9
Prepared by: EGBII; 8-13-11 GRAM POSITIVE BACILLI DANILO D. DEVEZA JR., M.D. Spore-Forming Gram Positive Bacilli Bacilli species - aerobic Clostridium species non aerobic Bacillus species A. Bacillus anthracis B. Bacillus cereus Bacilli species Large aerobic, gram positive rods in chains Most are saprophylic Most do not cause disease Important diseases: Anthrax, Food Poisoning Spores located in center Non-motile “Cut glass” appearance in culture Use nitrogen & carbon for energy Spores are resistant to environmental changes, dry heat, certain disinfectants 1. Bacillus anthracis Anthrax: primarily disease of herbivores Humans are infected incidentally Acquired by the entry of spores Injured skin (cutaneous anthrax) Mucous membrane (gastrointestinal anthrax) Inhalation (inhalation anthrax) PATHOLOGY Spores germinate in tissues at the site of entry Formation of edema & congestion Spread via lymphatics Three Anthrax toxin Protective antigen (PA) Edema factor (EF) Lethal factor (LF) PA binds to specific cell receptors: entry of EF & LF EF + PA = edema toxin LF + PA = lethal toxin (virulence factor) CLINICAL FINDINGS Humans: 95% (cutaneous), 5% (inhalation) Cutaneous anthrax 1-7 days: pruritic rash Papule → vesicle → necrotic ulcer Lesion has central black eschar Edema, lymphangitis, lymphadenopathy Healing by granulation & leaves a scar Incubation period: 6 weeks Inhalation (Wool sorter’s disease) Hemorrhagic necrosis & edema of mediastinum Pleural effusion Sepsis GI: bowel ulceration Brain: hemorrhagic menigitis DIAGNOSTICS Specimem: fluid from local lesion, blood & sputum Chains of large gram positive rods Blood agar: non-hemolytic gray to white colonies, with comma shaped outgrowths (Medusa head) TREATMENT Ciprofloxacin: recommended Penicillin G, Gentamycin, Streptomycin PREVENTION Proper disposal of animal carcasses Decontamination of animal products Protective handling of potentially infected materials Active immunization of domestic animals 2. Bacillus cereus Produce toxins that cause disease Spores germinate, vegetative cells produce toxins CLINICAL FINDINGS Emetic type (rice) Nausea, vomiting, abdominal cramps Self limiting Diarrheal type (meat dishes & sauces) Diarrhea w/ abdominal pain & cramps Enterotoxin : pre-formed or produced in the intestine Eye infections Organisms are introduced by foreign bodies Local & systemic infections Endocarditis, meningitis, osteomyelitis Presence of medical device (IV lines) & IV drugs Presence of B. cereus in stool is not diagnostic 10 5 bacteria or more per gram of food is diagnostic Culture: exhibit motility “swarmingTREATMENT Drug of choice: vancomycin or clindamycin Resistant to penicillins & cephalosporins Clostridium Species Clostridium botulinium Clostridium tetani Clostridium perfringens Clostridium difficile Clostridium Species Large, anaerobic, gram (+), motile rods Decompose proteins or form toxins or both Spore is place centrally, sub-terminally or terminally Produce large raised colonies (C. perfringens) Small colonies (C. tetani ) Many produce hemolysis on blood agar (C. perfringens: double zone) Ferment a variety of sugars Many digest proteins 1. Clostridium botulinium CHARACTERISTICS Causes Botulism Found in soil Produce toxins: Type A, B, E, occasionally F: human illness Among the most toxic substances Destroyed by heating for 20 mins at 100C PATHOGENESIS Illness is not infection INTOXICATION Toxins acts by blocking the release of acetylcholine at synapses & neuromuscular junctions FLACCID PARALYSIS CLINICAL FINDINGS Visual disturbances Dysphagia Speech difficulty Signs of bulbar paralysis Cause of death: respiratory paralysis or cardiac arrest

Sporeforming & Non-Spore Forming Bacteria

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Page 1: Sporeforming & Non-Spore Forming Bacteria

Prepared by: EGBII; 8-13-11

GRAM POSITIVE BACILLI DANILO D. DEVEZA JR., M.D.

