2016 Microbiology(BIO205W) EITable PDF

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Summary

2016 Microbiology(BIO205W) EITable...

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Table 1: Observations and results of observable characteristics, storage conditions, staining techniques, and biochemical tests used to identify the EI found in soil. Characteristics

Observations

Results

Colony Morphology 1. Color 2. Configuration 3. Margins 4. Elevation

Off white color on TSA plate 1. Off white/yellow color and slant with hairy figure and 2. Spindle configurations no elevation. 3. Filamentous margins 4. Flat

Growth Patterns

Straight line with branching spirals

Arborescent

Cell Morphology (simple stain)

Pink, rod-shaped cell

Streptobacillus

Gram Reaction

Purple cells

Positive

Capsule Stain

Red cells with dark blue back- Negative ground

Endospore Stain

Pink cells with green spores

Positive

Acid-fast Stain

Blue cells

Negative

Catalase

Did not bubble

Negative

Oxidase

Colorless, it did not turn purple

Negative

Carbohydrate Fermentation (glucose)

Turned yellow and bubbles were present

Positive (gas acid and gas were present)

Carbohydrate Fermentation (sucrose)

Turned yellow

Positive (acid)

Carbohydrate Fermentation (lactose)

No bubbles

Negative (no growth)

Carbohydrate Fermentation (mannose)

Turned yellow and bubbles were present

Positive (gas acid and gas were present)

Oxygen Requirements

Bacteria was present throughout tube and settled on top

Facultative anaerobe

Nitrate Reduction

No bubbles were present therefore reagents Nitrate A and B were added and immediately turned red

Positive reaction

Motility

Red cloudiness only near stab

Negative

Simmons’ Citrate

No color change

Negative

Hydrolysis

No color change

Negative

Kliger’s Iron agar

Precipitate was present

Negative

Gelatinase

Remained liquid even when placed in ice bath

Positive

Starch Hydrolysis

Stayed purple no clear zone

Negative

Lipid Hydrolysis

No growth

Negative

Casein Hydrolysis

No clear zone

Negative

Biofilm Test

No residue

Negative

Storage Conditions

Stored best in room temperature (22 degrees Celsius)

Room temperature was better a better storage for the EI

Table 2: The characteristics of Sporolactobacillus environmental isolate discovered in soil located in Flagstaff, Arizona compared and contrasted from five different bacteria in group 18 of Bergery’s Manual (Holt et. all 2000). 13.) Are bands present? If yes, what is the approximate size of each band? (in bp) Was the PCR successful? Lane 2: Yes there are bands present. There are two bands which may indicate that there are two different DNA sequences of different base pair sizes. The bands are about 250 bp and 400 bp in size. The PCR was successful on lane 2 because there are bands present.

Characteris- EI tics

Bacillus

Mycobacterium

Clostridium Sporolacto- Amphibabacillus ciellus

Cell Morphology

Rod

Rod

Rods and cocci

Rod

Rod

Rod

Facultative Anaerobe

Positive

Positive

Negative

Negative

Positive

Negative

Catalase

Negative

Positive

Positive

Positve

Negative

Negative

Gram Reac- Positive tion

Positive

Positive

Positive

Positive

Positive

Habitat

Soil

Soil, ground water

Soil

Feces, soil

Grass, manure

4/5

3/5

3/5

5/5

3/5

Soil

Correlation N/A to EI

Lane 3: Yes there is a band present. There is only one band which indicates there is only one DNA sequence and one base pair size. The band is approximately 275 bp in size. The PCR was successful because a band was present but there were not multiple bands. Lane 4: No there are no bands present. The PCR was unsuccessful because there were no bands present. 14.) Using #13 above as a model, explain your PCR results. Remember to include observations and results My EI had a band at 290 bp becausee we amplified the 16s RDNA so we should conclude it will remain at this size. It was successful because the bands were observed in the lanes.

Characteristics

EI

Bacillus anthracis

Bacillus cereus

Bacillus mycoides

Bacillus badius

Bacillus megaterium

Starch Hydrolysis

Negative

Positive

Positive

Positive

Negative

Positive

Casein Hydrolysis

Negative

Positive

Positive

Positive

Positive

Positive

Nitrate Reduction

Positive

Positive

Varies

Varies

Negative

Varies

Motility

Negative

Negative

Positive

Negative

Positive

Positive

Acid from Mannose

Positive

Negative

Negative

Positive

Negative

Negative

Correlation to EI

n/a

2/5

2/5

3/5

1/5

1/5

Table 3: Similarities and differences of the characteristics of five Bacillus species in table 5 of the Bacillus section in Bergey’s Manual compared to the unknown EI found in soil located in Flagstaff, Arizona...