
- Most common as a contaminant
- Motile
- Opportunistic
- ORN +
PANTOEA AGGLOMERANS
OLD NAME: ENTEROBACTER AGGLOMERANS
- Opportunistic
HAFNIA ALVEI
-
Opportunistic
65
OCDEN, Junalyn B.
SERRATIA
Serratia marcescens
Serratia liquifaciens
Serratia rubidaea
All are motile, chromogenic, red pigments due to the presence of:
1. Progigiosin: non-diffusible,no-water soluble
2. Pyrimine: diffusible, water-soluble
Enzymes:
1. Hydrolytic Enzymes: Lipase,DNAse,Gelatinase
PROTEUS
VF:
Pleomorphic
Hypermotile
Burnt gun powder odor
urease which causes UTI
a. Swarming phenomenon
Sps: Proteus vulgaris and mirabilis
Example:
b. Dienes Phenomenon:
-
Concentric zone, swarm toward each other without overlapping
MORGANELLA MORGANII
OLD NAME: Proteus morganii
Contains ureaseUTI
PROVIDENCIA
UREASE
CITRATE
66
OCDEN, Junalyn B.
Proteus alcalifaciens
Proteus stuartii
Proteus rettgeri
Proteus rustigianiis
Cause UTI
V
+
-
+
+
+
-
YERSINIA PESTIS
-
Short plump cocco bacilli

Grow best at 30 degrees celcius ; average: 25-32 degrees celcius
Non-motile
MOT:
Rat flea which is Xenopsylla cheopsis which causes:
Black Death/Plaque: Bubonic,Pneumonic, Septicemic
USE:
Stalactile Streamers
YYERSINIA ENTEROLITICA
-
Cause gastroenteritis
CIN
Pseudotuberculosis
Resides in the appendix of humans and
GIT of animals: Pigs, Dogs and Cats
ACID FAST BACILLI

GENUS: MYCOBACTERIUM
67
OCDEN, Junalyn B.
CHARACTERISTICS:
- NON- ENCAPSULATED, NON-MOTILE, FASTIDOUS, AEROBIC
- Contain Granules: MUCH GRANULES- cause beaded appearance
MYCOBACTERIUM TUBERCULOSIS
CHARACTERISTICS
VIRULENCE FACTORS
NONENCAPSULATED,
NON-MOTILE,
FASTIDOUS,
AEROBIC
Contains
SAPENTINE
CORD FACTOR
which can
assume X, Y, V,
L arrangement
RED BACTERIA;
BLUE
BACKGROUND
DISEASES
MYCOLIC ACID
1. Tuberculosis:
MOT: inhalation via Airborne (sneezing, coughing,
etc)
IP: several weeks to 2
months depending on the
resistance
TYPES OF TB:
1. Pulmonary tb:
- Affects the lower
lobe of the lungs
which can cause:
LESIONS:
1. Exudative Lesions:
- Made up of edema
fluid and edema
fluid is made up of
PMN or Polymorpho
Nuclear Cells and
Mononuclear cells
2. Granulomatous
Lesions:
- Formation of giant
tubercle bacilli
which consists of
mono,neutron histio
CYTES
- PROGRESS AND CAN
CAUSE:
68
LABORATORY DIAGNOSIS
Sputum analysis (early

in the morning)
Chest X-ray
*Apicogram
TREATMENT
Multiple Drug Theraphy
(MDT):
For light cases: 6-9
months
For severe cases: 9-12
months
1. Rifampin
INH(Isoniazid/Isonicot
inic hydrazide)
Pyrazinamide
Ethambutol: side
effect- red urine
2. Removal of
Ethambutol
PRIMARY COMPLEX OR TB IN
CHILDREN
- Same treatment
- Child to adult
transmission is not
applicable
OCDEN, Junalyn B.
3. Caseous Lesion:
cheese like
consistency and
emergence of:
S/S:
- Fever, chills, night
sweats, productive
cough (sputum:
coffee brown),
hemoptysis
(coughing out of
blood), rapid weight
loss
SECONDARY TUBERCULOSIS
-due to reinfection
- reactivation
Signs and Symptoms
Fever, weight loss, chills,
night sweats
But occurrence is more
rapid and it causes Extra
pulmonary
tuberculosis/Miliary tb
such as:
Prevention
BCG
(Bacillus Calmetter and
Guetin)
- Attenuated M.
bovis
Lab Diagnosis
Specimen: sputum
- Bronchial washing
- Broncho-alveoli or lavage
- Bronchial biopsy
In rare: urine
Occasions: Blood
69
OCDEN, Junalyn B.
1. Tb of the bones
2. Tb of the joints
3. Tb of the spine
specifically: Potts
disease
4. TB in the brain
(meningitis)
Rare cases:
5. Renal TB
6. Genital
Tests:
1. Direct detection test:
-stain
AFS: red bacteria ;blue background
*fluorochrome:
Use of :
1. Auramine O
2. Rhodamine B
*both have + result of luminescent yellow;
green bacteria; blue background
2. Digestion-Concentration Technique
- digest fats, mucus and destroy other Mycobacteria
except M. tuberculosis
3. Niacin Accumulation Test:
+ bacteria: Mycobacteria tuberculosis, Mycobacteria
bovis, Mycobacteria simiae, Mycobacteria marinum,
Mycobacteria chellonei
Cannot convert Niacin to Niacin Ribonucleotide
Niacin bacteria --- addition of: Yanogen
bromide, primary amide + result: no ability:
yellow; other color: negative
4.Nitrate Reduction:
Test:
+ bacteria to N.Reduction test:
Mycobacteria tuberculosis, Mycobacteria Aansasii,
Mycobacteria Szulgai, Mycobacteria fortuitum,
Mycobacteria flavescens, Mycobacteria terrae,
Mycobacteria trivial, Mycobacteria chellonei
All contain the enzyme Nitroreductase
70
OCDEN, Junalyn B.
5.Aryl Sulfatase
6. Tween 80 hydrolysis
7. Iron Uptake Test
8. Complex Media
Divided into 2:
a. Egg based media
Lowenstein-Jensen
Petragnani
ATS: American Thoracic Society
Dorset
b. Non- egg based media
- Middle brook 7H10, Middle brook 7H11
Mycobacteria tuberculosis 370C (I.P. 10-20 days)
Colonies: (either egg-based or non-egg based)
- Small, dry, scaly, warty ; Cauliflower colonies
- Eugonic growth luxurium growth enhanced by
glycerol
*Other species will exhibit dynamic growth: smooth,
scanty
9. Tuberculin test: skin test to show reactivity to
tuberculin antigen
Inject tuberculin antigens such as:
a. OT (Old Tuberculin)
b. PPD (Purified Protein Derivative)
*Prepared by: isolating Mycobacterium filtrates
treated by precipitation- make them non-pathogenic
Addition of Trichloro-acetic acid
71
OCDEN, Junalyn B.
a.a Mantoux Test: intracutaneous

a.b Von Pirquet
- rub tricholoro acetic acid on the skin of the patient
a.c. Vollmer Patch
- rub
a.d. Moro percutaneous test
- Mix PPD and OTLanolin then rub
a.e. Tine test:
antigen(OT or PPD) use Multiprong inoculators
Results: 10-12mm wheal; susceptible (48-72 hours
skin test)
MYCOBACTERIUM BOVIS
CN: BOVINE TUBERCLE BACILLI
DISEASE: GIT, TB
MOT: ingestion of contaminated cows milk
MYCOBACTERIUM AFRICANUM
-
respiratory infection
ATYPICAL MYCOBACTERIA
(Runyons Classification)
a. photochromogens:
produce: yellow-strong light
Mycobacteria kansasii, Mycobacteria simiae, Mycobacteria marinum, Mycobacteria szulgai
b. Scotochromogens:
- Bright yellow pigmented colonies: strong/dark light
Examples: Mycobacteria scrofulaceum, Mycobacteria gastrii, Mycobacteria flavescens, Mycobacteria xenopi
72
OCDEN, Junalyn B.
c. Non-photochromogens:
Examples: Mycobacteria intracellulate avium complex, Mycobacteria malmoense, Mycobacteria haemophilum,
Mycobacteria terrae, Mycobacteria gordonae, Mycobacteria trivial
d. Rapid growers:
Examples: Mycobacteria fortuitum, Mycobacteria chellonei
MYCOBACTERIUM LEPRAE
-
CN: Hansens Bacillus

Non encapsulated, Non-motile, Fastidous
Footpads of Armallicus, Rats
Palisade, cigar pocket, picket fence
Disease: Leprosy
MOT: skin, inhalation
IP:2-4 years
TUBERCULOID LEPROSY/ ANESTHETIC
Formatio Few hypo pigmented
n of
(skin lesions)
plaques
Sensory
Complete
loss
Ineffecti
Low
vity
Lepromi
Positive
n Test
Extracted lesions: treat lesions; inject to the patient
Prevention: BCG
Treatment:
MDT:
Rifampin
Dapsone (Diaminodiphenyl sulfone)
Clofazimine
LEPROMATOUS/NODULAR
Many hyperpigmented especially in cold areas
Patchy: the only part affected numbs
High
Negative
73
OCDEN, Junalyn B.
CLOSTRIDIUM
Habitat: GIT, soil, water, decaying matter
Incubation period: 1-3 days
CHARACTERISTICS
Anaerobic
Central/Eccentri
c swollen spore
Encapsulated
Non-motile
Box Shape
VIRULENCE FACTORS
1.
2.
3.
4.
Enzymes and toxins present:

