Microbiology 5: Obligate Intracellular and Gram Negative Cocci
Chlamydia trachomatis
Neisseria gonorrhea
Neisseria meningitidis
-obligate intracellular bacterium-parasitic, takes
-Theyre the ONLY common, important gram negative diplococci stain PINK
nutrients and ATP from host cytoplasm
*they have Lipooligosaccharide: LPS without the O-polysaccharidestill endotoxin
1) infect via Elementary body (EB)small dense,
*generally fastidious (need enriched media and more CO2) and tough to culture
infectious, NOT DIVIDE
2) becomes Reticulate body (RB) which replicates in
host cell (& larger)
3) daughters become EBs and exit (kill host)
Unique features (of its structure)?
-VERY SMALL (0.25 uM) and INTRACELLULAR (so hard to
detect, and hard to see w/microscope)
-inner+outer membrane BUT no peptidoglycan
Source/Transmission/ -generally person-to-person: 1) shed from urogenital mucosa and 2) transmitted in:
-colonize oropharynx of
Entry
a) sexual contact: [horizontal] via mucosa (genitals, anorectrum, oral)
healthy adolescents
*usually not fomitesNOT HARDY!
-spread via close-contact
-eye can be involved too via autoinoculation
and crowding
b) perinatal transmission: [vertical] mother to fetus (syphilis crosses placenta) or mother to baby @delivery
(cohabitation, kissing,
(gonorrhea, chlamydia)where baby tends to develop eye infections, and maybe even pneumonia
respiratory secretions)
Manifestation of
1) Non-focal mucosal (non-ulcerating) infection of genitals:
-fever, headache,
Disease
first F: vaginal discharge (mucoprurulent cervicitis) w/mild discomfort
photophobia,
M: urethritis (asymptomatic, discharge)
meningismus, possible
thenASCENDS:
acute shock
F: pelvic inflammatory disease (PID)inflammation of pelvic structures, tenderness, scarring, infertility,
low threshold for antibiotic treatment
Mepidymitis, prostatitis, [discharge, dysuria]
Note : Syphilis (Treponema) looks different, it shows a pattern of Focal infection (ulcerating) w/ local lesion and
lymphadenopathy: @skin+mucosa
2) Conjunctivitis from either autoinoculation or transmission to babies (opthalmia neonatorum w/Gonorrhea)
3) If @ anorectalproctitis (pain and discharge)
4) If @ pharynx--pharyngitis
Serotypes D-K are genital
-can progress to disseminated gonoccoal infection :
**also can cause reactive arthritis: bacteria arent @
-spread to blood (bacteremia)skin lesions
(papules, pustules, bulla) AND joints (supperative
joint, but inflamed joints
Reiters Syndrome=arthritis, conjunctivitis, urethritis
arthritis and swelling)
Serotype A-C cause trachoma
**high risk w/menstruation, complement deficiency
-eye infection, transmit person-to-person or via flies
**these strains appear resistant to human Abs
-inverts eyelid causing corneal scarring
**this is very rare
Serotype L causes lymphogranuloma venereum
Pathogenesis &
-Caused by HOST RESPONSE to PAMPs
-Caused by HOST RESPONSE TO Lipoligosaccharide
-Caused by HOST
Damage
-host response kills epithelial cells NOT pathogen
(LPS w/o O-polysaccharide, just lipid A+polysaccharide)
RESPONSE TO PAMPs
-order: PMNslymphocytesnecrosis & scarring
1) bind non-ciliated epithelial cells and endocytose
-invades blood to affect
A clear example: REACTIVE ARTHRITIS
meninges
Pathogenic Agent
Immune Response &
Pathogenic Evasion?
-evasion via intracellular growth
-NO immune protection from re-infection
-however, IFN-g seems to inhibit RB replication by
causing host to use tryptophan (depriving RB)
Diagnosis (Dx)
-PCR (aka nucleic acid amplification test or NAAT)
*test w/gonnorhea
-Culture is insensitive; gram stain doesnt work (no PG)
Treatment (Rx)
Prevention?
