BACT211

CLINICAL BACTERIOLOGY
BATCH
2021

COLLEGE OF MEDICAL LABORATORY SCIENCE-OUR LADY OF FATIMA UNIVERSITY (LAGUNA CAMPUS)

Enterobacteriaceae
General characteristics: •Microscopic examination:
•Macroscopic examination: - Gram (-) bacilli
•BAP and CAP
- Klebsiella & Enterobacter: large mucoid colonies •Fermentation of glucose, lactose, trehalose & xylose TSI: A/A
- E. coli: β-hemolytic •Indole (+) from tryptophan LIA: A/A or K/A
•EMB, MAC, HE, XLD •MR (+) VP (-) IMVC + +
- Ferment carbohydrate •Citrate (-)
•HE & XLD •H2S, Dnase, urease, PAD (-) --Phenylalanine deaminase
- Produce H2S
•Microscopic examination: A. UROPATHOGENIC E. COLI:
- Gram (-) non-spore-forming facultative anaerobic bacilli •Most common cause of UTI's in human
•Pyelonephritis in ICP
Classification: VIRULENCE FACTOR:
I. Escherichieae V. Klebsielleae 1. Pili
I. Escherichia I. Klebsiella V. Proteae • Adherence to epithelial cells
II. Shigella II. Enterobacter I. Proteus UPEC P pilus/pap pili, type 1 fimbriae Uropathogenic E. coli
II. Edwardsielleae II. Morganella DAEC Afa/Dr adhesions also cause GI infections Diffusely adherent E. coli
III. Pantoea
I. Edwardsiella III. Providencia 2. Cytolisins
III. Salmonelleae IV. Cronobacter • kill immune effector cells and inhibit phagocytosis and chemotaxis
I. Salmonella V. Hafnia VI. Yersinieae
3. aerobactins
IV. Citrobacteriaceae VI. Serratia I. Yersinia • Chelate iron
I. Citrobacter IV. Cronobacter
B. GASTROINTETINAL E. COLI:
Virulence factors: 5 major categories:
•Plasmids • ETEC: enterotoxigenic Escherichia coli
- Antimicrobial resistance • EIEC: enteroinvasive Escherichia coli
- Plasmid-mediated Extended spectrum β-lactamases (ESBLs) • EPEC: enteropathogenic Escherichia coli enterovirulent E. coli or
carbapenemases, cephalosporinases, or metallo-β-lactamases • EHEC: enterohemorrhagic Escherichia coli
inactivate extended-spectrum cephalosporins •enteroadherent diarrheogenic E. coli
•Antigens -EAEC: enteroaggregative Escherichia coli
- O antigen - DAEC: diffusely adherent Escherichia coli
•somatic antigen
•heat-stable antigen in cell wall Enterotoxigenic Escherichia coli : ETEC traveler’s diarrhea
- H antigen •one of the major causes of infant bacterial diarrhea
•flagellar antigen •106 to 1010 organisms Infective dose
•heat-labile antigen in flagella •stomach acidity
- K antigen - Inhibiting colonization
•capsular antigen - Achlorhydria: > risk Deficiency of HCl within the stomach
•heat-labile polysaccharide in capsule •Virulence factor:
•K1 antigen: E. coli • Fimbrae:attach to specific receptors on intestinal microvilli
•Vi antigen: S. enterica subsp. enterica serotype Typhi • LT (heat-labile toxin): A & B fragments
Hypersecretion of electrolytes & fluid in
- A: active portion; converts ATP to cAMP
intestinal lumen----- WATERY DIARRHEA
- B: moiety or binding portion to GM1 ganglioside of intestinal mucosal
• ST (heat-stable toxin): stimulates guanylate monophosphate

Enteropathogenic Escherichia coli : EPEC infantile diarrhea

•Virulence factor:
- Pathogenicity islands

Enteroinvasive Escherichia coli: EIEC dysentery

•direct penetration, invasion, and destruction of the intestinal mucosa
•Similar to shigella infection
•106 organisms
Escherichia coli Enterohemorrhagic Escherichia coli : EHEC hemorrhagic diarrhea, colitis & HUS
Characteristics: •watery diarrhea to bloody diarrhea (no leukocytes unlike shigella dysentery)
•Escherich in 1885 •Virulence factor:
•Colon microbiota •O157:H7 strain
- primary marker of fecal contamination in water quality testing

•Virulence factors:
-adhesive fimbriae and sex pili
-O, H & K antigens

•Medically important species:

