Gram-negative rods: Enterobacteriaceae Part II Common Organisms. Escherichia coli. Escherichia coli. Escherichia coli. CLS 418 Clinical Microbiology I
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1 Gram-negative rods: Enterobacteriaceae Part II Common Organisms Karen Honeycutt, M.Ed., MLS(ASCP) CM SM CM Session Enterobacteriaceae Antigens O somatic, part of cell wall (serogroup) Stimulates earliest antibody K capsular, heat labile (serotype) Vi (virulence) specific Salmonella Typhi K antigen H flagellar, heat labile (serotype) Latest antibodies formed Use specific antisera to identify organisms beyond species level Example: O157:H7 All Pictures Copyright University of Nebraska 2006 Most common facultative organism in stool Colony counts to determine fecal contamination of water E. coli inert/inactive group Biochemically very inert Nonmotile Can be biochemically mistaken for a Shigella (a stool pathogen) BAP: beta-hemolytic EMB: green metallic sheen Colony morphology MAC: dark pink diffuses around colony #1 etiologic agent of UTI - nephropathogenic Meningitis in 0-3 month age group Enterohemorrhagic produces Shiga toxin/verotoxin: example E. coli O157:H7 Ingest contaminated food (undercooked hamburger, unpastuerized apple juice & milk, leaf lettuce) Bloody diarrhea Children especially susceptible to HUS Enterobacteriaceae II Page 1
2 Enterohemorrhagic produces Shiga toxin/verotoxin: example E. coli O157:H7 Acute community acquired diarrhea: screen all stools submitted Rapid tests to detect toxin production: EIA, molecular (PCR) Specific screening agar: SMAC, agar with sorbitol (not lactose) E. coli O157:H7 is sorbitol neg (clear on SMAC) Gastroenteritis (not EHEC) 4 distinct syndromes caused by 4 distinct E. coli strains Usually not diagnosed by culture Identification Beta-hemolytic EMB: Green metallic sheen Abbreviated Id Not applicable for blood or gastrointestinal specimens GNR Oxidase negative Non-swarming, beta-hemolytic on BAP positive Agar Indole Citrate Abbreviated Id Not applicable for blood or gastrointestinal specimens GNR Oxidase negative Non-hemolytic BAP, lactose-fermenting positive PYR negative Shigella sp. Disease State Overt or primary pathogen (never NF) Bacillary dysentery or shigellosis Only human to human transfer 4 subgroups (A, B, C, D) Shigella sonnei (subgroup D) most common in US and least severe disease Enterobacteriaceae II Page 2
3 Shigella sp. Identification Always nonmotile! Shigella sp. Identification Biochemical identification should be confirmed with serogrouping Reportable disease LDC (-) & LDA (-) Salmonella sp. Overt or primary pathogen (not NF) Gastroenteritis: mild to fatal (dehydration) Ingestion of contaminated food, water, milk Chicken, eggs Contact with infected animals Reptiles Human to human transfer Usually doesn t invade bloodstream Salmonella Typhi Overt or primary pathogen (not NF) Typhoid fever Transmission: human to human Specimens: blood, urine then stool Salmonella sp. Identification Glucose F Red pigment Serratia sp. Not LF pathogen Serratia marcescens Hospitals Can produce a red pigment!! Lactose nonfermenter LDC + Enterobacteriaceae II Page 3
4 Citrobacter sp. Lactose variable LDC & LDA (-) C. fruendii: /- [LDC & LDA (-), ] LDC (-) & LDA (-) pathogens Klebsiella pneumoniae: destructive pneumonia Muciod colony due to capsule Always nonmotile! Klebsiella sp. Klebsiella sp.- Identification Muciod colony due to capsule Always nonmotile! Enterobacter sp. pathogen Muciod colony due to capsule ODC+ Proteus sp. pathogen Swarms on BAP, CHOC Strong urea hydrolyzer: creates alkaline ph in urine, causes renal calculi Proteus sp. - Identification Urea Glucose F LDA+ & ++ Enterobacteriaceae II Page 4
5 Proteus mirabilis Abbreviated Identification Proteus vulgaris Abbreviated Identification Swarming on BAP negative Ampicillin susceptible Swarming on BAP positive If ampicillin resistant, then ODC positive Morganella sp. pathogen LDA+, no H 2 S production LDA+ Summary - Enterobacteriaceae GNR, Glucose F, oxidase -, N0 3 to NO 2 growth: LF & NLF Normal flora, colonize hospital patients, opportunistic or primary/overt pathogens Summary - Enterobacteriaceae Identification clues Colony morphology: mucoid, swarming, beta-hemolytic, red pigment Lactose fermentation Nonmotile Deamination (LDA) positive VP + Enterobacteriaceae II Page 5
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