2) exit to subepithelium (killing cell in the process)
3) elicit strong inflammatory response via NEUTROPHILS
4) pus and more epithelial damage
-evasion via pilus variation:
-has different surface antigens encoded by diff
pilin genes; uses homologous recombo to SHUFFLE
-also makes IgA proteasemight it destroy antibodies?
-PCR (aka nucleic acid amplification test or NAAT)
*test w/chlamydia
-Gram stain in MEN (women have other flora)
-Culturable but fastidious
**cannot use B-lactams or aminoglycosides (no PG)
-3rd generation cephalosporins PLUS azithromycin or
-azithromycin (one dose) or doxycycline (1 week)
doxycycline (to avoid problem w/resistance)
-can also use erythromycin or fluorquinolones
-disseminated infections take longer to treat
-already resistant to fluoroquinolones
-for babies w/eye infection, use AgNO3 or antibiotics
Reducing transmission: 1) less contact, 2) less efficient (safe sex), 3) less time (treat quickly)
-evasion via capsule that
is hard to phagocytose
-immune response can
develop against capsule
-BUT: 13 dif serogroups
(w/dif carbohydrates)
serotype-spec. immunity
-conjugate vaccines for
common serotypes
-antibiotic prophylaxis if
in contact w/infected
Microbiology 6: Gram positive cocci
Pathogenic Agent
Staphylococcus aureus
-Gram positive cocci in grape-like CLUSTERS
coagulase+ catalase +
quite LARGE
-very hardy bacteria (but no spore)heat
resistant, grows in dry environment
-facultative anaerobe
Source/Transmission/ -found on surfaces and human SKIN as well as
Entry
nares/throat (endogenous; asympto carriers)
-person-to-person via direct contact or
aerosol droplets
-indirect contact via objects/fomites
-infection requires skin/mucosal breach, then
travels well in blood (hematogenous spread)
Note: exogenous infections is possible but less
common
Streptococcus pyogenes (Group A)
-Gram positive cocci in CHAINS
catalase
--hemolyticwide clear margin of hemolysis
-easily found in nares/throat (endogenous;
asympto carriers); transiently on SKIN
-person-to-person via aerosol drops or direct
contact (ideally nasal, not skin or pharynx)
-infection requires skin/mucosal breach, then
travels well in blood (adhesins help)
Streptococcus pneumoniae
-Gram positive cocci in PAIRS (diplococci)
catalase
--hemolyticthin GREEN margin (partial
lysis, Hb leakage)
-mucoid appearance in culture (capsule)
-anaerobic but can tolerate ambient O2 (lung)
-easily found in nares/throat (endogenous;
asympto carriers)
-person-to-person via aerosol droplets or
respiratory secretions
-often follows other upper respiratory tract
infection (esp smokers+immunocompromisd)
-if get to blood, cause bacteremia and spread
Manifestation of
Disease
1) Direct infection, destroy local tissue:
pyogenic infections and focal abscesses @
skin; spread to soft tissue, bone, lung, viscera,
endocarditis, pneumonia
2) Toxin-mediated diseases (distal sites):
-staphylococcal toxic shock syndrome
-scalded skin syndrome
-food poisoning
depends on serogroup---see below!
Immune Response &
Pathogenic Evasion?
A) Resisting phagocytosis via:
0. [some have a bacterial capsule and secrete
expolysaccharide slime]
1. protein Abind Ab to reduce opsonization
(i.e. avoid recognition)
2. catalaseconverts H2O2 to H2O,
neutralizing ROS killing power of neutrophils
(i.e. resist killing)
3. coagulase-->binds fibrin and form clots
-helpful to wall off abscesss from phago
*note that abseess center is necrotic
Pathogenesis &
Damage
B) Kill immune cells via pore-forming toxins:
Hemolysins (alpha toxin, lyse RBCs)
Leukocidins (lyse leukocytes)
Abscess formation facilitated by:
-collagen-binding adhesins (invade bones/CT)
-fibronectin-binding adhesins (invade
epithelia and endothelia)
-clumping factor/bound coagulase: converts
fibrinogen to fibrin to wall off abscess
*note that abseess center is necrotic
Also secrete degradative enzyme to invade
DAMAGE is from both immune response AND
toxins that destroy immune cells
1) Direct infection, destroy local tissue:
supperative diseases
- pharyngitis (strep throat), pneumonia,
pyoderma,
-@ fascial planes: necrotizing fasciitis
2) Toxin-mediated diseases (distal sites):
-streptococcal toxic shock syndrome
-scarlet fever
3) Autoimmune reaction:
Delayed, non-supperative
-acute rheumatic fever
-post-strep glomerulonephritis
A) Resisting phagocytosis via:
1. hyaluronic acid capsulecamouflage
because its a self-antigenno Abs!