A. Uropathogenic E. coli Enterohemorrhagic Escherichia coli : EHEC
B. Gastrointestinal E. coli • O157:H7 strain
C. Extraintestinal E. coli •MAC agar containing sorbitol (SMAC): stool culture Sorbitol is a sugar alcohol
- (-) ferment sorbitol in 48 hrs found in fruits and plants
Characteristics: - Colorless colonies with diuretic, laxative and
•Macroscopic examination: • 4-methylumbelliferyl-β-D-glucuronide (MUG) assay
•MAC: lactose fermenting colony with a surrounding area of precipitated bile salts - (+) produce β-glucuronidase cathartic property.
•EMB: green metallic sheen •ELISA & latex agglutination
•E. coli O157:NM
- (+) ferment sorbitol in 48 hrs
- non-motile, pink colonies

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 BACT211
CLINICAL BACTERIOLOGY
BATCH
2021

COLLEGE OF MEDICAL LABORATORY SCIENCE-OUR LADY OF FATIMA UNIVERSITY (LAGUNA CAMPUS)

Enteroadherent Escherichia coli

•DAEC EAEC: enteroaggregative Escherichia coli
- UTIs and pediatric diarrheal disease DAEC : diffusely adherent Escherichia coli

•EAEC
- Watery diarrhea
- Fimbriae adhere to HEp2 cells in “stacked-brick” pattern
HEp-2 cells possess well over 100 antigens in the nucleus
and cytoplasm that may have relevance for patients with
autoimmune disease. Enterobacter
Characteristics:
C. EXTRAINTESTINAL E. COLI •Macroscopic examination:
•septicemia and neonatal meningeal infection - MAC: lactose fermenting and mucoid colony
•Virulence factor: - Potassium cyanide broth
- capsular antigen K1 •Microscopic examination:
immunochemically identical to capsular antigen of N. meningitidis group B - Gram (-) bacilli
•Escherichia hermannii : yellow-pigmented organism in CSF, wound & blood •Fermentation of glucose, lactose, trehalose & xylose
•Escherichia vulneris: yellow-pigmented organism •Indole (-) from tryptophan
•MR (-) VP (+)
•Citrate (+)
•Urease & motility (+)
•H2S (-)
•Ornithine decarboxylase & lysine decarboxylase (+) EXCEPT E. gergoviae or E. cloacae

Characteristics:
isolated from wounds, urine, blood, and CSF

•Pantoea (Enterobacter) agglomerans

- septicemia resulting from contaminated intravenous fluid
Klebsiella - P. agglomerans HG XIII
Characteristics: yellow pigment, is primarily a plant pathogen
•Macroscopic examination:
- MAC: lactose fermenting and mucoid colony •E. gergoviae
- Potassium cyanide broth RT samples & blood

•Microscopic examination: •Cronobacter (Enterobacter) sakazakii

- Gram (-) bacilli Mucoid, yellow pigment
Neonatal meningitis & bacteremia from powdered milk formula
•Fermentation of glucose, lactose, trehalose & xylose
•Indole (-) from tryptophan E. hormaechei
•MR (-) VP (+) •blood, wounds, and sputum
•Citrate (+) E. asburiae
•Urease & motility (+) •biochemically similar to E. cloacae
•H2S (-) •blood, urine, feces, sputum, and wounds
E. cancerogenus
Characteristics: •formerly E. taylorae
•Microbiota of GIT •osteomyelitis after traumatic wound
•K. pneumoniae Enterobacter dissolvens & Enterobacter nimipressuralis
Virulence factor: •newly recognized species
- large polysaccharide capsule
protection against phagocytosis & antimicrobial absorption
- lower RTI among hospitalized patients & ICP

•K. oxytoca
•antibiotic-associated hemorrhagic colitis
•Biochemically similar with K. pneumonia EXCEPT indole (+)

•K. pneumoniae subsp. ozaenae

•atrophic rhinitis, a tissue-destructive disease restricted to the nose
•plasmid-mediated ESBLs

Characteristics:
•K. pneumonia subsp. rhinoscleromatis Rhinoscleroma is a chronic granulomatous
- Rhinoscleroma condition of the nose and other structures of the
upper respiratory tract.