Spore-Forming Gram Positive Bacilli Bacilli species - aerobic Clostridium species – non aerobic

Bacillus species

A. Bacillus anthracis B. Bacillus cereus Bacilli species

• Large aerobic, gram positive rods in chains • Most are saprophylic • Most do not cause disease • Important diseases: Anthrax, Food Poisoning • Spores located in center • Non-motile • “Cut glass” appearance in culture • Use nitrogen & carbon for energy • Spores are resistant to environmental changes, dry heat,

certain disinfectants

1. Bacillus anthracis • Anthrax: primarily disease of herbivores • Humans are infected incidentally • Acquired by the entry of spores

• Injured skin (cutaneous anthrax) • Mucous membrane (gastrointestinal anthrax) • Inhalation (inhalation anthrax)

PATHOLOGY

• Spores germinate in tissues at the site of entry • Formation of edema & congestion • Spread via lymphatics • Three Anthrax toxin

• Protective antigen (PA) • Edema factor (EF) • Lethal factor (LF)

• PA binds to specific cell receptors: entry of EF & LF • EF + PA = edema toxin • LF + PA = lethal toxin (virulence factor)

CLINICAL FINDINGS

• Humans: 95% (cutaneous), 5% (inhalation) • Cutaneous anthrax

• 1-7 days: pruritic rash

• Papule → vesicle → necrotic ulcer • Lesion has central black eschar • Edema, lymphangitis, lymphadenopathy • Healing by granulation & leaves a scar

• Incubation period: 6 weeks • Inhalation (Wool sorter’s disease)

• Hemorrhagic necrosis & edema of mediastinum • Pleural effusion

• Sepsis • GI: bowel ulceration • Brain: hemorrhagic menigitis

DIAGNOSTICS

• Specimem: fluid from local lesion, blood & sputum • Chains of large gram positive rods • Blood agar: non-hemolytic gray to white colonies, with

comma shaped outgrowths (Medusa head)

TREATMENT

• Ciprofloxacin: recommended • Penicillin G, Gentamycin, Streptomycin

PREVENTION

• Proper disposal of animal carcasses • Decontamination of animal products • Protective handling of potentially infected materials • Active immunization of domestic animals

2. Bacillus cereus • Produce toxins that cause disease

• Spores germinate, vegetative cells produce toxins

CLINICAL FINDINGS

• Emetic type (rice) • Nausea, vomiting, abdominal cramps • Self limiting

• Diarrheal type (meat dishes & sauces) • Diarrhea w/ abdominal pain & cramps • Enterotoxin : pre-formed or produced in the

intestine • Eye infections

• Organisms are introduced by foreign bodies • Local & systemic infections

• Endocarditis, meningitis, osteomyelitis • Presence of medical device (IV lines) & IV

drugs • Presence of B. cereus in stool is not diagnostic • 10

5 bacteria or more per gram of food is diagnostic

• Culture: exhibit motility “swarming”

TREATMENT

• Drug of choice: vancomycin or clindamycin • Resistant to penicillins & cephalosporins

Clostridium Species

Clostridium botulinium Clostridium tetani Clostridium perfringens Clostridium difficile Clostridium Species

• Large, anaerobic, gram (+), motile rods • Decompose proteins or form toxins or both • Spore is place centrally, sub-terminally or terminally • Produce large raised colonies (C. perfringens) • Small colonies (C. tetani) • Many produce hemolysis on blood agar (C. perfringens:

double zone) • Ferment a variety of sugars • Many digest proteins

1. Clostridium botulinium CHARACTERISTICS

• Causes Botulism • Found in soil • Produce toxins:

• Type A, B, E, occasionally F: human illness • Among the most toxic substances • Destroyed by heating for 20 mins at 100⁰C

PATHOGENESIS

• Illness is not infection • INTOXICATION • Toxins acts by blocking the release of acetylcholine at

synapses & neuromuscular junctions • FLACCID PARALYSIS

CLINICAL FINDINGS

• Visual disturbances • Dysphagia • Speech difficulty • Signs of bulbar paralysis • Cause of death: respiratory paralysis or cardiac arrest

Page 2: Sporeforming & Non-Spore Forming Bacteria

Prepared by: EGBII; 8-13-11

TREATMENT

• Antitoxins (A, B and E) • Supportive • Ventilation

2. Clostridium tetani • Characteristics

• Gram (+) bacilli, terminal spores • Obligate anaerobes, motile

• Reservoir • Soil/ feces of animals

• Transmission • Puncture wounds/trauma • Requires low tissue oxygenation (Eh)

• LOCALIZED

PATHOGENESIS

• Spores germinate in the tissues: tetanospasmin • A fragment: blocks NT release at inhibitory

synapses • B fragment: mediates binding to neuron and cell

penetration of A fragment • Carried intra-axonally to CNS • Binds to ganglioside receptors • Blocks release of inhibitory mediators (glycine and

GABA) at spinal synapses

• Excitatory neurons are unopposed → extreme muscle spasm

CLINICAL FINDINGS

• Incubation period: 4 – 5 days (up to weeks) • Tonic contraction of voluntary muscles • Spasm first in area of injury, then the muscles of the jaw • External stimuli may precipitate muscle spasm • Death: spasm of respiratory muscles

