MD-2021 | Manila Central University S.Y.

2018-2019

PATHOGENESIS
MICROBIOLOGY Toxigenic
AEROBIC GRAM-POSITIVE BACILLI: Diphtheria toxin inactivates EF-2 by catalyzing a reaction that
Corynebacterium, Listeria, Bacillus yields free nicotinamide + inactive adenosine diphosphate-EF-
Based on the lecture of Dr. DIzon | November 8, 2018 2 complex (ADP-ribosylation)
→ results to ARREST IN PROTEIN SYNTHESIS

GRAM-POSITIVE BACILLI 1. Diphtheria toxin’s receptor-binding domain B binding

host membrane
o Aerobic Gram-positive bacilli with high G + C content 2. Membrane bound toxin A +B enters endocytosis
and irregular shape 3. Catalytic subunit A is cleaved but held to the B
o Corynebacterium subunit by disulfide bonds. Endosome vesicle
o Aerobic Gram-positive bacilli with low G + C content acidifies; the disulfide bonds are reduced
and regular shape 4. The transmembrane domain facilitates passage of the
o Listeria and Bacillus catalytic A peptide through the vesicle membrane
5. The catalytic A domain ADP-ribosylates elongation
factor 2 (EF-2). This halts protein synthesis and kills
CORYNEBACTERIUM the cell
- Corynebacterium diphtheriae
Exotoxin effects
- Non-lipophilic
- Degeneration, fatty infiltration and necrosis of the
Fermentative
heart muscles, kidneys, liver and adrenals
C. ulcerans
- Nerve damage causing paralysis of the soft palate,
-cause disease similar to diphtheria
eye muscles and limbs
C. pseudotuberculosis - rarely cause disease
Non-fermentative
IN VIVO TEST FOR TOXIGENITY
C. auris
Done on guinea pig
- Lipophilic
C. jeikium • Mark skin into two 2 cm squared area
C. urealyticum • Inject IC 0.2 ml of pure culture of C.
- Other coryneform diphtheriae to square 1
Arcanobacterium haemolitycum - pharyngitis • After 5 hours inject 500 units of diphtheria
Rothia dentocariosa antitoxin IP
Stomatococcus mucilaginosis • Wait for 30 minutes
• Inject 0.2 mL of the remaining broth culture
• Intracutaneously into square 2
CORYNEBACTERIUM DIPTHERIAE
(Klebs-Loeffler bacillus) IN-VITRO TEST FOR TOXIGENICITY
➢ Subspecies – based on colony morphology, ELEK’S TEST
biochemical reaction, severity of the disease
o Gravis
o Mitis
o Intermedius
o Belfanti

Characteristics:
- High G + C
- Gram + clubbed or irregularly shaped, non-motile
bacilli; “Chinese-like characters” on smear
- Non-spore former

Metachromatic Granules
Babes-Ernst granules or Volutin granules
o Inorganic polyphosphates which serves as
depot for materials …
- strip than contains the antitoxin is placed into a pure culture
Virulence Factor of Corynebacterium diphtheriae
Exotoxin ➔ (+) reaction: formation of precipitate
o Inhibits elongation factor-2 (EF-2) o Ab from anti-toxin reacts with
o Heat-labile polypeptide antigen in the culture
o MW – 62, 000
o Lethal dose – 0.1 ug/kg

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 MD-2021 | Manila Central University S.Y. 2018-2019

