Agar m434 can also be used in place of chocolate agar base, which gives slightly better results than chocolate agar 4. Chocolate agar with vx, prepared media plates, 100 x 15. Specialized media enrichment broths encourage the growth of a particular type of. Place the membrane upon two ttc tergitol agar plates. Chocolate agar is a nonselective, enriched growth medium. Criterion urea agar base concentrate is a 10x concentrate for the preparation of christiansens medium and requires the supplementation of bacteriological grade agar cat. Chocolate ii agar gc ii agar with hemoglobin and isovitalex for the isolation and cultivation of fastidious microorganisms, especially neisseria and haemophilus species, from a variety of clinical specimens. It is a variant of the blood agar plate, containing red blood cells that have been lysed by slowly heating to 80c. R00 protective measures hands disposable vinyl gloves. Haemophilus are small, pleomorphic, gramnegative coccobacilli. Tryptic soy agar is a complex, nonselective, and nondifferential media similar to nutrient agar with the exception that it contains soy extract instead of yeast extract.
Agar agar is available as a powder, flakes, bars or strands. Selective media contain ingredients that inhibit the growth of some organisms but allow others to grow. Nn non nutrient amoeba saline agar s pe ci e dia r me. Precipitated hemoglobin may look as dark spots on the media but it does not have any impact on the media s performance. This media can be prepared as a broth with the exclusion of agar solidifying agent to form a broth.
It melts when heated and forms a jelly as it cools. Suitability of a chocolate agarbased medium for the transportation and long term storage of neisseria gonorrhoeae isolates at room temperature. To make solid media, a solidifying agent, agar, is required. A transparent mucilaginous substance, agaragar is used like gelatin, which it can replace in most recipes. Do not used media that shows signs of deterioration. The blood agar can be used with added phenolphthalein phosphate for the detection of phosphatase. If so, the media should be discarded and a new batch should be prepared. Agar of the same composition with the commercial agar can be made by following the correct procedures. Selective media for the isolation of actinobacillus pleuropneumoniae from the pig article pdf available in acta veterinaria 545 january 2004 with 587 reads how we measure reads. If you are going to use a commercially prepared cultured media be it chocolate agar or other types of media, make sure you perform quality control testing. Chocolate agar safety data sheet date of issue 27092007. The species listed are not an inclusive list and are provided to assist novice users in selecting compatible media and bacteria. Each batch of media prepared in the laboratory and each new manufacturers lot number of media should be tested using appropriate qc reference strains for sterility, the ability to support. For example, mannitol salt agar contains a high concentration of sodium chloride that inhibits the growth of most organisms but permits staphylococci to grow.
An enrichment medium should also be inoculated and one such as. First, a number of ingredients are added to water, including heart infusion brood, p. Apr, 2009 this is strictly a matter of taste and convenience. Test the medium with a laboratory stock strain of shigella species which is in the rphase. The centrifuged deposit should be inoculated on chocolate agar and incubated in a humidified atmosphere and 10%co 2, and on blood agar incubated aerobically. 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. Pdf selective media for the isolation of actinobacillus. The need for complex growth media has complicated routine susceptibility testing of haemophilus influenzae because of antagonism of certain antimicrobial agents by the medium or because of difficulties in interpretation of growth endpoints. Early in the 20th century, the american public health associa tion published the formula for a general purpose medium for the growth of a wide variety of.
Foods thickened with agaragar are firmer and melt less readily than those containing gelatin. Nnn can also be used to grow trypanosoma cruzi references. Muellerhinton agar an overview sciencedirect topics. In blood agar, blood is added to media after autoclaving heat sterilization. Darkening and clearing of the medium was usually associated with the species staphylococcus epidermidis, staphylococcus aureus, staphylococcus simulans, and streptococcus faecalis. Neisseria gonorrhoeae are the most fastidious of the neisseria species, require complex growth media and are highly susceptible to toxic substnces e. Summary and explanation chocolate agar was formulated in 1927 using peptones and yeast extract.
Chocolate agar with the addition of bacitracin becomes selective, most critically, for the genus haemophilus. Chocolate agar, a differential medium for grampositive cocci. This is strictly a matter of taste and convenience. Prepare the volume of tsa needed in a flask according to the instructions given on. It is recommended that the laboratory confirm the performance characteristics of this media. Typically, liquid media is prepared with agar, heated for sterilization purposes and then poured into petri dishes before the agar hardens. A hidden pitfall in the preparation of agar media undermines. Recommended for selective enumeration and identification of coliform organisms. Chocolate agar with vx, prepared media plates, 100 x 15 mm.
Improved medium for antimicrobial susceptibility testing. Chocolate agar intended use remel chocolate agar is a solid medium recommended for use in qualitative procedures for the isolation and cultivation of fastidious organisms, especially neisseria and haemophilus species. Selective media indicators are added to demonstrate hydrogen sulfide production, cho fermentation, ph. Nutrient media are basic culture media used for maintaining microorganisms, cultivating fastidious organisms by enriching with serum or blood and are also used for purity checking prior to biochemical or serological testing 5,6. Solid media generally contain agar at a concentration of 1. Novymacnealnicolle medium nnn is a culture medium used to grow leishmania needed when the amastigotes are not found in sufficient quantities by scraping the growth substrate. A pour plate is one in which a small amount of inoculum from broth culture is added by pipette to the centre of a petri dish. Media were prepared by autoclaving the phosphate buffer and agar either together t or separately s. In addition, if the specimen has been obtained from a neonate among whom e. Production of microbial medium from defatted brebra milletia. Water samples filter the specimen to be analyzed through two membranes. Nutrient agar difco media distributor pharmaceuticals. Differential media used to isolate fastidious specific diet organisms, determine hemolytic activity.
