INTRODUCTION TO MICROBIOLOGY AND  ALGAE (singular: alga)

PARASITOLOGY
- PHOTOSYNTHETIC EUKARYOTES
PREPARED BY SHADRACH EMNAS, RPh
- BOTH SEXUAL AND ASEXUAL REPRODUCTION
MICROBIOLOGY
- CELL WALLS COMPOSED OF CELLULOSE
 THE STUDY OF VERY SMALL LIVING
 VIRUS
ORGANISMS -- CALLED MICROORGANISMS
OR MICROBES - ACELLULAR

Naming & Classifying Microorganisms - MADE OF ONLY 1 TYPE OF NUCLEIC ACID, EITHER
DNA OR RNA
 SCIENTIFIC NOMENCLATURE ASSIGNS EACH
ORGANISM TWO NAMES - SURROUNDED BY PROTEIN COAT

- THE GENUS (FIRST NAME AND IS ALWAYS MULTICELLULAR ANIMAL PARASITES

CAPITALIZED)
- NOT STRICTLY MICROORGANISMS
- THE SPECIFIC EPITHET (SPECIES NAME,
HISTORY OF MICROBIOLOGY
NOT CAPITALIZED)

ex. Staphylococcus aureus 1665 - ROBERT HOOKE (ENGLISH)

[genus] [species]  OBSERVED A THIN SLICE OF CORK AND

REPORTED TO THE WORLD THAT LIFE'S
TYPES OF MICROORGANISMS SMALLEST STRUCTURAL UNITS WERE
“LITTLE BOXES” OR “CELLS”
 BACTERIA (singular: bacterium)
 CELL THEORY - ALL LIVING THINGS ARE
PROKARYOTES COMPOSED OF CELLS

-ENCLOSED IN PEPTIDOGLYCAN CELL WALLS ANTON VAN LEEUWENHOEK (DUTCH

MERCHANT & AMATEUR SCIENTIST)
- REPRODUCE BY BINARY FISSION
 PROBABLY THE FIRST TO ACTUALLY
 ARCHAEA OBSERVE LIVE MICROORGANISMS THRU
PROKARYOTES MAGNIFYING LENSES

- CELL WALLS LACK PEPTIDOGLYCAN RUDOLF VIRCHOW (GERMAN SCIENTIST)

- OFTEN FOUND IN EXTREME ENVIRONMENT  CONCEPT OF BIOGENESIS - CLAIM THAT

LIVING CELLS CAN ONLY ARISE FROM
 FUNGI (singular: fungus) PREEXISTING LIVING CELLS

EUKARYOTES

- TRUE FUNGI HAVE CELL WALLS MADE OF CHITIN GOLDEN AGE OF MICROBIOLOGY

- CAN REPRODUCE SEXUALLY OR ASEXUALLY LOUIS PASTEUR (FRENCH SCIENTIST)

