LKCR 3BMT 13

Bacteriology Prefinals Coverage

Biochemical Identification of Gram Negative Bacteria Principle Reagents/Indicator Presence of galactoside permease Depends on the Lactose Degradation and -galactosidase test/culture that cleaves lactose to medium monosaccharides Determines whether the organism is LLF or NLF Ortho-nitrophenyl(-galactosidase -Dhydrolyzes ONPG, ONPG galactopyranoside which is colorless, into test galactose and onitrophenol, a yellow compound Fermentation of glucose to pyruvic Methyl Red (MR) acid resulting in Methyl red test mixed acid products leading to a decrease in pH (4.5-5.5) Fermentation of glucose producing acetylmethylcarbinol Voges-Proskauer -naphthol, 40% (acetoin) and (VP) test KOH converted to diacetyl in the presence of oxygen and KOH Utillization of Citrate as a sole carbon Citrate Utilization source, producing NH3 test in Simmons and converted to Bromthymol blue Citrate Agar NH4OH that raises pH and takes the blue color Organisms that possess tryptophanase are Paracapable of Ehrlich Indole test dimethylaminodeaminating benzaldehyde tryptophan with formation of indole, pyruvic acid and Biochemical test (+) result LFs possess both enzymes LLFs possess galactosidase only (-) result NLFs do not possess either enzyme

LLF produce yellow pigmentation

Remains colorless

Red solution

Yellow solution

Red complex

Yellow color or copper-like

Prussian Blue color

Green color

Red color

Yellow color

 LKCR 3BMT 13

Bacteriology Prefinals Coverage

Motility test

Nitrate and nitrite reduction test

ammonia Growth pattern of the organism in reference to the inoculation line Ability of the organism to reduce nitrate to nitrite and further reduce nitrite to nitrogen gas Presence of cytochrome oxidase system Hydrolysis of urea into ammonia by the presence of urease and reacts to form ammonium carbonate that raises the pH and color changes the indicator to pink-red H2S production is indicated by a black precipitate Indole production is detected by a red ring/complex Motility is observed in the turbidity/ spreading from the inoculation line

use of semi-solid medium N,N-dimethyl-naphthylamine, sulfanilic acid, diazo red dye 0.5% or 1% tetramethyl-pphenylenediamine dihydrochloride

Movement away from stab line or hazy appearance Red color (presence of nitrite) Development of Lavander color on the moistened filter paper

Movement is confined on the stab line

No color change

Oxidase test

No color change

Urease test

Phenol red

Pink-red

No color change

Sulfide Indole Motility (SIM) Agar

Ehrlichs or Kovacs

Blackening of the butt, formation of red complex, movement away from the stab line/turbid medium

No color change and the movement is confined in the stab line

Triple Sugar Iron (TSI) agar

Ability of the organism to ferment glucose (butt), lactose and/or sucrose (slant);H2S production and gas production

pH indicator: phenol red H2S production indicator: Ferrous sulfate & sodium thiosulfate

Glucose fermentation: yellow butt Lactose and/or sucrose fermentation: yellow slant H2S production: black ppt Gas production: presence of crack, bubble or gas space Lysine Deamination: red slant

Glucose fermentation: orange butt Lactose and/or sucrose fermentation: orange/red slant H2S production: no black ppt Gas production: absence of crack, bubble or gas space Variable

Lysine Iron Agar (LIA) Slant

Ability of the organism to deaminate (slant) or

pH indicator: bromcresol purple

 LKCR 3BMT 13

Bacteriology Prefinals Coverage

decarboxylate (butt) lysine; fermentation of glucose (butt); H2S production

H2S production indicator: Ferrous sulfate & sodium thiosulfate

Lysine Decarboxylation And Glucose Fermentation: purple butt Glucose Fermentation only: yellow butt H2S production indicated by blackening of the medium

Reactions on Triple Sugar Iron Agar *from the BacteLab Finals pdf* Observations Yellow slant / Yellow Butt with presence of crack/ bubble/ gas space Yellow slant / Yellow Butt with presence of crack/ bubble/ gas space and black precipitate Red slant / red Butt Red slant / Yellow butt with presence of crack/bubble/gas space Red slant / Yellow butt with black precipitate Reactions Interpretations Lactose Fermenter Glucose fermenter Gas Producer No H2S Lactose fermenter Glucose fermenter Gas producer H2S producer Non-Lactose Fermenter Non-glucose fermenter No Gas No H2S Non Lactose fermenter Glucose fermenter Gas producer No H2S Non-lactose fermenter Glucose fermenter H2S producer No gas Non-lactose fermenter Glucose fermenter Gas producer H2S producer Representative organisms Escherichia (RLF) Enterobacter (RLF) Klebsiella (RLF) Serratia (LLF)

A/AG or A/A

A/AG+ or A/A+

Citrobacter freundii (LLF) Salmonella arizonae (LLF)

K/K

Pseudomonas aeruginosa Alcaligenes faecalis (These are not members of Family Enterobacteriaceae)

K/AG or K/A

Morganella

K/A+

Salmonella typhi

Red slant / Yellow butt with presence of K/AG+ or K/A+ crack/bubble/gas space and black precipitate

Salmonella Proteus Edwardsiella

 LKCR 3BMT 13

Bacteriology Prefinals Coverage

Yellow slant / Yellow butt

A/A

Red slant / Yellow butt

K/A

Lactose fermenter Glucose fermenter No gas No H2S Non-lactose fermenter Glucose fermenter No gas No H2S

Shigella sonnei (LLF) Yersinia enterocolitica (LLF)

Shigella Providencia

Reactions on LIA slant * from BacteLab Finals reviewer* Observations Reactions Interpretation Lysine deaminase negative Lysine decarboxylase positive Glucose fermenter No H2S Lysine deaminase negative Lysine decarboxylase positive Glucose fermenter H2S producer Lysine deaminase positive Lysine decarboxylase negative Glucose fermenter No H2S Lysine Deaminase negative Lysine decarboxylase negative Glucose fermenter No H2S Lysine deaminase negative Lysine decarboxylase negative Glucose fermenter H2S producer Representative Organisms Escherichia coli Enterobacter aerogenes Klebsiella pneumonia Serratia marcescens

Purple slant / dark purple butt

K/K

Reddish-purple slant / dark purple butt with black precipitate

K/K+

Salmonella typhi Salmonella arizonae Edwardsiella tarda

Reddish-purple slant / yellow butt

R/A

Proteus vulgaris Proteus mirabilis Providencia rettgeri Morganella morganii Enterobacter sakazakii Enterobacter cloacae Yersinia enterocolitica Shigella dystenteriae Citrobacter koseri

Purple slant / yellow butt

K/A

Purple slant / yellow butt with black precipitate

K/A+

Citrobacter freundii Salmonella paratyphi