- Cause digestion of collagen in :
muscle and sub-cutaneous tissues
Collagenase
Proteinase
Hyaluronidase- digest collagen
DNAse
TOXINS:
1. Alpha Toxin:
- Considered as lethal toxin and it
causes: cell lysis, hemolytic, tissue
necrosis
2. Theta toxin: hemolytic and necrotic
3. Enterotoxin
74
DISEASES
LABORATORY DIAGNOSIS
1. Gas gangrenc/
Anaerobi
myonecrosis:
- Through wounds
- Muscles will
undergo wasting
and it leads to
muscle
decomposition
resulting to a
decrease in size
of muscles and
difficulty in
contraction
Results:
- Foul odor,
discharge or pus
- Fever, shock,
toxemia,
hemolytic
anemia
2. Necrotizing
Enterocolitis
- Smooth(colon to
larhe intestine)
- Abnormal
peristaltic
movement
3. Food poisoning:
- Abdominal pain,
diarrhea,
1. BAM: double zone of

hemolysis
(alpha and beta hemolysis)
2. Cooked meat
medium:
- Chopped meat
glucose medium
1st reaction:
Saccharolytic Reaction :
- Reaction on sugars:
Results to :
- Glycogen
fermentation
- Productionn of acid
- Gas formation
- Intact meat
Proteolytic Reaction:
Use of : Robertsons
cooked meat medium
Resulting to:
- H2S formation:
Blackening and foul
odor due to sulfur
- NH3 formation- no
change
3. FT
4. Naglers Test:
- Measure alpha toxin
- Use: 5-10% egg yolk
in BA base
Result: Opacity or Opalescence
OCDEN, Junalyn B.
nausea,vomitting
White halo around

the colonies
5. Litmus milk test:
Result: formation of stormy clot
turbid; white to gray color
6. Reverse CAMP:
CLOSTRIDIUM DIFFICILE
CHARACTERISTICS
-
Anaerobic
Sub terminal
spore,ovoid
VIRULENCE FACTORS
DISEASES
1. Cytotoxins A and
B:
-related to
nosocomial
especially in
patients taking in:
--- Cephalospotin,
Clindamycin, Ampicillin
- They will cause
destruction of
normal flora
and C. difficile
becomes
resistant
1. Pseuudomembrano
us Colitis:
- Narrowing of
intestine
- A yellow
adherent
membrane
75
LABORATORY
DIAGNOSIS
1. LAT: LATEX
AGGLUTINATION
TEST
- Detect
Cytotoxin A and
B
2. CCFA:
CYCLOSERINE
CEFOXITIN
FRUCTOSE AGAR
- Components are
:
- Cycloserine:
inhibit growth of
OCDEN, Junalyn B.
G- bacteria
- Cefoxitin: inhibit
growth of G+
and G- bacteria
and
- Fructose
- Contain:
Neutral Red (pH
indicator)
Animal peptone
source of
protease
Result: golden yellow,
ground glass appearance
CLOSTRIDIUM TETANI
COMMON NAME: DRUMSTICK BACILLUS/ TACKHEAD BACILLUS
Incubation period: 4-5 days to several weeks( dependent on the number of
bacteria); entrance of bacteria to CNS
CHARACTERISTICS
VIRULENCE FACTORS
76
DISEASES
LABORATOR
Y
DIAGNOSIS
OCDEN, Junalyn B.
Terminal
Swollen Spore
Lollipop or
tennis racket
shape
Nonencapsulated
Motile
Anaerobic
Tetanus Toxin:
2 components:
o Tetanospamin- a
neurotoxin that inhibits
lysine resulting to
Spastic paralysis
o Tetanolysin- hemolytic
CLOSTRIDIUM BOTULINUM
CN: CANNED GOOD BACILLI
77
Tetanus:
infection of
devitalized
tissues( wound,
burn pts, injury
esp. 2nd to 3rd
degree)
COMPLICATIONS:
Respiratory Failure:
Clinical features are
mentioned above:
1. Lock Jaw or
Trismus (in the
lower jaw,
specifically the
masseter
muscle)
Resulting to Risus
Sardonicus or Sarcastic or
Sardonic Smile
2. Opisthotonos:
- Backward
arching
- Clenching of
fingers and toes
3. Arm Flexion
4. Leg Extension
Demonstratio
n of toxin
from the
patient
OCDEN, Junalyn B.
COMMON IN VACUUM SEALED PROCESS FOODS
HABITAT: SOIL
DIFFERENT STRAINS: A TO G; medically important: a,b ,e related to contaminated fish products
CHARACTERISTICS
VIRULENCE FACTORS
DISEASES
78
LABORATORY
DIAGNOSIS
OCDEN, Junalyn B.
-
Sub-terminal
swollen spore
Snow-shoe
appearance
Motile
Anaerobic
Nonencapsulated
Botulinum Toxin:
- A 1ug/kg of this toxin can
cause a disease
- Causes:
o Proteolysis of SNARE
PROTEINS such as
SYNTHOXIN, SNAP
25 and
SYNAPTOBREVIN-B
BOTULINUM:
May die due to:
- Cardiac Arrest
- Respiratory
Failure
Divided into 3 parts:
1. Classical Food
Borne:
IP: 18-24 hours
MOT: ingestion( spiced,
smoked, vacuum packed
or canned goods)
S/S:
- Diplopia:
double blurry
vision
- Dysphagia:
muscle are
relaxed thus,
difficulty in
swallowing
- Dryness: in the
oropharynx
- Dysphonia:
difficulty in
talking
- Abdominal
pain
- Constipation
2. Infantile Botulism:
- Ingestion of
milk
contaminated
with honey
derivatives
- In the 5th
79
1. Demonstration
of toxin on leftover food
Or blood from the
patient
2. Use of lab
animal:
- Specimen
from the
patient and
then, inject it
to the lab
animal
OCDEN, Junalyn B.
month: IB is
common since
mixed feeding
is done
S/S:
-
Diplopia:
double blurry
vision
Dysphagia:
muscle are
relaxed thus,
difficulty in
swallowing
Dryness: in the
oropharynx
Dysphonia:
difficulty in
talking
Abdominal
pain
Constipation
CAN INFECT
THE WHOLE
BABY
3. Wound botulism:
S/S:
- Diplopia:
double blurry
vision
- Dysphagia:
muscle are
relaxed thus,
difficulty in
swallowing
- Dryness: in the
80
OCDEN, Junalyn B.
-
oropharynx
Dysphonia:
difficulty in
talking
Abdominal
pain
Constipation
MOT: SKIN
CLOSTRIDIUM BUTYLICUM AND C. BARATIL: PRODUCES BOTULINUM TOXIN

81
OCDEN, Junalyn B.
GRAM CURVED BACILLI

FAMILY: VIBRIONACEAE
VIBRIO CHOLERAE
CN: KOMMAS BACILLUS
- Comma shaped bacillus
- Motile,monotrichous/polar flagella
- Aerobic-facultative anaerobic
- Oxidase+,NO3 reduction
MOT: ingestion
VF:
1. Chloragen: primary virulence factor, cause hyper secretion of water and chloride
2. Mucinase: function: attachment, 1st released before chloragen
DISEASE:
1. Asiatic cholera:
S/S: abdominal pain, vomiting, fever, RICE WATERY STOOL
DIVIDED INTO:
A. BIOTYPES:
CLASSICAL
LESS SEVERE
EL TOR
MORE SEVERE
TO IDENTIFY CLASSICAL FROM EL TOR:

Base it on:
Classica
l
El tor
- Hemolysis
- Chicken RBC agglutination
- Susceptibility to 50IU Polymyxin B
Suscep
Resis
82
OCDEN, Junalyn B.
tible
tant
- VP
- Grieg test= +result: hemolysis (use