 PROTOZOA (singular: PROTOZOAN)  SUPPORTED AND RESOLVED THE CONCEPT

OF BIOGENESIS
- REPRODUCE SEXUALLY OR ASEXUALLY
 FERMENTATION (YEASTS CONVERT THE
- MOVE BY PSEUDOPODS, FLAGELLA, OR CILIA SUGARS TO ALCOHOL IN THE ABSENCE OF
AIR)
PASTEURIZATION
 PASTEURIZATION'S SOLUTION TO SPOILAGE
PROBLEM WAS TO HEAT THE BEER AND
WINE JUST ENOUGH TO KILL MOST OF THE
BACTERIA THAT CAUSE THE SPOILAGE
 NOW COMMONLY, USED FOR MILK AND
SOME ALCOHOLIC BEVERAGES
PASTEUR
DEVELOPED VACCINES TO PREVENT:
 CHICKEN CHOLERA
 ANTRAX
 SWINE ERYSIPELAS
 RABIES IN DOGS, AND SUCCESSFULLY USED
THE VACCINE TO TREAT HUMAN RABIES
GERM THEORY OF DISEASES
MICROORGANISMS MIGHT CAUSE DISEASES
1860s - JOSEPH LISTER (ENGLISH SURGEON)
 APPLIED GERM THEORY TO MEDICAL
PROCEDURES
 USED PHENOL (CARBOLIC ACID) TO KILL
BACTERIA, BEGAN TREATING SURGICAL
WOUNDS WITH A PHENOL SOLUTION
 HIS FINDINGS PROVED THAT MICROORG.
CAUSE SURGICAL WOUND INFECTIONS
1876 ROBERT KOCH (GERMAN PHYSICIAN)
 DISCOVERED ROD-SHAPED BACTERIA NOW
KNOWN AS Bacillus anthracis (causes
anthrax)
 DISCOVERED Mycobacterium tuberculosis
(tuberculosis) and Vibrio cholerae (cholera)
 Koch developed methods of cultivating
bacteria on solid media.
R.J. PETRI, ONE OF KOCH'S COLLEAGUES, INVENTED
A FLAT GLASS DISC (NOW KNOWN AS PETRI DISH) IN
WHICH TO CULTURE BACTERIA ON SOLID MEDIA
FRAU HESS, THE WIFE OF ANOTHER COLLEAGUE OF
KOCH, SUGGESTED THE USE OF AGAR
KOCH'S POSTULATES
1. A particular microorganism must be found in all
cases of the disease and must not be present in the
healthy animals or human.
2. The microorganism must be isolated from the
diseased animal or human and grown in pure culture
in the laboratory.
3. The same disease must be produced when
microorg. from the pure culture are inoculated into
healthy susceptible laboratory animals.
4. The same microorganism must be recovered from
the experimentally infected animals and grown again
in pure culture
VACCINATION
 SMALLPOX VACCINE
MAY 4, 1789, YOUNG BRITISH PHYSICIAN EDWARD
JENNER EMBARKED ON AN EXPERIMENT TO FIND A
WAY TO PROTECT PEOPLE FROM SMALLPOX
 SMALLPOX EPIDEMICS - GREATLY FEARED
THE DISEASE PERIODICALLY SWEPT THRU EUROPE,
KILLING THOUSANDS, AND IT WIPED OUT 90% OF
THE NATIVE AMERICANS ON THE EAST COAST WHEN
EUROPEANS SETTLERS FIRST BROUGHT THE
INFECTION TO THE NEW WORLD.
ROOT WORD: VACCA - LATIN WORD FOR COW
BIRTH OF MODERN CHEMOTHERAPY
 CHEMOTHERAPY
