Gram Negative and Gram Positive Anaerobes

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Presentation transcript:

Gram Negative and Gram Positive Anaerobes Objectives and Key Concepts Know which anaerobic species cause which infections Identify key virulence factors important in pathogenesis of anaerobic bacteria Know which anaerobic pathogens cause infection as single agents vs those that are typically part of a mixed infection Know key microbiological features of anaerobic bacteria Know key signs and symptoms of infections caused by various Clostridia species Know which antibiotics that have good/excellent activity against anaerobes

Medically Important Anaerobes Gram positive rods Clostridium spp. Eubacterium Lactobacillus Actinomyces P. acnes Gram positive cocci Peptostreptococci Gram negative rods Bacteroides fragilis group Prevotella spp Porphyromonas spp Fusobacterium spp

Gram Positive Anaerobes (spore forming) Clostridium perfringens (single or mixed) Clostridium tetani (single) Clostridium botulinum (single) Clostridium difficile (single)

Clostridium perfringens Gas gangrene – toxin Intraabdominal infections- vegetative and toxin Food poisoning- toxin

Clostridium perfringens Structure: Gram positive Non-motile Encapsulated Forms spores Double zone hemolysis

“Box Car” appearance of Clostridium spp

Gas Gangrene Pathogenesis Trauma with devitalized tissue/muscle Spores of C. perfringens, C. novyi, C. septicum, C. ramosum germinate quickly Extremities, endometrium, abdominal wall Typing A-E – based upon combination of toxins produced by organism

Gas gangrene C. perfringens: Type A most common cause of human disease in U.S. Effects of toxins α toxin –lecithinase (phospholipase C), lysis of inflammatory cells, tissue destruction (muscle) β-toxin- enteritis necroticans (pig bel) i – necrosis and vascular permeability -ε toxin- Systemic, vascular permeability Θ toxin -Cardiotoxic

Gas Gangrene: Clostridium

GAS GANGRENE

Gas Gangrene Clinical Disease Rapid onset (6 - 72 hr) necrosis of muscle (myonecrosis), skin tense edema bullae formation gas formation (fermentation) Systemic shock hyper or hypothermia

Gas gangrene Diagnosis Treatment Clinical suspicion Gram stain culture Treatment debridement, debridement, debridement antibiotics (penicillin, B-lactam/inhibitor) clindamycin

Clostridium perfringens Food poisoning Heat resistant spores survive gravy, soups enterotoxin produced following germination of large numbers of organisms Clinical 8 - 24 hr after ingestion nausea, abdominal pain, diarrhea

C. perfringens (food poisoning) Diagnosis clinical culture (not needed) Treatment self limited disease supportive

Clostridium tetani Entry and Spread Tetanospasmin puncture wounds burns umbilicus local germination without necrosis Tetanospasmin neurotoxin (plasmid borne) blocks postsynaptic inhibition spinal motor reflexes (GABA) Spasmotic contractions

CLOSMIDIUM TETANI GRAM STAIN

Clostridium tetani Clinical Generalized Toxin trismus (lockjaw) risus sardonicus (increased tone orbicularis oris) opisthotonus (arm flexion, leg extension) Respiratory (obstruction, diaphragm) Toxin intraaxonal, transports cephalid over 2 weeks

OPISTHOTONUS

OPISTHOTONUS

Tetanus Localized Cephalic Neonatal rigidity of muscles at site of sporulation usually precedes generalized Cephalic cranial nerve musculature facial, ocular Neonatal failure of aseptic technique

Risus Sardonicus

Tetanus Diagnosis Treatment Clinical Human tetanus immunoglobulin Sedation Control of spasms supportive (airway)

Tetanus Prophylaxis Prevention 3 doses DTaP revaccination every 10 years (Tdap/Td) Prevention passive immunization HTIG for appropriate wound no previous vaccination > 5 years

Clostridium botulinum Ubiquitous organism (A-G serotypes, A, B, E, F associated with human disease) spores heat resistant (100 C x hours) Home canning fruits/vegetables, fish Rarely commercial sources Entry preformed toxin from contaminated food spores in honey (infants) 12 - 36 hrs after ingestion

Clostridium botulinum Toxin (bacteriophage borne) large single polypeptide (150-165 Da) cleaved by bacterial protease most potent toxin in nature blocks acetylcholine no transmitter release, paralysis synapse permanently damaged can travel axons heat labile

Botulism Clinical GI (nausea, dry mouth, diarrhea) descending paralysis (flaccid) cranial (III, IV, VI) symmetric neurologic effects afebrile patient conscious heart rate normal no sensory deficits Wound botulism

Botulism Diagnosis Prevention/Treatment Clinical (DDx myasthenia gravis, Eaton Lambert, tick paralysis, Guillain-Barre’) toxin assays (serum, stool, food) Prevention/Treatment avoid contaminated food heating of food (boiling) antitoxin (equine serum to A,B,E) supportive

Clostridium difficile Major hospital acquired pathogen Rare normal flora spores acquired in hospital antibiotic therapy usually precedes disease Toxin A - enterotoxin, inflammatory response, major toxin Toxin B - cytotoxic