•Raoutella (Klebsiella) ornithinolytica

- Indole & Ornithine decarboxylase (+)

•Raoutella (Klebsiella) planticola

•Raoutella planticola : difficult to distinguish from K. pneumoniae

•Klebsiella variicola : isolated from primarily sterile sites

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 BACT211
CLINICAL BACTERIOLOGY
BATCH
2021

COLLEGE OF MEDICAL LABORATORY SCIENCE-OUR LADY OF FATIMA UNIVERSITY (LAGUNA CAMPUS)

Citrobacter Providencia
Characteristics: Characteristics:
•GIT inhabitants •P. alcalifaciens, P. stuartii, P. rettgeri, P. rustigianii, and P. heimbachae. P.
•UTI & sepsis rettgeri
•MAC: resemble E. coli UTI & diarrheal disease among travelers
•Resemble Salmonella sp. EXCEPT ONPG(+) LDC(-) outbreaks in health care setting
•MR (+)
•Citrate (+) •P. stuartii
•Slow urease (+) - outbreaks in burn units
- Isolated in urines
•C. freundii •P. stuartii and P. rettgeri
•endocarditis in intravenous drug abusers - resistance to antimicrobials
•aortic valve replacement when antimicrobial therapy failed •P. alcalifaciens
•H2S (+) - Diarrhea in children
•P. rustigianii
•C. koseri
•nursery outbreaks of neonatal meningitis and brain abscesses Morganella
Characteristics:
•C. amalonaticus •M. morganii
•found in feces, blood and wounds •Subtypes:
- M. morganii subsp. morganii
- M. morganii subsp. sibonii.
Serratia •UTI & neonatal sepsis
Characteristics: •Motility (+)
•opportunistic pathogens associated with outbreaks in health care settings (UTI & RTI)
•bacteremic outbreaks in nurseries and cardiac surgery and burn units
•Slow lactose fermenters EXCEPT S. fonticola
•o-nitrophenyl-β-D-galactopyranoside (ONPG) test (+)
•Dnase (+)

•S. marcescens, S. rubidaea & S. plymuthica

•Produce prodigiosin: pink to red pigment in RT

•S. odorifera
•dirty, musty odor resembling that of rotten potatoes

Edwardsiella
Characteristics:
•E. tarda, E. hoshinae & E. ictaluri
•Human pathogen: E. tarda
Bacteremia & wound infections
•Citrate & urease (-)
•Lysine decarboxylase, H2S, indole (+)
Hafnia
Characteristics: Erwinia & Pectobacterium
•H. alvei Characteristics:
•Gastroenteritis •plant pathogens
•Delayed citrate (+) •Erwinia
- H. alvei and H. alvei biotype 1 Poor growth at 37° C
- H. alvei biotype 1 (-) growth in EMB & MAC
beer wort of brewerie
Salmonella
Proteus Characteristics:
Characteristics: •Gram (-) facultatively anaerobic bacilli
•Proteus, Morganella, and Providencia •MAC: clear, colorless, non–lactose fermenting colonies
•normal intestinal microbiota •XLD & HE: colonies with black centers
•opportunistic pathogens •Bismuth Sulfide Agar: black colonies with metallic sheen
•Proteus mirabilis & Proteus vulgaris •Non-lactose fermenter
•urine, wounds, and ear and bacteremic infections •Indole, VP, PAD, Urease (-)
•acute glomerulonephritis •H2S (+) EXCEPT S. paratyphi
•urease activity of P. mirabilis: struvite kidney stones •(-) growth potassium cyanide medium

•Proteus mirabilis & Proteus vulgaris Virulence factors:

•BAP: “swarming colonies” “burnt chocolate” - odor •fimbriae
- Adherence to GIT
•(+) deaminate the amino acid phenylalanine •Enterotoxin
•Non-lactose fermeter - traverse intestinal mucosa
•Urease (+)
•H2S (+)
•Proteus mirabilis: indole (-) & ornithine (+); OXK
•Proteus vulgaris: indole (+) & ornithine (-); OX2 & OX19
ferments sucrose TSI: A/A

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 BACT211
CLINICAL BACTERIOLOGY
BATCH
2021

COLLEGE OF MEDICAL LABORATORY SCIENCE-OUR LADY OF FATIMA UNIVERSITY (LAGUNA CAMPUS)

Antigen:
•Somatic O antigen
- Heat stable
- lipopolysaccharide in the outer membrane of the cell wall