2. M-protein (coiled-coil surface protein on
cell wall, has hypervariable end):
a. variable region (middle) binds
fibrinogen (complement cant access)
b. conserved region binds complement
control proteins (complement cant
opsonize around it opsonin)
**Can develop Ab for M-protein but because
of hypervariable region there are MANY types,
and type-specific immunity just isnt useful
B) Kill host immune cells via toxins:
Streptolysins (which cause hemolysis)
-Also has adhesins that help with invasion
-Also secretes degradative enzymes
-HA capsule can help with adhesions
DAMAGE is mostly from immune response
but also toxins and degrading enzymes
Direct infection, destroy local tissue:
@ respiratory tract (lobar pneumonia,
sinusitis, otitis media)chills, cough, sputum
OR if really bad, bacteremia and meningitis
4 stages of pneumonia:
1) alveoli fill with serous fluid since cell wall
stimulates inflammation
2) early consolidation or chemotaxis of
neutrophils+RBCs
3) late consolidation when neutrophils win
4) resolution when scavenging macs clean
up; lung goesback to normal
A) Resisting phagocytosis via:
-thick polysaccharide capsuleinhibits
antibody binding, so no complement
response OR antibody-mediated phagocyt)
**critical to survival in blood, also helps
cross BBB for meningitis
**strains w/o capsule are avirulent
**Can develop Ab for capsule antigens but
there are MANY serotypes
**Vaccine combines several common antigens
B) Kill host immune cells via toxins:
Pneumolysins (which bind cholesterol and
damages the host membrane)
DAMAGE is mostly from immune response as
there arent that many toxins:
Recall cell wall is a PAMP to stimulate
inflammation
As a catalase negative bacteria, this
allows the neutrophil to make ROS
this is inherently a bit damaging
Because there are few toxins, the lung
recovers pretty well after pneumonia
Special Consideration
Diagnosis (Dx)
Treatment (Rx)
Prevention?
Superantigens cross-link TCR and MHCII in generally, resulting in non-specific T-cell activation
and massive release of pro-inflammatory cytokines
-usually responsible for toxic shock syndrome
note that O2 is needed to form toxins!
Symptoms include: fever, sun burn-like rash, hypotension, conjunctivitis, vomiting,
diarrhea, high creatinine (kidney issue), systemic problems
septicemia if @blood
1) Toxic-shock syndrome is caused by
1) Toxic-shock syndromesee above
superantigen TSST-1 (assoc w/tampons,
2) Scarlet feverbegins as strep throat but
surgical abscess, lung infection)
then shows systemic rash; caused by
-enterotoxin serotypes A-X=like TSST-1
pyrogenic exotoxins (superantigens) SPE A/C
(this is not assoc w/tampons)
----------------------------------------------------------2) Scalded skin syndrome is caused by
Non-supparative immune diseases:
exfoliative toxins A and Bserine proteases
1) Acute post-strep glomerulonephritis:
that bind and break desmosomes
-hematurea, proteinurea
3) Food poisoning is caused by highly stable
-in the long runrenal failure
exotoxins w/superantigen properties even
Cause: antigen-antibody complexes deposit
w/o live bacteria
@glomerular basement membrane
complement-mediated renal injury
**antibiotics do NOT help with recurrence
(it still deposits)
Pretty suspectedCOMMON
Tough to know bacterial vs viral pharyngitis
-Gram stain sample from abscess
1st: rapid antigen swapimmunoassay for
-Culture sample from abscess (golden
carbohydrate antigen of GAS
colonies are characteristic)
2nd: throat culture on blood agar plates
-unlike other Staphpositive coagulase test
For immune disease, wont see bacteria:
-bacterial serotyping using PCR, or culture
-serology for IgG to antigens (streptolysin O,
and compare antibiotic response
DNase B)
-penicillins and cephalosporins that are B-oral penicillin (no resistance has developed
lactamase resistant
yet), or else erythromycin/macrolides
-MRSA is resistant to all B-lactams due to
alternative PBP; use vancomycin
**If @ skin, treat aggressively to avoid
-protein synthesis inhibitors to reduce toxins
invasion
Also: drain abscesses
Why do we treat group A streptococcal
Treat toxic shock syndrome w/lincosamide
infections?