LISTERIA
SCHICK’s TEST
TEST ARM CONTROL ARM - first discovered by Joseph Lister
Intracutaneous Heat inactivated - Food-borne (vegetables salads, sushi, cheese)
Monitor within 22-36hours .2 mL injected
10 to 15 mm diameter red No change occurs Epidemiology
flash - 5 to 10% in stools of normal adults
- 80% in raw poultry (major reservoir of infection for
Reactions for Schick’s Test: humans)
POSITIVE - Raw milk from infected cows (may escape normal
- 5 to 10 mm diameter; appears 4 to 7 days after the pasteurization process)
test is done. Control arm show no reaction - Most important specie: L. monocytogenes
- Susceptible - 1985 outbreak
o 49 persons (29% mortality) developed
PSEUDOPOSITIVE listeriosis from pasteurized milk
- Red colored inflammation (erythema) on both arms - In 1989
which disappears within 4 days o 90 deaths due to ingestion of contaminated
- Immune but hypersensitive to the toxin Mexican-style cheese
- In 1993
o 20% of hotdogs were contaminated
NEGATIVE - Another epidemic
- No reaction; patient is immune o 40 persons affected by eating coleslaw

COMBINED REACTION LISTERIA MONOCYTOGENES

- Same as pseudo-reaction but fades off in the control Characteristics:
arm but progresses in the test arm as typical positive ➢ Intracellular bacterium capable of growing and
- Susceptible and hypersensitive surviving inside the mononuclear phagocytes and
epithelial cells
DIPHTHERIA ➢ Resists the effects of freezing, drying and heat
Most common site of infection: pharynx ➢ Short gram-positive bacilli
➢ Tumbling motility at 22-28 deg C but not at 37 deg C
Manifestations: ➢ Catalase and esculin hydrolysis positive
- Erythema ➢ Survive temperature of 4 deg C under condition of low
- Presence of graying pseudo-membrane (made up of pH and high sold
dead tissues and inflammatory cells) ➢ Cold enrichment
o Pseudomembrane may infect the epiglottis ➢ Facultatively aerobic
causing respiratory distress ➢ Hemolysis (+) – beta hemolysis
- Enlargement of the lymph nodes “bull neck”
appearance Serotypes
- Fever, body malaise - 1/2a
- Involvement of the nose: erythema - 1/2b
- Skin: ulceration with large area of erythema - 4b
- 4b → causes food -borne infection
DIAGNOSIS
➢ Clinically Pathogenic determinants
➢ Gram-staining - Listeriolysin O (LLO)
➢ Culture isolation - Ability to induce their own
o Blood agar - Ability to overcome nonspecific IR mediated by
o Modified Tinsdale Agar phagocytes
o Cystine tellurite (black colonies) - Zinc- dependent phospholipase C --- its loss results in
impaired virulence
TREATMENT - Binds C1q – enhanced uptake by c1q-binding
➢ Antitoxin receptor son macrophages
o Needs to be neutralized - Possesses a LPS-like component in the cell wall →
➢ Penicillin/macrolide symptoms associated with endotoxins due to..
o To prevent reproduction of the organism
➢ Prevention Immune response
o Immunization (DPT) - Induce infected macrophage to secrete Il-12 which
▪ 3 doses starting 6 weeks old drives the differentiation of T cells towards Th1
▪ Booster: 1 year after the last dose, development → CMI
4 to 6 y/o then every 10 years

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 MD-2021 | Manila Central University S.Y. 2018-2019

