Boiled blood agar chocolate agar is an extremely rich culture medium and can be prepared by heating after the blood has been added. Potato dextrose agar pda is used for the cultivation of fungi. Composition, preparation, uses and types of hemolysis. Blood agar is a generalpurpose enriched medium used to grow fastidious organisms and to differentiate bacteria based on their hemolytic properties. Do not used media that shows signs of deterioration. Potato dextrose agar pda is a general purpose medium for yeasts and molds that can be supplemented with acid or antibiotics to inhibit bacterial growth. Both microorganisms grow on this selective media because they are gramnegative nonfastidious rods. In this article, im gonna explain the preparation of blood agar medium from the nutrient agar base check out the preparation of nutrient agar medium. Transfer thus prepared blood agar base to a 50c water bath. Blood agar consists of many ingredients but the primary ingredient is the blood, which came from a rabbit or a sheep. Blood agar is an enriched, bacterial growth medium.
Gentamicinblood agar for isolation of streptococcus pneumoniae from respiratory secretions article pdf available in journal of clinical microbiology 75. Nutrient agar contains beef extract, peptone and agar in water. Blood agar base media can be used with added phenolphthalein phosphate 7 for the. Solid media generally contain agar at a concentration of 1. Blood collection, medium preparation, and hemolysis. Some bacteria produce exoenzymes that lyse red blood cells and degrade hemoglobin.
This video has information about macconkey agar selective,differential primary plating medium microbiology. Media requirements for fastidious or infrequently isolated bacteria can be found in clsi document m45. Suspend 28 g of nutrient agar powder in 1 litre of distilled water. For example, it was used for studying irradiated escherichia coli, phages. In the us blood agar is usually prepared from tryptic soy agar or. Nutrient agar cm0003b and nutrient broth cm0001b from oxoid share almost the same medium composition. The blood agar medium is the simplest yet powerful enriched medium that supports the growth of almost all the microbes and extremely easy to prepare in the laboratory. The manner of preparation is the difference between them as chocolate agar requires lysis of the red blood cells when added to the molten agar base. This video has information about preparation of culture media. Blood agar bacterial growth medium science prof online. The key difference between blood agar and macconkey agar is that blood agar is an enriched differential medium used to isolate fastidious microorganisms and detect their hemolytic activity whereas macconkey agar is a selective and. Blood agar base infusion agar, with the addition of sterile blood, is used for the isolation, cultivation and detection of hemolytic activity of streptococci and other fastidious microorganisms.
This culture medium can be used without blood for setting up blood cultures and as a. The inclusion of blood as a nutritive supplement in culture media may pre date the use of agar. Oxoid blood agar base is a nonselective general purpose medium widely employed for the growth of pathogenic and nonpathogenic bacteria. Hardy diagnostics blood agar products are recommended for use as general purpose growth media for the isolation, cultivation, and differentiation of a wide variety of microorganisms. Examples includes crystal violet blood agar to select streptococcus pyogens from throat swabs, and kanamycin or neomycin blood agar to select anaerobes from pus. Prepare the volume of blood agar base needed in a flask according to the manufacturers instructions on the label of the dehydrated medium. Blood agar plates and hemolysis protocols american society of. The method for the preparation of basic microbiology media is given below. Pdf effect of blood agar from different animal blood on growth. Nutrient agar m001 intended use nutrient agar is used as a general purpose medium for the cultivation of less fastidious microorganisms, can be enriched with blood or other biological fluids. Blood agar composition, principle, preparation and uses online. Agar medium to be be poured into plates is sterilized in a flask, and then. It is treated and water and a number of other ingredients are added to sterilize the solution.
Blood agar base formulation has been used as a base for preparation of blood agar and to support good growth of a wide variety of fastidious microorganisms. As a result of rbc lysis, intracellular nutrients are released such as hemoglobin, coenzyme nicotinamide adenine dinucleotide, and hemin into the agar which is used by fastidious organisms like h. Preparation and use as heating all of the ingredients together is not advised, the medium is meant to be prepared in two parts as follows for a total of one liter mixture a. Reagent preparation liquefy bd tryptic soy agar bottled media by heating in an autoclave or steam cooker. Most tube media both broth and agar can be stored at room temperature for about two months.
Pdf gentamicinblood agar for isolation of streptococcus. The first ever developed solid differential medium for the growth of bacteria on the globe was macconkey agar medium mac medium, was formulated by and is named after a british bacteriologist, alfred theodore macconkey in the 20 th century besides being a differential solid medium, macconkey agar medium is a selective and indicator medium too and primarily used for the isolation of gram. Alternatively, the bottle may be placed into a jar containing water, which. Difference between blood agar and macconkey agar compare. Blood agar composition, preparation, uses vs chocolate. Differential identify hemolysis some bacteria secrete enzymes that lyse red blood cells hemolysins such that a clearing around the colony appears. Bd offers growth media backed by more than 180 years of combined bd difco and bd bbl expertise. Blood agar is a resuscitation medium according to 3. Autoclave the dissolved mixture at 121 degrees celsius for 15 minutes. Fastidious organisms, such as streptococci, do not grow well on ordinary growth media. Friday, 30 september 2005 author rebecca buxton information history the history of blood agar, as we know it today, is uncertain. Media are used to grow bacterial colonies millions of bacteria having arisen through the binary fission of a single progenitor.
