Produce TSA plates, TSA slants, and TSB which will be used in subsequent lab periods.
Understand how to make media, how to sterilize it, and how to distribute it in different formats. As a medium for blood cultures BHI with 10% defibrinated sheep blood Useful for isolation and cultivation of Histoplasma capsulatum and other fungi BHI with 0.\) To prepare inoculum suspensions for antibiotic susceptibility testing, microdilution, minimal inhibitory concentration (MIC)Ģ. Some common modifications and its usage are described below: Modified medium Usage BHI with Fildes Used for cultivating capsular strains of Haemophilus influenzae BHI with Hemin and Viatmin K Primarily used for anaerobic organism cultivation and the recovery of fastidious anaerobic organisms. Addition of one or more growth supplements facilitates the growth of the desired organism. Various modifications of BHI serve various processes. Since nutritional requirements of organisms vary, some strains may fail to grow or grow poorly in this medium. For the culture of yeasts or fungi, a prolonged incubation period may be required to obtain good growth. If desired, a loopful of the culture broth can then be sub-cultured onto an appropriate solid medium to observe colony morphology and further tests can be performed from the isolated colonies. Interpretation of Results: After the incubation period examine tubes for turbidity which is an indication of growth. For anaerobic cultures, examine tubes after 48 hours. Reincubate tubes an additional 24 hours if required. For aerobic and others, examine tubes after 24 hours and at 48 hours. For anaerobic organisms, incubate tubes anaerobically at 35☌ with loose caps. For fastidious organisms such as H aemophilus species incubate in a CO2-enriched atmosphere at 35☌ with a loose cap. For aerobic organisms, incubate aerobically at 35☌ with loose caps. For anaerobic organisms, prior to inoculation, pre-reduce the broth by placing the tubes in an anaerobic environment overnight at room temperature and then inoculate lightly. Lightly inoculate the broth using the test sample or with the organism of interest (for aerobic and others). Allow medium to adjust to room temperature prior to inoculation. Prepared tubed or bottled medium needs to store in the dark and below 20☌. Storage conditions and Shelf lifeĭehydrated medium can be stored at 10-30☌ and use before the expiry date on the label. Please refer to the instruction given by the manufacturer. NOTE: The composition and method for preparation of media vary according to the manufacturer. Store the culture medium at 2-8☌ sealed in plastic bags to reduce the chances of contamination. Label the side of each tube with the date of preparation and batch number. Sterilize by autoclaving at 15 Ibs (121° C) for 15 minutes. Dispense into bottles or tubes as desired. 
Heat if necessary to dissolve the medium completely.Suspend 37.0 g of medium in 1 liter of distilled water.Procedure for preparation and storage of BHI Broth:
Ingredients Gms / Litre Calf brain, infusion from 200 Beef heart, infusion from 250 Proteose peptone 10 Dextrose 2 Sodium chloride 5 Disodium phosphate 2.5 Final pH ( at 25☌) 7.4☐.2 The quantity of the ingredients may vary depending upon the manufacturer and the addition of some selective growth supplements however, the basic ingredients remain the same. Composition of Brain Heart Infusion (BHI) Broth Disodium phosphate helps in maintaining the buffering action of the medium whereas sodium chloride maintains the osmotic equilibrium of the medium. The medium contains proteose peptone and infusions from calf brain and beef heart which serve as sources of carbon, nitrogen, essential growth factors, amino acids and vitamins. Uses of Brain Heart Infusion Broth Principle.Composition of Brain Heart Infusion (BHI) Broth.
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