Microbiology

- Discovered highly resistant bacterial structure, later

known as endospore, in the infusion of hay.
Albert Ludwig Sigesmund Neisser (1855 – 1916)
INTRODUCTION TO MICROBIOLOGY - Discovered the causative agent (pathogen) of
- The Greek word, “mikros” means small; and “bio” gonorrhea, a strain of bacteria that was named in his
means life honour (Neisseria gonorrhoeae)
- the study of all living organisms that are too small to be Ignaz Philipp Semmelweis
visible with the naked eye - Described as the “savior of mothers”
Evolution of Microbiology Selman Abraham Waksman (1888 – 1973)
discovery of microbiology in the 17th - Researched into decomposition of organisms that live
century: in soil enabled the discovery of streptomycin and several
Robert Hooke (1635 – 1703 other antibiotics.
-First use a microscope, Discovered cell, the basic unit John Snow (1813 – 1858)
of a living organism - “Father of Field Epidemiology” conducted studies of
Antonie Van Leeuwenhoek (1632 -1723) cholera outbreaks both to discover the cause of disease
-Observe microorganisms, using a microscope of his and to prevent its recurrence
own design, creating the single-lens microscope - Developed geographic distribution of cases, is called a
- The “Father of Microbiology. spot map.
Edward Jenner (1749-1823) Luc Antoine Montagnier (1932)
- English physician was the first to prevent small pox. - French virologist
Louis Pasteur (1822-1895) - Discovered of the human immunodeficiency virus
- Father of modern microbiology/ father of bacteriology; (HIV) – 1983
introduced pasteurization. Significance to the Nursing Profession
Joseph Lister (1827-1912)
- Introduced aseptic techniques for control of microbes
by the use of physical and chemical agents.
- The “Father of Antiseptic Suger
Robert Koch (1843 – 1910)
- all, isolated anthrax bacillus (Bacillus anthracis, the
cause of anthrax) in 1876
- perfected the technique of isolating bacteria in pure
culture
Koch’s four postulates are:
-The organism causing the disease can be found in sick
individuals but not in healthy ones.
-The organism can be isolated and grown in pure culture.
- The organism must cause the disease when it is
introduced
into a healthy animal.
Paul Ehrlich (1854 – 1915)
- In 1904, found that the dye Trypan Red was active
against trypanosome that causes African sleeping
sickness and could be used therapeutically, referred as
“magic bullet”
Alexander Fleming (1881 – 1955)
- In 1929 discovered the first antibiotic penicillin
Hans Christian Gram (1853 – 1938)
- Inventor of the Gram staining technique was a
pioneering biologist who devised the system of
classification which led to as many as 30,000 formally
named species of bacteria.
Theodor Escherich (1857 – 1911)
- Discovered the bacterium which he called “bacterium
coli commune” and which was later to be called
Escherichia coli in 1919
John Tyndall (1820 – 1893)
 - A thin flexible thread like object or very
Classification of Pathogens long thin cylindrical single cells
Bacteria - Simple, single-celled organisms, no true Mycotic - diseases caused by fungi that invades the
nucleus. (unicellular) tissues.
Grow in many environments; Candida – yeast like fungus that may infect a
Grouped into: weakened host; it may be in the skin, oral/mouth,
a) Anaerobic - Can grow in the absence of oxygen. - Ex: digestive tract, vaginal tract and lungs
Clostridium tetani – a tetanus Protozoa - Single-cell, animal like organism
b) Aerobic - Requires oxygen for growth. - Ex: - Found in soil, body of water from moist grass to mud
Mycobacterium tuberculosis – lungs puddles to sea
c) Facultative Anaerobes - Can grow with or without Four (4) main division includes:
oxygen. - Ex: Escherichia coli – lower intestine 1) Amoeboid (Amebas/Amoeba)
Bacteria are classified by their shape and how they 2) Ciliates
cluster together(arrangement):
3) Flagellates
A. Cocci – spherical, ovoid or round
4) Sporozoa
a) Streptococci - Gram-positive bacteria arranged in
chains, like a string of beads. Helminths
b) Staphylococci – Gram-positive arranged in large 1) Flatworms
clusters 2) Acanthocephali
c) Diplococci - Can be gram-positive or negative; round 3) Roundworms
the bacterium that typically occurs in the form of Algae
two joined cells. Group of oxygenic, phototrophic microbes which has a
B. Bacilli – Gram-positive, straight, slender rods, cigar- nucleus; can generate oxygen through photosynthesis
shaped with tapering ends. NORMAL FLORA OF THE HUMAN BODY
- All endospore-forming bacteria are bacilli. - The human microbiome (or human microbiota)
C. Endospores - Bacteria that produce resistant forms
microorganisms that reside on the skin, saliva, and oral
that can tolerate long periods of dryness or other adverse
mucosa, conjunctiva, and gastrointestinal tracts.
