Medical Microbiology: Bacteriology I

Macroscopic (classification)

colony morphology, color, smell

Microscopic (classification)

shapes, Gram +, Gram -, Endospores, capsule, flagellum, staining

Phenotypic (classification)

aerobic vs anaerobic, biochemical characteristics, selective growth conditions (temperate, growth media)

Antigenic (classification)

Direct detection, indirect detection, immunofluorescence, enzyme-linked immunosorbent assay (ELISA), Wester Blot

Molecular (classification)

polymerase chain reaction (PCR)

Gram +

thick layer of peptidoglycan

Gram -

thin layer of peptidoglycan, outermembrane

Endospores

only gram +, vegetative state, needs signaling to get out

flagellum

movement

direct detection

use conjugated anti-target antibody

indirect detection

use conjugated secondary antibody

immunoflorescence

microscopic detection of antigens

enzyme-linked immunosorbent assay (ELISA)

detect pathogen--> active infection Capture --> detect presence of particular antigen antibody detection --> start with antigen to detect antibody quick test

Western Blot

each band represents a different viral protein often used to confirm ELISA results if you have known antibodies --> use to detect virus look at immune responses

Polymerase chain reaction (PCR)

amplify particular sequence of DNA mimicking primer and DNA replication can use for detection, sequencing (mutations), and RFLP analysis

If sequence is present in PCR...

amplification

If sequence is NOT present...

no amplification

Restriction fragment length polymorphisms (RFLP)

enzymes come from bacteria and recognize and cut DNA sequences to get particular patterns DNA only (can be PCR product DNA) DNA digestion with restriction enzymes producing DNA patterns

Transformations (mechanism)

[Acquisition of foreign DNA] donor cells --> cell lysis --> DNA enters the recipient cell and integrates into DNA

Transductions (mechanism)

[Acquisition of foreign DNA] transducing phage containing donor genomic DNA --> cell lysis --> phage infects recipient cell; donor integrates into recipient DNA

Transpositions (mechanism)

[Acquisition of foreign DNA] moving and relocating in genome uses transposon with inverted repeats

Conjugation (mechanism)

[Acquisition of foreign DNA] free plasmid moves from donor to recipient cell via sex (F) pilus integrated plasmid (episome) promotes transfer of genomic DNA, which integrates into recipient DNA

Capsule

produced outside cell; slippery so things can't bind or penetrate no opsonization, no MAC, no phagocytosis

Biofilm

grow into giant, multicellular structure that surrounds pathogen

Spores

inert object is protected from the immune system and antibiotics until signaled almost impossible to get rid of

proteases

destroy host proteins

variable surface proteins

antigenic variation

Adhesion

bind to receptors (proteins, sugars) on the surface of target cell

Endotoxin

part of the pathogen structure Ex: LPS on Gram -

Exotoxin

product produced and released by the pathogen Ex: A-B

Superantigens

trigger over release of cytokines by CD4+ T cells

Intoxication

large quantity of toxin taken in; immediate effects

Infection

accumulation of toxin in infected individual; slower effects

Bacteriostatic agent

prevent growth (put in stasis) Ex: fridge/freezer

Bactericidal agents

destroy bacteria Ex: cleaning wipes, bleach, detergents

Step 1 of Koch's Postulate

Bacteria must be present in every case of the disease

Step 2 of Koch's Postulate

Bacteria must be isolated from host with disease and grown in pure culture

Step 3 of Koch's Postulate

specific disease must be reproduced when a pure culture of the bacteria is inoculated into a health susceptible host

Step 4 of Koch's Postulate

bacteria must be recoverable from the experimentally infected host

Antibiotics

disruption of cell wall; inhibition of protein synthesis; inhibition of nucleic acid synthesis; antimetabolite

CD8+ T cells

Capsules cannot protect bacteria from which of the following? -CD8+ T cells -Opsonization via antibodies -Opsonization via complement proteins -Destruction via the membrane attack complex

Detergents

Which of the following would not be an example of a bacterial static process? -Freezing -Refrigeration -Detergents -Using antibiotics that prevent the synthesis of new cell walls

All of the other answers can be used to protect a bacterial pathogen from

Which of the following are mechanisms by which bacteria can protect themselves from immune responses? -Biofilm formation -Spore formation -Encapsulation -All of the other answers can be used to protect a bacterial pathogen from immune responses.