sheeps blood)
B. SEROGROUPS:
Ogawa- aka Original J; Philippines
Inaba- Middle/Intermediate; Japan
Hikojima- India
LAB DIAGNOSIS:
SPECIMEN: STOOL
1. DARKFIELD MICROSCOPY: perform to observe darting motility
2. CULTURE:
a. APW: ALKALINE PEPTONE WATER
- Enrichment broth
b. TCBS: THIOSULFATE CITRATE BILE SALT AGAR
Colonies: 2-4 mm, yellow, opaque with transparent periphery
c. TTGA: _______________________________
- Dark centered colonies with cloudy zones
d. TGA: _________ GELATIN AGAR
- Flat transparent with a cloudy halo
e. GA: GELATIN AGAR
Colonies: transparent with opaque halo
3. STRING TEST +
a. Drop of colony
b. Mix with .5% sodium desoxycholate
c. Mix,lift
4. CHOLERA RED TEST +
+ red
5. 0/129 SUSCEPTIBILITY TEST
83
OCDEN, Junalyn B.
VIBRIO PARAHAEMOLYTICUS
- Comma shaped bacillus
- Motile,monotrichous/polar flagella
- Aerobic-facultative anaerobic
- Oxidase+,NO3 reduction
DIFFERENCE: halophilic salt loving bacteria
VF:
1. Enterotoxin
DISEASE:
1. Gastroenteritis:
- Ingestion of contaminated seafoods
- Watery to bloody diarrhea, vomiting, nausea, headache
LAB DIAGNOSIS:
1. TCBS: + result: BlueGreen colonies
2. Kanagawa Blood Agar: Normal Blood Agar but with an increased concentration of salt
- Exhibit Kanagawa phenomenon
Cause: hemolysis
+ result: hemolysis
AEROMONAS HYDROPHILA
- Water loving; either in sea/fresh water
- Motile
DISEASE:
1. Cellulitis: inflammation of the connective tissue
Specifically 2 regions: papillary and reticular
2. Wound infections
3. Diarrhea: caused by Cholera like toxin
4. Septicemia
5. UTI
6. Ear infection
PLESIOMONAS SHIGELLOIDES
OLD NAME: AEROMONAS SHIGELLOIDES
Habitat: surface water and soil
84
OCDEN, Junalyn B.
DISEASE: gastroenteritis
CAMPYLOBACTER
Campylobacter fetus
CHARACTERISTICS:
- Curved bacilli
- S shaped, gull wing
- Motile, monotrichous/polar flagella
DISEASES:
1. Meningitis
2. Pneumonia
3. Arthritis
4. Septicemia
5. Proctitis
6. Infertility/Sterility
Campylobacter jejuni
VF: Enterotoxin
DISEASE: Gastroenteritis
IP: 1-10 days
S/S: watery/bloody diarrhea, vomiting, abdominal pain
DIFFERENCE OF FETUS FROM JEJUNI
LAB DIAGNOSIS:
BASED ON GROWTH
FETUS
25 degrees celcius
37 degrees celcius
45 degrees celcius
JEJUNI
+
+
+
+
85
OCDEN, Junalyn B.
HIPPURATE HYDROLYSIS
FETUS +
JEJUNI
SUSCEPTIBILITY TO:
Nalidixic
Cephalothin
Fetus
Resistant
susceptible
Jejuni
Susceptible
Resistant
CAMPYLOBACTER SPECIES
MEDIA:
1. Skirrows Medium:
Use: blood agar base (lysed horse blood)
Add:
- Vancomycin, Trimethoprim, Polymyxin B
2. Butzlers
Use: FT (incorporate with agar 3% sheeps blood:
Add:
- Bacitracin, Novobiocin, Colistin,Actidione, Cephalothin
3. Blasers aka Campy BAP
- Easiest at 37 degrees celcius
- Brucella agar base with sheeps blood
Add:
- Vancomycin, Trimethoprim, Polymyxin B, Cephalothin, Amphotherium B
Colonies:
- Dry, flat, gray and irregular
- Moist, rounded, convex , glistening and entire
HELICOBACTER PYLORI
CN: CAMPYLOBACTER PYLORI
DISEASE: Gastritis, Peptic Ulcer
S/S: GIT bleeding, pain in the upper abdomen
86
OCDEN, Junalyn B.
Treatment: Omeprazole
NON-FERMENTATIVE G BACILLI
MAC
Hugh-Leifson oxidation: fermentation medium
PSEUDOMONAS
-
Grow on MAC
Non-lactose fermenter
Aerobic
Oxidase +
Motile(mostly)
PSEUDOMONAS AERUGINOSA
Characteristics:
- See aboce
- Monotrichous
- Pigments:
o Pyocyanin: blue
o Pyoverdin: green
o Pyorubin: Red
o Pyomelanin: brown /black
VF:
1. Endotoxin: for signs and symptoms
2. Exotoxin: cause septicemia
3. Slime Layer: for attachment
DISEASE:
1. Septicemia: causes this dse to immunocompromised
Cause Nosocomial Infection
87
OCDEN, Junalyn B.
2. UTI
3. Infections: patients with cystic fibrosis (genetic)- difficulty in sweating
LAB DIAGNOSIS:
MAC: non lactose fermenters thus, the + result: colorless
BAM: beta-hemolytic
Blue colonies if exposed to woods light
Odor: grape like odor/fruity
PSEUDOMONAS MALLEI
AKA: BURKHOLDERIA
-
Non-motile
Zoonotic- horses
Cause: Glanders/Farcy- thickening of the superficial lymphatics of horses (lymph nodes)
Man:
PSEUDOMONAS PSEUDOMALLEI
- Motile
- Lophotrichous
DISEASE:
Glander like disease/VTB or Viatnamese Time Bomb:
2 types:
1. Acute VTB:
S/S: high fever, bloody purulent sputum, sepsis, death
2. Chronic VTB:
S/S: pneumonia like, lung abscess
XANTHOMAS MALTROPHILA
- Motile, lophotricus
- Non-fermentative
MAC:
Yellow-yellow green colonies
88
OCDEN, Junalyn B.
ALACALIGENES FAECALIS AND ALCALIGENES ODORANS

-
Motile
Peritrichous flagellar arrangement
Opportunistic
ACINETOBACTER
Gram cocco bacilli (shorter) in pairs
Non-motile
4 SPECIES:
ACINETOBACTER CALCOACETICUS- soil

ACINETOBACTER CALCOACETICUS VAR ANITRATUS/ACINETOBACTER BAUMANNII
ACINETOBACTER LWOFFII- NF: of the skin
ACITNETOBACTER JOHNSONII: NF: of the skin
MAC: bluish tint; opportunistic
MORAXELLA
- Non-motile
- Small G- cocco bacilli to diplococcic
- Found in respiratory tract
MORAXELLA LACUNATAL/MORAXELLA LIQUIFACIENS
Cause: Conjunctivitis
MORAXELLA NON-LIQUIFACIENS
Cause: endophthalmitis
MORAXELLA OSICENSIS AND MORAXELLA PHENYLPYRUVICA
Cause: Sinusitis, Conjunctivitis, Bronchitis, Endocarditis, Septicemia
FLAVOBACTERIUM MANINGOSEPTICUM
89
OCDEN, Junalyn B.
- Non-motile
Disease:
Meningitis, Pneumonia in adults, neonatal meningitis
MISCELLANEOUS AND FASTIDOUS G BACILLI

HAEMOPHILUS
-
Non-motile
Non-encapsulated/Encapsulated
Fastidious: need
o Factor X- hemin/hematin ;source: blood; heat stable
o Factor V- aka NAD(Nicotinamide Adenine Dinucleotide or COE1; NADP (P=Phosphate) or COE2
HAEMOPHILUS INFLUENZAE
CN: Pfeiffers bacillus
-
NE/E ; dividen into 6 (A-E)

Most pathogenic HIB: cause severe respiratory disease
HAEMOPHILUS PARAINFLUENZAE
Disease: SBE (sub acute Bacteria Endocarditis)
HAEMOPHILUS AEGYPTIUS
CN: Koch-Weeks Bacillus
Cause: pink eye conjunctivitis
HAEMOPHILUS DUCREYI
CN: Ducreys Bacillus
- Pleomorphic
- School of fish appearance
Cause: Chancroid /Sof Chancre- painful genital lesions
LAB DIAGNOSIS:
90
OCDEN, Junalyn B.
1. CAM: moist and gray
2. Staphylococcal Streak Technique
- Presumptive test to detect Haemophilus Influenzae
a. Streak staph
b. Overlay suspected colonies
c. Pinpoint/dew drop around staph
Satellite phenomenon
Small transparent and convex
3. Casmans Agar/HIA(Haemophilus Isolation Agar)
4. Fildes Enrichment Agar: aka Levinthals
EMB aka Levines;ratio = 6:1
5. Tests
X
H. influenzae
H. aegyptius
H.ducreyi
H.
parainfluenzae
Y
+
+
+
+
+
+
-
Porphyrin
+
-
6. Porphyrin Test:
+ result: red
BORDETELLA
- Non-motile, aerobic, encapsulated
3 species:
BORDETELLA PERTUSSIS
CN: Bordet-Gengou Bacillus
VF: Capsule
91
OCDEN, Junalyn B.
DISEASE: Pertussis/Whooping Cough
MOT: inhalation
IP: 7-10 days
3 PRIMARY STAGES:
1. Catarrhal: risky stage
- Insidious; abrupt
- Experience: colds only for 1-2 weeks
Progress to:
2. Paroxymal
- Give off distinctive whoop; forcible expiratory cough
- Inhale=inspiratory
3. Convalescence
Other species:
Bordetella parapertussis (milder), Bordetella bronchioseptica
Motility
BGA(Bourdet
Gengou Agar)
Browning of
peptone
Citrate
Urease
Nitrate Reduction
Oxidase
Bordetella
pertussis
Pearly
sheen(looks like
mercury like
droplets)
+
Bordetella
parapertussis
Large, dull,
brown
Bordetella
bronchioseptica
+
large
+
+(24 HOURS)
-
LAB DIAGNOSIS:
Method:
- Cough plate technique
92
(4 HOURS)
+
-
OCDEN, Junalyn B.
- Nasopharyngeal aspirate
Culture:
1. BGA: Bordet Gengou Agar
Other name: PBGA( Potato Blood Glycerol Agar)
- Smooth, transparent, convex, pearly sheen, narrow zone of hemolysis
2. Regan-Low Agar
BRUCELLA
- Non-motile, Non-encapsulated, Aerobic
- Found in Animals
- Grow slowly on culture, do not ferment carbohydrates
IP: 3-7 DAYS
CO2
Brucella
abortus(Bangs
Bacillus)
Brucella suis
Brucella canis
Brucella
melitensis
Found in cattle
Pigs
Dogs
Sheep and
Goats
Basic Fuchsin
+
V
+
Culture:
1. Brucella Agar/Castaneda Medium
- Pinpoint,s mooth, convex, glistening
Test:
1. Abortus/Bangs Ring Probe Test
+ blue line/ring at the surface of the agar
Diseases:
Brucellosis: 2 types:
IN ANIMALS:
- Cause of sterility in animals
93
Thionine
+
+
+
OCDEN, Junalyn B.
IN MAN:
- Brucella melitensis
- Ingestion of goats/sheeps milk
- Undulant/Malta/Intermittent fever
FRANCISELLA TULARENSIS
- Small Gram cocco bacilli
- Non-motile
MOT: bites of vectors(ticks,mites,lice)
DISEASE:
Tularemia
Tick Fever/Rabbit Fever
Type of fever for both:
Ulcero - grandular type
ACTINOBACILLUS ACTINOMYCETENOMITANS
- Small G cocco bacilli
- Morse code appearance
DISEASE:
1. Periodontitis
2. Endocarditis
CULTURE:
MGB: Malachite Green Bacitracin
TSBV: Trypticase Soy Agar with Serum Bacitracin Vancomycin
PASTEURELLA MULTOCIDA
- Small Gram bacilli
Cause: wound infection from bites and scratches of cats and dogs
BAM: 1-2mm, smooth, moist-mucoid, greenish discoloration, non-hemolytic
LEGIONELLA PNEUMOPHILA
- Very small Gram cocco bacilli
MOT: inhalation
Contaminates freezers and aircon
94
OCDEN, Junalyn B.
Temperature can be too cold/too hot: warm-hot
DISEASES:
1. Legionnaires disease/Atypical pneumonia/Broad street; cough and a non purulent sputum
2. Pontiac Fever: URTI, mild cough, sore throat
LAB DIAGNOSIS:
1. Dieteries Silver Impregnation
2. BCYE(Buffered Charcoal Yeast Extract)
3. Feedy and Gorman
- Flat, entire and rounded
STREPTOBACILLUS MONILIFORMIS
- Gram bacilli in chains
- Pleomorphic(shape)
DISEASE:
1. Rat Bite fever/Havernill fever
- High fever, reddish brown rashes lymphadenopathy
LAB DIAGNOSIS:
1. Thioglycollate with Ascitic Fluid:
Cotton balls/Puff balls/Bread crumbs
SPIRILLUM MINOR
Cause:
Rat bite fever/Soduku
Same signs and symptoms (see above)
MYCOPLASMA
1. Mycoplasma pneumoniae
- PPLO- Pleuropneumonia like organism
- Smallest free living bacteria(.2-.3um)
- Wall-less
- Sterols in the cell membrane
RESISTANT TO:
- Penicillin
- Cephalospotin
95
OCDEN, Junalyn B.
CULTURE:
Fried-egg appearance
- Raised center, thin edges
1. Cold Agglutination Method
Disease:
1. PAP(Primary Atypical Pneumonia)
- Cough, purulent sputum
2. Tracheobronchitis: obstruction of windflow
3. Pharyngitis
2. Mycoplasma hominis
Cause: PID
3. Ureaplasma urealyticum- cause urethritis
CHLAMYDIA
- Obligate intracellular cell
- RNA and DNA
- Prokaryotic
Have replicative cycle:
a. Elementary body: extracellular, small, inert(inactive),infective
b. Reticulate/Initial body: large,non-infective body
Chlamydia
Chlamydia
Chlamydia
trachomatis
psirtaci
pneumoniae
Host
Man,mice
Man,birds
Man
Disease
a. Trachoma
Psi
Pneumonia,
- Conjuncivitis Hacosis/Ornithosi bronchitis,
with scarring s/Parrot fever
pharyngitis, flu
b. Inclusion
-red rashes, fever like illness
conjunctivitisconjunctivitis
with pus
c. LGV (Lympho
96
OCDEN, Junalyn B.
Elementary
bodies
Inclusion body
Granuloma
Venereum)white
discharge,
inflammation of
lymph nodes
Round, vacuolar
Dense, variable
Dense,round
Halberstatder
Prowazek
Levinthal-ode
lillie
Round
LAB DIAGNOSIS:
1. Giemsa,Gimenez,Machiavella Stain
2. Cell cuture
3. Frie-skin test (LGV)
SPIROCHETES
- Thin-walled
- Helical
- Motile
- Cork-screw
- Endoflagella
Genus:
1. Treponema- found in tight rigid cells
Treponema pallidum sub specie pallidum
DISEASE: Venereal Syphilis/Great Pox/Italian Disease
4 STAGES:
1. Primary: hard chancre- oral and genital lesions, painless
2. Secondary Stage:
Maculopopular rash- red rashes in palm and sole
Condylomata lata- white patches
3. Latent Stage:
97
OCDEN, Junalyn B.
All Signs and Symptoms will disappear