conditions
D. Curved rods - They include bacteria, fungi bacteria, fungi
- Ever-present phenotype bacteria Types of Normal Flora
1) Vibrios- short rods with a slight curvature like a 1. Resident flora - Fixed types of microorganisms
comma; gram-negative and highly motile. regularly found in a given area at a given age.
2) Spirilla - Long and wavelike cells resembling a - If disturbed, it reestablishes itself.
corkscrew; gram negative bacteria 2. Transient flora - consists of non-pathogenic or
3) Spirochetes potentially pathogenic microorganisms
- Spiral-shaped capable of waving and Two types of flora in the Skin:
twisting motion 1. Resident Flora
Other types of bacteria include: - Staphylococcus- epidermis
D. Flagella Are of the many types of bacteria that are -Staphylococcus aureus
capable of -Staphylococcus
swimming rapidly by means of threadlike - Diphtheroid
appendages. -Propionibacterium acnes
E. Mycoplasma Ex: Mycoplasma pneumonia (walking - Yeast- candida
pneumoniae) 2. Transient Flora
F. Chlamydia Ex: Chlamydia trachomatis (common -Micro coccus
sexually transmitted disease) -Bacteria
Virus Normal flora of Digestive System
- Smallest of all pathogens; replication within the host
1. Microflora of Oral Cavity
Fungi
- Can be single-celled or very complex multicellular - Aerobic and anaerobic bacteria
Organisms Mouth/ Oral cavity - Lactobacilli, staphylococcus,
Yeasts – single-cell forms streptococcus, Bacteroid, Corynebacterium, Nocardia -
- which needs food, warmth, and moisture Teeth: Streptococcus mutants, hemolytic streptococci,
to thrive. fusobacterium
Molds – filamentous forms Small Intestine:
1. Duodenum: gram-positive cocci and rod - Foods, floors, equipment, contaminated
2. Jejunum: gram-positive, gram-negative bacterium water
can be seen, Lactobacilli Diphtheroid, Enterococcus 3rd Link: Portal of Exit
faecalis, Yeast- Candida albicans - Via
3. Ileum – PH – alkaline, anaerobic gram-negative - Bodily fluids
bacteria are mainly entero bacterial. - Coughing, sneezing, diarrhea
Large Intestine: - Larges microorganism in the body. - Seeping wounds
Anaerobic grampositive, non-sporing bacteria - Tubes, IV lines
Normal flora Respiratory Tract 4th Link: Mode of Transmission
Upper respiratory tract - Nasal passage, pharynx, - Contact
esophagus - staphylococcus, streptococcus, diphtheroid, - Direct – touching, kissing, sexual contact
Bacteroides, Micrococci, Haemophilus etc. - Indirect – contact with a fomite
Lower respiratory tract. Trachea, Bronchi, Lungs - It - Droplet: Cough, sneeze
is mostly sterile - Because the mucous membranes of the - Airborne: Via air conditioning, sweeping
lungs remove any microbes. 5th Link: Portal of Entry
MEDICAL & SURGICAL ASEPSIS (microbial -Eye, nares, mouth, vagina, cuts, scrape
control) -Wounds, surgical sites, IV or drainage tube sites
Asepsis - Absence of germs or microorganism - Clinical -Bite from a vector 6th Link: Susceptible Host
condition where infectious agent is spread throughout -Person with inadequate defense
the body of the individual from localize side infection Nurse’s role in
and manifest symptoms of organ damage.  Containing nosocomial infections
Medical Asepsis - is any practice that reduces the - Clean, disinfect, sterile
number and spread of microorganisms.  Controlling/ eliminating reservoirs
Surgical Asepsis - is the process that eliminates - Bathing, dressing changes, patent drainage systems
completely all microorganisms and their spores from the  Controlling the portal of exit
surface of an object. - Cover mouth/nose, wear mask, client teaching
Transmission–based Precautions for Use with Specific  Controlling transmission
Types of Patients - Do not share equipment, proper handling of linens,
Airborne Precaution (Droplet nuclei Smaller than 5 handwashing, PPE
microns)  Controlling portal of entry
Infection/Condition - Maintain skin integrity, position changes, proper
- Measles, wiping techniques
- chicken pox (varicella)
- pulmonary or laryngeal TB
Barrier Protection
- Private room with negative-pressure
- Mask or N-95
Droplet precautions (droplets larger than 5 microns)
Infection/Condition
- Diphtheia (pharyngeal),
- rubella, pneumonia or scarlet fever,
- pertussis, mumps, M. pneumonia,