Bd supplies bd difco and bd bbl brand media formulations and ingredients in dehydrated form. Ancillary reagents and laboratory equipment as required. Blood agar composition, principle, preparation and uses. Agar slant tubes are sterilized and then the rack is tilted to allow the agar to solidify in a slanted fashion. It is a highly nutritious medium generally used as a basal medium for preparing blood agar by supplementation with blood. The exceptions are blood agar, chocolate agar, kligler iron agar, lowensteinjensen agar, litmus milk broth, sim, and. In situations where preparation is uneconomic in time, prepared, sterilized media liquid and solid are available from the major school science equipment suppliers.
Blood agar, chocolate agar, loefflers serum slope etc are few of the enriched media. Add above components 40 gm, except sheep blood, to distilleddeionized water and bring volume to 950. Meningitis lab manual storage and shippingneisseria cdc. The exceptions are blood agar, chocolate agar, kligler iron agar, lowensteinjensen agar, litmus milk broth, sim, and thioglycollate broth, all of which should. Macconkey agar selective,differential primary plating. This study compared citrated sheep blood agar csba and huba with defibrinated horse blood agar and defibrinated sheep blood agar dsba for the isolation and antibiotic susceptibility testing of reference and clinical strains of streptococcus pneumoniae, streptococcus. The composition of chocolate agar and the blood agar is the same and the only difference is while preparing chocolate agar, the red blood cells are lysed. Blood agar is almost identical to nutrient agar except that it contains five to ten percent sheep, rabbit, or horse blood. Heat this mixture while stirring to fully dissolve all components. The concept of isolating microorganisms in culture media has. It can also be used as generalpurpose media without the addition of blood.
Because it is a highly nutritious medium it can also be used as a general purpose growth media without adding blood. Blood agar base infusion agar himedia laboratories. The specimen to be cultivated should be streaked onto a chocolate agar plate or onto the agar slant. Blood agar base is a highly nutritious medium generally used as a basal medium for preparing blood agar by supplementation with blood. It is useful for cultivating fastidious organisms and for determining the hemolytic capabilities of an organism. An interesting method of adding blood to agar media is described in. The main difference between them is that nutrient agar contains a solidifying agent, agar powder that causes the medium to solidify in room temperature, whereas nutrient broth remains in liquid form. Bacteriological media come an a wide range of types. Blood agar base media can be used with added phenolphthalein phosphate 7 for the detection of phosphate producing staphylococci, with. Blood agar is prepared by adding 510% by volume to a basal medium such as nutrient agar or other blood agar bases. Broths and agar deeps are dispensed into tubes and then sterilized.
The addition of blood permits the interpretation of hemolytic activities. Human blood agar huba is widely used in developing countries for the isolation of bacteria from clinical specimens. As a bacterial culture medium, citrated sheep blood agar. Heat with frequent agitation and boil for 1 min to completely dissolve. It can also be used for preparation of salmonella typhi antigens 9.
Bacteria can produce different types of hemolysins. It will cause the agar to breakdown and become unusable. Blood agar and chocolate agar are quite similar but they differ in preparation. All broth media should be stored in an appropriate container at 4. Use mueller hinton agar for routine testing of nonfastidious organisms. Blood agar is a type of growth medium trypticase soya agar enriched with 5% sheep blood that encourages the growth of bacteria, such as streptococci, that otherwise wouldnt grow. Preparation of media and cultures microbiology online. By standardizing the media employed, the lowdust granule mixtures guarantee that the correct enrichment will be present for optimal cell viability and growth. Tryptic soy agar base with 5% sheep blooddehydrated microbiology culture media cultivate and isolate microorganisms for researching purposes. Preparation, production, storage and performance testing of culture media. Pdf quality control of culture media in a microbiology. Composition, principle, preparation and uses of blood agar. Blood agar is a generalpurpose enriched medium used to grow fastidious.
P33 is not intended to be used for the diagnosis of human disease. Blood agar is a bacterial growth medium that can distinguish normal from. Preparation of blood agar medium from nutrient agar base. Blood agar contains general nutrients and 5% sheep blood. Blood agar base for the isolation and cultivation of various fastidious microorganisms find msds or sds, a coa, data sheets and more information. Blood agar ba are enriched medium used to culture those bacteria or microbes that do not grow easily. It is recommended for plate count methods for foods, dairy products and testing cosmetics. Original article a suitable blood agar containing human. It is also used as a base for the preparation of a variety of special media. Target media were blood agar, chocolate agar, macconkey agar, sorbitol macconkey agar smac, campylobacter agar, and selective media for pathogenic neisseria spp. Preparation of macconkey agar medium mac in laboratory. Prepare the blood agar base as instructed by the manufacturer.
Once the nutrient agar has been autoclaved, allow it to cool but not solidify. Blood contains inhibitors for certain bacteria such as neisseria and. Blood agar composition, preparation, uses and pictures. Summary and explanation infusion agar is an allpurpose medium which has been used for many years as a base for the preparation of blood agars. The prepared media is distributed in different ways, depending on the form one is making. Nutrient agar is a complex medium because it contains ingredients with contain unknown amounts or types of nutrients. Blood agar plate ba nutrient agar with 5% sheep blood cultivation of fastidious and non fastidious bacteria. Mueller hinton agar w 5% sheep blood should be used to test strains that fail to grow satisfactorily on plain mueller hinton, such as streptococus spp. Potato dextrose agar pda principle, uses, procedure. Since 1935, bd bbl prepared media products have brought to the microbiology laboratory the highest levels of quality and performance. Blood agar and macconkey agar are two different media which belong to the category of differential media.
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