- IFN-y may activate the listericidal mechanisms od o Do not use refrigerated foods beyond due
macrophages dates
o Keep fridge clean
CAUSES of LISTERIOSIS ➢ Food Temperature
RISK FACTORS o Must be below 5 deg C
➢ Older adults >50 y/o o Keep hot foods hot (above 60 deg C) and
➢ Pregnant women cold foods cold
➢ People with weakened immune systems, organs
transplant patients on immunosuppressive drugs
BACILLUS
HOST CELLS: M cells and Peyer’s patches
o Reheat stored foods
Adhesions molecules are also found in the blood-brain barrier
- Gram positive; spore former
and in the fetal-placental barrier
- May produce hemolysis (B. cereus)
- Liquefies gelatin (inverted fir tree appearance)
Target cells: macrophages and epithelial cells
- Aerobic or facultative
- Has a receptor called E-cahedrin which cause
activation of Rho- GTPase → WAsp (Wiskott-Aldrich)
Species:
→ Arp 2/3 complex → ActA (Actin-assemble inducing
➢ B. anthracis
protein) → actin polymerization → para cytophagy
➢ B. cereus
Paracytophagy
BACILLUS ANTHRACIS
- mediated by filopods; exotoxin will have double
- No hemolysis in blood agar
membrane before it will be released by the LLO,
Phospholipase B to cause infection on the body
Virulence determinants
2 plasmids which codes for:
LISTERIOSIS
- Capsule
- Incubation period 3 to 7 days
o Has poly-D-glutamic acid
- Clinical forms
- Toxin
o Perinatal
ANTHRAX TOXIN
▪ Early onset syndrome
COMPONENTS FUNCTION
• Granulomatis
EF (edema factor) Inactive cAMP activated by
infantiseptica – early onset
calmodulin
• Late onset syndrome –
LF (lethal factor) Causes pulmonary edema
meningitis
and death in rats; cytolytic
o Adults
for macrophages
▪ Meningoencephalitis
▪ Etc. PA (protective antigen) Required for the binding of
both EF and LF to the host
Diagnosis cell
➢ Clinical
➢ Culture EF + PA = edema toxin increases cAMP in PMN thus inhibiting
phagocytosis
Treatment LF + PA = severe pulmonary edema; cytolysis of macrophages
➢ Ampicillin
➢ Erythromycin Causes Anthrax
➢ Trimethoprim sulfamethoxazole ➢ Cutaneous 95%
o 1 to 12 days incubation period
Prevention o Malignant pustule
➢ Hints in food handling o 20% mortality if not treated with appropriate
o Wash hands antibiotics
o Wash raw fruits and vegies o 1% if treated
o Cook all foods of animal origin ➢ Woolsorter’s Disease (respiratory/inhalational)
o Don’t use the same boards and knives for o Common in people working in sheep’s farm
raw and cooked food o Incubation period 2 days to 6-8 weeks
➢ Food storage o Meningitis
o Defrost food by placing it on the lower o CXR: mediastinal widening, bloody fluids
shelves of the fridge or use a microwave around the lunds
➢ Store Food with Car o 90% lethal if untreated; 50% if treated
o Keep food covered ➢ Gastric Disease
o Unfinished foods must be place in fridge o Incubation period 1 to 6 days
within one hour o Oropharyngeal or intestinal
▪ Nausea, diarrhea, abdominal pain
o >40% mortality

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 MD-2021 | Manila Central University S.Y. 2018-2019

➢ Injection Anthrax
o Usually in drug addicts who use needles
o 1 to 2 days incubation after injection
o Inflammation, abscess, may progress to
sepsis even without extensive local infection
o 30% mortality

*anthrax is an animal disease which is accidentally

transmitted to humans (usually from cattle)

*B. anthracis was used long ago for bioterrorism

Diagnosis
➢ Direct smears
➢ Blood culture
➢ Guinea pig test

Treatment
➢ Ciprofloxacin
➢ Gentamicin

Prevention
➢ Disposal of animal carcass in burning pits or deep
burial lime pits
➢ Decontamination of animal products
➢ Protecting clothing and gloves
➢ Active immunization of both animals and humans
o Ideal vaccine: PA toxin (binds both EF and
LF)

BACILLUS CEREUS
- Beta hemolytic
- Facultative anaerobe
- Cause food-borne infection → enterotoxin
- Necrotizing fasciitis

Source of Infection
➢ Uncooked rice
o Chinese/Japanese restaurant syndrome
➢ Meat and meat products
➢ Cream or pudding

Foodborne infections
- Due to the enterotoxin
o Emetic – heat labile
o Diarrheal type – heat stable

Prevention
➢ Prompt refrigeration of boiled rice and dried foods

Management
➢ No need to give antibiotic because the causative
molecule is an enterotoxin not the bacteria itself
➢ Rehydrate patient

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©icacomedian 4