Chain of Infection
1st Link: Presence of Pathogens
- Infectious Agents
- Normal flora that become pathogenic
2nd Link: Reservoir
- refers to the source of pathogens
- Where pathogens live and multiply
- May be living
- Humans, animals, insects
- May be non-living
Microscopy – science of investigating small objects and Nomarski Interference contrast (NIC)
structures using a microscope - Developed by Georges Nomarski 1952
Simple Microscope - Uses only one lens or group of - 2 beams of light instead of one
lenses in one unit to magnify objects - Appears 3 dimensional
- Ex: jewelry eyepieces, pocket magnifiers, reading - Higher resolution
glasses - Colors due to prisms
Compound microscope - Uses two types of lens to Fluorescent Microscope
magnify object - Two types of lenses a) Eyepiece lens b) - Uses 1 ultraviolet light and 2 fluorescent dyes called
Objective lens – create resolved image then magnified fluorochromes.
by eyepiece lens for viewing; with different zoom levels - Micro-organisms fluoresces or appears to shine against
of magnification dark background
3 types of Microscope Confocal Microscope
 Light microscope – 1,000x - Other names:
 Electron microscope – 100,000x  Confocal Laser Scanning Microscope (CLSM)
 Atomic force microscope  Laser Confocal Scanning Microscope (LCSM)
1. Light microscope - Micro-organisms is stained with fluorescent dye to
- Compound light microscope - Uses visible light and a emit light. - Scanned by laser in planes and region.
system of lenses to magnify images of small objects BACTERIA AND DISEASE
- There are two basic types of magnifying lens system: Pollution - Undesirable substance in water, air or soil;
surroundings
a) Eyepiece (ocular) – contains ocular lens b) Object
Contamination - Presence or organisms outside body,
lenses – positioned above microorganisms
water, food and other biological substances
Bright-field Microscope Infection - Invasion by pathogenic microorganisms -
- Illuminates field evenly Bot synonymous to disease
- Magnifies 1000 to 1500x Disease - Undesirable relationship bet host and pathogen
- Specimen appears dark with low contrast, so object - Interruption in normal functioning Pathogen -
needsstaining Organism that invades and causes damage, injury
- Use for bacteria and fungi Pathogenicity – Pathogenic – ability of organism to
Darkfield Microscope cause disease
- 1,000x  Non-virulent form – no surrounding outer capsule of
- Uses reflected light instead of transmitted light polysaccharide
- opaque disc condenser that blocks light
 Virulent form – surrounding outer capsule
Phase-contrast Microscope
Incidence of a Disease
- 1,000x
- Refers to the number of person in a population who
- Introduced by Frits Zernike, dutch physicist 1934
acquired the disease at a particular point of time
- Produces high-contrast images of specimen (with
 Incidence Rate – number or new cases
contrast-enhancing optical technique
2. Electron Microscope  Prevalence rate – number of new and old cases
- Uses a beam of accelerated electrons as a source of Occurrence of a Disease
illumination, and magnets to focus the beam. Epidemic
- Higher resolving power than light microscopes and - Rapid spread of infectious disease to large number of
canreveal the structure of smaller objects people
2 types: in a given population within a short period of time
1. Transmission electron microscope Endemic
(TEM) - Constantly present in a particular place
- Original electron microscope
- 2-dimensional; black and white Sporadic
image - Occurs occasionally
- Magnifies 200,000x Pandemic
2. Scanning electron microscope - Present worldwide
- 3D structure; black and Duration of Disease
white Acute
- Magnifies it 10,000x - Shows rapidly for short period of time
3. Atomic force Microscope - A type of high-resolution Chronic
scanning probe microscope that has a resolution that you - Occurs slowly; occurs long period of time
can measure in fractions of a nanometer. Latent
Differential Interference Contrast Microscope - Inactive for long time but can become active again
Stages of Infectious Disease and on the other end they bind with various white cells
1. Incubation period that effectively block and destroy the antigen
