Medical Microbiology I

SPM 623 01

Enrique Rosario Aloma, DPM,

PhD
 INTRODUCTION TO MEDICAL
MICROBIOLOGY
• Viruses- smallest infectious particle
– Cannot be seen with light microscope
– Cellular parasites
• Bacteria
– Prokaryotic organisms
– Unicellular organisms, no nuclear
membrane
– Reproduce by asexual division

2
Commensal & Pathogenic
Microbial Flora in Humans
• What is Medical Microbiology?

• Microorganisms also play a critical role in

human survival
– participate in the metabolism of food products
– protects against infections with highly virulent
microorganisms
– stimulates the immune response
 INTRODUCTION TO MEDICAL
MICROBIOLOGY
• Fungi
– Cellular structure more complex than
bacteria
– Eukaryotic organisms
• Parasites
– Eukaryotic, unicellular & multicellular
– Protozoa, tapeworms, arthropods

4
Microbial Flora in Humans

The normal flora in humans is the

population of bacteria that has
colonized certain areas of the body
and has developed a commensal
relationship.
Terms to Know

• COMMENSALISM – relationship of
organisms of different species in
which neither is harmful to the other
& one gains some benefit such as
protection or nourishment
• PATHOGEN – microorganisms which
cause disease in a host
Terms to Know
• COLONIZATION – when one of more
species populate an area
Normal Commensal
Population
• Consists of mainly bacteria, fungi and
some parasites
• Live on the surface of the skin and all
mucosal membranes
• Bacteria live on these surfaces and
protect us from colonization with
pathogenic microbes
Terms to Know

• CARRIER STATE – pathogens that

transiently colonize apparently
healthy individuals
• DISEASE - occurs when the
interaction between microbe and
human leads to a pathologic process
characterized by damage to the
human host
Propensity for causing
disease
• Strict pathogens vs Opportunistic
pathogens
Colonization and Disease

• Infection vs colonization
Factors affecting Microbial
Flora
• Main factors affecting normal flora
– Age
– Diet
– Hormonal state
– Overall health
– Personal hygiene
- Physiology
- pH and nutrients
- Resistance vs. susceptibility
Microbial Flora

• Normally Sterile- no organisms present

• Body fluids
– Blood
– Cerebrospinal fluid
– Urine
– Pleural fluid
• All but superficial tissues (this includes
cornea)
DISINFECTION, STERILIZATION,
AND ANTISEPSIS
• Sterilization - total destruction of all
microbes including the more resilient forms such
as bacterial spores, mycobacteria, non-enveloped
viruses, and fungi
• Disinfection - destruction of most organisms,
although the more resilient microbes can survive
some disinfection procedures
• Antisepsis - use of agents on skin or other
living tissue to inhibit or eliminate the number of
microbes; no sporicidal action
• Germicide - chemical agent capable of killing
microbes; spores may survive
Sterilization – Physical
Sterilants
• Filtration – removing bacteria and fungi
from air with high efficiency particulate
air (HEPA) filters
– Unable to remove viruses and some small
bacteria
• Moist heat & Dry heat (autoclave)
– most commonly used in hospitals
– Indicated for most materials unless heat
sensitive or toxic or volatile chemicals
Moist Heat Sterilization

• Boiling water inefficient because

maintaining a temperature greater
than 100 degrees C difficult
• Boiling vegetative organisms kill
them, but spores remain viable
• Autoclave – device that creates
steam under pressure for sterilization
Moist Heat Sterilization

• achieve higher
temperature
(121 to 132
degrees C)
• Rate of killing
rapid
• 15 to 30 minutes
Chemical Sterilant or
Disinfectant
• Although some agents are used both
for sterilization and disinfection, the
difference is the concentration of the
agent and duration of treatment
• The types of agents that are used are
determined by the nature of the
material to be disinfected and how it
will be used
Disinfection and Sterilization