TETANUS

• Trismus, risus sardonicus, opisthotonus

DIAGNOSIS: Clinical *

TREATMENT

• Hyperimmune human globulin (TIG) to neutralize toxin • + Metronidazole or Penicillin • Spasmolytic drugs (diazepam), debride, delay course

PREVENTION

• DTP, DTaP, Td • TIG • Proper wound care

Wound Management

3. Clostridium perfringens CHARACTERISTICS

• Encapsulated • Non motile • Double hemolysis • Ferment CHO • Reservoir: Soil and human colon • INVASIVE

TOXINS

• Alpha toxin (Lecithinase) • Necrotizing & hemolytic effect

• Theta toxin • Necrotizing & hemolytic effect

• DNase & Hyaluronidaes • Digest collagen

• Enterotoxin • Alters cell membrane, disrupting ion transport

PATHOGENESIS

• Spores germinate under anaerobic conditions in tissues • Distention of tissues &interference of blood supply,

presence of toxins • Spread of infection • Tissue necrosis • Hemolytic anemia • Severe toxemia

• Gas gangrene (clostridial myonecrosis) • Mixed infection • Toxigenic & proteolytic clostridia • Various cocci & gram negative bacteria

CLINICAL FINDINGS (Gas Gangrene)

• Spreads in 1-3 days • Crepitation & subcutaneous tissues & muscles • Foul smelling discharge • Necrosis, fever • Toxemia → shock → DEATH

CLINICAL FINDINGS (food poisoning)

• Enterotoxin forms in GUT • Diarrhea without vomiting or fever in 6-18 hours • Resolves in 1-2 days

DIAGNOSIS

• Clinical • Laboratory

• Culture • Chopped meat • Growth + gas • BAP • Target or double zone of hemolysis

DISEASES

• Gas gangrene (myonecrosis) • Anaerobic cellulitis • Food poisoning (alpha toxin) • Necrotic enteritis (fire in the bowel)

Page 3: Sporeforming & Non-Spore Forming Bacteria

Prepared by: EGBII; 8-13-11

• Clostridial endometritis

TREATMENT

• Surgical debridement: most important • Antibiotics: Penicillin • Food poisoning: supportive

4. Clostridium difficile PSEUDOMEMBRANOUS COLITIS

• Drug resistant C. difficile produce toxins • Toxin A: enterotoxin • Toxin B: cytotoxin

• Pseudomembranes/ microabscesses in bowels • Watery/ bloody diarrhea • Clindamycin, Cephalosporins, Amoxicillin, Ampicillin

TREATMENT

• Discontinue offending antibiotic • Metronidazole or vancomycin

ANTIBIOTIC-ASSOCIATED DIARRHEA

• Administration of antibiotics leads to mild to moderate diarrhea

• Less severe than pseudomembranous colitis • 25% associated with C. difficile

DIAGNOSIS

• Toxins (stools) • Endoscopic exam

Non-Spore Forming

Gram Positive Bacilli Listeria Erysipelothrix Actinomycetes

1. Listeria monocytogenes • Gram positive • Short rod • Catalase positive • Beta hemolysis on BA • Tumbling motility at 22 – 28

oC

• Diffrentiates from diptheroids • Widespread of disease in human & animals • Important food-borne pathogen

• Survive in refrigerator temperature (4oC)

• Survive low pH • Survive high salt conditions • Overcome food preservation and safety barriers

• Antigenic Classifications • 1/2a • 1/2b • 4b: causes most food-borne outbreaks

PATHOGENESIS

• Ami, Fbp A, Flagellin protein • Bind to host cells & virulence • Internalin A

• Interacts with E-cadherin: cell well protein in epithelial cells

• Promote phagocytosis • Listeriolysin O: enzyme

• Lyses membrane, bacteria escape to cytoplasm

• Act A: surface protein • Induces host cell actin polymerization • Propels bacteria to cell membrane • Cause formation of Filipods

• Filipods • Ingested by epithelial cells,

macrophages & hepatocytes • Lifecycle begins abain

RESERVOIR

• Widespread: animals (gastrointestinal and genital tracts), • Unpasteurized milk products • Plants and soil • Cold growth: soft cheeses, deli meats, cabbages

(coleslaw)

PERINATAL HUMAN LISTERIOSIS

• Early onset syndrome • Granulomatosis infantseptica • Infection in utero • Neonatal sepsis, pustular & granulomas • Death; before / after delivery