Operons

Which of the following is not a mechanisms that allows bacteria to take in foreign DNA? -Transduction -Conjugation -Operons -Transformation

All of the other answers can be used to help identify a bacterial pathogen.

Which of the following observations can be used to help identify a bacterial pathogen? -Antigenic -Phenotypic -Molecular -Microscopic -All of the other answers can be used to help identify a bacterial pathogen.

ELISA

Antibodies can be used to help identify a bacterial pathogen in which methods? -PCR -ELISA -RFLP analysis -All of the other answers use antibodies to help identify a bacterial pathogen.

The use of DNA as the genetic material**??

Which of the following is not different between human and bacterial cells? -The composition of the ribosomes -The average size of a cell -The use of a cell wall -The use of DNA as the genetic material

All of the other answers can be used to help identify a bacterial pathogen.

Which of the following can be used to help identify a bacterial pathogen? -All of the other answers can be used to help identify a bacterial pathogen. -Selective growth -Enzymatic activity -Aerobic vs anaerobic growth

normal flora and environment

Where is Staphylococcus found?

Staphylococcus: Transmission

person to person; fomite

Often observed with Staphylococcus

disruption of normal flora (makes spaces for new things to grow) insertion of foreign bodies half-life: days to weeks

Staphylococcus: Immune Avoidance

capsule, biofilm, slime layer, Protein A

Staphylococcus: Virulence Factor

adhesions (slime layer)

Staphylococcus: Toxins

cytotoxins endotoxins superantigens

Staphylococcus: Treatment

Antibiotics --> resistance

Methicillin-resistant Staphylococcus aureus (MRSA)

What is an example of an antibiotic-resistant form of Staphylococcus?

Staphylococcus: Control

ubiquitous clean/sterilize medical equipment minimize exposure

Streptococcus A (S. pyogenes)

strep throat, cellulitis, scarlet fever, necrotizing fasciitis

Strep A: Transmission

person to person, fomite

Strep A: immune avoidance

capsule M proteins: Block C3b binding C5a peptidase (S. pyogenes): block inflammation

Strep A: M&F proteins

adhesion cell invasion (disruption of tight junctions)

Strep A: exotoxins

enterotoxins (A-B toxins) Superantigens [Ex: streptolysin (lyse RBS) --> hemolytic]

Strep A: Treatment

antibiotics

Streptococcus B (S. agalactiae)

neonatal infections, normal flora getting in the wrong place

Strep B: Transmission

normal flora of vaginal tract (birth) location is the issue

Strep B: Pathogenesis

In newborns: bacteremia (bacteria in blood) pneumonia meningitis (in brain)

Strep B: Treatment

antibiotics test women for it

Strep B: Control

minimize exposure and treatment

S. pneumonia (pneumonia)

causes infection in the lungs diplococci Control: vaccine against capsule

normal flora of GI tract

Where is Enterococcus found?

Enterococcus: Transmission

person to person, fomite

disruption of normal flora

Enterococcus is often observed...

Enterococcus: Virulence factors

wide range of growth: aerobic and anaerobic, high salt, broad pH biofilm antibiotic resistance

Enterococcus: Treatment

antibiotics --> resistance

Bacillus: Epidemiology

spore forming stable in environment aerosol transmission (anthracis)

Anthracis (anthrax): Transmission

ingestion (livestock) cutaneous- most common (skin) inhalation

Anthracis (anthrax): immune avoidance

capsule

Anthracis (anthrax): toxins

Exotoxin: plasmid encoding 3 genes

edema toxin

PA + LF = ?

lethal toxin

LF + EF = ?

Anthracis (anthrax): Treatment

antibiotics --> resistance vaccination --> toxoid (animals)

Cereus

bigger problem --> seen in humans a lot

Cereus: Transmission

ingestion (food born pathogen)

Cereus: Virulence Factors

exotoxin

Cereus: heat stable form

emeric (vomiting)

Cereus: heat liable

diarrheal

Bacillus cereus

food poisoning

Cereus: Treatment

Pepto-Bismol for symptoms antibiotics --> resistance

Cereus: Prevention

minimize exposure: cook/store food properly

Emeric Form: Disease onset

~2 hours

Emeric Form: Disease Duration

8-10 hours

Diarrheal Form: Disease onset

~9 hours

Diarrheal Form: Disease duration

20-36 hours

Listeria: Epidemiology

grows @ 4 C, high salt concentration, and wide pH range facultative intracellular pathogen (can exist outside of cell) lives inside macrophage --> pt of immune response doesn't work

Listeria: Transmission

contaminated food/dairy

Listeria: Virulence factors

cell invasion escapes lysosome

Listeria: Disease

can cause spontaneous abortions diarrhea meningitis (rare)

Listeria: Treatment

self-limiting antibiotics --> resistance (for severe cases)

Listeria: Prevention

minimize exposure: cook food properly; avoid raw dairy (soft cheeses and milk), raw vegetables, undercooked meat, etc.