• Microbes also destroyed during

disinfection, but some do survive
• Often the terms are used
interchangeably, but they are not the
same
DISINFECTION, STERILIZATION,
AND ANTISEPSIS
• STATIC versus CIDAL activity
• STATIC (bacteriostatic, fungistatic)
– INHIBITS GROWTH
– INCOMPLETE KILLING
• -CIDAL (bactericidal, virucidal,
sporocide, fungicidal)
– KILLS
– IRREVERSIBLE
DISINFECTION, STERILIZATION,
AND ANTISEPSIS
• SEPSIS
– THE PRESENCE OF PATHOGENIC ORGANISMS IN TISSUE
• ANTISEPSIS
– USE OF AGENTS (ANTISEPTICS) TO KILL OR INACTIVATE
MICROORGANISMS ON ANATOMICAL SURFACES
• “COMPATIBLE” WITH HUMAN TISSUE
– ALCOHOLS
– IODOPHORS
– CHLORHEXIDINE
– TRICLOSAN
Diagnostics

• Brightfield (light) (most used)

• Darkfield
• Phase-contrast
• Fluorescent
• Electron – not used in routine clinical
microbiology
Diagnostics

• Direct smear examination vs staining

Diagnostics

• Acid fast staining:

• Uses weak acids to stain pathogen

• Used to stain acid fast organisms

wax-like surface.
Diagnostics

• Blood agar vs chocolate agar:

• Blood agar = blood + basal medium

• Chocolate agar = blood + heated

basal medium
Diagnostics

• Thioglycollate broth:

• Will support the growth and

differentiate between aerobe and
anaerobe bacteria
Diagnostics

• Different types of pathogenic detection

• Microscopy
• Direct Antigen Detection
• Nucleic Acid Detection
• Culture
• Serology – antibody response
• Molecular
Diagnostics

• PCR

• Fastest detection method: antibody

detection
Diagnostics

• Serology: using body fluids,

immunoessays

• Polyclonal: recognizes several epitopes

• Monoclonocal: recognizes specific

epitopes
Immune response

• Barriers are the first line of defense

against pathogens:
• Skin, mucosal surfaces, hair, gastric
acid, bile from the liver
• Barriers stop pathogens from
entering the body
Immune response

• Innate vs adaptive immune system

(specific produces antibodies)
• Innate: rapid, triggered first, does not
adapt
• Adaptive: triggered by the innate
immune response, responsible for
memory and for the evolution of the
immune system
Immune response

• Innate immune system:

• All healthy individuals are born with it.
• Has 2 components: cell component
and soluble factor component.
• Cell component: phagocytes and NKC
• Soluble factor component:
complement system, stimulators
Immune response
• Cytokines: hormone-like proteins that
regulate the immune system
• Interferon: a protein produced in response
to infections
• Chemotaxis: cellular movement regulated
by concentration gradient if certain
cytokines
• Chemokines: cytokines responsible for
stimulating and regulating chemotaxis
Immune response
• Adaptive immune system: B and T cells.
• All cells are born in the bone marrow.
• B Cells are born and mature in the bone
marrow.
• T cells are born in the bone marrow, but
mature in the Thymus.
• Bone Marrow and the Thymus are the
primary lymphoid organs
Immune response
• Secondary Lymphoid Organs - peripheral
lymphoid organs
• Lymph nodes
• Spleen
• Mucosa-associated lymphoid tissue (MALT)
• Gut-associated lymphoid tissue (Peyer’s
patches)
• Bronchus-associated lymphoid tissue
(tonsils, appendix)
Immune response

• The secondary lymphoid organs are

where the B and T lymphocytes
reside and where the information
picked up by APCs is presented.
• Where proliferation of lymphocytes
occur, which is represented by
swollen glands
Immune response

• Phagocytosis: engulfing cells

• Pinocytosis: absorbing fluid
• Macropinocytosis: engulfing soluble
components such as proteins and
enzymes
Immune response

• Neutrophils: fastest cell to arrive at

infection site, short lived, attracted
by chemokines.
• What they can do: phagocytosis,
exposure to lysozyme and lactoferrin
• Major component of pus
Immune response
Immune response

• M-CFU differentiate into Neutrophils

or Monocytes.