- THE TREATMENT OF DISEASE BY USING CHEMICAL
SUBSTANCES
- THE TERM ALSO COMMONLY REFERS TO CHEMICAL
TREATMENT OF NONINFECTIOUS DISEASES SUCH AS
CANCER
BIRTH OF MODERN CHEMOTHERAPY
CHEMOTHERAPEUTIC AGENTS PREPARED FROM
CHEMICALS IN THE LABORATORY ARE CALLED
SYNTHETIC DRUGS
CHEMICALS PRODUCED BY BACTERIA AND FUNGI TO
ACT AGAINST OTHER MICROORGANISMS ARE
CALLED ANTIBIOTICS
THE FIRST SYNTHETIC DRUGS
 PAUL EHRLICH (GERMAN PHYSICIAN)
SPECULATED ABOUT A MAGIC BULLET THAT COULD
HUNT DOWN AND DESTROY A PATHOGEN WITHOUT
HARMING THE INFECTED HOST
 DISCOVERED SALVARSAN
SALVARSAN
 AKA: ARSPHENAMINE, COMPOUND 606
 ARSENIC DERIVATIVE
 TREATMENT FOR SYPHILIS (causative agent:
Treponema pallidum)
 THE FIRST MODERN CHEMOTHERAPEUTIC
AGENT
A FORTUNATE ACCIDENT
 ALEXANDER FLEMING (SCOTTISH PHYSICIAN
& BACTERIOLOGIST)
ALMOST TOSSED OUT SOME CULTURE PLATES THAT
HAD BEEN CONTAMINATED BY MOLD
FORTUNATELY, HE TOOK A SECOND LOOK AT THE
CURIOUS PATTERN OF GROWTH ON THE
CONTAMINATED PLATES
AROUND THE MOLD WAS A CLEAR AREA WHERE
BACTERIAL GROWTH HAD BEEN INHIBITED
PENICILLIN
FROM THE MOLD Penicillium
USED AGAINST MANY BACTERIAL INFECTIONS
CAUSED BY Staphylococci AND Streptococci
MODERN DEVELOPMENTS IN MICROBIOLOGY
 BACTERIOLOGY -STUDY OF BACTERIA
-BEGAN WITH VAN LEEUWENHOEK'S FIRST
EXAMINATION OF TOOTH SCRAPINGS
 MYCOLOGY -STUDY OF FUNGI
-INCLUDES MEDICAL, AGRICULTURAL AND
ECOLOGICAL BRANCHES
 PARASITOLOGY -STUDY OF PROTOZOA
AND PARASITIC WORMS
 IMMUNOLOGY -STUDY OF IMMUNITY
-ACTUALLY DATES BACK IN WESTERN CULTURE TO
JENNER'S FIRST VACCINE IN 1796
-VACCINES ARE NOW AVAILABLE FOR NUMEROUS
DISEASES, INCLUDING MMR, CHICKENPOX,
TETANUS, POLIO, HEPA B, DENGUE, ETC.
 VIROLOGY -STUDY OF VIRUS
-ORIGINATED DURING THE GOLDEN AGE OF
MICROBIOLOGY
MICROBES AND HUMAN DISEASE
 NORMAL MICROBIOTA OR FLORA*
-THE NORMAL MICROBIOTA NOT ONLY DO US NO
HARM, BUT ALSO IN SOME CASES CAN ACTUALLY
BENEFIT US
-ANATOMICAL AREAS: SKIN, CONJUNCTIVA, GI
TRACT, URETHRA AND BLADDER, FEMALE
REPRODUCTIVE SYSTEM, ORAL CAVITY, ETC
 INFECTIOUS DISEASES
-WHEN PATHOGENS INVADE A SUSCEPIBLE HOST
-EX: MALARIA, DIPHTHERIA, CHOLERA
 EMERGING INFECTIOUS DISEASES (EIDs)
-DISEASES THAT ARE NEW OR CHANGING AND ARE
INCREASING OR HAVE THE POTENTIAL TO INCREASE
IN INCIDENCE IN THE NEAR FUTURE
-EX: WEST NILE ENCEPHALITIS, MAD COW DISEASE,
EBOLA HEMORRHAGIC FEVER, HANTAVIRUS
PULMONARY SYNDROME, AIDS/HIV,
MICROSCOPY TOTAL MAGNIFICATION