• Late onset syndrome • Meningitis: birth to 3

rd week of life

• Caused by serotype 4b • High mortality rate

ADULTS

• Listeria meningoencephalitis • Immunocompromised • Insidious to fulminant

• Bacteremia

• Focal infections

TREATMENT

• Ampicillin, Erythromycin, IV Trimethoprom-sulfamethoxazole

• Ampicillin + Gentamycin – recommended

PREVENTION

• Precautions with food may reduce incidence

2. Erysipelothrix rhusopathiae Distinguishing Characteristics:

• Gram positive bacilli, non-branching • Catalase (-), Oxidase (-), Indole (-) • Alpha hemolysis on BA • Produced H2S on TSI (Butt Black)

• Differentiated from L. monocytogenes • Distributed in land & sea animals • Transmitted by direct inoculation • Greatest risk: fishermen, fish handlers, butchers, those in

contact with animal products ERYSIPELOID/ SEA FINGER/ WHALE FINGER

• Icubation Period: 2-7 days • Raised lesion w/ violaceous color • Pain & swelling • No pus

TREATMENT

• Penicillin G

3. Actinomycetes • Aerobic • Large diverse group of gram positive bacteria • Form chains or filaments • Categories:

• Acid fast positive: Mycobacteria • Weakly positive: Nocardia & Rhodococcus • Acid fast negative: Streptomyces

&Actinimadura

4. Nocardia Human infection: N. nova complex

• N. farcinica • N. asteroides type IV • N. brasiliensis

Page 4: Sporeforming & Non-Spore Forming Bacteria

Prepared by: EGBII; 8-13-11

DISTINGUISHING CHARACTERISTICS

• Aerobic • Gram-positive branching rods • Catalase positive • Partially acid-fast • Produce urease • Can digest paraffin

• Inhalation of bacteria

• Not transmitted from person to person • Opportunistic infection

• Corticosteroid treatment, immunosuppression, organ transplantation, AIDS, TB

• Begins with lobar pneumonia • Mimic TB • Granulona & caseation are rare

• Abscess formation: usual pathologic process • Spread to CNS, skin, kidney & eyes

TREATMENT

• Trimethoprim-Sulfamethoxazole • Treatment of choice

• Amikacin, imipenem, minocycline, linezolide & ceftaxime • Surgical drainage

Actinomycetoma - mycetomal disease affecting the skin and connective tissue Mycetoma (Madura Foot)

• Most Common cause: • N. asteroides, N. brasiliensis, Streptomyces

somaliensis & Actinomadura madurae • Localized, slowly progressive chronic infection • Begins in subcutaneous tissues & spreads to adjacent

tissues • Destructive & painless

ANAEROBE BACTERIOLOGY

Definition of Terms

Aerobic Bacteria – require oxygen, and will not grow in the absence of oxygen

Anaerobic Bacteria – do not use oxygen for growth & metabolism but obtain their energy from fermentation

reactions

Capnophilic Bacteria – require CO2 for growth

Facultative Anaerobes – can grow either oxidatively or use fermentation reactions to obtain energy

FACTORS THAT INHIBIT GROWTH OF ANAEROBES BY OXYGEN

• Toxic compounds are produced • e.g. H2O2 , Superoxides

• Absence of catalase & superoxide dismutase • Oxidation of essential sulfhydyl groups in enzymes without

sufficient reducing power to regenerate them

ANAEROBIC BACTERIA OF CLINICAL IMPORTANCE

Bacilli Gram-negative

• Bacteriodes fragilis • Prevotella

melaninogenica • Fusobacterium

Gram-positive • Actinomyces • Lactobacillus • Clostridium

Cocci Gram-positive

• Peptostreptococcus • Peptococcus

Gram-negative • Veilonella

PATHOGENESIS OF ANAEROBIC INFECTIONS

• Polysaccharide capsule • Ability to induce abscess formation

• Lipopolysaccharide • Endotoxin: lack lipopolysaccaride structures with

endotoxic activity • Not directly produce clinical signs of sepsis

• Enzymes • Proteases, Neuraminidases, Cyclolysins • Cause hemolysis of erythrocytes • Damage and destroys tissues

• Superoxide dismutase (some bacteria) • Can survive in the presence of oxygen for days

DIAGNOSIS OF ANAEROBIC INFECTIONS

• Foul-smelling discharge (pus) • Short-chain fatty acid products of anaerobic

metabolism • Infection in proximity to a mucosal surface

• Anaerobes are part of normal flora • Gas in tissues

• Production of CO2 and H2 • Negative aerobic cultures

• Involve mixture of organisms • Form closed spaced infections or burrowing through tissues

(Lungs, brain, pleura, pelvis) • Most are susceptible to penicillin G

• Except: Bacteroides, some Prevotella species • Favored by reduce blood supply, necrotic tissues, low Eh

• Interfere with delivery of antibiotics • Uses special collection methods & transport media

