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 sorters 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 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

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 sorters 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 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 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