• Monocytes can further differentiate

into Macrophages or Dendritic Cells
Immune response

• Macrophages: come from Monocytes,

unlike Neutrophils they have
mitochondria and are long lived.
• What they can do: phagocytosis,
antigen presentation to T cells and
secretion of cytokines
Immune response

• Dendritic cells: come from Monocytes,

more specialized than macrophages,
main function is to collect and process
information to stimulate the adaptive
immune system.

• They are the BRIDGE between the

innate and adaptive immune system
Immune System

• Immature dendritic cells

• vs.

• Mature dendritic cells

Immune response

• Lymphocytes: B and T cells

Immune response
• T lymphocytes:
• acquired their name because they develop in the
thymus
• T cells have the following two major functions in
response to foreign antigen:
1. Control, suppress (when necessary), and activate
immune & inflammatory responses by cell-cell
interactions & by releasing cytokines
2. Directly kill virally infected cells, foreign cells (tissue
grafts), and tumors
Immune response

• B lymphocyte:

• PRODUCE ANTIBODIES
Immune response
• Macrophage: Pac Man
• phagocyte which is activated by
interferon-γ & then becomes
efficient at killing phagocytized
microbes and producing cytokines
• Lymph Node: Police department
• Repository for B and T cells
Immune response
• CD4 T cell: Desk sergeant/dispatch officer
• CD4 T cell is presented with the microbial
problem by antigen presenting cells, it tells
other cells to take care of the problems by
producing cytokines
• CD8 T cell: "Cop on the beat"/patrol officer
• CD8 T cell gets activated in the lymph node &
then moves to the periphery to patrol for virus
infected or tumor cells
• it then grabs the perpetrator and inactivates it
Immune response

• B cell: Product design and building

company
• Pre-B cells and B cells alter the DNA
of their immunoglobulin genes to
produce the plans for a specific
immunoglobulin
Immune response
• Plasma cell: Factory
• plasma cell is an immunoglobulin-producing
factory with a small office (nucleus) and many
assembly lines (ribosomes) for making antibody
• Mast cell: Activatable chemical warfare unit
• mast cell has Fc receptors for IgE that will trigger
the release of histamines and other agents upon
binding to an allergen signal
Immune response

Macrophages: M1 vs M2

Different functions depending on

stimulation pathway
MI – Th1 pathway
M2 – Th2 pathway
Immune response
• M1 macrophages –
• “Classical Activation” – mediated by IFN-γ produced by NK cells and CD4 and CD8 T cells
• Part of TH1 response
• Able to kill phagocytosed bacteria
• Produce cytokines and enzymes
• Reinforce local inflammation by producing cytokines
Immune response
• M2 macrophages
• “Alternatively Activated” by cytokines IL-4 and IL-13
• Part of the TH2 response
• Anti-parasitic responses
• Wound repair
• Form granulomas to surround chronic infections such as unresolved mycobacterial infections
Immune response

• Dendritic cells: immature vs mature

• Immature DC pick up information
• Mature run to lymphoid organs to
present the information
• Information is picked up using their
dendrites and TLR.
• Receptors recognize PAMPs
Immune response
• Natural killer cells
• Great against viral infected cells and
tumor cells.
• Responsible for ADCC
• Large lymphocytes, similar to T cells.
• Stimulated by IFN, interleuking 2 and TNF
• Stimulate differentiation of Monocytes
and stimulate the Th1 pathway
Immune response

• NKC cannot identify specific antigens

like T cells.

• They are regulated by the MHC 1,

which all normal cells have. If the
MHC 1 is affected or modified, then
they attack.