METHODS USED TO PRODUCE ANAEROBIOSIS

• Gas Pak Jar, Brewer Jar, Torbal Jar • Cooked meat medium / Chopped cooked meat medium

sealed with petrolatum

• Anaerobic glove box & chamber PRAS Medium

• Pre-Reduced Anaerobically Sterilized medium

• e.g. Roll Tube of Hungate

• Thioglycollate medium • Aerobic • Anaerobic • Microaerophilic

• Resazurin Kanamycin-Vancomycin-Colistin (KVC) test

• Antibiotic Disks for the Presumptive Identification of Anaerobes

KVC PATTERN

• RRR- B. fragilis • SRS

• Fusobacterium • B. ureolyticus • Veillonella

• SSR- Clostridium, gram (+) cocci • RSR- Porphyromonas, P. anaerobius • RRS- Prevotella

RESPIRATORY TRACT

• Prevotella melaninogenica, Fusobacterium &

Peptostreptoccocus • Periodeontal infections, perioral abscess, sinusitis,

mastoiditis • Saliva aspiration: necrotizing pneumonia, lung

abscess & empyema CENTRAL NERVOUS SYSTEM

• Brain abscess, subdural empyema, septic thrombophlebitis • Originate from respiratory tract, spread hematogenously

INTRA-ABDOMINAL & PELVIC INFECTIONS

• Flora of colon: B. fragilis, Clostridia, Peptostreptococcus

• Infection due to perforated bowel • Prevotella originate from female genital organs

SKIN & SOFT TISSUES

• Anaerobes & aerobes: synergistic infections • Gangrene, necrotizing fasciitis, cellulitis

Anaerobic Infections

TREATMENT • Surgical drainage: most important • Antimicrobial Therapy

• Penicillin G • Clindamycin

Page 5: Sporeforming & Non-Spore Forming Bacteria

Prepared by: EGBII; 8-13-11

• Preferred for infections above diaphragm • Metronidazole

Page 6: Sporeforming & Non-Spore Forming Bacteria

Prepared by: EGBII; 8-13-11

Spore-Forming Gram Positive Bacilli Bacilli species - aerobic Clostridium species – non aerobic

A. Bacilli species – Spore forming bacilli

• Large aerobic, gram positive rods in chains • Most are saprophylic

• Most do not cause disease • Important diseases: Anthrax, Food

Poisoning

• Spores located in center • Non-motile • “Cut glass” appearance in culture

• Use nitrogen & carbon for energy • Spores are resistant to environmental

changes, dry heat, certain disinfectants

1. Bacillus anthracis

• Anthrax: primarily disease of herbivores • Humans are infected incidentally

• Acquired by the entry of spores • Injured skin (cutaneous anthrax) • Mucous membrane

(gastrointestinal anthrax) • Inhalation (inhalation anthrax)

PATHOLOGY

• Spores germinate in tissues at the site of

entry • Formation of edema & congestion • Spread via lymphatics

• Three Anthrax toxin • Protective antigen (PA) • Edema factor (EF)

• Lethal factor (LF) • PA binds to specific cell receptors:

entry of EF & LF

• EF + PA = edema toxin • LF + PA = lethal toxin (virulence

factor)

CLINICAL FINDINGS

• Humans: 95% (cutaneous), 5% (inhalation) • Cutaneous anthrax

• 1-7 days: pruritic rash

• Papule → vesicle → necrotic ulcer

• Lesion has central black eschar

• Edema, lymphangitis, lymphadenopathy

• Healing by granulation & leaves

a scar • Incubation period: 6 weeks • Inhalation (Wool sorter’s disease)

• Hemorrhagic necrosis & edema of mediastinum

• Pleural effusion

• Sepsis • GI: bowel ulceration

• Brain: hemorrhagic menigitis

DIAGNOSTICS

• Specimem: fluid from local lesion, blood & sputum

• Chains of large gram positive rods • Blood agar: non-hemolytic gray to white

colonies, with comma shaped outgrowths (Medusa head)

TREATMENT

• Ciprofloxacin: recommended • Penicillin G, Gentamycin, Streptomycin

PREVENTION

• Proper disposal of animal carcasses • Decontamination of animal products • Protective handling of potentially infected

materials • Active immunization of domestic animals

2. Bacillus cereus • Produce toxins that cause disease

• Spores germinate, vegetative

cells produce toxins

CLINICAL FINDINGS

• Emetic type (rice) • Nausea, vomiting, abdominal

cramps • Self limiting

• Diarrheal type (meat dishes & sauces)

