Proteus Mirabilis
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Investigative Journal
10/05/05
Journal # 1
Today I was given my unknown broth and began my journal.
Unknown broth
I received unknown broth #34, a tube of distilled water, and a TSA plate of media.
I recorded the number of my unknown, #34, onto my Descriptive Chart.
I labeled my TSA plate, my tube of unknown broth, and my distilled water with my name, date, lab section, media name, and my unknown number.
Growth characteristics from broth
I observed the broth for growth characteristics. I characterized the surface as membranous because there was a thin film of growth at the top. The subsurface I characterized as turbid because it was very cloudy. After agitating the tube, I was able to characterize the sediment as viscid.
I recorded the broth growth characteristics onto my Descriptive Chart. (Brown, pg 226)
Surface: Membranous
Subsurface: Turbid
Sediment: Viscid
Four quadrant streak
I rolled my broth tube to get a good dispersion of the organism.
Next, using four quadrant streak techniques for isolated colonies, I stroked my TSA plate. (method A, Brown, p 73)
I labeled, inverted, and stored the TSA plate in the incubator at 37 C for 24-48 hours.
Gram stain and morphological characteristics of my unknown
In order to define the morphology and Gram reaction of my unknown, I followed the procedure for making a smear from my unknown (Brown, page 84). I used the aseptic procedure for organism removal, (Brown, page 85) and Gram stained using five loopfuls of my broth (Brown pg 96). I then stored my unknown broth and distilled water in the refrigerator rack.
I observed my Gram stained slides with my microscope (Brown pg 6-8) and recorded morphological characteristics (Brown pg 45, 95) of the gram stained broth onto my descriptive chart. I stored the slides in my tables slide storage box.
Gram Reaction: Negative ( Pink)
Cell Shape: Rod ( Brown pg 45)
Arrangement: Individual(Sneath pg1033)
10/10/05
Journal #2
Today I observed my TSA plate.
Cultural Characteristics from TSA plate.
I observed TSA plate for cultural characteristics by using an isolated colony (Brown 228). I recorded on my Descriptive Chart, and stored the plate in the refrigerator.
Configuration: Filamentous
Margins: Lobate
Elevations: Convex
Colony Size: 2mm
I created working and reserve stock from my streak plate.
First I located an isolated colony.
Using the steps of aseptic procedure (Brown pg 64), I inoculated from the center of my isolated colony, to two nutrient agar slants. (Brown pg 221)
I labeled one slant 37 C and stored in the incubator to incubate at body temperature for 24-48 hours.
He other, I labeled 24 C and placed it in the cabinet near the autoclave to incubate at room temperature for 24-48 hours.
Next, I stored my TSA plate in the refrigerator
10/12/05
Journal # 3
I distinguished between working and reserve stock, observed characteristics, and gram stained.
Determination of working and reserve stock,
First, I examined the two slants. I looked for growth on the slants and determined which slant showed better growth (Brown pg 221). The slant stored in the incubator had better growth. I selected the slant with the better growth for my reserve stock and the other slant I used for my working stock. Also, I knew that the optimal temperature which my organism would prefer to grow was 37 C (Brown pg 221).
I stored my two slants in the refrigerator with the distilled water and broth.
Cultural characteristics from agar slant
I looked at my slant from the working stock (Brown pg 225-226) and recorded the characteristics on the descriptive chart.
Form: Rhizoid
Color: Buff
Opacity: opaque
Gram Stain Slant and Broth
I gram stained ( Brown pg 220) my two slants to assure purity
Test elimination check list
Since I my gram reaction was negative rod, I could eliminate the following tests:
Catalase production test
Oxidase production test
Starch hydrolysis test
Casein hydrolysis test
Fat hydrolysis test
Litmus milk reaction test
Tests: SIM, Urea hydrolysis, Simmons Citrate, MR-VP, Gelatin, KIA and FTM
First, I gathered my materials from the refrigerator, and labeled the tubes with my name, date, section #, unknown #, and the name of the test I was performing. I inoculated the following tests with my working stock:
I inoculated it by stabbing to a depth of 75% if the media and incubated for 24-48 hours. (Brown pg 222)
Urea Hydrolysis:
I inoculated it heavily with a needle by stabbing