Also use IVIG (in hopes that someone has IgG 1) help kids go back to school; reduce spread
against TSST-1/2)
2) criticallyprophylaxis for ARF to avoid
-support for shock/hypotension
damage to heart
Hard to develop vaccinenot all have
Hard to develop vaccineconsider HA
capsule; protein A, catalase, coagulase=inside capsule being a self-antigen
--------------------------------------------2) Acute rheumatic fever
-polyarthritis, carditis, chorea,
subcut nodules
-in the long runscarring and
heart failure
Cause: autoimmune damage to
similar host tissues; some host
similarities to M-proteins
**use antibiotics as prophylaxis!
Pretty suspectedCOMMON
-Gram stain sputum (see neutrophils and
lancet-shaped diplococci)
-Culture on blood agar for hemolysis (green)
-penicillin, macrolides, and fluoroquinolones
-still MANY PEOPLE DIE---antibiotics lyse
bacteria, and this can worse inflammation
(like EHEC)
-also, problems w/antibiotic resistance and
transformation of DNA
Vaccines need to address antigenic diversity:
(from capsules):
1) polysaccharide vaccine: 23-valent for
capsule polysacch B-cell response only
2) conjugate vaccines: multi-valent for
capsule polysacch linked to protein carrier
*improve immune response by stim. T-cells
Microbiology 7: Mycobacterium tuberculosis, Borrelia burgdorferi
Mycobacterium tuberculosis
-all Mycobacteria are acid fastdue to mycolic acid (waxy FA on cell wall), resistant to drying,
chemicals, and phagocytosis
stains as red rods on blue background
-slow-growing, obligate aerobe
-facultative intracellular pathogen
Source/Transmission/ -person-to person via inhalation of aerosol droplet nuclei
Entry
**cough, sneeze, talknote that droplet nuclei can be DRY
**air in room remains infectious after person leaves!
-this is VERY EFFICIENT: tiny nuclei bypass the lining of large airways to go straight to alveoli
-large respiratory secretions and fomites too (but rare)
-infects 1/3 of world (crowded areas), but only 10% get clinical disease
-outcome/risk depends on timing, age, immunocompetence (AIDS patients will get
active TB, drug/alcohol abusers)
-however, issue of dormancy/latency
Pathogenic Agent
Manifestation of
Disease
Primary TB:
-droplet nuclei multiply @ alveoli (generally in middle lung areas of high airflow)
-some are ingested by macs but grow well intracellularly
-attract more lymphos+macs such that infected macs disseminate to lymph nodes
-in general, grows until body mounts immune response (hence characteristic cavities in
lung lobes)
1) If mount proper T-cell mediated immune response (positive TST, IFN-g, macrophages kil),
control infection; could be latent though
2) If NOT develop immune response primary progressive TB
w/ progressive pneumonia; possible growth elsewhere; can be fatal
Risk: elderly, <5 yrs old, HIV w/reduced CD4 count
Latent/reactivated TB: (only 10%, usually < 2 yrs)
-commonly @ apex of lung, where lesions become necrotic
-undergo caseous necrosis and merge into larger lesions; center of necrosis is liquid and
leaks into bronchi
-again, creation of lung cavities, great for proliferation
-pulmonary disease w/fever, weight loss, non-productive cough which can become
productive/blood
-extrapulmonary: enlarged LN, pleural effusion, osteomyelitis, meningitis (immune response,
not bacterial invasion)
Risk: HIV w/reduced CD4 count, malnourished, meds (steroids), immunocompromised
Pathogenesis &
Damage
DAMAGE is primarily from HYPERSENSITIVE host IMMUNE response
-cell-mediated immune response involving CD4 T-cells making IFN-g
-TB-specific lymphocytes grow to activate more macs to kill
-leads to hypersensitivity (T-cells) but also cellular immunity (macs)
Borrelia burgdorferi
-spirochete
-inner membrane, peptidoglycan, LOOSE
outer membrane, and NO LPS!