• Diarrhea w/ abdominal pain & cramps

• Enterotoxin : pre-formed or

produced in the intestine • Eye infections

• Organisms are introduced by

foreign bodies • Local & systemic infections

• Endocarditis, meningitis,

osteomyelitis • Presence of medical device (IV

lines) & IV drugs • Presence of B. cereus in stool is not

diagnostic • 10

5 bacteria or more per gram of food is

diagnostic • Culture: exhibit motility “swarming”

TREATMENT

• Drug of choice: vancomycin or clindamycin

• Resistant to penicillins & cephalosporins

B. Clostridium Species

Clostridium botulinium Clostridium tetani

Clostridium perfringens Clostridium difficile

Clostridium Species • Large, anaerobic, gram (+), motile rods • Decompose proteins or form toxins or both

• Spore is place centrally, sub-terminally or terminally

• Produce large raised colonies (C.

perfringens) • Small colonies (C. tetani) • Many produce hemolysis on blood agar (C.

perfringens: double zone)

• Ferment a variety of sugars • Many digest proteins

1. Clostridium botulinium CHARACTERISTICS

• Causes Botulism • Found in soil • Produce toxins:

• Type A, B, E, occasionally F: human illness

• Among the most toxic

substances • Destroyed by heating for 20

mins at 100⁰C

PATHOGENESIS

• Illness is not infection

• INTOXICATION • Toxins acts by blocking the release of

acetylcholine at synapses &

neuromuscular junctions • FLACCID PARALYSIS

CLINICAL FINDINGS

• Visual disturbances

• Dysphagia • Speech difficulty • Signs of bulbar paralysis

• Cause of death: respiratory paralysis or cardiac arrest

TREATMENT

• Antitoxins (A, B and E)

• Supportive • Ventilation

2. Clostridium tetani • Characteristics

• Gram (+) bacilli, terminal spores • Obligate anaerobes, motile

• Reservoir

• Soil/ feces of animals • Transmission

• Puncture wounds/trauma

• Requires low tissue oxygenation (Eh)

• LOCALIZED

PATHOGENESIS

• Spores germinate in the tissues: tetanospasmin

• A fragment: blocks NT release at inhibitory synapses

• B fragment: mediates binding to

neuron and cell penetration of A fragment

• Carried intra-axonally to CNS

• Binds to ganglioside receptors • Blocks release of inhibitory mediators

(glycine and GABA) at spinal synapses

• Excitatory neurons are unopposed → extreme muscle spasm

CLINICAL FINDINGS

• Incubation period: 4 – 5 days (up to weeks)

• Tonic contraction of voluntary muscles • Spasm first in area of injury, then the

muscles of the jaw

• External stimuli may precipitate muscle spasm

• Death: spasm of respiratory muscles

Page 7: Sporeforming & Non-Spore Forming Bacteria

Prepared by: EGBII; 8-13-11

TETANUS

• Trismus, risus sardonicus, opisthotonus

DIAGNOSIS: Clinical *

TREATMENT

• Hyperimmune human globulin (TIG) to neutralize toxin

• + Metronidazole or Penicillin

• Spasmolytic drugs (diazepam), debride, delay course

PREVENTION

• DTP, DTaP, Td • TIG • Proper wound care

Wound Management

3. Clostridium perfringens CHARACTERISTICS

• Encapsulated • Non motile • Double hemolysis

• Ferment CHO • Reservoir: Soil and human colon • INVASIVE

TOXINS

• Alpha toxin (Lecithinase) • Necrotizing & hemolytic effect

• Theta toxin

• Necrotizing & hemolytic effect • DNase & Hyaluronidaes

• Digest collagen

• Enterotoxin

• Alters cell membrane, disrupting ion transport

PATHOGENESIS

• Spores germinate under anaerobic conditions in tissues

• Distention of tissues &interference of blood

supply, presence of toxins • Spread of infection • Tissue necrosis

• Hemolytic anemia • Severe toxemia

• Gas gangrene (clostridial myonecrosis)

• Mixed infection • Toxigenic & proteolytic clostridia • Various cocci & gram negative

bacteria

CLINICAL FINDINGS (Gas Gangrene)

• Spreads in 1-3 days • Crepitation & subcutaneous tissues &

muscles • Foul smelling discharge • Necrosis, fever

• Toxemia → shock → DEATH

CLINICAL FINDINGS (food poisoning)

• Enterotoxin forms in GUT • Diarrhea without vomiting or fever in 6-18

hours • Resolves in 1-2 days

DIAGNOSIS

• Clinical

• Laboratory • Culture • Chopped meat

• Growth + gas • BAP • Target or double zone of

hemolysis

DISEASES

• Gas gangrene (myonecrosis) • Anaerobic cellulitis

• Food poisoning (alpha toxin) • Necrotic enteritis (fire in the bowel) • Clostridial endometritis