Corynebacterium diphtheriae

a species of bacterium that causes *diphtheria*

Diphtheria: Epidemiology

normal flora facultative anaerobes

Diphtheria: Exotoxin (virulence factor)

result of transduction regulated expression "A" blocks eukaryotic translation --> no protein synthesis

Diphtheria: Treatment

antibiotics passive immunization against exotoxin

Diphtheria: Prevention

vaccination: Diphtheria toxoid --> DPT vaccine

Clostridium

Gram + rod Genus: difficile, perfinges, tetani, botulinum Environmental: soil, water, sewage, normal flora Spore forming --> stability Exotoxins --> neuro toxins obligate anaerobes

obligate anaerobes

only live in the absence of oxygen

Clostridium difficile: Transmission

Growing: normal flora (spread with antibiotic usage) Spores: hospital acquisition

Clostridium perfinges: Transmission

Growing: contaminated food (meat) [growth at 4-60 C] Spores: cutaneous

Clostridium tetani: Transmission

Spores: cutaneous

Clostridium botulinum: Transmission

Spores: contaminated food (canned food, infant formula, dairy)

difficile

Clostridium: Which has A-B exotoxin

perfinges, tetani, and botulinum

Clostridium: Which has neurotoxin

Clostridium tetani: Virulence factors

interferes with the ability to communicate with muscles

Clostridium botulinum: Virulence factors

interfere with receptors

botox only has toxin and cannot reproduce

Botulinum vs botox

Clostridium difficile: Disease

Mild: diarrhea Severe: Colitis

Clostridium perfinges: Disease

Mild: cellulitis Severe: Myonecrosis (necrosis of muscle)

Clostridium tetani: Disease

Mild: tetanus (muscle spasms) {localized} Severe: tetanus (muscle spasms) {systemic or neonatal}

Clostridium botulinum: Disease

Severe: botulism {paralysis and respiratory arrest}

Clostridium difficile: Treatment

antibiotics fecal transplant

Clostridium perfinges: Treatment

antibiotics debridement

Clostridium tetani: Treatment

antibiotics debridement passive immunization (toxin)

Clostridium botulinum: Treatment

antibiotics passive immunization (toxin)

Clostridium difficile: Prevention

proper antibiotic usage hygiene

Clostridium perfinges: Prevention

Cooking of food

Clostridium tetani: Prevention

toxoid: DTaP

Clostridium botulinum: Prevention

proper canning food preparation (heat)

Myobacterium

rods weakly gram + --> Gram Neutral Acid fast lipid rich cell wall

Acid Fast

test that you can run to see if it has outer membrane structure

lipid rich cell wall

slow growth resistant to many disinfectants (don't have structures) resistant to some antibiotics

Tuberculosis: Epidemiology

~1/3 of the world population is infected restricted to human

Tuberculosis: virulence factors

intracellular pathogen

Intracellular Pathogen (tuberculosis)

prevents fusion to lysosome resistant to nitric oxide-mediated killing immune-mediated control but not clearance (chronic infection) Granuloma

Granuloma

little prison cells; how it is contained and sealed up, but not destroyed formed in lungs

Tuberculosis: Disease

respiratory infection: blood in sputum

Those people don't have the ability to form granulomas

Why is the respiratory infection of tuberculosis systemic in immunocompromised people?

Tuberculosis: Treatment

controlled in most healthy individuals antibiotics --> very complex because of slow growth and multiple drug resistance

Tuberculosis: Transmission

aerosol

Tuberculosis: Prevention

screening: TB test Vaccination (BCG live attenuated)

Tuberculosis vaccination...

is not in the U.S. or with HIV positive individuals

Immunocompromised Populations

Transplant recipients Age Immunosuppressant infractions (i.e. AIDS) or genetics Immunotherapies

Transplant recipients

immunosuppressants block organ rejection

Age: very young

developing immune system

Age: old

declining immune system

Immunotherapies

not a global decline, compromised specific parts of the immune system