PREPARED BY SHADRACH EMNAS, RPh  FORMULA:

UNITS OF MEASUREMENT OBJECTIVE LENS MAGNIFICATION x OCULAR LENS

MAGNIFICATION
MICROMETER (μm) 10-6 m
 MOST OCULAR LENS: 10x
NANOMETER (nm) 10-9 m
 OBJECTIVE LENSES:
MICROSCOPE
-10x (low power)
 simple microscope -40x (high power)
-only one magnifying lens -100x (oil immersion)
-late 1600s, Leeuwenhoek used simple other terms
microscopes to observe many tiny objects (bacteria
and protozoa)  RESOLUTION

-AKA RESOLVING POWER

-THE ABILITY OF LENSES TO DISTINGUISH FINE

DETAIL AND STRUCTURE

 REFRACTIVE INDEX

-MEASURE OF THE LIGHT-BENDING ABILITY OF A

MEDIUM
 compound microscope
OTHER PARTS OF MICROSCOPE
-more than 1 magnifying lens

 compound light microscope

-have a built-in light bulb

-used in today's laboratories (contain two

magnifying lens system)

BASIC PARTS OF LIGHT MICROSCOPE

 illuminator/light source
 condenser - where light passes thru/directs
the light rays thru the specimen
 objective lenses - the lenses closest to the
specimen
 ocular lens/eyepiece - image of specimen is
magnified again
DARKFIELD MICROSCOPE TAKES ADVANTAGE OF fluorescence - THE ABILITY
OF SUBSTANCES TO ABSORB SHORT WAVELENGTHS
USED FOR EXAMINING LIVE MICROORGANISMS
OF LIGHT (ULTRAVIOLET) AND GIVES OFF LIGHT AT A
THAT EITHER ARE [1] INVISIBLE IN THE ORDINARY
LONGER WAVELENGTH (VISIBLE)
LIGHT MICROSCOPE, [2] CANNOT BE STAINED BY
STANDARD METHODS, [3] OR ARE SO DISTORTED BY FLUOROCHROMES - FLUORESCENT DYES
STAINING THAT THEIR CHARACTERISTICS THEN
CANNOT BE IDENTIFIED

*brightfield - light microscope

CONFOCAL MICROSCOPY

CONFOCAL MICROSCOPY USES A PINHOLE

APERTURE, IT ELIMINATES THE BLURRING THAT
PHASE-CONTRAST MICROSCOPE OCCURS WITH OTHER MICROSCOPE

PERMITS DETAILED EXAMINATION OF THE INTERNAL AS A RESULT, EXCEPTIONALLY CLEAR TWO-

STRUCTURES IN LIVING MICROORG. DIMENSIONAL IMAGES CAN BE OBTAINED

DIFFERENTIAL INTERFERENCE CONTRAST

MICROSCOPE

DIC MICROSCOPE

SIMILAR TO PHASE-CONTRAST MIC.

USES 2 BEAMS OF LIGHT INSTEAD OF 1

THE IMAGE IS BRIGHTLY COLORED AND APPEARS

NEARLY 3-D

ELECTRON MICROSCOPY

A BEAM OF ELECTRONS IS USED INSTEAD OF LIGHT

TRANSMISSION ELECTRON MICROSCOPE (TEM)

HAS A VERY TALL COLUMN, AT TOP OF WHICH AN

FLUORESCENCE MICROSCOPE
ELECTRON GUN FIRES A BEAM OF ELECTRONS
DOWNWARD
SCANNING ELECTRON MICROSCOPE (SEM)
HAS A SHORTER COLUMN AND INSTEAD OF BEING
PLACED INTO THE ELECTRON BEAM, THE SPECIMEN
IS PLACED AT THE BOTTOM OF THE COLUMN
ELECTRONS THAT BOUNCE OFF THE SPECIMEN ARE
CAPTURED BY DETECTORS
PREPARATION OF SPECIMENS FOR LIGHT
MICROSCOPY
STAINING - COLORING THE SPECIMEN WITH A DYE
SMEAR - A THIN FILM OF MATERIAL CONTAINING
THE MICROORG. SPREAD OVER THE SLIDE SURFACE
BASIC DYES:
CRYSTAL VIOLET, METHYLENE BLUE, MALACHITE
GREEN, SAFRANIN
MORE COMMONLY USED THAN ACIDIC DYES
SIMPLE STAINS
AN AQUEOUS OR ALCOHOL SOLUTION OF A SINGLE
BASIC DYE
PURPOSE: TO HIGHLIGHT THE ENTIRE MICROORG.
SO THAT CELLULAR SHAPES AND BASIC STRUCTURES
ARE VISIBLE
DIFFERENTIAL STAINS
GRAM STAIN
DEVELOPED BY HANS CHRISTIAN GRAM
CLASSIFIES BACTERIA INTO TWO LARGE GROUPS:
GRAM POSITIVE
GRAM NEGATIVE
GRAM STAIN - PROCEDURE
1. SMEAR IS COVERED WITH BASIC DYE USUALLY
CRYSTAL VIOLET; IMPARTS ITS COLOR TO THE CELLS -
IT IS REFERRED TO AS A PRIMARY STAIN.
2. AFTER A SHORT TIME, PURPLE DYE IS WASHED
OFF, AND SMEAR IS COVERED WITH IODINE (A
MORDANT - INCREASES AFFINITY OF THE DYE TO THE
CELLS). WHEN IODINE IS WASHED OFF, BOTH G+
AND G- BACTERIA APPEARS DARK VIOLET OR PURPLE
3. SLIDE IS WASHED WITH ALCOHOL OR AN
ALCOHOL-ACETONE SOLUTION. THIS IS A
DECOLORIZING AGENT, WHICH REMOVES THE
PURPLE FROM THE CELLS OF SOME SPECIES BUT NOT
THE OTHERS.
4. ALCOHOL IS RINSED OFF, SLIDE THEN STAINED
WITH SAFRANIN, A BASIC RED DYE. SMEAR IS
WASHED OFF AGAIN, BLOTTED DRY AND EXAMINED
MICROSCOPICALLY.
GRAM STAIN DIAGRAM
EASY TO REMEMBER - VIAS
V - VIOLET [CRYSTAL VIOLET] [PRIMARY STAIN]
I - IODINE
[MORDANT]
A - ALCOHOL/ACETONE [DECOLORIZER]
S - SAFRANIN [COUNTERSTAIN]
RESULTS
ACID-FAST STAIN
USED TO IDENTIFY:
1. ALL BACTERIA IN GENUS Mycobacterium
M. tuberculosis, M. leprae
2. GENUS Nocardia
ACID-FAST STAIN PROCEDURE
1. RED DYE CARBOL FUCHSIN IS APPLIED TO THE
SMEAR
2. SLIDE IS GENTLY HEATED FOR SEVERAL MINUTES
(HEATING ENHANCES PENETRATION AND
RETENTION OF DYE - SERVES AS MORDANT
CELL STRUCTURE AND FUNCTION
PREPARED BY SHADRACH EMNAS, RPh
SIZE, SHAPE & ARRANGEMENT OF BACTERIAL CELLS
BASIC SHAPES
COCCUS (SPHERICAL); PLURAL: COCCI
BACILLUS (ROD-SHAPED); PLURAL: BACILLI
SPIRAL
 COCCI
USUALLY ROUND, BUT CAN BE OVAL, ELONGATED
OR FLATTENED ON ONE SIDE

