Pseudomonas sp.
Scientific classification:
Kingdom: Bacteria
Phylum: Proteobacteria
Class: Gamma Proteobacteria
Order: Pseudomonadales
Family: Pseudomonadaceae
Genus: Pseudomonas
Hydrocarbonoclastic bacterium Pseudomonas sp is capable of degrading various types of hydrocarbons. The successful use of Pseudomonas bacteria in environmental bioremediation of hydrocarbon pollution requires an understanding of the mechanisms of interaction between the bacterium Pseudomonas sp with hydrocarbon compounds.
Pseudomonas sp is a rod-shaped bacterium, is gram negative, have a flagellum, no capsule. Form a blue pigment that seeped into the seed consists of substances: flouresens green color water soluble and insoluble pyocianin greenish blue color in the chloroform. The bacteria are only describing glucose and grow on all types of media.
In a small number of these bacteria live as normal flora of human and animal intestinal tractus, also found on healthy human skin. Infection occurs in:
1. When bekteri enters the body of its resistance decreases, for example: chronic diseases.
2. Pathogen Pseudomonas aeruginosa are usually together when other germs, mixed infections with other bacteria (coccus pyogen) or with one of Enterobacteriaceae bacteria, such as: burns.
Germs are spread by dust and air. At the hospital Pseudomonas become contaminants such as: the surgical instrument will cause infection and it is very dangerous because the patient was weak.
The medium used for growth of bacteria Pseudomonas sp
1. Selenit Enrichment Broth
This media is used as a medium for fertilizer by Pseudomonas sp.
Selenit inhibit the growth of coliform and Enterococcus bacteria during the first 6-12 hours of incubation, while Salmonella, Proteus, and Pseudomonas are not hampered its growth.
Composition:
Peptone 5 grams
4 grams of lactose
Na selenit 4 grams
Dikalium hygrogen phosphate 3.5 gram
Potassium dihydrogen phosphate 6.5 grams
How to manufacture:
Mix 23 g / liter (if necessary heating temperature should not exceed 60 ° C), if the media will be stored for long periods sterile filter and placed in a suitable container.
Never in an autoclave, if there is a red sediment on selenit the media can not be used further. pH 7.0 ± 0.1.
2. Pseudomonas Selective To Base (Cetrimide order)
This media is used as a medium for the isolation and differential for Pseudomonas aeruginosa of various materials.
Most of the compounds in this medium inhibits the growth of accompanying microbial flora so that the original concentration of inhibitor (0.1%) is reduced to minimize interference with the growth of Pseudomonas. Pigment production in Pseudomonas aeruginosa no inhibition when grown on this medium.
Composition:
Peptone from gelatin 20 grams
Magnesium chloride 1.4 grams
Potassium sulfate 10 grams
Cetrimide 0.3 grams
Agar-To 13.6 grams
Glycerol 10 ml
How to manufacture:
Mix 45.5 g / liter add 10 ml glycerol / liter, sterilized in an autoclave. Pour into sterile Petri dish.
For cetrimide is a kind of gelatin used for the selective isolation of gram-negative bekteri, Pseudomonas aeruginosa. As the name suggests this medium containing cetrimide, which is a selective agent against microbial flora alternate. Cetrimide also increase the production of pigments such as pyocyanin and Pseudomonas fluorescens, which showed blue-green and yellow-green characteristics respectively. Cetrimide agar is widely used in testing cosmetics, pharmaceutical and clinical specimens to test for the presence of Pseudomonas aeruginosa.
3. For Pseudomonas and Pseudomonas F Base For Base P
This media is used for the isolation and differentiation of Pseudomonas based on the formation of pyocyanin and pyorubin or fluorescein.
Pseudomonas To stimulate the formation of P Base pyocianin and reduce fluorescein, whereas Pseudomonas F Base To stimulate the production of fluorescein and reduce pyocianin. Simultaneously using both culture media allows rapid identification when prminilary from Pseudomonas species, because some strains can only synthesize pyocyanin, some only able to synthesize fluorescein and there that produce both pigments.
Composition:
a. Pseudomonas To F Base
Peptone from casein 10 grams
Peptone from meat 10 grams
Magnesium sulfate 1.5 grams
Di-potassium hydrogen sulfate 1.5 grams
Agar-agar 12 grams
Glycerol 10 ml
b. Pseudomonas To P Base
Peptone from gelatin 20 grams
Magnesium chloride 1.4 grams
10 grams of potassium phosphate
Agar-agar 12.6 grams
Glycerol 10 ml
How to manufacture:
Mix 10 ml of glycerol / liter to 35 grams of Pseudomonas To F Base / liter or 44 grams of Pseudomonas For Base P / liter. Sterilize in an autoclave, then poured in sterile petri dish.
Procedure and evaluation:
Surface inoculation of culture media suspected to contain Pseudomonas so that individual colonies grown.
Incubation: 1 week at 37 ° C
Check the growth of bacteria after 24,48,72 hours and then after 6 days.
Only Pseudomonas aeruginosa can grow in order for P Base with Pseudomonas colonies surrounded by blue zones because of the formation pyocianin green to red to brown or dark zones due to the production pyorubin. Colored pigments can be extracted with chloroform. Pseudomonas aeruginosa Pseudomonas appears in order for F Base as colonies surrounded by a zone of greenish-yellow to yellow resulting from the production of fluorescein. If pycyanin also synthesized, bright green color of the resulting fluorescence in ultraviolet light.
