Experimental procedure and evaluation pseudomonas cfc selective agar. Pseudomonas isolation agar base, used for the selective isolation and identification of p. If growth is observed on pseudomonas f agar, fluorescein production is determined by visual examination of the. The test isolate should be in pure culture and 1824 hours old. Pseudosel agar or cetrimide agar is used for the selective isolation and presumptive identification of pseudomonas aeruginosa. Pseudomonas putida will be incubated in a 150 ml of lb broth at 37 celsius with shaking 100 rpm. Can nutrient agarbroth be used for pseudomonas aeruginosa in. Pseudomonas agar base for microbiology sigmaaldrich. Cultivation 24 hours in an aerobic atmosphere, 37c. Cetrimide agar is recommended in the united states pharmacopoeia xxvi 1 and european pharmacopoeia iv 2 for use in microbial limit tests. Pseudomonas p agar is used to isolate, and detect pigment production by pseudomonas aeruginosa. Pseudomonas f agar is used to detect pigment production by pseudomonas species.
The medium is modification of kings a medium 1 and contains magnesium chloride and potassium sulphate for improved pigment production and recovery of pseudomonas aeruginosa while strongly suppressing klebsiella, proteus and providencia spp. This medium is very useful for isolating pseudomonas spp. A new medium, pseudomonas selectiveisolation agar, was. The base medium is a modification of kings medium a which uses magnesium and potassium salts to enhance production of the pigments pyocyanin green and fluorescein detected by u.
Pseudomonas agar p is mentioned in the fda bacteriological analytical. Thirtynine strains of 10 different species of pseudomonas. A novel chromogenic medium for isolation of pseudomonas. Biofilm detection was tested by tube method tm, congo red agar method cra. Pseudomonas isolation agar base m406 composition ingredients gms litre peptone 20. Pseudomonas agar base is a modification of kings a medium 5 which contains magnesium chloride and potassium sulphate to enhance pigment production. If growth is observed on pseudomonas f agar, fluorescein.
The ratio of peptones in peptospecial is conducive of fluorescein production by pseudomonas. Pseudomonas isolation agar includes irgasan, a potent broadspectrum antimicrobial not active against pseudomonas spp. An opportunistic, nosocomial pathogen of immunocompromised individuals, p. Cfc pseudomonas agar, base, cn pseudomonas agar, base, cetrimide nalidixic acid agar, base, pp pseudomonas agar, base. Pseudomonas agar p, also known as tech agar, is recommended for enhancement of pyocyanin production by pseudomonas aeruginosa. Pseudomonas isolation agar is prepared according to a slight modification of the medium a formulation of king, ward and raney. Pseudomonas species also typically give a positive result to the oxidase test, the absence of gas formation from glucose, glucose is oxidised in oxidationfermentation test using hugh and leifson of test, beta hemolytic on blood agar, indole negative, methyl red negative, vogesproskauer test negative, and citrate positive. Add several drops of chloroform to the growth and observe for a bluegreen color. Glycerol serves as an energy source and also helps to promote pyocyanin production. Cled cysteine, lactose, and electrolytedeficient agar is a differential culture medium for use in isolating and enumerating bacteria in urine from the suspected cases of urinary tract infection. Pancreatic digest of casein 10gl, proteose peptone no. Their ease of culture in vitro and availability of an increasing number of pseudomonas strain. Technical information pseudomonas agar medium for detection. The optimal composition of the medium per litre was bacto peptone 10 g, gel.
Pseudomonas agar base is designed so that by the addition of the appropriate supplement sr0102 or sr0103 the medium becomes selective for pseudomonas aeruginosa or pseudomonas spp. Cetrimide agar composition, principle, uses, preparation. Pseudomonas aeruginosa pseudomonas aeruginosa is a gramnegative, rodshaped, motile organism polar flagella which characteristically produce watersoluble pigments which diffuse through the medium. Goto and enomoto 2 formulated cetrinix supplement for the selective isolation of pseudomonas aeruginosa from clinical specimens. Using the oxidase test and cetrimide agar to identify. Pseudomonas can, in rare circumstances, cause communityacquired pneumonias, as well as ventilatorassociated pneumonias, being one of the most common. Pseudosel agar or cetrimide agar is used for the selective isolation and presumptive identification of pseudomonas aeruginosa the medium was first developed by lowburry and is a modification of tech agar developed by king et al. It is the most common cause of infections of burn injuries and of the external ear otitis. The organism can utilize an array of nutrients and will thrive in a variety of environments, including medical equipment such as catheters, infusion fluids, disinfectants, and cosmetics. Pseudomonas isolation agar medium for pseudomonas aeruginosa. Some pseudomonas strains elaborate both pigments, while others elaborate only one of the two.