3. SLIDE IS COOLED AND WASHED WITH WATER DIPLOCOCCI - IN PAIRS AFTER DIVIDING

4. SMEAR THEN TREATED WITH ACID-ALCOHOL STREPTOCOCCI - CHAINLIKE PATTERNS

(DECOLORIZER) STAPHYLOCOCCI - GRAPELIKE CLUSTERS OR BROAD
5. SMEAR IS THEN TREATED WITH METHYLENE BLUE SHEETS
(COUNTERSTAIN)

ACID-FAST STAIN - EASY TO REMEMBER

C - CARBOL FUCHSIN [PRIMARY STAIN]

H - HEAT [MORDANT]

A - ACID-ALCOHOL [DECOLORIZER]

M - METHYLENE BLUE [COUNTERSTAIN]

RESULTS

 BACILLI

SINGLE BACILLI - SINGLE RODS

DIPLOBACILLI - IN PAIRS AFTER DIVISION

SPECIAL STAINS STREPTOBACILLI – CHAINS
NEGATIVE STAINING FOR CAPSULES

PRESENCE OF CAPSULE = ORGANISM'S VIRULENCE

ENDOSPORE (SPORE) STAINING

ENDOSPORE = SPECIAL RESISTANT, DORMANT

STRUCTURE WITHIN A CELL THAT PROTECTS A
BACTERIUM FROM ADVERSE ENVIRONMENTAL
CONDITIONS
FLAGELLA STAINING

BACTERIAL FLAGELLA (SINGULAR: FLAGELLUM) ARE

STRUCTURES OF LOCOMOTION (MOVEMENT)
  SPIRAL  FIMBRIAE AND PILI

VIBRIOS - LOOK LIKE CURVED RODS
SPIRILLA - HELICAL SHAE; LIKE A CORKSCREW, RIGID
BODIES
SPIROCHETES - HELICAL AND FLEXIBLE
FIMBRIAE - HAVE A TENDENCY TO ADHERE TO EACH
OTHER AND TO SURFACES
PILI - LONGER THAN FIMBRIAE, INVOLVED IN
MOTILITY AND DNA TRANSFER
CONJUGATION (SEX) PILI - DNA TRANSFER