-MUST scavenge building blocks from HOST
-main reservoir=infected rodents
-Ixodes tick nymphs/larvae as vectorbites
rodent, carries bacteria in gut
-adults hang onto deer
-tick bites humanif not removed by day 3,
spirochetes replicate in tick
-hosts protease plasmin in blood helps
spirochetes disseminate
**high risk areas=Northeast, Upper Midwest
**common in spring-summer-fall
A multistage disease:
0. Inoculation (2-7 days, goes to dermis)
1. Localized infection (days to weeks)
-bulls eye rash (erythema migrans) @ bite
-mild, flu-like symptoms
2. Disseminated infection (wks to months)
-bacteria transiently enter blood and spread
to other tissues
-intermittent symptoms of arrhythmias,
multiple erythema migrans, meningitis
3. Late stage (months to years)
-long-term chronic manifestations in joints,
CNS, or skin pauciarticular arthritis,
encephalopathy (rare, but pain/numb,
memory issues)
DAMAGE is mostly from immune response
**no known toxins, mostly bystander injury
@ stage 1: erythema migrans reflects
Immune Response &
Pathogenic Evasion?
Special Consideration
Diagnosis (Dx)
Treatment (Rx)
Prevention?
**delicate balance of killing bacteria and avoiding hyper
PROBLEM: granuloma (APC+lymphos) formed w/acute inflammation as a way of walling off
the bacteria
-tubercle center become necrotic (caseous necrosis),unable to fully kill so bacteria grow !!l
-granulomas also lead to SCARRING
-like Chlamydia, they survive well inside the host immune cells
-TB specifically prevents acidification of the phagolysosome
In HIV patients, treatment with drugs that increase CD4 T-cell levels can exacerbate TB
symptoms due to restoration of immune response
1) Acid-fast stain of sputum for active TB
2) Culture (slow but best) and 3) PCR (faster) for active TB
*2+3 help w/testing for antibiotic susceptibility
4) Tuberculin skin test (TST) to test for immune response to TB
(active AND latent TB, false negatives are common for immunocompr)
-measure of APC recruitment @ skin site of antigen injection (PPD)delayed
hypersensitivity rxn
**cannot use if got BCG vaccine
5) Interferon- release assay also tests for immune response to TB
(active AND latent TB, avoids problem of BCG/other mycobacteria)
-measures IFN-g immune response by stimulating persons T-cells w/TB protein
*4 and 5 are similar
For active TB, need to use combination antimicrobial therapy:
-many TB cells in lungs, each is a little different and resists dif drugsone strain likely not
resist two antibiotics, so hit with multiple to kill more bacteria
Examples: isoniazid, rifampin, ethambutol, etc.
Treat for 6 to 9 months
May be prophylactic for latent TB
For latent TB (fewer bacteria)use single drug for 9 months
Overall, problems with resistancehard to treat
Airborne precautionsfor small, dry particles that travel in air
**use private hospital room w/neg pressure
**provider wears N-95 respirator; patient wears surgical masks
inflammation @ skin
@ stage 2: bacteria @ vessel walls (some
direct BV damage) but lots of inflammation
@ stage 3: can trigger autoimmunity
prophylaxis w/long-term antibiotics>
-fairly adaptablewhen going from tick to
human, changes surface protein structure
and gene expression profile
-makes it hard to develop immuntiy
-hard to culture spirochetes; also few
-serology for antibodies against protein
antigens (need both to make a positive
diagnosis)
1st) ELISAuse protein antigens to
detect patient antibodies
(false positives are common)
2nd) Western Blotsmear antigens on
gel, run patient antibodies against it;
need several antibodies to be positive
(this can be expensive)
- hard to do w/early diseasejust use
clinical criteria!