Persons are highly infectious
+ sero test
4. Tertiary Stage:
Signs and symptoms will occur ,plus:
- Gummas- tumor in any organ (tumor-abnormal growth of cells)
- Neurosyphilis
- Cardio Syphilis
MOT: sexual,congenital/transplacental/direct inoculation to blood
IP:3-90 days;average: 3 weeks
Test:
1. Darkfield Microscopy
2. Serology
a. Non Treponemal
- Use cardiolipin(reagent) from beef heart and add serum= (positive result) flocculation
LABORATORY
98
OCDEN, Junalyn B.
MICROBIOLOGY
NOTES
OCDEN, JUNALYN B. (2015)
Microscope- instrument to magnify an image of specimens
Types:
1. Light Microscope- background is illuminated, dark-filled, phase contrast, differential
interference, fluorescence
- Eyepiece or Ocular: Further enlarge of image 5x-20x
- Virtual Image- because the image is inverted/upside down/reverse
- Total Magnification: product of the magnifying power of the lenses
99
OCDEN, Junalyn B.
-
Numerical Aperture: pertain to the light gathering ability of objectives, determined

through the size of objectives.
2. Electron Microscope
- TEM: Transmission Electron Microscope
- SEM: Scanning Electron Microscope
- Scanner- .10; MPO-.25;HPO-.65;OIO-1.25- wet using
- As magnification increases , aperture increases ; direct relationship
Refractive Index: the refractive index of the glass is the same to
the refractive index
*Avoids refraction
- Measure of the relative velocity of light as it passes through a medium
Example of Light Microscopes:
1. Phase contrast- works by _____________ the differences between the refraction index and
surrounding medium.
RI- cleare/denser image
2. Dark-filled microscopes- illuminating only the specimen but not the field.
- Resolution or Resolving Power: ability to distinguish fine details or structures within.
- Minimum distance between two objects wherein these objects could be seen as two separate
entities.
- Working distance:
o Distance between objective and specimen
o In mm
o Scanner- 18.5 mm; MPO-10.6mm;HPO-0.6mm; OIO-.13mm
o Magnification increases as working distance decreases; indirect relationship
- Contrast: the number of visible shades seen in the specimen
- Stain the slide before focusing
Illumination System: light source
- Condenser: focus the light
- Iris diaphragm: regulate light
- Filter : select what light trace with a wavelength of 410nm.-a clearer image; 700 nmwavelength of daylight
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Support System: base, arm, stage, observative tube/body tube , revolving nose piece
Adjustable System: Focusing knob: Coarse- broad; Fine- precise, Y-axis & X-axis feed knob, Diopter
ring, Interpupillary Distance, Condenser lever, Iris diaphragm aperture ring, Light Adjustment
Knob
Parfocality- ability to remain in focus when shifting objective
EXPERIMENT 3
MICROBIAL CONTROL
-introduced in the 1800s by Semmelweiss and Joseph Lister
Reason:
- 10% who get hospitalized die because of Nosocomial Infections. (Hospital acquired
infections)
- 25% of mothers will die after delivery due to contamination.
Sterilization- complete absence of all forms of microbial life including endospores. 2
groups: Genus bacillus & Genus clostridium.
Example:
- Commercial sterilization( esp. canning industry)
*Clostridium botulinum = canned good bacilli
*Botulism- no contraction of muscles, difficulty in swallowing
- it cannot remove the endospores of thermophiles (- most are non-pathogenic)
Disinfection
- reducing the number of harmful microorganisms
- cannot destroy endospores
- can destroy: vegetative pathogens or non-spore forming bacteria
*Disinfectant- for non-living things
*Antiseptic- for living tissues
*Degerming- mechanical removal of microbes in a limited area
Sanitation
- More on in the food industry
- use of chemical agents to reduce microbes in food handing equipments.
Reason:
- Meet public health standards
- Reduce bacterial contamination
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METHODS TO CONTROL MICROBIAL GROWTH:
A. PHYSICAL denature or alter the structure of microorganism
1. Use of heat:
2 types: Moist Heat (denature by coagulation) and Dry Heat(have an oxidizing effect)
*TDP- Thermal Death Point- pertains to the lowest temperature at which microbes in a liquid
suspension will be killed in 10 minutes.
Example: E. coli- 370C420C- 1015 mins.
*TDT- Thermal Death Time- minimal length of time at which all microbes will be killed at a given
temperature.
*DRT- Decimal Reduction Time- minimal length of time at which 90% of microbes will be killed at a
given temperature.
MOIST HEAT:
1. Boiling
- 1000C at sea level
- Kills: vegetative bacteria, fungi & some viruses except those with endospores.
Example: Hepa A- survives boiling at 30mins. And bacteria with endospores can survive 20 hours.
2. Autoclave- steam under pressure
3. Pasteurization
- LTH- 63-650C or classical
- UHT- Ultra High Temperature- 1400C at 3 sec.- cool in vacuum.
DRY HEAT:
1. Direct Flaming- expose instruments directly on a flame until its red hot. Esp.,
inoculating hoods and needles and can also be called Flame Sterilization.
2. Incineration- until it forms ashes, applicable to disposable instruments.
3. Hot air sterilization- place instruments for 2 hours at 1700C.
-
2. Filtration
Removal of microbes as liquids/gases would now pass through a screen like material.
For heat sensitive materials
Examples: vaccines, enzymes, antibiotics, culture media
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1. HEPA - High Efficiency Particulate Air Filter
Used in: Operating Room & Burn Units- removes bacterial contamination.
2. Membrane Filter- uniform pore size(0.22-0.45 micrometer)
Cannot be filtered: spirochetes, Mycoplasma, viruses
1.1 micrometer pore size for large proteins are virus
3. Low Temperature
3 types:
1. Refrigeration(0-70C)
-lower metabolic rate of microorganisms thus they cannot reproduce and cannot
release toxins
2. Freezing (lower than 00C)
2 methods: Flash Freezing & Slow Freezing
3. Freeze drying or Lyophilization- killing for the purpose of preservation.
4. Dessication- extreme dryness; dry-out
Example of those who can survive even if it is dry;
Endospores- esp. Clostadium and Bacillus (decades); Neisseria gonorrheae (one day); Mycobacterium
tuberculosis (months)
Problem: most viruses are resistant
5. Osmotic pressureHigh salt concentration= increase osmotic pressure
High sugar concentration=increase osmotic pressure= hypertonic solution
Bacteria will die- Plasmolysis because of the absence of water