TREATMENT

• Surgical debridement: most important • Antibiotics: Penicillin

• Food poisoning: supportive

4. Clostridium difficile PSEUDOMEMBRANOUS COLITIS

• Drug resistant C. difficile produce toxins

• Toxin A: enterotoxin • Toxin B: cytotoxin

• Pseudomembranes/ microabscesses in bowels

• Watery/ bloody diarrhea • Clindamycin, Cephalosporins, Amoxicillin,

Ampicillin

TREATMENT

• Discontinue offending antibiotic • Metronidazole or vancomycin

ANTIBIOTIC-ASSOCIATED DIARRHEA

• Administration of antibiotics leads to mild to moderate diarrhea

• Less severe than pseudomembranous colitis

• 25% associated with C. difficile

DIAGNOSIS

• Toxins (stools) • Endoscopic exam

Non-Spore Forming Gram Positive Bacilli

Listeria Erysipelothrix Actinomycetes

1. Listeria monocytogenes • Gram positive • Short rod • Catalase positive

• Beta hemolysis on BA • Tumbling motility at 22 – 28

oC

• Diffrentiates from diptheroids

• Widespread of disease in human & animals

• Important food-borne pathogen

• Survive in refrigerator temperature (4

oC)

• Survive low pH

• Survive high salt conditions • Overcome food preservation and

safety barriers

• Antigenic Classifications • 1/2a • 1/2b

• 4b: causes most food-borne outbreaks

PATHOGENESIS

• Ami, Fbp A, Flagellin protein

• Bind to host cells & virulence • Internalin A

• Interacts with E-

cadherin: cell well

protein in epithelial cells

• Promote phagocytosis • Listeriolysin O: enzyme

• Lyses membrane,

bacteria escape to cytoplasm

• Act A: surface protein

• Induces host cell actin polymerization

• Propels bacteria to cell

membrane • Cause formation of

Filipods

• Filipods • Ingested by epithelial

cells, macrophages &

hepatocytes • Lifecycle begins abain

RESERVOIR

• Widespread: animals (gastrointestinal and

genital tracts), • Unpasteurized milk products • Plants and soil

• Cold growth: soft cheeses, deli meats, cabbages (coleslaw)

PERINATAL HUMAN LISTERIOSIS

• Early onset syndrome

• Granulomatosis infantseptica • Infection in utero • Neonatal sepsis, pustular &

granulomas • Death; before / after delivery

• Late onset syndrome

• Meningitis: birth to 3rd

week of life

• Caused by serotype 4b

• High mortality rate

ADULTS

• Listeria meningoencephalitis • Immunocompromised

• Insidious to fulminant • Bacteremia • Focal infections

TREATMENT

• Ampicillin, Erythromycin, IV Trimethoprom-sulfamethoxazole

• Ampicillin + Gentamycin – recommended

PREVENTION

• Precautions with food may reduce

incidence

Page 8: Sporeforming & Non-Spore Forming Bacteria

Prepared by: EGBII; 8-13-11

2. Erysipelothrix rhusopathiae Distinguishing Characteristics:

• Gram positive bacilli, non-branching • Catalase (-), Oxidase (-), Indole (-)

• Alpha hemolysis on BA • Produced H2S on TSI (Butt Black)

• Differentiated from L.

monocytogenes

• Distributed in land & sea animals • Transmitted by direct inoculation

• Greatest risk: fishermen, fish handlers, butchers, those in contact with animal products

ERYSIPELOID/ SEA FINGER/ WHALE FINGER

• Icubation Period: 2-7 days

• Raised lesion w/ violaceous color • Pain & swelling • No pus

TREATMENT

• Penicillin G

3. Actinomycetes • Aerobic • Large diverse group of gram positive

bacteria

• Form chains or filaments • Categories:

• Acid fast positive: Mycobacteria

• Weakly positive: Nocardia & Rhodococcus

• Acid fast negative: Streptomyces

&Actinimadura

4. Nocardia Human infection: N. nova complex

• N. farcinica • N. asteroides type IV • N. brasiliensis

DISTINGUISHING CHARACTERISTICS

• Aerobic • Gram-positive branching rods

• Catalase positive • Partially acid-fast • Produce urease

• Can digest paraffin

• Inhalation of bacteria

• Not transmitted from person to person

• Opportunistic infection • Corticosteroid treatment,

immunosuppression, organ transplantation, AIDS, TB

• Begins with lobar pneumonia

• Mimic TB • Granulona & caseation are rare

• Abscess formation: usual pathologic

process • Spread to CNS, skin, kidney & eyes

TREATMENT

• Trimethoprim-Sulfamethoxazole • Treatment of choice

• Amikacin, imipenem, minocycline,

linezolide & ceftaxime • Surgical drainage

Actinomycetoma - mycetomal disease affecting the skin and connective tissue Mycetoma (Madura Foot)

