VIII : GRAM - FERMENTATIVE BACILLI

F

Gram -

FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES is gram + (t/f)

T

FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES is Gram – enteric coccobacilli, short, plump bacilli (t/f)

F

Non-spore Formers

CPON NO GF FA

FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES is spore forming (t/f)

F

FACULTATIVE ANAEROBES

CPON NO GF FA

FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES is facultative aerobe (t/f)

T

Antigenic - once infected produce = Ab

FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES is considered to be antigenic (t/f)

1. Somatic / O / Cell wall Antigen

2. Envelope / K / Capsule Antigen

3. Flagellar / H Antigen

KOH

Antigenic structure of FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

what are the antigenic structure

Somatic / O / Cell wall Antigen

Antigenic structure of FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

HEAT STABLE, lipopolysaccharide

Somatic / O / Cell wall Antigen

Antigenic structure of FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

For E. coli and Shigella serotyping

164 types

Antigenic structure of FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Somatic / O / Cell wall Antigen

- For E. coli there are how many types O antigen and specific type may be associated with a particular disease (i.e. SEROTYPE 0111- Diarrhea in infants; SEROTYPE 0157- Verotoxin production)

T

Antigenic structure of FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Somatic / O / Cell wall Antigen

For E. coli there are 164 types of O antigen and specific type may be associated with a particular disease (t/f)

SEROTYPE 0111- Diarrhea in infants

SEROTYPE 0157- Verotoxin production

Antigenic structure of FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Somatic / O / Cell wall Antigen

For E. coli there are 164 types of O antigen and specific type may be associated with a particular disease

– Diarrhea in infants

- Verotoxin production

Flagellar / H Antigen

Antigenic structure of FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Protein in nature and heat labile

Flagellar / H Antigen

Antigenic structure of FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

- Found only among motile enteric

Flagellar / H Antigen

Antigenic structure of FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

- For Salmonella serotyping

Envelope / K / Capsule Antigen

Antigenic structure of FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Consists of capsular POLYSACCHARIDE

T

Antigenic structure of FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Envelope / K / Capsule Antigen is HEAT LABILE (t/f)

K₁ antigen-

Antigenic structure of FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Envelope / K / Capsule Antigen

- E. coli

Vi antigen

Antigenic structure of FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Envelope / K / Capsule Antigen

- S. typhi

large, moist, gray colonies except Klebsiella and Enterobacter

BAP & CAP of FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Klebsiella and Enterobacter

BAP & CAP of FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES is large, moist, gray colonies except

E. coli

All are gamma hemolytic except

Klebsiella & Enterobacter

EK

All are non-encapsulated except

Shigella

All are GLUCOSE FERMENTERS and often with gas production, aerogenic except

Shigella, Klebsiella, & Yersinia pestis

SKY

All are motile (peritrichous) at 37C except

Yersinia pestis

All Yersinia are motile except

25 °C

All Yersinia are motile except Yersinia pestis at which temperature

Shigella dysentery

All are CATALASE POSITIVE except

Plesiomonas

All are CYTOCHROME OXIDASE NEGATIVE except

Erwinia & Enterobacter agglomerans

All are NITRATE REDUCERS except

Salmonella

Shigella

Yersinia

SSY

Most are commensal flora of the intestinal tract except

MacConkey agar

some are with pili or fimbriae of FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES which grows well on

1. trimethoprim-SXT

2. aminoglycosides

3. 3rd generation cephalosporins

   TA3

Drug of choice of FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

1. Pili

2. Some may possess PLASMID; resistance to antibiotics

3. Some may possess ANTIGENS that may be used to identify other groups

Virulence Factors of FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

antibiotics

Virulence Factors of FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

1. Some may possess PLASMID; resistance to -

identify other groups

Virulence Factors of FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

1. Some may possess ANTIGENS that may be used to -

GN Broth

Other Media of FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

For Salmonella

Selenite Broth

Other Media of FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

ennrichment broth for Salmonella and Shigella

Tetrathionate broth

Other Media of FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

enrichment for Salmonella

CIN (Cefsulodin Irgasan Novobiocin)

Other Media of FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

for Yersinia

Brilliant Green Agar

Other Media of FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

for other Salmonella except S. typhi

Brilliant Green

Other Media of FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Inhibitory agent of Brilliant Green Agar

Phenol Red

Other Media of FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

pH indicator of Brilliant Green Agar

• Eosin Y

• Methylene Blue

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Inhibitor of Eosin-methylene blue (EMB)

Lactose

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

CHO of Eosin-methylene blue (EMB)

• Eosin Y

• Methylene Blue

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Indicator of Eosin-methylene blue (EMB)

Pink-purple

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Fermentator of Eosin-methylene blue (EMB)

Colorless

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Non-Fermentator of Eosin-methylene blue (EMB)

• Crystal Violet

• Bile Salts

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Inhibitor of MacConkey agar (MAC)

Lactose

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

CHO of MacConkey agar (MAC)

Neutral Red

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Indicator of MacConkey agar (MAC)

Pink

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Fermentator of MacConkey agar (MAC)

Colorless

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Non-Fermentator of MacConkey agar (MAC)

Bile Salts

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Inhibitor of Xylose Lysine Desoxycholate (XLD)