- The time interval between entry of microorganism and - Two identical strands of heavy chains and two
the appearance of 1st signs and symptoms identical strands of light chains
2. Prodromal period Types of Immunoglobulin (GAMED)
- Mild symptoms of disease which are nonspecific IgG
(fever, cough, colds, malaise) - 75% of total immunoglobulin
3. Period of Illness - Appears in serum and tissues
- Period of maximal invasion/Disease Is most acute IgA
during this period - 15% of total immunoglobulin
- Fulminant infection - Appears in body fluids
- If the disease is not successfully overcome, patient IgM
may die in short period of time - 10% of total immunoglobulin
- Carrier state - Appears mostly in intravascular serum
- Does not show s/s but still continues to shed IgE
infecting microorganism - 0.004% of immunoglobulins
4. Period of Decline/ defervescence - Appears in serum
- sign and symptoms start to subside. IgD
- patient may become vulnerable to secondary infections - 0.2% of immunoglobulins
(not fully recovered) - Appears in small amount in serum
5. Period of Convalescence Types of T Cells
- patient regains strength, body returns to its pre-disease 1. Helper T cells
normal condition. (Complete recovery) - Secrete cytokines that attract and activate B cells
Routes of Transmission - Produce different types of cytokines
Transmission – the pathway of causative agents from 2. Cytotoxic T cells
source to infection of susceptible host - Direct attack microorganism
1. Contact - Called killer cells
 Direct – person to person - Releasing cytolytic enzymes and cytokines
 Indirect – from the reservoir to a susceptible host 3. Suppressor T cells
 Droplet – spread in droplet nuclei Travels <1 meter - Suppressing cytotoxic and helper T cells
2. Vehicle Transmission - Preventing the cytotoxic cells
- Transmission of the organism through media such
as food, water, air.
3. Vectors - Animals that carry microorganism from one
host to another

Antigens
- Substance which when introduced parenterally into the
body stimulates the production of an antibody which
reacts specifically and in an observable manner. Types
of Antigens
1. Autoantigens - Usually, a normal protein or complex
protein - Ex: are a person own self antigen
2. Alloantigen’s - Are antigen found in different
members of the same species (the red blood cell antigen
A and B are examples)
3. Heterophile antigen - identical antigens found in the
cells of different species
Immunoglobulins
- also referred to as antibodies or Ig; Y shaped molecules
that connect on one end to invading microbes (antigens)
Vaccine - An immunobiological substance designed to a) Diplobacilli
produce specific protection against a given disease b) Streptobacilli
Vaccination – administration of antigenic material to c) Palisades
stimulate an individual immune system to develop d) Coccobacillus
adaptive immunity to a pathogen 3. Spirillum (plural: Spirilla) - Spiral shape and curved
Types of Vaccine bacteria - Can be gently curved shape to a corkscrew like
1. Live attenuated vaccines - Prepared from live
generally attenuated organisms Glycocalyx
2. Inactivated or killed vaccines - Organisms killed by - Thick layer of material located outside cell wall
heat or chemicals - Safe but generally less efficacious - Produced by cell membrane and secreted outside cell
than live wall
3. Toxoids - Certain organisms produce exotoxins, these - Made of polysaccharide or polypeptide
toxins are detoxicated and used in the preparation of - Outermost covering of some bacteria - Slimy,
vaccines gelatinous material
4. Cellular fractions - Vaccines that are prepared from Types of Glycocalyx
extracted cellular. a) Capsule – strong attached
5. Combinations - If more than one kind of immunizing - contains polysaccharides
agent is included in the vaccine it is called a mixed or b) Slime layer – loosely attached
combined vaccine. - Not highly organized, not firmly attached to the
6. Polysaccharide Vaccines - A unique type of cell wall
inactivated subunit vaccine composed of long chains of Biofilm
sugar molecules that make up the surface capsule of - Aggregate of bacteria held together by a mucus like
certain bacteria matrix of carbohydrate that adheres to a surface.
