Proteus Mirabilis

Investigative Journal

10/05/05

Journal # 1

Today I was given my unknown broth and began my journal.

Unknown broth

a. I received unknown broth #34, a tube of distilled water, and a TSA plate of media.

b. I recorded the number of my unknown, #34, onto my Descriptive Chart.

c. 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

d. 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.

e. I recorded the broth growth characteristics onto my Descriptive Chart. (Brown, pg 226)

i. Surface: Membranous

ii. Subsurface: Turbid

iii. Sediment: Viscid

Four quadrant streak

f. I rolled my broth tube to get a good dispersion of the organism.

g. Next, using four quadrant streak techniques for isolated colonies, I stroked my TSA plate. (method A, Brown, p 73)

h. 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

i. 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.

j. 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 table's slide storage box.

i. Gram Reaction: Negative ( Pink)

ii. Cell Shape: Rod ( Brown pg 45)

iii. Arrangement: Individual(Sneath pg1033)

10/10/05

Journal #2

Today I observed my TSA plate.

Cultural Characteristics from TSA plate.

a. 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.

i. Configuration: Filamentous

ii. Margins: Lobate

iii. Elevations: Convex

iv. Colony Size: 2mm

b. I created working and reserve stock from my streak plate.

i. First I located an isolated colony.

ii. 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)

iii. I labeled one slant 37 C and stored in the incubator to incubate at body temperature for 24-48 hours.

iv. He other, I labeled 24 C and placed it in the cabinet near the autoclave to incubate at room temperature for 24-48 hours.

v. Next, I stored my TSA plate in the refrigerator

10/12/05

Journal # 3

A. I distinguished between working and reserve stock, observed characteristics, and gram stained.

B. Determination of working and reserve stock,

a. 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).

b. I stored my two slants in the refrigerator with the distilled water and broth.

C. Cultural characteristics from agar slant

a. I looked at my slant from the working stock (Brown pg 225-226) and recorded the characteristics on the descriptive chart.

i. Form: Rhizoid

ii. Color: Buff

iii. Opacity: opaque

D. Gram Stain Slant and Broth

a. I gram stained ( Brown pg 220) my two slants to assure purity

E. Test elimination check list

a. Since I my gram reaction was negative rod, I could eliminate the following tests:

i. Catalase production test

ii. Oxidase production test

iii. Starch hydrolysis test

iv. Casein hydrolysis test

v. Fat hydrolysis test

vi. Litmus milk reaction test

F. Tests: SIM, Urea hydrolysis, Simmons Citrate, MR-VP, Gelatin, KIA and FTM

a. 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. SIM

1. I inoculated it by stabbing to a depth of 75% if the media and incubated for 24-48 hours. (Brown pg 222)

ii. Urea Hydrolysis:

1. I inoculated it heavily with a needle by stabbing