MacConkey agar
MacConkey agar is a selective and differential solid culture medium widely used in clinical and environmental microbiology to isolate and identify Gram-negative enteric bacteria, particularly those from the Enterobacteriaceae family. Developed by British bacteriologist Alfred Theodore MacConkey at the turn of the 20th century as an improvement on earlier bile salt media, with initial description in 1900 and detailed formulation in 1905, it inhibits the growth of Gram-positive organisms while allowing differentiation of lactose-fermenting bacteria (which produce pink or red colonies due to acid production lowering the pH) from non-fermenters (which form colorless colonies).[1][2] In practice, MacConkey agar is essential for processing clinical specimens like stool, urine, and wound swabs to detect pathogens such as Escherichia coli, Salmonella spp., and Shigella spp., aiding in the presumptive identification based on colony morphology, color, and growth characteristics.[3][4] It is also employed in food and water testing by agencies like the FDA to isolate coliforms and other indicators of fecal contamination.[5] While highly effective for its intended purpose, limitations include potential overgrowth by fastidious non-enteric Gram-negatives and the need for confirmatory biochemical tests for definitive identification.[3]Composition and Preparation
Key Ingredients
MacConkey agar is formulated with a precise combination of nutrients, selective agents, indicators, and gelling components to support the growth of Gram-negative bacteria while enabling differentiation based on lactose fermentation. The standard composition, as specified by manufacturers such as Oxoid (Thermo Fisher Scientific), includes gelatin peptone at 17.0 g/L as the primary nitrogen source, providing essential amino acids and peptides for bacterial metabolism.[6] Additional nitrogen sources consist of casein peptone at 1.5 g/L and meat peptone at 1.5 g/L, which supplement the medium to support the growth of fastidious enteric organisms.[6] Lactose serves as the key fermentable carbohydrate at 10.0 g/L, allowing for the differentiation of lactose-fermenting bacteria through acid production.[6] Bile salts, present at 1.5 g/L, act as selective agents by disrupting the cell membranes of Gram-positive bacteria, thereby inhibiting their growth and favoring Gram-negative enterics.[6] Sodium chloride is added at 5.0 g/L to maintain osmotic balance and approximate physiological salinity for optimal bacterial growth.[6] The pH indicator neutral red is incorporated at 0.03 g/L, which remains colorless above pH 6.8 but turns red in acidic conditions resulting from lactose fermentation.[6] Crystal violet, at 0.001 g/L, enhances the selective inhibition of Gram-positive bacteria by interfering with their cell wall synthesis.[6][7] Agar, ranging from 13.5 to 17 g/L depending on the formulation, provides the solidifying agent to create a gel matrix for colony formation.[6] The medium is prepared in distilled water to a total volume of 1 L and sterilized by autoclaving at 121°C for 15 minutes.[6] Variations in the exact recipe exist among manufacturers; for instance, BD Difco specifies pancreatic digest of gelatin (equivalent to gelatin peptone) at 17 g/L and combined meat and casein peptones at 3 g/L, with the remaining components matching the standard proportions.[8]| Ingredient | Amount (g/L) | Function |
|---|---|---|
| Gelatin peptone | 17.0 | Primary nitrogen source, supplies amino acids for growth |
| Casein peptone | 1.5 | Supplementary nitrogen for fastidious organisms |
| Meat peptone | 1.5 | Supplementary nitrogen for fastidious organisms |
| Lactose | 10.0 | Fermentable carbohydrate for differentiation |
| Bile salts | 1.5 | Selective inhibition of Gram-positive bacteria |
| Sodium chloride | 5.0 | Osmotic balance |
| Neutral red | 0.03 | pH indicator (red below pH 6.8) |
| Crystal violet | 0.001 | Enhances Gram-positive suppression |
| Agar | 13.5–17.0 | Solidifying agent |