Section 1 identification of the substancemixture and of. Incubate the plate in ambient air caps loosened for slants at 3337c for 1824 hours. A new medium for the detection of fluorescent pigment production by. The medium contains pigmentenhancing components and the selective agents, triclosan 4 which selectively inhibits nonpseudomonads. Cetrimide also enhances the production of pseudomonas pigments such as pyocyanin and pyoverdine, which show a characteristic bluegreen and yellowgreen colour. Pseudomonas agar medium for detection of fluorescein is used for the detection of fluorescein production by pseudomonas species, and is in accordance with united states pharmacopoeia 2008. Staphylococcus aureus and pseudomonas aeruginosa biofilm. Pseudomonas p agar, resulting in a variety of hues from blue to red. Technical information pseudomonas agar medium for detection of fluorescein product code.
Pseudomonas aeruginosa produces a variety of watersoluble pigments. Pseudomononas agar base is a modification of kings a medium 1 which contains magnesium chloride and potassium sulphate to enhance pigment production. The base medium is a modification of kings a medium 1 in which magnesium chloride and potassium sulphate are present to enhance pigment production. Goto and enomoto 2 formulated cetrinix supplement for the selective isolation of pseudomonas aeruginosa from. Colonisation rates are estimated at 80% in adults and 60% in the cf population overall. Pseudomonas selective agar base untuk mikrobiologi granucult find msds or sds, a coa, data sheets and more information. Jul 05, 2016 this video lesson demonstrates how to identify a bacterium as pseudomonas aeruginosa by using the oxidase test and selective cetrimide agar. Selectively isolate pseudomonas species from a range of samples with thermo scientific oxoid pseudomonas agar base dehydrated. Typical composition gliter peptone from gelatine 16. Pseudomonas isolation agar is used for the isolation of pseudomonas aeruginosa and other pseudomonas spp. Using the oxidase test and cetrimide agar to identify pseudomonas. Iso 720 and iso 16266 granucult find msds or sds, a coa, data sheets and more information. Pseudomonas agar f and p tech agar hardy diagnostics.
Pseudomonas aeruginosa forms fluorescent green colonies while the growth of klebsiella, proteus, and providencia is strongly suppressed enhanced pigment production aids identification. The members of the genus demonstrate a great deal of metabolic diversity and consequently are able to colonize a wide range of niches. A new selective medium for isolating pseudomonas spp. Food and drug administration 10903 new hampshire avenue silver spring, md 20993 1888infofda 18884636332 contact fda. Storage requirements jun 12, 2018 cetrimide agar composition, principle, uses, preparation and colony morphology last updated. Pseudomonas aeruginosa morphology, growth characteristics and. Pseudomonas aeruginosa growing on macconkey agar macconkey agar is a selective medium used for the isolation of nonfastidious gramnegative rods, particularly members of the family enterobacteriaceae and the genus pseudomonas, and the differentiation of lactose fermenting from lactose nonfermenting gramnegative bacilli. Page 1 of 7 sds 9222 rev 0 november 2015 section 1 identification of the substancemixture and of the company 1. It consists of 350 micrograms of nitrofurantoin per ml and 2 micrograms of crystal violet per ml in a nutrient agar base. The best known are pyocyanin bluegreen, pyoverdine yellowgreen, fluorescent, and pyorubin redbrown, produced by a small proportion of strains. A nonselective or selective agar plate may be used for inoculation of the test isolate. Pseudomonas is a genus of gramnegative, gammaproteobacteria, belonging to the family pseudomonadaceae and containing 191 validly described species.
