Plated Culture Media:

Acronym

Description

Uninoculated state and Results

XLD

Xylose Lysine Deoxycholate Agar 1. Yellow colonies with black centre of xylose fermenting and hydrogen sulphide producing Gram negative enteric bacilli like: a. Citrobacter freundii b. Proteus mirabilis c. Proteus vulgaris 2.

uninoculated

yellow colonies without black centre of xylose fermenting BUT non-hydrogen sulphide producing Gram negative bacilli like: a. Escherichia coli b. Serratia marcescens c. Klebsiella pneumoniae d. Enterobacter aerogenes beta haemolytic domee. Providencia rettgeri shaped colonies Highly selective and differential medium which is used to isolate and differentiate negative enteric bacilli, especially Salmonella and Shigella species based on xylose fermentation and lysine decarboxylation inhibitors: deoxycholate and citrate pH indicator: phenol red carbohydrates: xylose, dextrose and sucrose H2S indicator: Ferric ammonium citrate with thiosulphate Other components: amino acid lysine

pin head colonies Staphylococcus aureus

Acronym

Description

Uninoculated state and Results

BAP

Blood Agar Plate an enriched medium that can be used to isolate both gram positive and gram negative cocci and bacilli. The enriching substance is preferably unheated 5% defibrinated sheep blood. this is also classified as differential culture medium because it differentiates organisms based on their haemolytic patterns exhibited by the medium. Alpha-haemolytic- if colony is surrounded by an incomplete zone of haemolysis wich is translucent and green in colour. Beta-haemolytic- if colony is surrounded by a complete zone of haemolysis which is clear and transparent Gamma-haemolytic- if colony is not surrounded by any zone of haemolysis; there is not change in the surrounding medium uninoculated

large gray mucoid colonies of gram negative enteric bacilli

Acronym

Description

Uninoculated state and Results

TCBS

Thiosulphate Citrate Bile Salts Sucrose Agar - a highly selective medium which isolate and differentiate the sucrose fermenting from non-sucrose fermenting species of Vibrio. - inhibitors: thisulphate, citrate, bile salts, and high pH - indicator: bromthymol blue - carbohydrate: sucrose

Uninoculated

yellow colours of sucrose fermenting Vibrio like Vibrio cholerae and Vibrio alginolyticus SSA Salmonella-Shigella Agar a highly selective and differential medium which is used to differentiate species of Salmonella and Shigella inhibitors: brilliant green, bile salts, citrate pH indicator: neutral red uninoculated hydrogen sulphide indicator: ferric ammonium citrate with sodium thiosulphate carbohydrate: lactose

Acronym

Description

Uninoculated state and Results

MSA

Mannitol Salt Agar both selective and differential culture medium used to isolate and differentiate mannitol fermenting from non-mannitol fermenting Staphylococcus inhibitor: 7.5% NaCl pH indicator: Phenol red carbohydrate: mannitol Pink colonies of non-mannitol fermenting Staph like: Staphylococcus Epidermidis Staphylococcus lugdunensis Staphylococcus saprophyticus Yellow colonies of mannitol fermenting Staph like Staphylococcus aureus

unioculated

Acronym

Description

Uninoculated state and Results

BSA

Bismuth Sulphite Agar a highly selective medium for the isolation of Salmonella typhi inhibitors: bismuth sulphite and brilliant green carbohydrate: glucose uninoculated

beta haemolytic domeshaped colonies

pin head colonies Staphylococcus aureus

Acronym

Description

Uninoculated state and Results

CAP

Chocolate Agar Plate an enriched culture medium that can be used to grow both gram positive and gram negative cocci and bacilli. the enriching substance is heated blood, preferably 5% defibrinated sheep blood uninoculated

Acronym

Description

Uninoculated state and Results

EMB

Eosin Methylene Blue Agar both a selective and differential culture medium which is used to isolate and differentiate the lactose-fermenting from the nonlactose fermenting gram negative bacilli inhibitors: eosin and methylene blue pH indicators: eosin and methylene blue carbohydrate: lactose uninoculated