Diffuse Hemorrhagic colitis and pseudomembrane formation

PSEUDOMEMBRANEOUS COLITIS

C. difficile Clinical BI (Nap1) Strain pseudomembranous colitis diarrhea (watery, severe, bloody) abdominal pain leukocytosis fever toxic megacolon BI (Nap1) Strain Increased Toxin A production Higher mortality, especially elderly Up to 50% of isolates

C. difficile Diagnosis PCR- New standard-amplifies toxin region 90+ % sensitivity, 98% specificity Detection of toxin A in stool- not recommended alone for definitive diagnosis-combine with PCR ELISA 85 - 90% sensitivity (single assay) 95% sensitive (two assay) Culture not routinely recommended Sigmoidoscopy/colonoscopy

C. difficile Treatment Stop antibiotics if possible Vancomycin (oral only) Fidaxomycin- primary and relapse colon resection Lactobacillus Fecal Transplant (FMT) - > 2 episodes following treatment

Clostridium septicum (single) Nontraumatic myonecrosis Bacteremia- likely result of break in integrity of bowel mucosa High association with colon cancer, leukemia Fulminant disease with high mortality Clostridium sordelli (single) Abortion (11%), pregnancy (18%), IVDA (22%), other (43%) Profound leukocytosis 50-100% mortality

Other Anaerobic Gram positive Rods Actinomyces- non spore forming (single or mixed) Filamentous hyphae Sulfur granules (yellow sand colonies) Non- acid fast Clinical Slow growing, supporative Typically oral associated Cervicofacial, thoracic, pelvic or intraabdominal Diagnosis Culture, but slow growing Treatment Penicillin best Clindamycin or erythromycin

Propionobacterium acnes (single) Slow growing anaerobe Contaminant in blood cultures Cause of infection: Prosthetic devices or hardware Opportunistic infections Acne Treatment – if indicated Penicillin as well as many other agents except metronidazole!!

Anaerobic Gram positive cocci Peptostreptococcus (single or mixed) Normal flora of mouth, GI tract, pelvis Contiguous infections (usually mixed) Intra-abdominal, endometritis, pulmonary Brain abscess Diagnosis –culture Treatment – debridement and penicillin, metronidazole, cabapenems, clindamycin

Gram Negative Anaerobes B. fragilis group (mixed) Non-spore forming, non-motile Requires enriched media to grow Normal inhabitant of GI tract (colon) Provide protection from invasion (?)

Bacteroides fragilis group B. fragilis B. thetaiotaomicron P. distasonis B. ovatus B. caccae B. uniformis B. vulgatus B. eggerthiae B. merdae B. stercoris

B. fragilis group Pathogenesis Hallmark is abscess formation Almost always mixed infections (aerobes and anaerobes Abscess: Inflammatory response to infection Encapsulated “pus” PMN’s Fibrin, fibrinogen Debris, necrotic tissue

B. fragilis group Virulence factors Polysaccharide capsule Adherence to peritoneal cavity Resists phagocytosis Resistance to T cell and humoral immunity Oxygen tolerance Super oxide dismutase Catalase Toxins : Defective endotoxin – not associated with sepsis

B. fragilis group Most commonly recovered from clinical infections Intraabdominal (3/2 ratio anaerobe/aerobe) female genital tract (3/1 ratio anaerobe/aerobe) aspiration pneumonia (mixed) Empyema (mixed) otitis media ?? brain abscess (mixed) Skin and Soft Tissue infections (3/1 ratio)

Peritonitis

Pathogens from 900 Intraabdominal Infections

Fournier Gangrene

Liver Abscess

Liver Abscess: Microbiology

Organisms Isolated from Diabetic Foot Infections

Prevotella spp (mixed) Residents of mouth, GI tract, and Pelvis Gram negative strict anaerobes Require enriched media (blood containing) Non-motile Non-encapsulated Virulence – unknown Pathogensis – abscess formation

Prevotella spp Female genital tract infections Oral, pleuropulmonary P. bivia, P. disiens Oral, pleuropulmonary P. oris, P. buccae, P. oralis Oral cavity, urogenital, GI tract P. melaninogenica, P. corporis, P.denticola, P. intermedia, P. loescheii, P. nigrescens

Brain Abscess

Other Gram negative Anaerobes of Clinical Importance Porphyromonas (mixed or single) oral, periodontal infections Fusobacterium (mixed or single) F. nucleatum aspiration pneumonia, lung abscess, empyema, chronic otitis media, sinusitis, brain abscess liver abscess F. necrophorum Lemierre’s syndrome/ post anginal sepsis widespread metastatic infection Highly virulent – potent endotoxin

Antibiotic Therapy for Anaerobes Non-C difficile Bacteroides Most Active: Metronidazole (100%), Ampicillin/sulbactam (96%), piperacillin/tazobactam (99%), carbapenems (99.5%), chloramphenicol (100%) Formerly Active: cefoxitin (75-90%), clindamycin (60%), piperacillin (60%)

Antibiotic Therapy for Anaerobes Fusobacterium Most Active: metronidazole, amp/sulb, pip/tazo, clindamycin, cefoxitin Active: piperacillin (?), penicillin (>80%) Peptostreptococus Most Active: Penicillin, clindamycin, amp/sulb, pip/tazo, metronidazole (some resistant strains)