• Xylose

• Lactose

• Sucrose

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

CHO of Xylose Lysine Desoxycholate (XLD)

Phenol Red

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Indicator of Xylose Lysine Desoxycholate (XLD)

Yellow

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Fermentator of Xylose Lysine Desoxycholate (XLD)

Colorless

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Non-Fermentator of Xylose Lysine Desoxycholate (XLD)

Bile Salts

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Inhibitor of Hektoen enteric agar (HEA)

• Salicin

• Lactose

• Sucrose

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

CHO of Hektoen enteric agar (HEA)

Bromthymol blue

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Indicator of Hektoen enteric agar (HEA)

Yellow

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Fermentator of Hektoen enteric agar (HEA)

Colorless

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Non-Fermentator of Hektoen enteric agar (HEA)

• Bile salts

• Brilliant Green

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Inhibitor of Salmonella Shigella (SSA)

Neutral Red

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Indicator of Salmonella Shigella (SSA)

Lactose

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

CHO of Salmonella Shigella (SSA)

Red

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Fermentator of Salmonella Shigella (SSA)

Colorless

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Non-Fermentator of Salmonella Shigella (SSA)

Brilliant Green

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Inhibitor of Bismuth Sulphite Agar (BSA)

Glucose

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

CHO of Bismuth Sulphite Agar (BSA)

Bismuth Sulfite

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Indicator of Bismuth Sulphite Agar (BSA)

Salmonella : Black

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Fermentator of Bismuth Sulphite Agar (BSA)

Salmonella : Black

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Non-Fermentator of Bismuth Sulphite Agar (BSA)

• Bile salts

• pH 8.6

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Inhibitor of Thiosulfate-Citrate-Bile-Salt Sucrose Agar (TCBS)

Sucrose

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

CHO of Thiosulfate-Citrate-Bile-Salt Sucrose Agar (TCBS)

• Thymol Blue

• Bromothymol Blue

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Indicator of Thiosulfate-Citrate-Bile-Salt Sucrose Agar (TCBS)

Yellow

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Fermentator of Thiosulfate-Citrate-Bile-Salt Sucrose Agar (TCBS)

Green / Colorless

Media Used for FAMILY ENTEROBACTERIACEAE/ ENTEROBACTERALES

Non-Fermentator of Thiosulfate-Citrate-Bile-Salt Sucrose Agar (TCBS)

1. TSI (Triple Sugar Iron)

2. LIA (Lysine Iron Agar)

3. IMVC Test

DIAGNOSTIC TEST FOR GRAM (-) ENTERIC BACILLI

TSI (Triple Sugar Iron)

Uses TSI dispensed as slant and butt

1 part glucose (butt)

10 parts of sucrose & lactose (slant)

Composition of TSI (Triple Sugar Iron)

Fermentable CHO

Phenol Red

Composition of TSI (Triple Sugar Iron)

pH indicator

Ferrous sulfate/ Sodium Thiosulfate

Composition of TSI (Triple Sugar Iron)

H2S indicator

Yellow (acid)

Composition of TSI (Triple Sugar Iron)

Possible Results in Acid Urine

Red (K)

Composition of TSI (Triple Sugar Iron)

Possible Results in Alkaline Urine

Black color

Composition of TSI (Triple Sugar Iron)

Possible Results of H2S production

splitting of media & pulling away / cracks

Composition of TSI (Triple Sugar Iron)

Possible Results of Gas production

K/K

TSI (Triple Sugar Iron)

If slant/butt is Red/ Red report it as

No sugar was fermented

TSI (Triple Sugar Iron)

If slant/butt is Red/ Red interpret it as

NON- FERMENTERS

TSI (Triple Sugar Iron)

If slant/butt is Red/ Red classify it as

K / A

TSI (Triple Sugar Iron)

If slant/butt is Red / Yellow report it as

Glucose (+); lactose, sucrose (-)

TSI (Triple Sugar Iron)

If slant/butt is Red / Yellow interpret it as

Non-lactose Fermenters

TSI (Triple Sugar Iron)

If slant/butt is Red / Yellow classift it as

A/ A

TSI (Triple Sugar Iron)

If slant/butt is Yellow / Yellow report it as

Glucose (+); both or either lactose and sucrose

TSI (Triple Sugar Iron)

If slant/butt is Yellow / Yellow interpret it as

LACTOSE FERMENTERS

TSI (Triple Sugar Iron)

If slant/butt is Yellow / Yellow classify it as

18 - 24 hrs

TSI (Triple Sugar Iron)

Rapid Lactose Fermenters (A/A) hours

• E. coli

• Klebsiella

• Enterobacter

EKE

TSI (Triple Sugar Iron)

Rapid Lactose Fermenters (A/A) are

48 hrs

TSI (Triple Sugar Iron)

Late Lactose Fermenters (A/A) hrs

1. Citrobacter

2. Serratia

3. Salmonella arizonae

4. Shigella sonnei

5. Y. enterocolitica

CSSSY

TSI (Triple Sugar Iron)

Late Lactose Fermenters (A/A) are

1. Proteus

2. Providencia

3. Edwardsiella

4. Morganella

5. All Salmonella except S. arizonae

6. All Shigella except S. sonnei

7. All Yersinia except Y. enterocolitica

PPEM

TSI (Triple Sugar Iron)

Non-Lactose Fermenters (K/A) are