Resistant in Osmotic Pressure: Staphylococcus, molds, yeast
Halophilic- salt-loving
6. Radiation
A. Ionizing Radiation
Examples; X-ray, electron beams, gamma rays- emits wavelength at less than 1nm.
How will it kill microbes? It will dislodge electrons from atoms
Ions + Peroxide and it will cause mutation

Where to use IR? Disposable medical supplies
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Disadvantages: capable of penetrating the skin causing genetic mutation to an individual.
B.Ultraviolet radiation- non-ionizing radiation will produce thymine dimmers and it will cause
microbial mutation. But it cannot penetrate: paper, cloth and glass.
Wavelength:
C.Microwave (1mm-1m)
- it will kill microbesin the presence of water
-release heat absorbed by water kills microorganisms
Esp. Vegetative bacteria
Cooking pork in microwave Trichinosis; causative agent: Trichinella spiralis
*Asepsis- absence of contamination
*Sepsis- presence of bacterial contamination
*Aseptic technique- methods in removingor avoiding contamination esp. among patients,
instruments and medical personnels
*Bacteriostatic- agent that will inhibit the growth of microbes.
*Germicidal- agent that acts as Bactericide, fungicide, viricide(does not kill viruses but it
inactivates them)
*Sporicide- destroys endospores(protection) & fungal spores(help bacteria reproduce)
B. CHEMICAL METHODS:
- Most in the form of disinfectants
1. Phenols & Phenolics: kills microorganisms by destroying or disrupting the cell membrane of
microbes.
Example: Carbolic Acid (additive in throat spray)- local anaesthetic
Disadvantage; strong irritating odor & skin irritant
Phenolics: chemical derivative of phenols
TYPES:
1. Cresols
Example: Lysol- comes from coal tar & effective disinfectant
2. Biphenols
Example: Phischex- kills staphylococcus and streptococcus bacteria
Advantage:
1. They are stable
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2. They remain active even in the presence of organic compounds
Factors affecting Microbial Control:
1. Number of Microbes
2. Type of Microbe
3. Environmental Influence
4. Effectiveness of an anti-microbial
5. Time of exposure depends on what method are you using
3. Persists for a long time
2.Halogens- effective in compound form/ single form
A. Iodine:
a. Tincture of Iodine- Iodine in an alcoholic solution, used for disinfecting inanimate
objects
- Stains skin, clothing and considered as skin irritant
b. Iodophors- Iodine with several compounds, used as skin antiseptic esp. in surgery ;
Example: Betadine & Isodine
B.Chlorine- must always be water+Cl HOCl(Hypochlorous acid)- drinking water, pools, sewage
Disadvantage: ineffective in the presence of organic compounds
*NaOCl- Sodium Hypochlorite- active ingredient of Bleach
*Chloramines-Cl+ammonia- used as a less effective germicidal
3.Alcohols
How? Disrupt the cell membrane
Type: Vegetative bacteria, fungi
- It cannot kill naked viruses (example: Parvovirus B19- causes hemolytic anemia; AdenovirusURT; Rotavirus- Gastroenteritis; Coxsackie- causes URTI)
- Can kill bacteria with endospores:
Example:
1. Ethanol (Ethyl Alcohol)- drinking alcohol with optimum concentration of 70%
2. Isopropanol( Isopropyl Alcohol)- rubbing alcohol
4.Heavy Metals- considered to have oligodynamic action
- small amount are sufficient to kill microbes effectively
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Example:
1. Silver- 1% Silver Nitrate- used to prevent the occurrence of opthalmia neonatum (from:
neonate; results to purulent conjunctivitis)
*Credes method- dropping of Silver Nitrate in the eye.
2. Mercury- merthiolate (disinfect skin wounds) & Mercurochlome
3. Copper- Copper Sulfate- anti microbial
It can destroy: algae in pools and fishtanks
4. Selenium- anti- fungal properties
5. Zinc- Zinc Chloride- compound for mouthwash- anti bacterial; zinc chloride- paints
5.Quarternary Ammonium Compounds AKA QUATS
- most effective disinfectant
How? By destroying the bacterial structures
Example: Zephiran and Cepacol- both can be found in lab spray bottles
Exception: resistant = pseudomonas
Disadvantages:
1. It will form foams
2. Easily inactivated by soaps . Why? Because soaps are anionic detergents.
3. Easily inactivated by organic compounds.
6.Aldehydes:
How? By forming covalent cross links with several functional groups
Example:
1. Formaldehyde (37% aqueous)- preserve biological specimen, disinfection & antiseptic,
neutralize bacteria & viruses in vaccines.
2. Glutaraldehyde(Cidex)
- Effective disinfectant & sterilizing agent
- Soaking solution in hospitals
- Denature microorganisms
- Bactericidal, tucerculocidal, viricidal, and also sporadical
- Soak instruments for 3-10 hours
7.Gaseous Sterilizers- chamber lined with chemicals (ethylene oxide)
- used to disinfect large equipments
Disadvantage: Explosive in pure form
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8.Peroxygens
*Ozone (O3)- highly reactive form of oxygen expose to Ultraviolet Light
-very reactive in chlorine
-disinfect pools
-neutralize the taste and odor from water
Disadvantage: less stable & more expensive
*Hydrogen Peroxide (H2O2)- effective as a disinfectant
Disadvantage: not suitable for large wounds because Hydrogen Peroxide is immediately acted on as
catalase Water+Oxygen
-can be used as sporadicidal (not in room temperature) but at high temperature.
*Benzoyl Peroxide
*Peracetic acid- effective liquid sporicide
- kills bacteria and fungi in 5 mins
- it kills endospores & viruses in 30 minutes.
-used to clean food handling equipments because it doesnt leave any toxic residues.
Clean Catch Method or Mid-Stream Method
After urine collection: it must be at least one hour

If process is delayed: refrigerate for 18-24 hours at 40C
1. Grams Stain
2. Culture
3. Colony Count: assess bacteriuria from contamination
Example: less than 10,000/ml contaminant
More than 100,000/ml- bacteriuria
a. Pour Plate
b. Filter paper strip method
c. Calibrated loop method- 0.001ml of urine placed in a culture media -(non-selective
BAM/BAP;CLED: Cysteine Lactose Electrolyte Deficient; PLA- Purple Lactose Agar; MAC with
C.V)
d. Incubate at 350C overnight
e. Count the colonies and multiply to 1000
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D. STOOL
Suspecting: Gastroenteritis ; Salmonella, Shigella, Campylobacter, Vibrio
Amount: 5g or thumb size or pea size
Container: Sterile, wide mouth leak proof container
Utmost 2 hrs.
If delayed: Give a transport medium such as:
APW- Alkaline Peptone Water (for Vibrio)
Amies
Stuarts
Cary-Blair
Substitute: 33mmol/L Glycerol Phosphate buffer
Problem in defecation:
- Use swab (2-3 sterile swab)
- Introduce to the liquid media
- Culture
- Gram Stain is not routinely done
- Culture Media: MAC, EMB (Eosin Methylene Blue), SSA (Salmonella Shigella Agar), Skirrows
(for Campylobacter)
E. THROAT CULTURE
- collect in the uvula
Suspecting: Strep. pyogenes, Strep. pneumoniae, Klebsiella pneumoniae
Steps:
1. Bacterial smear
2. Liquid media (TSB)
3. Incubate
4. Prepare smear
5. Grams stain
F. SPUTUM
Suspecting: Myco. tuberculosis, Strep. pyogenes, Strep. pneumoniae, Klebsiella pneumoniae
- Best to collect early in the morning
Method of collection:
1. Deep expectoration
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2. Lung biopsy
3. Transtracheal Aspiration (near windpipe)
4. Bronchial lavage
Prepare specime:
2 slides method
Stain:
1. Grams Stain (Strep. pneumoniae and Klebsiella)
Culture
*NQ-Neufeld-Quellung Test- capsular swelling test
2. AFS (Myco. Tuberculosis)
- Do not culture
Proceed with: Digestionconcentrationdecontamination
G. GENITAL TRACT
Suspecting: STI-> N. gonorrhea, Treponema pallidum
Method:
Swabbing (narrow tip)
Examples: Charcoal treatment tip/swab; pacron tips;Calcium Alginate; Regular Swab
Rotatory (manner of withdrawing the swab)
Liquid broth
Urethral opening (90% of N. gonorrhea)
Cervix
Anal canal(proctitis)
--discharge
Grams Stain (Gram diplococci)
Culture
Biochemical tests
H. WOUNDS, ABSCESS, PURULENT EXUDATES
Caused by: Traumatic Wounds (Clostridium), Surgical Wound (Staph. aureus), animal bites
(Pasteurella)
Can have:
Inflammation of lymph node
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Drainage
Use Pasteur pipette
Grams Stain Culture
Abscess, Wounds Exudates;
Syringe needles
Aspirate containerProcess
LAB METHODS:
I.
DIRECT METHOD:
- Examining microorganism in natural and living state
a. Wet mount/ Saline mount
b. Hanging drop-*motility
II. INDIRECT METHOD:
- Fixed stained stain
Steps:
1. Smear preparation (2-3 cm diameter)
2. Air dry
3. Heat fix
4. Stain: artificially coloring the specimen with the use of dyes
a. Positive staining:
Bacteria stained; Background- unstained
Basic dyes:
C.V
Safranin
CF
Methylene
b. Negative Staining:
Stained- background; unstained- bacteria
Acidic dyes:
Nigrosine, India Ink
Subtypes: few capsule
Grams Stain:
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1. Crystal Violet:
Crystal Violet- 2g
Ethanol(95%)-20ml
Ammonium oxalate- 0.8 g
Distilled water- 100ml
2. Grams Iodine:
Potassium iodide- 2g
Iodine Crystals- 1g
3. Acid alcohol:
Acetone
95% Ethanol
4. Safranin:
Safranin- 4g
Ethanol- 200ml
Update/ introduced by: Mesaros, Army, Strekonski, Leon

