MacConkey Agar- Composition, Principle, Uses,

Preparation and Colony Morphology

Last updated: June 11, 2018 by Sagar Aryal
MacConkey agar (MAC) was the first solid differential media to be formulated which was
developed at 20th century by Alfred Theodore MacConkey. MacConkey agar is a selective and
differential media used for the isolation and differentiation of non-fastidious gram-negative rods,
particularly members of the family Enterobacteriaceae and the genus Pseudomonas.
Composition of MacConkey Agar

Ingredients Amount
Peptone (Pancreatic digest of gelatin) 17 gm
Proteose peptone (meat and casein) 3 gm
Lactose monohydrate 10 gm
Bile salts 1.5 gm
Sodium chloride 5 gm
Neutral red 0.03 gm
Crystal Violet 0.001 g
Agar 13.5 gm
Distilled Water Add to make 1 Liter
Final pH 7.1 +/- 0.2 at 25 degrees C.
Principle of MacConkey Agar

MacConkey agar is used for the isolation of gram-negative enteric bacteria and the
differentiation of lactose fermenting from lactose non-fermenting gram-negative
bacteria. Pancreatic digest of gelatin and peptones (meat and casein) provide the essential
nutrients, vitamins and nitrogenous factors required for growth of microorganisms. Lactose
monohydrate is the fermentable source of carbohydrate. The selective action of this medium is
attributed to crystal violet and bile salts, which are inhibitory to most species of gram-positive
bacteria. Sodium chloride maintains the osmotic balance in the medium. Neutral red is a pH
indicator that turns red at a pH below 6.8 and is colorless at any pH greater than 6.8. Agar is the
solidifying agent.
Uses of MacConkey Agar

1. MacConkey agar is used for the isolation of gram-negative enteric bacteria.

2. It is used in the differentiation of lactose fermenting from lactose non-fermenting gram-
negative bacteria.
3. It is used for the isolation of coliforms and intestinal pathogens in water, dairy products
and biological specimens.
Preparation of MacConkey Agar

1. Suspend 49.53 grams of dehydrated medium in 1000 ml purified/distilled water.

2. Heat to boiling to dissolve the medium completely.
3. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes.
4. Cool to 45-50°C.
5. Mix well before pouring into sterile Petri plates.
Result Interpretation on MacConkey Agar

Lactose fermenting strains grow as red or pink and may be surrounded by a zone of acid
precipitated bile. The red colour is due to production of acid from lactose, absorption of neutral
red and a subsequent colour change of the dye when the pH of medium falls below 6.8.
Lactose non-fermenting strains, such
as Shigella and Salmonella are colourless and transparent and typically do not alter appearance
of the medium. Yersinia enterocoliticamay appear as small, non-lactose fermenting colonies
after incubation at room temperature.
Colony Morphology on MacConkey Agar

Organism Colour Remarks

red/pink non-mucoid
Escherichia coli

pink mucoid
Aerobacter aerogenes
 red minute, round
Enterococcus species

pale pink opaque

Staphylococcus species

green-brown fluorescent growth

Pseudomonas aeruginosa

Limitations of MacConkey Agar

1. The colonial characteristics described give presumptive identification only of the isolated
organisms. It is necessary to subculture and carry out confirmation tests for final
identification.
2. Some strains may be encountered that grow poorly or fail to grow on this medium.
3. Incubation of MacConkey Agar plates under increased CO2 has been reported to reduce
growth and recovery of a number of strains of Gram-negative bacilli.
4. Some strains of Proteus may swarm on this medium.
References

1. Austin Community College, 5930 Middle Fiskville Rd., Austin, Texas

2. ASM Microbe Library: MacConkey Agar Plates Protocols
3. Thermo Fisher Scientific Inc., Dehydrated Culture Media: MacConkey Agar
4. Acumedia Manufacturers: MacConkey Agar
5. HiMedia Laboratories Pvt. Ltd, Technical data: MacConkey Agar
6. Hardy Diagnostics: MacConkey Agar
7. Science Prof Online (SPO): MacConkey Agar
8. Bacteriological Analytical Manual, 8th Edition, Revision A, 1998.
9. Collin County Community College District.
10. Microbe Online
11. Wikipedia
CategoriesCulture Media
12 thoughts on “MacConkey Agar- Composition, Principle, Uses, Preparation and Colony
Morphology”

1. David Kator
July 17, 2018 at 2:37 PM | Reply
Thank you sir for your excellent notes, it really guides me in the treatment plant where i’m working as a
microbiologist in the lab. please did you have notes on other media, their composition, uses, preparation
and their appearances /colour after culturing.
2. Dr. Manjunath Ramanna,
January 21, 2017 at 7:42 AM | Reply
Dear Sir,
I salute you for the nice explanations in Microbiology you regularly upload in Facebook. I share & use
your blogs for teaching my Uunder graduate students. I am sure, someday some one will recognize your
writing in blogs & offer you a good post & position. Pls continue your service to Microbiology
 Thank You Sir,

Dr. Manjunath Ramanna

Assistant Professor of Agricultural Microbiology,
University of Horticultural Sciences, Bagalkot, Karnataka, India
3. Spencer
January 20, 2017 at 11:09 PM | Reply
Aerobacter aerogenes?
4. Dan
January 20, 2017 at 6:57 PM | Reply
If MAC is for non-fastidious Gram-negative bacilli, why would Enterococcus and Staphylococcus give a
reaction on this medium? Shouldn’t they show no growth? It may be that Enterococcus and
Staphylococcus ferment lactose, but wouldn’t the crystal violet and bile salts inhibit their growth? Just
curious.
5. Samiksha J
January 12, 2017 at 8:45 PM | Reply
I am samiksha working as a microbiologist in ferm we are using macconkeys agar for water pathogen
testing some times we found some submerged precipitation like something but not grow when
subculture, in macconkeys agar plate after 60 to 72 hrs of incubation at 30 to 35 degrees incubation.
Please suggest .
6. Giba
December 20, 2016 at 3:09 PM | Reply
Pls what is the colour of salmonella spp on MacConkey agar
 Soni
March 16, 2017 at 7:38 PM | Reply
Colourless or cream
7. Jennifer Gaita
July 4, 2016 at 12:50 PM | Reply
Asante sana (thank you much) for your post. Am a foods, nutrition & dietetics student currently on
practicum in a dairy factory. Initially couldn’t understand this MacConkey that i encountered in the
microbiology lab but now i can understand it’s uses. For this am thankful.
8. okojie pamela
May 12, 2016 at 10:56 PM | Reply
This is remarkably helpful. Thank you so much. Am a student of food and technology, working on the
microbiology of pap was like been in the dark cos I was just following manual instructions without having
a full understanding of my project work but now I know better. Thank you very much
9. nirupama rao
May 5, 2016 at 1:48 AM | Reply
should I know brief description of
mycoplasma PPLO
10. Erytmyst
March 15, 2016 at 6:23 PM | Reply
Your example of vulgaris and aureus look like they have been switched to me
11. JAYANTA ROY
February 15, 2016 at 6:27 PM | Reply
Peptone-20gm
Agar-15gm
Vile solt-5gm