Hemorrhagic Coli Agar is general media used for the isolation of Hemorrhagic colitis type of Escherichia coli. Hemorrhagic colitis is a type of gastroenteritis in which certain strains of the bacterium Escherichia coli infect the large intestine and produce a toxin that causes bloody diarrhea and other serious complications.
The media is composed of tryptone, sorbitol, sodium chloride, bile salt, bromocresol purple, and agar. Tryptone provides carbon, protein, and nutrient sources required for the growth of microorganisms. Sorbitol provides selectivity and Hemorrhagic coli (E.coli O157:H7) does not ferment sorbitol and produces colorless colonies.
Sodium chloride maintains the osmotic balance of the medium. Bile salts inhibit accompanying gram-positive bacteria. Homogenize 10 grams of the test sample in 90 ml Peptone Water (prepare 1:100 dilutions). Pipette 1 ml aliquots through disposable 100 µm pre-filter and add to 10 ml Peptone Water filtered through Hydrophobic Grid Membrane Filters.
Place the filters onto Hemorrhagic Coli Agar and incubate at 43°C for 16-20 hours. Replicate colony growth onto other HGMFs using an HGMF replicator. Incubate replicates on HC Agar at 43°C for 16-20 hours and test original filters with conjugated antibody.
Use: Recommended for isolation and enumeration of Escherichia coli with an enzyme-labeled monoclonal antibody.
