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Sporeforming & Non-Spore Forming Bacteria

Sporeforming & Non-Spore Forming Bacteria

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Sporeforming & Non-Spore Forming Bacteria
Sporeforming & Non-Spore Forming Bacteria

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Published by: Ernie G. Bautista II, RN, MD on Oct 06, 2012
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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

• 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

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

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

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)

1. Clostridium botulinium

2. Bacillus cereus
• Produce toxins that cause disease • Spores germinate, vegetative cells produce toxins

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 Prepared by: EGBII; 8-13-11

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 5 • 10 bacteria or more per gram of food is diagnostic

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

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

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)

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

3. Clostridium perfringens
CHARACTERISTICS • Encapsulated • Non motile • Double hemolysis • Ferment CHO • Reservoir: Soil and human colon • INVASIVE

Prepared by: EGBII; 8-13-11

Clostridial endometritis •

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

• 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

Focal infections

TREATMENT • Ampicillin, Erythromycin, IV Trimethopromsulfamethoxazole • Ampicillin + Gentamycin – recommended PREVENTION • Precautions with food may reduce incidence

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

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

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

Non-Spore Forming
Gram Positive Bacilli Listeria Erysipelothrix Actinomycetes

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

1. Listeria monocytogenes
• • • • • • • Gram positive Short rod Catalase positive Beta hemolysis on BA o Tumbling motility at 22 – 28 C • Diffrentiates from diptheroids Widespread of disease in human & animals Important food-borne pathogen • Survive in refrigerator temperature (4oC)

4. Nocardia
Human infection: N. nova complex • N. farcinica • N. asteroides type IV • N. brasiliensis

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

• • •

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 glove box & chamber

PRAS Medium • Pre-Reduced Anaerobically Sterilized medium • e.g. Roll Tube of Hungate • Thioglycollate medium • Aerobic • Anaerobic • Microaerophilic • Resazurin

ANAEROBIC BACTERIA OF CLINICAL IMPORTANCE Bacilli Cocci Gram-negative Gram-positive • Bacteriodes fragilis • Peptostreptococcus • Prevotella • Peptococcus melaninogenica Gram-negative • Fusobacterium • Veilonella Gram-positive • Actinomyces • Lactobacillus • Clostridium 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

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

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

Prepared by: EGBII; 8-13-11

• Preferred for infections above diaphragm Metronidazole

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

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

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 5 • 10 bacteria or more per gram of food is diagnostic • Culture: exhibit motility “swarming” TREATMENT • Drug of choice: vancomycin or clindamycin • Resistant to penicillins & cephalosporins •

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

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

Alters cell membrane, disrupting ion transport

• • •

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

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

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

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

Non-Spore Forming Gram Positive Bacilli Listeria Erysipelothrix Actinomycetes

1. Listeria monocytogenes
Gram positive Short rod Catalase positive Beta hemolysis on BA o Tumbling motility at 22 – 28 C • Diffrentiates from diptheroids • Widespread of disease in human & animals • Important food-borne pathogen • Survive in refrigerator o temperature (4 C) • 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 Ecadherin: cell well • • • • •

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 rd • Meningitis: birth to 3 week of life • Caused by serotype 4b • High mortality rate ADULTS • Listeria meningoencephalitis • Immunocompromised • Insidious to fulminant • Bacteremia • Focal infections TREATMENT • Ampicillin, Erythromycin, IV Trimethopromsulfamethoxazole • Ampicillin + Gentamycin – recommended PREVENTION • Precautions with food may reduce incidence

4. Clostridium difficile
PSEUDOMEMBRANOUS COLITIS • Drug resistant C. difficile produce toxins • Toxin A: enterotoxin • Toxin B: cytotoxin

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

• • •

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

• •

Absence of catalase & superoxide dismutase Oxidation of essential sulfhydyl groups in enzymes without sufficient reducing power to regenerate them

• • •

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

ANAEROBIC BACTERIA OF CLINICAL IMPORTANCE Bacilli Cocci Gram-negative Gram-positive • Bacteriodes • Peptostrept fragilis ococcus • Prevotella • Peptococc melaninogenica us • Fusobacterium Gram-negative Gram-positive • Veilonella • Actinomyces • Lactobacillus • Clostridium 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

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

PRAS Medium • Pre-Reduced Anaerobically Sterilized medium • e.g. Roll Tube of Hungate • Thioglycollate medium • Aerobic • Anaerobic • Microaerophilic • Resazurin

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

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

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

FACTORS THAT INHIBIT GROWTH OF ANAEROBES BY OXYGEN • Toxic compounds are produced • e.g. H2O2 , Superoxides

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

Prepared by: EGBII; 8-13-11

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