Pink to purple with dark centre colonies (fish-eye) suggestive of Enterobacter aerogenes

colourless non-lactosefermenting colonies on EMB plate. Salmonella typhi, Shigella dysenteriae, Providencia sp. Morganella sp.

purple with greenish metallic sheen, suggestive of Escherichia coli

Acronym

Description

Uninoculated state and Results

HEA

Hektoen Enteric Agar both selective and differential culture medium used to isolate and differentiate the lactose fermenting from non-lactose fermenting gram begative enteric bacilli inhibitors: bile salts pH indicator: bomthymol blue hydrogen sulphide indicator: ferric ammonium citrate with Na thiosulphate carbohydrates: lactose, sucrose, salicin orange colonies without black centrelactose fermenting, non-hydrogen sulphide producing gram negatice enteric bacilli like E. coli, K. pneumoniae and E. aerogenes

uninoculated

orange colonies with black centre- lactose fermenting, hydrogen sulphide producing gram negative enteric bacilli like Citrobacter freundii

Acronym

Description

Uninoculated state and Results

MAC

MacConkey Agar both selective and differential culture medium which is used to isolate and differentiate the lactose fermenting from the non-lactose fermenting gram negative bacilli inhibitor: crystal violet and bile salts pH indicator: neutral red carbohydrate: lactose Pink to red colonies of lactose fermenting gram negative enteric bacilli yellow to colourless colonies of lactose fermenting Gram negative enteric bacilli

uninoculated

MHA

RED PIGMENTED COLONIES OF Serratia marcscens in Mueller Hinton Agar

IMVC Test Indole Test Set-up

Acronym

Description

Uninoculated state and Results

Tryptone Broth the liquid medium is the substrate in the INDOLE TEST which is used to identify the gram negative enteric bacilli based on the ability of organisms to produce the enzyme tryptophase. this enzyme decomposes this tryptone broth producing indole that subsequently forms a red coloured complex upon the addition of Kovacs reagent or paradimethyl amino benzaldehyde reagent. Kovacs reagent para-dimethyl amino benzaldehyde reagent

positive: red negative: yellow

Acronym

Description

Uninoculated state and Results

Methyl Red

The liquid medium is also known as peptone glucose broth and is used to identify the gram negative enteric bacilli used on the ability of the organisms to ferment glucose pyruvic acid by one of two pathways: mixed acid fermentation pathway, which is the Methyl red test and the butylene glycol pathway which is tested by the Vogues Proskauer test. Set-up: if the organism would ferment glucose via the mixed acid fermentation pathway, more acids are produced like lactic, acetic, formic, succinic acids, which decrease the pH of the medium to 4.4 or lower, hence, upon the addition of the indicator, methyl red the broth becomes red in colour. if the organism would not use this pathway, the pH of the medium increases to 5.5 or higher, ence upon the addition of the indicator, methyl red, the broth becomes yellow in colour.

Vogues Proskauer Test

if the organism would ferment glucose via the butylene glycol pathway, an intermediate product, acetyl methyl carbinol or acetoin which is neutral is converted to diacetyl upon the addition of the VP reagent. B (40% KOH with 0.3% creatine) in the presence of VP-reagent A (5% alpha-naphthol in abs.methyl alcohol). Diacetyl is red in colour. Negative is yellow in colour.

Acronym

Description

Uninoculated state and Results

Coagulase Test plasma.

-differentiates pathogenic S. aureus from the non pathogenic S. epidermidis and saprophyticus. slide test: plasma + specimen positive: visible clumping negative: no visible clumping tube test- plasma+ specimen positive: visibleclot or coagulation negative: no clot or coagulation pathogenic S. aureus - positive Non-pathogenic S. saprophyticus/epidermidis- negative

catalase test

catalase test uses 3% h202. the positive reaction is indicated by bubbling or effervescence positive: all Staph sp. all gram negative enteric bacilli except Shigella dysenteriae negative: all strep sp.

Acronym

Description

Uninoculated state and Results

Acronym

Description

Uninoculated state and Results

Acronym

Description

Uninoculated state and Results

Acronym

Description

Uninoculated state and Results