Xylose Lysine Deoxycholate Agar Plate

Principle And Interpretation

Enterobacteriaceae is a family of gram-negative, non-spore-forming bacilli that contains more than 100 species of bacteria that normally inhabit the intestines of humans and animals. Members forming part of the normal intestinal microflora are referred to as coliforms. The clinically significant genera of Enterobacteriaceae include Cedecea, Citrobacter, Edwardsiella, Enterobacter, Escherichia, Ewingella, Hafnia, Klebsiella, Kluyvera, Proteus, Salmonella, Shigella and Yersinia (1).

The Salmonellae are the most complex of all the Enterobacteriaceae. Human Salmonella infections are most commonly caused by ingestion of food, water or milk, contaminated by human or animal excreta (2). A large number of media have been developed for the selective isolation and identification of enteric bacilli including Salmonella.

Xylose Lysine Deoxycholate Agar is a selective as well as differential medium formulated by Taylor (3-7) for the isolation and identification of enteric pathogens especially Shigellae from stool samples. It is also used for pharmaceutical testing and non-sterile product testing for the detection (or absence) of Salmonella after enrichment in Rappaport Vassiliadis Salmonella Enrichment Broth (MH1491) in accordance with the harmonized method of USP/EP/BP/JP/IP (8,9,10,11,12).

Deoxycholate, ferric ammonium citrate and sodium thiosulphate are selective agents that inhibit gram-positive microorganisms. Essential nutrients, growth factors for growth of microorganism are provided by yeast extract. Xylose, sucrose and lactose are the fermentable sugars in this medium. Xylose is fermented by almost all the enteric bacteria except Shigellae, which enable the differentiation of Shigellae from Salmonellae. Salmonellae metabolize the xylose and decarboxylate lysine and thus change the pH to alkaline and mimic Shigellae reaction. However to prevent this reaction by lysine positive coliforms, lactose and sucrose are added in excess to produce acid and hence nonpathogenic H2S producers do not decarboxylate lysine. Sodium thiosulphate helps in reactivation of sulphur containing compounds and prevents the desiccation of these compounds during storage. It also forms the substrate for enzyme thiosulphate reductase, which breaks it to form H2S. Thiosulphate and ferric ammonium citrate are the H2S indicators in the medium. Sodium thiosulphate is also inactivator of halogens, mercurial and aldehyde and can minimize its toxicity in the testing sample, if any during microbial limit tests. Sodium chloride maintains the osmotic equilibrium in this medium. Phenol red is the pH indicator. Degradation of fermentable sugars proceed concurrently and generates acids, which cause pH indicator to give various shades of colour, causing a color change in the colonies and in the medium from red to yellow on prolonged incubation. Hydrogen sulfide production results in colonies with black centers under alkaline conditions, which can be inhibited by acid production by carbohydrate fermentation. Alkaline condition causes the color of the medium to change back to red. This medium is an ideal medium for screening samples containing mixed flora of enteric pathogens as recovery of Salmonellae and Shigellae is not conspicuous by even profuse growth of other species (13,14).