Thermo scientific remel pseudomonas f agar flo agar is used for the differentiation of pseudomonas aeruginosa from other pseudomonas species based on fluorescein production. Cetrimide agar is used as a selective medium for the isolation of pseudomonas aeruginosa from pus, sputum and drains, etc. Pseudomonas can, in rare circumstances, cause communityacquired pneumonias, as well as ventilatorassociated pneumonias, being one of the most common agents isolated in several studies. Pseudomonas agar p base for microbiology find msds or sds, a coa, data sheets and more information. The medium was first developed by lowburry and is a modification of tech agar developed by king et al. Pseudomonas cn selective agar ready to use plates 55 and 90 mm. A new medium, pseudomonas selective isolation agar, was developed to isolate pseudomonas spp. Macconkey agar contains the dye crystal violet well as. Lowbury and collins 6 studied cetrimide as a selective agent. Clinical isolates were identified as per standard microbiological procedure. Pseudomonas colonies into the medium on which they grow. Pyocyanin is a blue, phenazine pigment that diffuses into the agar medium to give the surrounding medium a blue coloration. Cled agar supports the growth of all potential urinary pathogens, and a number of contaminants such as diphtheroids, lactobacilli, and micrococci.
Recommended for selective isolation of pseudomonas species. Select the center of a wellisolated colony and streak onto pseudomonas f agar. Cetrimide agar is a type of agar used for the selective isolation of the gramnegative bacterium, pseudomonas aeruginosa. Pseudomonas aeruginosa morphology, growth characteristics. Colonies of pseudomonas aeruginosa and staphylococcus aureus yellow on tryptic soy agar. Irgasan is incorporated into the medium to inhibit the growth of many grampositive and gramnegative microorganisms other than pseudomonas spp. I want to select a suitable time period for the incubation until bacterial growth reached the. Pseudomonas cepacia cfc agar, or pseudomonas aeruginosa cn agar. Selective media for pseudomonas pseudomonas sigmaaldrich. Streak from the liquid enrichment on a solid medium of the same composition, as observed following lab 2. These are not pure cultures and a variety of colony morphologies are visible.
Pseudomonas aeruginosa is a gramnegative, rodshaped, motile organism polar flagella which characteristically produce watersoluble pigments which diffuse through the medium. Section 1 identification of the substancemixture and of the. Antibiotic susceptibility test of biofilm producing bacteria was performed by using the kirbybauer disc diffusion technique. Summary pseudomonas aeruginosa is an environmental isolate commonly found in soil, water, food, and in many manmade products. This video lesson demonstrates how to identify a bacterium as pseudomonas aeruginosa by using the oxidase test and selective cetrimide agar. Can nutrient agarbroth be used for pseudomonas aeruginosa. Student streaks on pseudomonas isolation agar as observed following lab 3. Cetrimide also enhances the production of pseudomonas pigments such as pyocyanin and pyoverdine, which show a.
Pseudomonas aeruginosa is the most common cause of infections of burn injuries and of the outer ear otitis externa, and is the most frequent colonizer of medical devices e. Pseudomonas isolation agar also includes glycerol as an energy source, while peptone provides nutrients necessary for bacterial growth. Section 1 identification of the substancemixture and of the company 1. The medium is made selective for pseudomonas aeruginosa by the addition of x107 c. Pseudomonas aeruginosa colonies on tryptic soy agar. June 12, 2018 by sagar aryal cetrimide agar is a selective and differential medium used for the isolation and identification of pseudomonas aeruginosa from clinical and nonclinical specimens.
If pigment production is questionable small amount of bluegreen color confirm the reaction by extraction with chloroform. Cfc pseudomonas agar, base cn pseudomonas agar, base cetrimide nalidixic acid agar, base pp pseudomonas agar, base pseudomonas agar base may be used for the selective detection of pseudomonas aeruginosa pathogens in mineral water sample that are mainly responsible for food and waterborne infections. Pseudomonas aeruginosa produces a number of water soluble iron chelators, including the yellowgreen or yellowbrown fluorescent pyoverdin. Pseudomonas aeruginosa is a common encapsulated, gramnegative, rodshaped bacterium that can cause disease in plants and animals, including humans.