Question

In: Biology

You have isolated an organism from the urine of Patient X, a 24 year old female....

You have isolated an organism from the urine of Patient X, a 24 year old female. It is showing gamma hemolysis on blood agar and the colonies are large with an entire margin and are white in color. You perform a gram stain with the results pictured below.

You perform a catalase test and it produces bubbles. You next perform an oxidase test and it is oxidase negative.

1. What is the next step to take in identifying the organism?

2. If your test is positive, what is the genus and species of the organism?

3. If the test is negative, what further test needs to be done to further confirm the negative result?

4. If the result is truly negative, what further testing needs to be run on this particular specimen? In other words, what organism of this type can cause urinary tract infections in sexually active females and how is it tested? (You will need to do some research to find out these results.)

Solutions

Expert Solution

  • Blood agar is a solid medium containing RBCs, which is used to detect bacteria which produce enzymes resulting in rupture of the blood cells. This process is also termed hemolysis. The degree of hemolysis is used to distinguish one bacteria from another. Hemolysis can be divided into alpha, beta, and gamma.

    Alpha hemolysis produces a green discoloration surrounding a bacterial colony growing on the agar. This type of hemolysis represents a partial decomposition of the hemoglobin of the red blood cells. Alpha hemolysis is characteristic of Streptococcus pneumonia.

    Beta hemolysis represents a complete breakdown of the hemoglobin of the red blood cells. There is a clearing of the agar around a colony. Beta hemolysis is characteristic of Streptococcus pyogenes and some strains of Staphylococcus aureus.

    Gamma hemolysis is a lack of hemolysis in the area around a bacterial colony. A blood agar plate displaying gamma hemolysis actually appears brownish. Gamma hemolysis is a characteristic of Enterococcus faecalis.
  • Enterococcus faecalis are generally catalase negative; they don't synthesize haem or porphyrin. However, when grown in haem-containing medium, they can synthesize hemoproteins. Some strains are known to give a weakly positive catalase test, which is attributed to the presence of some non-haem catalase (pseudocatalase). Enterococcus haemoperoxidus is known to give a positive catalase reaction when cultivated on blood agar, but not when grown on blood-free medium. Some strains of E. fecalis too give such a reaction.
  • Oxidase negative normally means the bacterium does not contain cytochrome c oxidase and, therefore, either cannot use oxygen for energy production with an electron transfer chain or employs a different cytochrome for transferring electrons to oxygen.

    Enterococci are typically Oxidase negative.

  • Species identification:
    16s rRNA sequencing can be done. Acid production from 1% (w/v) mannitol, sorbitol, sorbose, L-arabinose, D-ribose, sucrose and raffinose can be tested in basal M17 broth (Terzaghi and Sandine 1975) (lacking lactose), using bromocresol purple as pH indicator. Deamination of arginine can be tested in Thornley's semi-solid arginine medium (Smibert and Krieg 1981). Motility can be using phase contrast microscopy (Smibert and Krieg 1981). Pigment production on tryptic soy agar. Tolerance to tellurite can be tested on GM17 agar containing 0á04% (w/v) potassium tellurite. Utilization of pyruvate tested using pyruvate broth (Facklam and Sahm 1995).


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