Treat with antibiotis (doxycycline,
amoxicillin, cephalosporins)
Treat for longer if later stage (neurological
or arthritic issues)begin w/tougher
antibiotics but can switch to oral once
controlled
Microbiology 8: Introduction to Viruses
Poliovirus
Pathogenic Agent
-icosahedral, unenveloped virion
-single-stranded, positive-strand RNA genome
Part of Picornaviridae family (pico=small)
-3 serotypes, IDd by antibodies against capsid proteins (VP1, VP2, VP3all common so
VACCINE)
Enterovirus D68
-another RNA virus that infects CNS
-found in 1960s; saw cytopathic effect in monkey
kidney cells
Source/Transmission/
Entry
Manifestation of
Disease
Pathogenesis &
Damage
Immune Response
Diagnosis (Dx)
Treatment (Rx)
Prevention?
Transmitted via shedding in fecesfecal oral!
Life cycle:
Especially in warm weather
1. enter cell via endocytosis or just deliver genome into cell
-ingested and replicates in mucosa of pharynx and gut
2. uncoat, then translate (+)RNA to polyprotein
(tonsils, Peyers patches)
3. viral-encoded proteases cut this into 7 proteins for RNA
-spread via lymphatics to extraneural tissues
replication and protein cleavage; 3 proteins for capsid
-via viremia (@blood, symptoms aka prodrome), gets to CNS
assembly
(blood and nerve fibers)
4. viral enzyme replicase copies (+)RNA into (-)RNA, then
-targets motor neurons (anterior horn of SC)
progeny (+), which are encapsulated
Consider: needs to cross BBBvia glial cells?
Or intestinal part of vagus nerve?
Poliomyelitis=inflammation of CNS gray matter
Causes a polio-like illness acute flaccid myelitis,
Severity ranges from:
just like polio
1) Inapparent infection (90-95%)
Based on clinical case:
2) Abortive polio (4-8%)fever, sore throat, headache
-headache, fever, sore throat, tender neck
3) Aseptic meningitis (non-paralytic, 1%)more headache, higher fever, stiff neck,
-progressive weakness in arms, myalgia, cant walk
CSF pleocytosis
-cardiac arrest
4) Paralytic polio (0.1%) begin w/stage 2+vomiting (for a few days) 3 (5
-kids showed CSF pleocytosis aka more
days)flaccid limb paralysis (due to nerve damage, possible encephalitis)
lymphocytes @ CSF
-showed lesions in grey matter (anterior horn and
-Incubation period of 7-14 days
Worst kind=bulbar poliomyelitis (@medulla oblongata, hurts respiratory muscles)
brain stem)
-targets anterior spinal cord motor neurons; leads to necrosis: it inhibits host cells
**inflammation of anterior spinal cord and
macromolecular synthesis, which kills cell and leads to lysis
neuronal degeneration
**Viral replication in itself generally involves lysis
-body produces neutralizing antibodies against capsid proteinsthis is serotype-specific
Can be found in cerebrospinal fluid (CSF)
Dx in CSF using RT-PCR + sequencing
-recent outbreak in 2014 is a novel strain
Not in reading material
1) Oral Polio Vaccine (live, attenuated virus, OPV)grown in
2) (Parenteral) Inactivated poliovirus vaccine (IPV) grown in
culture until lose neurovirulence (attenuating mutations affect monkey kidney cells, inactivate infectivity w/formaldehyde
translation of viral RNA)
-works on all 3 serotypes
-works for all 3 serotypes (trivalent)
-need multiple inoculations+boosters
-multiple doses needed
Fxn: wont protect gut (no IgA) but protect against viremia (IgG
Fxn: produce circulating antibody (IgG) to neutralize + local
neutralization)
immunity via IgA
(+) strong humoral immune response
-attenuated virus replicates well in gut (leading to response)
(+)thought to be a safer vaccine (dead)
but not nervous system
(+) incorporate with other vaccines (DTP)
(+) mucosal and humoral immunity
(-) must be parenteral injection
(+) life-long immunity
(-) no mucosal immunity
(+) easy to give
(-) QC needed for potency, inactivation
(+) cheap
(-) more expensive
(-) virus could mutate and be active
(-) needs to stay cold for transport
