1. Practical No.17
Proteus & Pseudomonas

2. Proteus ; Gram-ve , actively motile pleomorphism rods.
Causes UTI, cystitis, wound infections.
Four speices ; Proteus morgani, Proteus vulgaris, Proteus mirabilis and Proteus rettgeri are distinguished from each other on the bases of fermentation of maltose, mannitol, sucrose and the production of indole.
All Proteus species are indole +ve except Proteus mirabilis.
All Proteus do not ferment mannitol except Proteus rettgeri (late fermenter).

3. Labrotory diagnosis: Specimen; urine, exudates, swabs.
Smear; G-ve bacilli.

4. Culture;
MacConkey…Proteus appear as a non- lactose fermenter. (colorless colonies).
EMB…. There is no metallic sheen on EMB.
S.S agar….Proteus appear as pale colonies with black centers.
Blood agar…highly motile ( swarming), therefore they produce swarming spread overgrowth on blood agar plate .swarming is characterized by expanding rings (waves) over the surface of blood agar.

5. Laboratory diagnosis
*Blood agar  Swarming.

6. Biochemical tests; 1- I M Vi C test _ _ _ + + ­­­­­- +
_ _ _
­­­­­
2-TSI ; K / A Gas +ve H2S +ve
Or ; A / A Gas +ve H2S +ve

7. 3- Urease test: + ve
Converting the slant of urea agar from yellow color to pink- purple color due to the utilization of urea by urease enzyme(produced by proteus) and the formation of ammonia converting the medium into an alkaline pH and producing a pink purple color by a change in the phenol red indicator.

8. Laboratory diagnosis
*Urea agar (urease test)  Pink (+); Yellow (-).

9. Serology:
certain serotypes of proteus have cross-reacting Ags with some Rickettsias, It is purely coincidental, but serves as a useful clinical tool to determine if a person has been infected with rickettsia .
Mixing the serum of a patient suspected of having a rickettsial disease with Ag from Ab Ag will agglutinate ,indicating +ve results; ( weil- felix test).

10. Typhus group rickettsiae reacts with P
Typhus group rickettsiae reacts with P. vulgaris OX19, and scrub typhus reacts with P. mirabilis OXK. The spotted fever group rickettsiae reacts with P. vulgaris OX2 and OX19, to varying degrees, depending on the species.
The Weil-Felix Test can be done as either a slide or a tube test. The antigens necessary (OX2, OX19, and OXK) can be obtained commercially.

11. Pseudomonas
A genus of Gram-negative, nonsporeforming, rod- shaped bacteria. Motile species possess polar flagella. They are strictly aerobic. Members of the genus Pseudomonas cause a variety of infective diseases .Pseudomonas aeruginosa is the most significant cause of hospital-acquired infections. The spectrum of clinical disease ranges from urinary tract infections to septicemia, pneumonia, meningitis, and infections of postsurgical and posttraumatic wounds.

12. Pseudomonas pigments;
pyocyanin …. Blue color
pyoverdin….green (fluorescent) color
Pyorubin ….. reddish brown color
Pyomelanin …. Black color
Some strains of pseudomonas however do not produce any ofthese pigments.

13. Genus Pseudomonas
Pseudomonas
Pyocyanine Pigment

14. Genus Pseudomonas
Pseudomonas
Pyocyanine Pigment

15. Pseudomonas aerogenosa:
a common isolate from wounds, burns and urinary tract infections and from many other accumulations of pus . Also commonly found in otitis. Its presence may be indicated by a distinctive blue or green color of the pus or infected site.

16. Laboratory diagnosis;
Specimen: urine , pus, blood, CSF, sputum, swab.
Culture:
MacConkey…… Pseudomonas appears as a non – lactose fermenter.
EMB......there is no metallic sheen on EMB agar.
Nutrient agar.... Pseudomonas aerogenosa produces a blue– green pigment and a fruity aroma. Milk agar may be added to nutrient agar to give a white background.

18. Nutrient agar  Demonstrate pigment production.
Laboratory diagnosis
Nutrient agar  Demonstrate pigment production.

19. Laboratory diagnosis
Blood agar spreading flat pigmented, sometime with Beta- hemolysis, mucoid, confluent growth.

20. Biochemical tests: Oxidase test, catalase test all are positive.
Laboratory diagnosis
Biochemical tests: Oxidase test, catalase test all are positive.
Catalase +ve
Oxidase +ve

21. Biochemical tests: 1-I M Vi C - - - +
2- TSI: no change k/k Gas –ve H 2S –ve
3- oxidase + ve

22. Thank You