Brown Brenn:
G+ : Blue
G- : Red
Background: Yellow
Consideration:
Smear preparation: not too thick
Heat fixation: do not prolong
Decolorization: at least 15 seconds
Over decolorize: predominant G+ bacteria
Under decolorize: predominant G- bacteria
AFS:
- Mycolic acid cell wall or Hydroxymethoxy Acid Acid Fast Microorganism
*Difficult since hydroxymethoxy acid is present
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*[Tergitol #7] Detergent is required to allow the stain to penetrate the cell but once stained
AF org. are resistant to decolorization by mineral acids.
Hot Method/Ziehl Neelsen
1. Carbol fuschin (.3%)
Phenol- 50g
95% Ethanol- 100ml
Basic Fuschin- 3ml
Distilled water- depends on the concentration
2. Acid Alcohol
HCl- 30ml
Ethanol- 1L
3. .3% Methylene Blue
Methylene Blue- 3g
Distilled water- 1L
*Nocardia- slightly AF
Cold Method : Kinyoun
Increase concentration of carbol fuschin
1. CArbol Fuschin
- Liquid Phenol/ Tergitol #7 (wetting agent)
(surface active ingredient)- increase =80g
Ethanol- 200ml
Basic Fuschin- 40g
Distilled water- 1L
2. Acid Alcohol:
HCl- 30ml
Distilled water- 970 ml
3. NO METHYLENE BLUE.
Substitute: Malachite green- .3g
1L distilled water
REPORT Acid Fast Bacilli:
Pappenheims:
-different Mycobaterium laticola & Myco. smegmatis (blue)
- Myco. tuberculosis: Red
-
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1. Carbol fuschin
2. Rosolic Acid in Alcohol
3. Meth. Blue
Baumgartens Method:
Diluted alcohol fuschin
Myco. leprae (Red); Myco. tuberculosis (Blue)
Special Stains:
1. Flagellar Stains:
Common stain: Pararosalinine (Basic Fuschin) and Tannic Acid or Potassium Aluminum (Coat and
thicken flagella)
Methods: Grays, Leifsons, Fisher-conn
2. Spore stains: use steaming process
a. Acetic Acid method
Components: Carbol fuschin, 5% HAc, Meth. Blue
*Red Spore;Blue cell
b. Wirtz-Conklin/ Schaeffer Fulton
5% safranin, 5% malachite green
Spore- green; bacteria- red
c. Dorners
1. Carbol fuschin
2. Nigrosine (10%)
Spore red;colorless to utmost gray bacteria
Background- dark gray
3.Capsular Stains:
- Hiss, Gins, Welchs, Negative/Indirect/Relief (Nigrosine; Burns India inkwhen used alone it
can stain Cryptococcus & spirochetes)
4.Metachromatic Granules/ Volutin Granules
Bacteria- blue; granule- red
LAMB: Loeffers Alkaline Meth. Blue
Neissers Method, Alberts, Ljubinsky
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Biosafety- Safety from infectious agents
- Section of environment safety that suggest safety
Purpose:
To reduce risk of hazards
Levels:
1 Biosafety Level 1 (BSL)- microorganisms that do not consistently cause disease in healthy
individual.
Example: E. coli K12
S. cerevisiae
Polyomavirus
*Know basic laboratory techniques:
- Flame sterilization
- Heat fixation- kill without altering the morphology, prevent specimen from being washed off.
FOLLOW STANDARD MICROBIOLOGICAL PRACTICES
Wear PPE
Hand washing after handling agents and before leaving the lab
Even after removing surgical gloves
Disinfection of the working area- every after handling infectious dse.
No eating, drinking, smoking, handling of contact lenses and pipetting by mouth.
Sharps- leak/puncture proff container
Broken glass- Broken glass container
6. Do not recap, bend, break needles fish out
7. Disposal of biological wastes:
1.
2.
3.
4.
5.
Solid plated medium: inoculate the specimen

Get a Colony and describe to perform biological reaction
Dispose
Sterilize in the autoclave at 1200C 15 psi 20 minutes
Biohazard bags
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-lysol culture media
Let it stand for 10-15 mins
Liquid biological waste
Pour Lysol/bleach
Let it stand for 10-15 mins
Pour at the sink
2. BSL 2
- Moderate potential hazards
-Transmitted through Contact(direct or not), Ingestion ( Salmonella typhi), Puncture (Needle Stick
Injury), Blood borne pathogens (HEPA)
Things to observe basic laboratory techniques
- Extreme sharp precaution
- Observe SMP
- Proper training in handling infectious agents
- Use of safety equipment
3. BSL 3
- organisms that would cause serious dse.
Example: Mycobacterium tuberculosis, Hanta virus, Yellow fever virus Yersinia pestis (Black
plaque)
-BSL 2 and 1 + containment in the lab.strict access
---double-door entry (swinging
4. BSL 4
- fatal organism with no treatment/ vaccines
Example: Ebola virus, AIDS, Marburg
Under BSL 3+ use of positive pressure ventilated suits/ moon suite
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PRINCIPLE OF BIOSAFETY
-
Containment- safety equipments only

No spread to external environment, lab and pesonnels
A. Primary Containment:
- Protects personnel, lab environment
Things to consider;
- Proper lab techniques
- SMP
- Safety Equipments
B. Secondary Containment:
- Protection of the external environment
Observe :
-Facility design ( no wood, carpet or rag)- tiles
-Filters must be present
-Windows must be properly opened
-Use of safety equipments
- having biosafety cabinets:
a. Open fronted class I and class II
- protection from agents
b. Class II- open fronted, above it (Hepa Filter)
c. Gas tight class
EXPERIMENT 5
CYTOLOGY
BACTERIAL GROWTH CURVE
METHODS OF DETERMINING MICROBIAL GROWTH
-
Cell counting
Increase in number: Binary Fission , asexual reproduction of cells
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Generation Time/ doubling Time/ double Time
- Time required for bacterial cells to divide at regular intervals.
- Can be expressed as:
GT= Generation Time
A. DIRECT METHOD:
1. Plate Method/ Viable Cell count:
Count the number of living cells
-
CFU (Colony Forming Units) no exact value

Assume that all living bacterial cells will reproduce and divide.
POUR PLATE METHOD:
2. Filtration
The bacteria is retained in a membrane filter transferred into a culture media , incubated
and bacterial growth would be counted similar to the plate method.
Membrane Filter: .22-.45 um
3. MPN (Most probable number)

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-dealing with liquid media
- count colonies based on statistical estimation
4. Direct Microscopic Count
B. INDIRECT METHOD:
1. Turbidity- use of spectrophotometer
Clear =Increases
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Turbid =decreases
2. Metabolic Activity:
Check the number of metabolic product such as Carbon dioxide
Directly proportional to the number of bacterial chromosomes
3. Dry weight
Measure / count filamentous microorganism (ex: fungi)
InoculateIncubate Weight- Subtract 2nd weight from 1st weight= weight of filamentous
microorganism
EXPERIMENT 6
SPECIMEN COLLECTION
Primary Site: Timing, Volume, Sterile, Label
Label the body of the container

Name, Age, Gender, time of collection, date , test
Urine: 1 hr
Feces: 1hr-2hr
*Universal Precaution
A. Blood: sterile
Bacteremia- presence of bacteria in blood
Things to consider;
1. Timing of blood collection: before anti-biotic is administered and before the onset of
fever and chills.
How many? 3 blood cultures
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2. Reason why 2-3 blood cultures:
To reduce missing out a transient bacteria
*1ml of blood=few bacteria
How will you collect?
- 1 hour interval
-
3. Volume
Adult: 10 ml
Children: 2-5 ml
Neonate/ Infant: 1-2 ml
4. Skin disinfection:
- Must be meticulously prepared
Different disinfectants:
Blood culture: 70 percent alcohol, 0.5 % Chlorhexidine in 70% ROH, 10& Porvidone
Tincture of Iodine: wipe with 70% ROH to lessen skin irritation
5. Anti- coagulant:
SPS: Sodium Polyanethol Sulfate
Inhibit anti-bacterial of serum and phagocytes
Clot- trap microorganisms
No SPS
With anti coagulant but no SPS
No coagulant, No SPS, No broth

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-
Let it clot
6. Incubate:
7 days, can incubate in 2 weeks
7. Choice of broth:
TSB- Tryptic Soy Broth
All purpose liquid media
8. Quantity of Broth:
Volume of broth:
50 ml: 10X the volume of blood
9. Broth preparation:
-screw-capped conical tube
(should not be tightly screwed)
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10.
Examine: on a daily basis
B. CSF:
Appearance: Normal: clear, no RBC, can have WBC but around 3 per cubic meter (mm3)
Why collect? When the patient is suspected to have:
-Nisseria meningitis, S. pneumoniae, E. coli, Hemophilus IB, Strep. agalactiae(for animals:
Non Pathogenic, for humans: pathogenic; causes Bovine Mastitis; inflammation of the mammary
gland), Mycobacterium tuberculosis
Common = peptidoglycan
How to collect?
-
Procedure: side lying position