It is recommended that prepared media plates be ordered no more than 1 to 2 weeks before use, as media plates are highly perishable. Agaragar is also known as kanten, its japanese name, agar is the malay word for. Chocolate agar base, with the addition of supplements, gives excellent growth of the gonococcus without overgrowth by contaminating organisms. Since many pathogens require carbon dioxide on primary isolation, bd columbia agar with 5 % sheep blood plates should be incubated in an aerobic atmosphere containing approximately 3 to 10 % co2. Rphase shigellae should grow satisfactorily on macconkey agar. Bd chocolate agar gc ii agar with isovitalex and bd chocolate agar blood agar no. Chocolate agar contains less agar than other solid media therefore streaking should be done carefully to avoid gouging into the agar quality control after checking for correct ph, colour, depth, and sterility, the following organisms are used to determine the growth performance of the completed medium. Chocolate agar choc or chocolate blood agar cba, is a nonselective, enriched growth medium used for isolation of pathogenic bacteria. The addition of blood permits the interpretation of hemolytic activities.
Boiled blood agar chocolate agar is prepared by heating after the blood has been added to the blood agar base. A highly nutritious medium used for the isolation and cultivation of fastidious microorganisms, especially neisseria and haemophillus species from a variety of clinical specimens. Different types of agar are needed for the cultivation of different types of microorganisms. Physical and chemical properties physical state solid. Reactions incurred on chocolate agar by grampositive cocci were correlated with species identity.
The blood agar medium is prepared in a twostep process. Of those expensive media, peptone agar is the one that currently used for different purposes in microbial culture. The specimen to be cultivated should be streaked onto a chocolate agar plate or onto the agar slant. Whats the difference between chocolate agar medium and. Examples of enriched media include sheep blood agar and chocolate heated blood agar. Gonococci are not able to grow on common blood agar. Thermo scientific chocolate agar medium print share thermo scientific chocolate agar medium. Use them within 1 month of receipt for best results. The blood agar base is a highly nutritious medium that supports the growth of fastidious organisms. The treatment and control of gonorrhea is complicated by increasing resistance to a variety of antimicrobial agents, including penicillin, tetracycline, and fluoroquinolones.
Summary and explanation bacto gc medium base was designed in 1947. Chocolate agar definition of chocolate agar by medical. Pp0100 chocolate agar with 5% defibrinated horse blood e. Product information and quality control sheet chocolate agar intended use chocolate agar is an enriched medium for the isolation and cultivation of fastidious microorganisms, especially neisseria and haemophilus species from a variety of clinical specimens. Preparation and sterilization of culture media should be done with great care to avoid contamination of unwanted microorganisms. Chocolate agar supplemented with 10% sterile defibrinated blood is suited for isolating haemophilus and neisseria species. Although agar agar will not dissolve in cold water, it does absorb moisture, becoming soft and swollen. Learn vocabulary, terms, and more with flashcards, games, and other study tools. An agar plate is a petri dish that contains agar as a solid growth medium plus nutrients, used to culture microorganisms. Enterococcus faecalis is part of our normal intestinal flora and found in some probiotic foods. Chocolate agar is used for the isolation of fastidious organisms, such as haemophilus influenzae, when incubated at 3537c in a 5% co2 atmosphere. The choice may slightly change the morphology of colonies, so if this is an important phenotypic characteristic, it might matter. Blood agar is an enriched medium in which nutritionally rich whole blood supplements the basic nutrients. Basic practical microbiology why microbiology matters.
Under the right conditions he can cause endocarditis, meningitis, bacteremia and. Enterococcus faecalis colonies on blood agar flora. Nnn modified medium twin pack m681 nnn modified medium twin pack is used for cultivation of leishmaniae and trypanosomes. Bd chocolate agar gc ii agar with isovitalex bd chocolate. Chocolate agar with vx, prepared media plates, 100 x 15 mm, pack of 10. It is also used as a base for the preparation of a variety of special media.
This medium can be made selective by the addition of antibiotics. Only partial lysis of blood beta hemolytic show lysis and complete digestion of blood cells around colony gamma hemolytic lack of hemolytic activity. Pp0100 chocolate agar with 5% defibrinated horse blood a highly nutritious medium enriched with horse blood, where the blood has been chocolated by heating the medium to 60c. Suitable for the isolation of most pathogens including the most fastidious and is particularly useful for the cultivation of haemophilus spp. All lot numbers of chocolate agar have been tested using the above quality control organisms and have been found to be acceptable. The media is further enriched with suplex polyvitex that provides vitamins, amino acids, coenzymes, glucose and other factors which improve the growth of neisseria and haemophillus species. Chocolate agar is used for growing fastidious respiratory bacteria, such as haemophilus influenzae and neisseria meningitidis. Transfer the molten agar to suitable vessels and sterilize at 15 psi for 15 minutes. Muellerhinton agar is the most commonly used media for zoi testing of nonfastidious organisms and was recommended by bauer, kirby, et al.
This organism is available as a cultiloop precautions prolonged incubation may lead to confusing results. This organism is oxidasepositive, and growth is best obtained on chocolate agar or thayer martin agar. Pp0100 chocolate agar with 5% defibrinated horse blood. Pdf suitability of a chocolate agarbased medium for the. Agar agar is a mild laxative and can provoke allergic reactions in some people.
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