 THE CELL WALL

 SHAPES BACTERIAL CELL WALL - PEPTIDOGLYCAN
MONOMORPHIC - MAINTAINS A SINGLE SHAPE GRAM POSITIVE CELL WALLS - THICK, RIGID WALL;
PLEOMORPHIC - MANY SHAPES, NOT JUST ONE CONTAIN TEICHOIC ACIDS

 STRUCTURES GRAM NEGATIVE CELL WALLS - FEW LAYERS

FLAGELLA - LONG FILAMENTOUS APPENDAGES FOR

MOVEMENT

ATRICHOUS - LACK FLAGELLA

PERITRICHOUS - DISTRIBUTED OVER THE ENTIRE

CELL

MONOTRICHOUS - SINGLE FLAGELLUM AT ONE POLE

LOPHOTRICHOUS - A TUFT OF FLAGELLUM AT ONE

POLE

AMPHITRICHOUS - FLAGELLA AT BOTH POLES

MICROBIAL GROWTH EX. BISMUTH SULFITE AGAR (USED TO ISOLATE THE
GRAM-NEGATIVE SALMONELLA TYPHI; IT INHIBITS
PREPARED BY SHADRACH EMNAS, RPh GRAM-POSITIVE BACTERIA
PHYSICAL REQUIREMENTS SABOURAUD'S DEXTROSE AGAR (HAS A pH of 5.6;
 TEMPERATURE USED TO ISOLATE FUNGI THAT OUTGROW MOST
BACTERIA AT THIS pH)
PSYCHROPHILES - COLD LOVING

MESOPHILES - MODERATE TEMPERATURE

THERMOPHILES - HEAT LOVING

 pH AND OSMOTIC PRESSURE

3. differential media - make it easier to distinguish
ACIDOPHILES - ACID TOLERANT colonies of the desired organism from other colonies
grwoing on the same plate.
HALOPHILES - ADAPTED TO HIGH SALT
CONCENTRATION EX. BLOOD AGAR

 OXYGEN

OBLIGATE AEROBES - REQUIRE OXYGEN TO LIVE

FACULTATIVE ANAEROBES - HAVE THE ABILITY TO

CONTINUE GROWING IN THE ABSENCE OF OXYGEN

OBLIGATE ANAEROBES - DOESNT REQUIRE OXYGEN

TO LIVE

4. ENRICHMENT CULTURE - favor the growth of a

CULTURE MEDIA
particular microbe but not the others; increase
1. CHEMICALLY DEFINED MEDIUM numbers of desired microbes to detectable levels.

- EXACT CHEMICAL COMPOSITION IS KNOWN.

EX. CHEMICALLY DEFINED MEDIUM FOR GROWING A

TYPICAL CHEMOHETEROTROPH, SUCH AS Escherichia
coli

CONSTITUENTS: glucose, ammonium phosphate,

NaCl, water, potassium phosphate, MgSO4

2. selective media - designed to suppress the growth

of unwanted bacteria and encourage the growth of
the desired microbes
BACTERIAL GROWTH

 PHASES OF GROWTH

1. LAG PHASE - CELLS DO NOT IMMEDIATELY

REPRODUCE IN A NEW MEDIUM

PERIOD OF LITTLE OR NO CELL DIVISION

2. LOG PHASE - CELLS BEGIN TO DIVIDE AND ENTER A

PERIOD OF GROWTH, OR LOGARITHMIC INCREASE

CELLULAR REPRODUCTION IS MOST ACTIVE DURING

THIS PERIOD

3. STATIONARY PHASE

NUMBER OF MICROBIAL DEATH BALANCES THE

NUMBER OF NEW CELLS, AND THE POPULATION
STABILIZES

THE PERIOD OF EQUILIBRIUM

4. DEATH PHASE

LOGARITHMIC DECLINE PHASE

POPULATION IS DIMINISHED