Insert the Spinal TAP/ Lumbar puncture/Ventricular Puncture b/n Lumbar 3 and Lumbar 5.
Needle: 17 or 18 gauge
4. Volume: 5-10 ml
5. 2 tubes: one is vented and the other is non-vented
6. Take note of the appearance: Clear, hazy, furulent, yellowish (icteric) fibrin webs,
pellicles
7. If Cloudy/ Purulent: Examine immediately and do not centrifuge
8. Other types: Centrifuge for 5-10 minutes at a speed of 10,000 rpm
Undergo 3 tests: cytological chemical, microbiology

9. In cases of delay: Refrigerate for an hour
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More than one hour= disintegrate
10. Gram stain- most have peptidoglycans
If TB is suspected: AFS
Neufeld-Quellung capsular swelling to identify Strep. Pneumoniae and Klebsiella
pneumoniae
11. Culture Media
MAC ( Maconkey) Agar
Reverse Camp Christle-Alkins-Munch-Petersen Agar
CAM- Chocolate Agar Medium can be used to isolate and grow Haemophilus influenzae
C. URINE:
Most common specimen
Common pathogens isolated:
E.coli- UTI
Proteus mirabilis
Proteus vulgaris
Enterobacter
Morganella
Providencia
Strep. Faecalis
UTI
Cystitis
Pyelonephritis (nephron) kidneys
*Collection Method:
Suprapubic aspiration- just above the belly button (for very old patients)
Use of Catheter/ Catheterization
CULTURE MEDIA
Culture: growth of microorganisms in a culture media
Types:
1. Pure/Axonic: 1 specie
2. Mixed: 2 or more specie
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3. Stock: pure (study, research, industry)
Culture Media: anything that contains nutritional and environmental supplements
TYPES:
A. CONSISTENCY:
1. Solid Media: contain 2-3% agar
2. Semi-solid media: also called as Gelatinous media; .5-1% agar
- used to check motility of microorganisms.
*motile: if there is cloudiness or haziness after incubation
*non-motile: no haziness or clear
3. Liquid or Broth Media:
-no agar; always be placed in a test tube
B. COMPOSITION:
1. Synthetic/ Defined Media:
- Exact chemical composition of media is known
2. Non-synthetic/ Undefined Media:
- Chemical composition of one component is not exactly known
3. Tissue Culture Media:
- Contains living cells:
o WI38: normal human skin
o Hep2: Human epithelial cancer cells
o Hela cells: contains human cervical cancer cells
C. How is it dispensed:
1. Plated Media: placed on a Petri dish
2. Tube media: placed on test tubes
D. USE:
1. Simple/ All purpose Media
- Allows the growth of wide variety of microbes
Example: Nutrient agar, BAM, TSA (Tryptic Soy Agar)
2. Enrichment media:
- Promote the growth of pathogens while inhibiting the growth of normal flora.
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Example:
Most are in liquid form
FI Fluid Thioglycollate
BHI- Brain Heart Infusion
TSB- Tryptic Soy Broth
Selenite
Tetrathionate
3. Enriched Media: contains nutritive supplements to allow the growth of fastidious bacteria.
Example: BAM, CAM
4. Differential media: differentiate microorganisms that are growing together in one culture
media through their cultural characteristics.
Example:
BAM: can differentiate through hemolysis/hemolytic patterns
MSA: differentiates ria Mannitol fermenters from non-mannitol fermenters

MaConkey: contains pH indicators that can identify lactose (color red)
5. Selective Media: promote the growth of wanted microorganism that inhibits the growth of
unwanted microorganism.
How? Since the selective media contains inhibiting agents such as:
a. Dyes:
Example:
- EMB (Eosin Methylene Blue)
Inhibit the growth of Gram+ bacteria
- BSA (Bismuth Sulfate Agar)
Same function with the EMB
b. Bile salts:
c. High concentration of NaCl ~ MSA
Concentration: 7.5%-10%
Specific for Staphylococcus aureus
d. Chloral hydrate
functions to prevent the swarming
e. Alcohol
of proteus
f. Sodium tellurite
allows only the growth of
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g. Sodium azide
h. Antibiotics:
Corynebacterium diptheriae
-inhibits the growth of Gram bacteria
Vancomycin (VCN)- inhibits growth of G+ bacteria

Colistin (CN)- inhibits the growth of G- bacteria
Nystatin: inhibits fungal growth
i. pH:
RogosasTomato Juice agar: Growth of Lactobacillus acidophilus
SDA (Saborauds Dextrose Agar)- inhibit bacterial growth; allows fungal growth. pH=5.6
6. Selective/Specific: for the growth of one type or a specific type of a bacteria
Example:
Petragnanis media; for the growth of Mycobacterium tuberculosis.
TM: Thayer-Martin: for bacteria Neisseria
Dieudonnes media: for the growth of Vibrio Cholerae
Mac Bride: for the growth of Listeria monocytogens
EXPERIMENT 11
Inoculation: Introduction of microorganism in a media
Techniques:
1. Liquid/Broth:
Insert inoculating needle at the acute angle
2. Agar slant streak:

- Streak in zigzag manner from top-bottom
3. Agar butt:
- Needle->Stab->2/3 of the culture media
4. Agar slant and butt:
126
OCDEN, Junalyn B.
-
Stab and streak
*TSI- Triple Sugar Iron

5. Placed Streak Methods:
a. Simple Streak:
- 1 drop of the inoculums on the farthest side
- Streak , forming parallel lines
b. Radial Streak:
- Same 1st step
- Streak in a concentric fashion
c. Multiple Streak
- Use of petri dish with 4 divisions
- Separate streak per division using simple streak
d. Overlap:
- Same 1st step
- Rotate : 60 degrees angle
- Streak
e. Interrupted Streak;
- Same 1st step
- Streak and stop at the middle
- Tilt for 180 degrees
- Streak
f. Multiple Interrupted:
- Divide culture media into 3
- Streak the 1st division and stop at the apex
- Flame sterilize
- Turn plate around 900; let loop cool
- Touch the apex once or twice
- Streak
- Flame sterilize
- Turn at 900
- Flame sterilize; touch the apex
- Streak the last division
g. Quadrant Streak:
- Division of 4
127
OCDEN, Junalyn B.
-
1st quadrant4th quadrant (more diluted compared to the 1st)
EXPERIMENT 12
INCUBATION
3 METHODS:
1. Aerobic incubation:
- Simplest method of incubation
2. Capnophilic incubation:
- Requires 5-10% Carbon
- Use of candle jar
3. Anaerobic incubation:
Types:
A. Use of Anaerobic jar:
- Removal of O2 in the chamber with the reaction to H2 added to the system in the presence
of catalysts.
2 techniques:
a. Disposable Gas Generator Envelope
Example: Gas Pack
Colorless plastic package blue (basis for the absorbance of O2)
b. Evacuation Replacement Technique (ER-Tech)

-
With the use of vacuum pump

O2 must be replaced by the following:
Nitrogen- 85%
H2- 10%
CO2- 5%
B. Use of reducing Media

- Example of culture media containing chemical component will combine or dissolve O2 to
remove O2.
- Examples:
128
OCDEN, Junalyn B.
*Cooked Meat Media Sealed with petroleum: help absorb O3
*Thioglycollate: also known as __
- allow the growth of aerobes, anaerobes and microaerophiles
*PRAS Media (Pre reduced Anaerobically Sterilized Media)

C. Alkaline Pyrogallol
- Pyrogallic acid must combine with alkaline substances such as NaOH; result: help in the
absorption of O2
- From colorless blue
D. Anaerobic Glove box
- Gas generator envelope/ Alkaline pyrogallol
EXPERIMENT 13
COLONIAL MORPHOLOGY
Colony
-
Group of microorganisms/ loop of microbes
A. Describe the FORM

