Bacterial culture Media


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  Notes
 
 
1 : Bacterial Culture Media basics Dr.T.V.Rao MD
2 : Major Contribution to Culture Media
3 : Agar - Agar Frau Hesse’scontribution
4 : Agar – Agar Solid medium is made by adding Agar Agar is obtained from Sea weeds New Zealand agar is more Agar contain long chain poly saccharides.Inoranic salts and protein like substance Melts at 980c and sets at 420c
5 :
6 : Agar - Agar Complex polysaccharide Used as solidifying agent for culture media in Petri plates, slants, and deeps Generally not metabolized by microbes Liquefies at 98°C Solidifies ~42°C Dr.T.V.Rao MD’s ‘e’ learning series
7 :
8 : Media and Culture Media: Nutrients (agar, pH indicators, proteins and carbohydrates) used to grow organisms outside of their natural habitats Culture: The propagation of microorganisms using various media
9 : Culture media Used to grow bacteria Can be used to: Enrich the numbers of bacteria Select for certain bacteria and suppress others Differentiate among different kinds of bacteria
10 : Culture and Medium Culture is the term given to microorganisms that are cultivated in the lab for the purpose of identifying and studying them. Medium is the term given to the combination of ingredients that will support the growth and cultivation of microorganisms by providing all the essential nutrients required for the growth (that is, multiplication) in order to cultivate these microorganisms in large numbers to study them.
11 : Specific Media Defined media are media composed of pure ingredients in carefully measured concentrations dissolved in double distilled water i.e., the exact chemical composition of the medium is known. Typically, they contain a simple sugar as the carbon and energy source, an inorganic nitrogen source, various mineral salts and if necessary growth factors (purified amino acids, vitamins, purines and pyrimidines
12 : Need for Culture Media It is usually essential to obtain a culture by grwoing the organism in an artificial medium. If more than one species or type of organism are present each requires to be carefully separated or isolated in pure culture. Several organism need the determination of Antibiotic sensitivity pattern for optimal antibiotic selection
13 : Basic requirements of culture media Nutrients - Energy source - Carbon source - Nitrogen source Mineral salts – Sulphate, phosphates, chlorides & carbonates of K, Mg & Ca. A suitable pH – 7.2 – 7.4 Accessory growth factors - Tryptophan for Salmonella typhi - X & V factors for H. influenzae
14 : Pouring the Culture Plates
15 : Petri dish with Media Plate: provide large surface for isolation and observation of colonies Using a sterile loop or a sterile swab streak your sample on the petri plate Important let your sterilized loop cool before you pick up your sample
16 : Classification of Culture media Based on the consistency: Liquid -- Peptone water, Nutrient broth Semisolid -- Nutrient agar stabs Solid -- Blood agar, Serum agar Based on Oxygen requirement: -- Aerobic medium -- Anaerobic media
17 : Aerobic Media Simple media Complex media May be Synthetic or Defined Medium - Enriched media - Differential media - Enrichment media - Selective media Semisyntetic Medium - Sugar media - Transport media
18 : Aerobic media Liquid media - Peptone water(1% peptone +0.5%Nacl + 100 ml water) - Nutrient broth ( peptone water + 1% meat extract Solid media - Nutrient agar (nutrient broth + 2% Agar) Use: To grow non-fastidious microorganisms Simple media- consists of only basic necessities
19 : Liquid Medium Difficulat to identify all types of organisms Suitable for isolation of bacteria from Blood culturing and water analysis
20 : Peptone Peptone contain partially digested proteins Proteases Polypeptides Aminoacids Inorganic salts Phosphates Potassium and Magnesium Riboflavin Meat exract called as Lab lemco
21 : Nutrient Agar Contain 2% agar added to Nutrient agar commonly used Concentration can be increased to 6% to prevent swarming Can be reduced to 0’5%
22 : Pigment producing Staphylococci
23 : Complex media Nutrient agar + 5 to 10% sheep blood Melt the sterile nutrient agar by steaming, cool, to 450 c Add the blood aseptically with constant shaking Mix the blood with molten nutrient agar thoroughly but gently avoiding froth formation Immediately pour in to the Petri dishes or tubes and allow to set Enriched media: Blood agar Use: To cultivate all the fastidious organisms
24 : Enriched Medium To culture medium Blood serum or egg are added to medium eg Blood agar Chocolate agar Egg
25 : Different types of hemolysis on Blood Agar
26 : Other Enrichments – Chocolate Agar Several organic materials are added to the basic constituents of the Medium such as Blood, yeast, yeast extract etc
27 : Chocolate agar
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