• Most Common cause: • N. asteroides, N. brasiliensis,

Streptomyces somaliensis &

Actinomadura madurae

• Localized, slowly progressive chronic infection

• Begins in subcutaneous tissues & spreads to adjacent tissues

• Destructive & painless

ANAEROBE

BACTERIOLOGY

Definition of Terms

Aerobic Bacteria – require oxygen, and will not grow in the absence of oxygen

Anaerobic Bacteria – do not use oxygen for growth & metabolism but obtain their energy from fermentation

reactions

Capnophilic Bacteria – require CO2 for

growth

Facultative Anaerobes – can grow either oxidatively or use fermentation reactions to obtain energy

FACTORS THAT INHIBIT GROWTH OF ANAEROBES BY OXYGEN

• Toxic compounds are produced • e.g. H2O2 , Superoxides

• Absence of catalase & superoxide dismutase

• Oxidation of essential sulfhydyl groups in enzymes without sufficient reducing power to regenerate them

ANAEROBIC BACTERIA OF CLINICAL IMPORTANCE

Bacilli Gram-negative

• Bacteriodes

fragilis • Prevotella

melaninogenica • Fusobacterium

Gram-positive • Actinomyces • Lactobacillus • Clostridium

Cocci Gram-positive

• Peptostrept

ococcus • Peptococc

us Gram-negative

• Veilonella

PATHOGENESIS OF ANAEROBIC INFECTIONS

• Polysaccharide capsule

• Ability to induce abscess formation

• Lipopolysaccharide • Endotoxin: lack

lipopolysaccaride structures with endotoxic activity

• Not directly produce clinical signs of sepsis

• Enzymes • Proteases, Neuraminidases,

Cyclolysins • Cause hemolysis of

erythrocytes • Damage and destroys tissues

• Superoxide dismutase (some bacteria) • Can survive in the presence of

oxygen for days DIAGNOSIS OF ANAEROBIC INFECTIONS

• Foul-smelling discharge (pus) • Short-chain fatty acid products

of anaerobic metabolism • Infection in proximity to a mucosal

surface • Anaerobes are part of normal

flora • Gas in tissues

• Production of CO2 and H2 • Negative aerobic cultures • Involve mixture of organisms

• Form closed spaced infections or burrowing through tissues (Lungs, brain, pleura, pelvis)

• Most are susceptible to penicillin G • Except: Bacteroides, some

Prevotella species • Favored by reduce blood supply,

necrotic tissues, low Eh • Interfere with delivery of

antibiotics • Uses special collection methods &

transport media METHODS USED TO PRODUCE ANAEROBIOSIS

• Gas Pak Jar, Brewer Jar, Torbal Jar • Cooked meat medium / Chopped

cooked meat medium sealed with petrolatum

• Anaerobic glove box & chamber PRAS Medium

• Pre-Reduced Anaerobically Sterilized medium

• e.g. Roll Tube of Hungate

• Thioglycollate medium • Aerobic

• Anaerobic • Microaerophilic • Resazurin

Kanamycin-Vancomycin-Colistin (KVC) test

• Antibiotic Disks for the Presumptive Identification of Anaerobes

KVC PATTERN

• RRR- B. fragilis • SRS

• Fusobacterium

• B. ureolyticus • Veillonella

• SSR- Clostridium, gram (+) cocci • RSR- Porphyromonas, P. anaerobius • RRS- Prevotella

RESPIRATORY TRACT • Prevotella melaninogenica,

Fusobacterium & Peptostreptoccocus • Periodeontal infections,

perioral abscess, sinusitis, mastoiditis

Page 9: Sporeforming & Non-Spore Forming Bacteria

Prepared by: EGBII; 8-13-11

• Saliva aspiration: necrotizing pneumonia, lung abscess & empyema

CENTRAL NERVOUS SYSTEM

• Brain abscess, subdural empyema, septic thrombophlebitis

• Originate from respiratory tract, spread hematogenously

INTRA-ABDOMINAL & PELVIC INFECTIONS

• Flora of colon: B. fragilis, Clostridia,

Peptostreptococcus • Infection due to perforated bowel • Prevotella originate from female genital

organs SKIN & SOFT TISSUES

• Anaerobes & aerobes: synergistic infections

• Gangrene, necrotizing fasciitis, cellulitis Anaerobic Infections

TREATMENT • Surgical drainage: most important • Antimicrobial Therapy

• Penicillin G • Clindamycin

• Preferred for

infections above diaphragm

• Metronidazole