Consider the:
a. Size: measure the diameter (mm)
Terms: punctiform : tiny colonies
129
OCDEN, Junalyn B.
b. Shape:
Circular
Irregular- no definite shape
Filamentous
Rhizoid- darker or thicker than filamentous , hair-like projections
c. Surface:
Smooth easily emulsified with NSS
Shiny
Glistening- sparkling
Rough/Granulated- not easily emulsified in NSS
Dull
Wrinkles/ shriveled
d. Texture/consistency:
Butyrous- butter like
Viscid/Viscous
Dry
Mucoid: clue- most are encapsulated,
Or perform string test:
Brittle
B. ELEVATION: at side view
Flat
Raised
Convex
Pulvinate- cushion
Umbonate- knob like protuberance
C. MARGIN EDGE:
Entire
Undulate colonies
Lobate
130
OCDEN, Junalyn B.
Curled- wave like patterns
Filamentous
D. HEMOLYSIS:
- Alpha- hemolysis
o Incomplete/partial destruction of RBC
Change in the cell membrane/ colonies to greenish
- Beta- hemolysis:
o Complete destruction of RBC
o There is clearing of colonies
E. COLOR
F. AMOUNT:
Scanty, Abundant, Few
G. PIGMENT PRODUCTION:
- Non diffusible present within the colony
- Diffusible- can scatter
H. DENSITY:
Transparent, Translucent, Opaque
I. Odor:
Example: Grape-like odor pseudomonas
EXPERIMENT 14
BIOCHEMICAL REACTIONS
Cell membrane contain: enzyme systems
Different biochemical test:
1. Carbohydrate Fermentation systems:
- With the use of lactose/broth
131
OCDEN, Junalyn B.
Cracks=fermentation
Fermentation has 3 end products:
1. Gas
2. Acid production:
TSI- present changes in the pH indicator
3. Neutral products:
Acetone, ethanol, Acetoin aka Acetyl-methyl-carbinol
TSIA Agar: Triple Sugar are Glu-Lac-Suc ratio is 1:10:10
pH indicator: phenol red; if acidic = yellow
Source of sulfur: Sodium thiosulfate
H2S indicator: Ferric sulfate, Ferric ammonium citrate
Alternative to TSI: KIA (Krigler Iron Agar)
2. Catalase Test:
Hydrolyses 3% H2O2 to Water and Oxygen
Positive result: rapid bubble formation or effervescence
3. Oxidase test:
How to detect Cytochrome oxidase(combine with O2) .. Act on aromatic amines(produce)
colored compound
How will you know if it is positive with oxidase test?
Reagents used:
P-amino-di-methyl-aniline-monohydrochloride
Pink to red to blue/purple
Tetra- methyl-p-phenylene diamine-di-hydrochloride
Result: blue/black
4. Nitrate Reduction Test:
Nitrate (converted)Nitrite + (2 reagents)
Result: Red Diazonium dye(Red colored component) specifically, p-sulfobenzene-azo-alphanaphthalamine
5. IMVIC- used to differentiate enterobacteria
132
OCDEN, Junalyn B.
Divided in to 4 tests: I, M, V and C
a. Indole:
Media: tryptophan/ tryptone broth media
Principle: Bacteria that contain tryptophanase
How?
2 processes
b. Methyl Red:
Media: MR-VP (Methyl Red-Voges Proskauer)
Substances: Clark-Lubs Dextrose Broth
MR VP (would produce)strong acids (pH: below 4.5)
What are formed?
c. Voges Proskauer:
Media: MR VP
Substance: Clark-Lubs Dextrose Broth
MR VP (produces) acetoin (chief end product of glucose metabolism from Butylene
Glycol Pathway
133
OCDEN, Junalyn B.
Acetoin + 40% KOH,O2 Acetoin is broken down into simple products which is di
acetyl + alpha-naphtol
Positive result: red
d. Citrate Utilization Test:
Media: Simmon Citrate Agar Slant
pH indicator: bromthymol blue
if color green: neutral
if acidic, slant is color yellow or blue: alkaline
Bacteria that can utilize citrate as source of carbon can also utilize ammonium phosphate
as their source of nitrogen with the production of ammonia to ammonium hydroxide.
Positive result: Green Prussian Blue
6. Urease Test :
Stuarts Urea Broth/ Christensens Urea Agar
Bacteria containing urease:
pH indicator: phenol/red
Positive result: red color
Negative result: yellow
7. Carboxylase Test:
Media: Moehers decarboxylase
Amino acids: Lysine, Ornithine, Arginine
134
OCDEN, Junalyn B.
Purple= alkaline ; Yellow =acidic

8. PAD (Phenylalanine Deaminase Test)
Phenylalanine present in bacteria
Add 10% Ferric Chloride
Conversion to Phenyl pyruvic acid
Positive Result: Green color
SEROLOGICAL REACTIONS
Skin tests:
Methods:
1. Intradermal injection of Antigen
- Produce localized delayed Hypersensitivity
Experience
Allergic Reaction
Sensitivity to Leprosy:
a. Anesthetic: leprosy on the skin
b. Tuberculoid: change in color of skin
Dicks- if the person scarlet fever
Shicks test- diphtheria
2.
Example
Antibody/ Antitoxin:
Administer if the person is developing rasher
of tests:
Schultz-Charlton scarlet fever
Determine specific strain
PHAGE TYPING
135
OCDEN, Junalyn B.
-
Postive result: lysis/ clearing / plaque
EXPERIMENT 15- AST
Antimicrobial test: inhibiting activity of an agent towards the microbe

Anti microbial: microbial product in low concentration (ug/ml)
Purpose: guide for clinicians
Help to predict and monitor the effectiveness of a treatment
*Susceptible*Resistant*Intermediate- some bact. Are inhibited, some are resistant
TYPES:
1. AGAR DIFFUSION METHOD/ KIRBY BAUER:
INOCULUM
MEDIA
ANTIBIOTICS
INCUBATION
READING
AMOUNT OF
BACTERIA: 1X108
cells/mL
- Compare
the
turbidity
with
MacFarlan
d
(BaCl2 and H2SO4
BaSO4)
TSB
Size: 6.5mm
350C -370C
For 18-24
hours
Diameter of
zone of
inhibition
INTERPRETATIO
N
Base it on
the table of
NCCLS
(National
Community
Clinical
Laboratory
Standard)
2. BROTH DILUTION METHOD:

- Not routinely done in the lab.
INOCULUM
MEDIA
ANTIBIOTICS
INCUBATION
READING
1.5x106
cells/mL
Mueller
Hinton Broth
Liquid
consistency:
350C -370C
For 18-24
MIC- MINIMUM
INHIBITORY
136
INTERPRETATIO
N
Base it on
the table of
OCDEN, Junalyn B.
- compare it
to 0.5
MacFarland
standard
a serial two
fold anti
biotic
hours
CONCENTRATION
: lowest
concentration
of anti
microbial
that can kill
all microbes
NCCLS
(National
Community
Clinical
Laboratory
Standard)
MBC- MINIMUM
BACTERICIDAL
CONCENTRATION
: lowest
concentration
at which an
anti
microbial can
kill 99.99%
of the
microbes
3. AGAR DILUTION METHOD:
- Done during researches
INOCULUM
MEDIA
ANTIBIOTICS
1X108 cells/
mL (compare
to
MacFarland)
3.8x108cells/
mL(compare
to 1.0
MacFarland)
MHA
Liquid
consistency:
cook the
agar
together
with the
anti biotic:
1 part
antibiotic
is to 9
parts molten
agar
INCUBATION
READING
350C -370C
For 18-24
hours
Based on MIC
137
INTERPRETATIO
N
Either no
growth/
Growth
OCDEN, Junalyn B.
4. NOVEL TESTS:
Advantage: use less amount of anti biotic in shorter time
Example:
A. MICRODILUTION SUSCEPTIBILITY TEST
B. COLORIMETRIC MICRODILUTION
- Developed to avoid misinterpretation ; inclusion of dyes
1. MABA (MICRO ALAMAR BLUE ASSAY)
Function: oxidation reduction indicator
Blue to pink(indicate actual growth or no growth)
- Further evaluated through the use of the spectrophotometer
-
2. MTT
Water soluble tetrozolium salt
Use:
[3-(4,5 dimethyl thiazole-2-yl)2,5 diphenyl tetrazolium bromide]
Thiazoyl blue- converted into formazan crystal; result: yellowpurple
C. E-TEST : Epsilometer
Use of strips (specifically: PDM epsilometer)
Distance from antibiotic to the edge of the plate= 15mm
Distance of one antibiotic to the other=24mm
4-5 strips on 1 agar
Graduation of the strips read base on the chart interpret
138

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OCDEN, Junalyn B.

Microbiology Notes (2015)

Food microbiology- example: yakult (Lactus bacillus), yogurt

Joseph Lister: father of Aseptic Surgery

c. Incubatory- transmit disease during the incubation period. Example: Hepa A

Exception: Not all microorganisms grow inside the body.

Walter and Fany Hesse

Paul Erlich- discovered Salvarsan/ 6C6- treatment for syphilis.

Alexander Fleming- discovered penicillin from a fungi (penicillum notatum)

Kingdom: made up of two or more phylum

Acid Fast Bacilli- red bacteria; blue background

2 cocci- diplococci- produces a horizontal line in 1 plane

a. Cell membrane- semi-permeable membrane

b. Cell Wall- rigid structure that encloses the cell membrane

the porins contains transmembrane channels (antibiotics)

Cortex: made up of modified peptidoglycan

Bacteria Growth Curve:

Purely dead cells

Relationship between the log number cells and time

2. Hfr: High Frequency for recombination

3. F prime cell (F cell)

VI. HOST MICROBE RELATIONSHIP

SYMBIOSIS: relationship where one organism is dependent from the other

Through the openings- pores, follicles

- Definite: 10-14 days

Bacteria containing adhesin:

Extra genital gonorrhea

Pathogens will alter the shape to resist destruction by anti bodies

Provoke Immune system

-it will release a lot of

All cocci are G+ except: Neisseria, Branhamella, Veilonella

Non spore forming: Corynebacterium, Listeria, Erysiphelothrix

Aerobic to Facultative Anaerobic

Streptococci in chains; in pairs (S. pneumoniae)

Classification of streptococci in chains:

a. Alpha-hemolytic aka Green Streptococcus/ Viridans

(found in oral cavity)

STREPTOCOCCUS PNEUMONIAE/ DIPLOCOCCUS PNEUMONIAE/STREPTOCOCCUS LANCOELATUS

5. HIPPURATE HYDROLYSIS TEST:

+ result: red color indicating Streptococcus agalactiae

6. LAP: LEUCINE AMINO PEPTIDASE

Fastidious: need substance such as pyridoxal(add Vitamin B6), thiol

B. EXTRA GENITAL GONORRHEA

of CAM with enrichment supplements:

b. Modified Thayer Martin:

f. JEMBEC (John E. Martin Biological Environmental Chamber)

1. RIA (radio Immuno Assay)

4. Particle Agglutination Method:

Fever, stiff neck, vomiting , head ache and formation of petechiae

*Waterhouse Friedrichsen Syndrome (WFS):

dessiminated intravascular coagulation

**13 serogroups: 5 medically important: A,B,C,Y, W135

1. Non spore formers:

Immunodiffusion test to verify if a certain strain of corynebacterium is toxigenic

Serogroups: gravis, mitis, belfanti, intermedius

GRAM + BACILLI SPORE FORMERS

c. Protein Somatic antigen: destroy

d. Anthrax toxin: produced by pXo1

CN: Colons Bacillus

ETEC (ENTERO TOXIGENIC ESCHERICHIA COLI)

Disease: Travellers diarrhea/ Childhood diarrhea

EIEC (ENTERO INVASIVE ESCHERICHIA COLI)

Colonize large intestine species ; electrolytes of the large intestine