•H flagellarantigen
- Heat labile
- Phase I flagellar antigen: specific phase;occur in few strains
determine the immunologic identity of certain serotype Antigens:
agglutinate only with homologous antisera
•(+) O and K antigen
- Phase II flagellar antigen: non-specific phase; occur in may strains
•(-) H antigen
agglutinate only with heterologous antisera
•Capsular K antigen (Vi)
Infections:
•For ID of Salmonella serotype Typhi & Salmonella serotype Choleraesuis
•dysentery
•24 to 48 hours after ingestion of the organisms
Infections:
•Acute gastroenteritis or food
•Typhoid fever Yersinia
- most severe form of enteric fever Salmonella serotype Typhi Characteristics:
- enteric fevers Salmonella paratyphi and choleraesuis •Y. pestis
•Non-typhoidal bacteremia •Causative agent of plaque disease of rodents transmitted to humans by fleas
•Carrier state following Salmonella infection •3 forms:
•MOT: ingestion of contaminated food, water and milk •Bubonic/ glandular form
•from bite of an infected flea
Infections: •symptoms appear 2 to 5 days after infection
1. Gastroenteritis •high fever with painful regional lymph nodes known as buboes
•One of the most common forms of “food poisoning” •Septicemic form
•From poultry, milk, eggs, and egg products as well as to handling pets •bacteria spread to the bloodstream
•Infective dose: 106 bacteria •Pneumonic form
•8 to 36 hours after ingestion of contaminated food •occurs secondary to bubonic plague or septicemic form
•TOC: chloramphenicol, ampicillin & trimethoprim-sulfamethoxazole •Primary infection through inhalation Fatality rate: 100% if untreated

2. Enteric fevers Characteristics:

•Salmonella Typhi: typhoid fever •Y. pestis
•Only host: human •Gram (-) short plum bacillus
•Salmonella serotypes Paratyphi A, B, and C and Salmonella serotypeCholeraesuis •Methylene blue or Wayson stain:
•9 to 14 days after ingestion of the organisms •bipolar staining
•“safety-pin” appearance
•Preferred growth temperature: 25° C to 30°

Characteristics:
•Y. pseudotuberculosis & Y. enterocolitica
•sporadic cases of mesenteric lymphadenitis in humans
3. Bacteremia
•Typhimurium, Paratyphi, and Choleraesuis Y. enterocolitica
4. Carrier state •Natural reservoir: ____
•Site of chronic carriage: gallbladder •MOT: direct contact with household pets
•Organisms secreted in the feces continuously or intermittently Ingestion: contaminated pork & vacuum-packed deli meat, beef, lamb, chicken,
•Treatment: chocolate milk and water
-antimicrobial therapy •Gastroenteritis
- cholecystectomy •Sepsis associated with the transfusion of contaminated packed RBCs
•Mimics acute appendicitis

Yersinia
Characteristics:
Y. enterocolitica Bull’s eye colony
•Gram (-) coccobacilli with bipolar staining
•Optimal growth temperature: 25° C to 30° C
•grows better with cold enrichment
•Motility (+) at 25° C not at 35° C
•Cefsulodin-irgasan-novobiocin (CIN)
- cefsulodin, irgasan, novobiocin, bile salts, and crystal violet (inhibitors)

Shigella Characteristics:
Characteristics: Y. pseudotuberculosis
•Escherichieae with E. coli •Pathogen of guinea pigs
•Cause: bacillary dysentery •Natural reservoir: birds
presence of blood, mucus, and pus in the stool •Causative agent of : caseous swellings called pseudotubercles
•Not GI microbiota •typical-looking plague bacillus
•Japanese microbiologist Kiyoshi Shiga •Motility at 18° C to 22° C
•Motility (-) •Urease (+)
•(+) gas from glucose EXCEPT S. flexneri •Ferment rhamnose
•Urease, H2S, LDC (-)
•(-) use of acetate or mucate as carbon source
•susceptible to disinfectants & high concentrations of acids and bile

Characteristics: •S. sonnei

•Ornithine decarboxylase (+)
•ONPG (+) •slowly ferments lactose
•MAC: delayed fermentation; pink colonies on MAC only after 48 hours of inc.

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 BACT211
CLINICAL BACTERIOLOGY
BATCH
2021

COLLEGE OF MEDICAL LABORATORY SCIENCE-OUR LADY OF FATIMA UNIVERSITY (LAGUNA CAMPUS)

Enterobacteriaceae
Specimen collection:
•No special considerations required
•transport media: Cary-Blair, Amies or Stuart media

Direct Microscopic Examination:

•gram (-) bacilli
•Stool sample: not helpful but may reveal if GI infection is Invasive process

Culture
•35°C to 37°C for 18-24 hours of incubation
•1-5°C: Serratia & Yersinia
•45° C to 50° C: E. coli

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 BACT211
CLINICAL BACTERIOLOGY
BATCH
2021

COLLEGE OF MEDICAL LABORATORY SCIENCE-OUR LADY OF FATIMA UNIVERSITY (LAGUNA CAMPUS)

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