REFERENCES
E. Merck, Darmstadt. 1984. Handbook of Culture Media Merck.
http://en.wikipedia.org/wiki/Cetrimide_agar downloaded on March 16, 2011
http://id.wikipedia.org/wiki/Pseudomonas downloaded on March 16, 2011
Scientific classification:
Kingdom: Bacteria
Phylum: Proteobacteria
Class: Gamma Proteobacteria
Order: Pseudomonadales
Family: Pseudomonadaceae
Genus: Pseudomonas
Hydrocarbonoclastic bacterium Pseudomonas sp is capable of degrading various types of hydrocarbons. The successful use of Pseudomonas bacteria in environmental bioremediation of hydrocarbon pollution requires an understanding of the mechanisms of interaction between the bacterium Pseudomonas sp with hydrocarbon compounds.
Pseudomonas sp is a rod-shaped bacterium, is gram negative, have a flagellum, no capsule. Form a blue pigment that seeped into the seed consists of substances: flouresens green color water soluble and insoluble pyocianin greenish blue color in the chloroform. The bacteria are only describing glucose and grow on all types of media.
In a small number of these bacteria live as normal flora of human and animal intestinal tractus, also found on healthy human skin. Infection occurs in:
1. When bekteri enters the body of its resistance decreases, for example: chronic diseases.
2. Pathogen Pseudomonas aeruginosa are usually together when other germs, mixed infections with other bacteria (coccus pyogen) or with one of Enterobacteriaceae bacteria, such as: burns.
Germs are spread by dust and air. At the hospital Pseudomonas become contaminants such as: the surgical instrument will cause infection and it is very dangerous because the patient was weak.
The medium used for growth of bacteria Pseudomonas sp
1. Selenit Enrichment Broth
This media is used as a medium for fertilizer by Pseudomonas sp.
Selenit inhibit the growth of coliform and Enterococcus bacteria during the first 6-12 hours of incubation, while Salmonella, Proteus, and Pseudomonas are not hampered its growth.
Composition:
Peptone 5 grams
4 grams of lactose
Na selenit 4 grams
Dikalium hygrogen phosphate 3.5 gram
Potassium dihydrogen phosphate 6.5 grams
How to manufacture:
Mix 23 g / liter (if necessary heating temperature should not exceed 60 ° C), if the media will be stored for long periods sterile filter and placed in a suitable container.
Never in an autoclave, if there is a red sediment on selenit the media can not be used further. pH 7.0 ± 0.1.
2. Pseudomonas Selective To Base (Cetrimide order)
This media is used as a medium for the isolation and differential for Pseudomonas aeruginosa of various materials.
Most of the compounds in this medium inhibits the growth of accompanying microbial flora so that the original concentration of inhibitor (0.1%) is reduced to minimize interference with the growth of Pseudomonas. Pigment production in Pseudomonas aeruginosa no inhibition when grown on this medium.
Composition:
Peptone from gelatin 20 grams
Magnesium chloride 1.4 grams
Potassium sulfate 10 grams
Cetrimide 0.3 grams
Agar-To 13.6 grams
Glycerol 10 ml
How to manufacture:
Mix 45.5 g / liter add 10 ml glycerol / liter, sterilized in an autoclave. Pour into sterile Petri dish.
For cetrimide is a kind of gelatin used for the selective isolation of gram-negative bekteri, Pseudomonas aeruginosa. As the name suggests this medium containing cetrimide, which is a selective agent against microbial flora alternate. Cetrimide also increase the production of pigments such as pyocyanin and Pseudomonas fluorescens, which showed blue-green and yellow-green characteristics respectively. Cetrimide agar is widely used in testing cosmetics, pharmaceutical and clinical specimens to test for the presence of Pseudomonas aeruginosa.
3. For Pseudomonas and Pseudomonas F Base For Base P
This media is used for the isolation and differentiation of Pseudomonas based on the formation of pyocyanin and pyorubin or fluorescein.
Pseudomonas To stimulate the formation of P Base pyocianin and reduce fluorescein, whereas Pseudomonas F Base To stimulate the production of fluorescein and reduce pyocianin. Simultaneously using both culture media allows rapid identification when prminilary from Pseudomonas species, because some strains can only synthesize pyocyanin, some only able to synthesize fluorescein and there that produce both pigments.
Composition:
a. Pseudomonas To F Base
Peptone from casein 10 grams
Peptone from meat 10 grams
Magnesium sulfate 1.5 grams
Di-potassium hydrogen sulfate 1.5 grams
Agar-agar 12 grams
Glycerol 10 ml
b. Pseudomonas To P Base
Peptone from gelatin 20 grams
Magnesium chloride 1.4 grams
10 grams of potassium phosphate
Agar-agar 12.6 grams
Glycerol 10 ml
How to manufacture:
Mix 10 ml of glycerol / liter to 35 grams of Pseudomonas To F Base / liter or 44 grams of Pseudomonas For Base P / liter. Sterilize in an autoclave, then poured in sterile petri dish.
Procedure and evaluation:
Surface inoculation of culture media suspected to contain Pseudomonas so that individual colonies grown.
Incubation: 1 week at 37 ° C
Check the growth of bacteria after 24,48,72 hours and then after 6 days.
Only Pseudomonas aeruginosa can grow in order for P Base with Pseudomonas colonies surrounded by blue zones because of the formation pyocianin green to red to brown or dark zones due to the production pyorubin. Colored pigments can be extracted with chloroform. Pseudomonas aeruginosa Pseudomonas appears in order for F Base as colonies surrounded by a zone of greenish-yellow to yellow resulting from the production of fluorescein. If pycyanin also synthesized, bright green color of the resulting fluorescence in ultraviolet light.
REFERENCES
E. Merck, Darmstadt. 1984. Handbook of Culture Media Merck.
http://en.wikipedia.org/wiki/Cetrimide_agar downloaded on March 16, 2011
http://id.wikipedia.org/wiki/Pseudomonas downloaded on March 16, 2011