7. Recombinant Vaccines - Vaccine antigens may also
be produced by genetic engineering Flagella (singular: flagellum)
BACTERIA MORPHOLOGY - Whip-like structures
3 basic shapes of bacteria - Thread like structures
1. Coccus (plural: cocci) - Made of protein sub unit flagellin
- Shapes: Oval, Round and Spherical - Project from the capsule
- Ex: staphylococcus and streptococcus Types of flagella:
Arrangement of Bacterial Cocci: Monotrichus – single polar (Vibrio cholerae)
a) Diplococci  Lophotrichous– tuft at one end (Bartonella
- In pairs bacilliforms)
- Ex: Neisseria gonorrhoeae (gonorrhea) and  Amphitrichous – at both ends of the bacteria
Neisseria meningitides (meningococcal) (Spirillum serpens)
b) Streptococci  Peritrichous – all around the bacillus (Escherichia
- In chains coli)  Atrichous – without flagellum
- Ex: streptococci pyogenes (strep throat) and Ribosomes
Streptococci mutans (tooth decay) - site of protein synthesis - target site of some antibiotic
c) Staphylococcus Nucleoid
- Irregular, grape-like cluster - no true nucleus - does not contain nuclear membrane
- Ex: staphylococcus aureus (skin infection, food Mesosome
poisoning) - for secretion of substance (chromosomes) by bacteriu
d) Sarcina/ Octad
- Cube-like packet of 8 cocci Granules or Inclusion bodies
- Ex: Sarcina aurantiaca (normal flora of skin/ GIT – - found in certain bacteria
body odor) Endospores/ Spores - Composed of dipicolinic acid -
e) Tetrad Resistant to cold, heat, drying, chemical and radiation
- Cubelike packet of 4 cocci Cytoplasm
- Ex: Aerococcus (found in urine – cause UTI, - semifluid gelatinous nutrient matrix
septicemia bacteremia, endocarditis) - insoluble H2O

2. Bacillus (plural: bacilli) BACTERIAL GROWTH REQUIREMENT

- Shapes: rod shape and cylindrical Environment/ Physical Requirement
- Ex: Escherichia coli and Salmonella water/ moisture
Arrangement of bacteria bacilli: oxygen
osmotic condition Generation Time - the time it takes for binary fission to
occur. - varies from one bacterial species to another &
Temperature also depends on the growth condition
Oxygen Requirement
Obligate Aerobes Steps in the Pathogenesis of Infectious Disease
- Have absolute or obligate need for oxygen 1. Portal of Entry
Obligate Anaerobes - Cannot multiply if any oxygen is 2. Attachment
present, often killed even if with traces of oxygen 3. Multiplication
Facultative Anaerobes - Grows with oxygen but can also 4. Invasion or spread
grow without it 5. Evasion of host defenses
Microaerophiles 6. Damage to hot tissues
- Require small oxygem (2%-10%) for aerobic resp
Aerotolerant Anaerobes - Can grow with oxygen Characteristic of Viruses
presence but do not use it to transform energy  Lacks enzymes for energy production
Bacterial Growth Rate  Unable to replicate (multiply) on their own,
1.Lag Phase replication is directed by genome once in a host
 Absorbs nutrient cell Viruses are classified access to:
 Prepare for cell division  type of genetic material - DNA or RNA
 No increase in number  shape of capsid
2. Log Phase/ Logarithmic / Exponential Phase  size of capsid
 Multiplies rapidly & double in each generation time  number of capsomeres
 Cells are metabolically active  presence/absence of envelop
3. Stationary Phase  type of host that it infects
 Equilibrium / Growth slows down  type of disease it produces
 The culture is at its greatest population density  target cell
 The number of bacteria that are dividing equal the  immunologic / antigenic properties
number of dying ANTIMICROBIAL AGENTS
4. Death (or Decline) Phase  Chemotherapy * Antibacterial Agents
 Concentration of waste product increase while the  Antifungal Agents * Antiprotozoal Agents
nutrient decreases  Antiviral Agents * Anthelminthic Agent
 Sporulation may occur in some bacteria  Antimicrobial Chemotherapy
 Antibiotic
Nutritional Requirement
1 Carbon
 Autotroph/ Lithotroph
 Heterotroph
 Phototroph
 Chemotroph
2. Nitrogen, Sulfur and phosphorus
 Nitrogen & phosphorus
 Nitrogen & sulfur - for synthesis of protein
3. Magnesium, Potassium, Calcium, iron and trace
Elements
4. Growth factor

Bacterial Reproduction
- Bacteria are prokaryotic cells…
Binary Fission - The division of one cell into two cells,
after DNA replication & the formation of a separating
membrane & cell wall DNA
Replication - before it divides in half, its chromosome
must be duplicated first.
Binary Fission - parent cell splits in half to become 2
daughter cells.