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March 30, 2022

General Microbiology Spring 2022 Final Lab Repo

Biology

Please look at the grading rubric. 

Paper should be in APA style.

For the Results part include very table from the reports provided in the right order. 

Reference minimum is 8 (please use scholar journals), and has to alphabetically organized. 

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    FinalLabReportGradingRubricSpring2022.pdf

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    Spring2022ClassActivity1.docx

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    ClassActivity2.docx

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    ClassActivity3.docx

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    ClassActivity4.docx

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    ClassActivity5.docx

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    ClassActivity6.docx

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    ClassActivity7.docx

General Microbiology Spring 2022 Final Lab Report Grading Rubric

Section Portion Explanation Points

Introduction

2-3 paragraphs (7 points)

Background information Description of why isolation/identification of bacteria is important (1) and importance of microbiology is addressed (1). Should include citations here to support all information addressed (1)

3

Purpose of the experiment stated

Student states the purpose behind conducting the experiment 1

Summary of methodology addressing all parts of the experiment.

Should include:

1 Serial dilution, viable titer, isolation, colony morphology, cellular morphology, biochemical tests, selective differential media, API, Bergey’s, and Kirby Bauer

There should be NO mention of their bacterial unknown’s identity in the introduction

Research Questions that encompass ALL parts of the experiment

There should be a minimum of 2 questions in question format that address identification of an unknown bacteria and antibiotic sensitivity.

2 Students can choose to address multiple questions for each part of the experiment but must encompass all parts, not just some.

Results

No more than 8 tables (no

more than 10 pages)

2-3

paragraphs for written portion

(23 points)

Week 3:

Table showing three dilution plates (0.25) with population survey (including unique colonies) and general colony morphology descriptions of the colonies on the plate (0.25) Students address and fully explain if the patterns in dilution among the three plates make sense. (0.5) Descriptions of chosen colony (only one included- five colonies and two isolation streaks were conducted in class but only one was chosen) (0.5) and gram stains 40x and 100x pictures with cellular morphology included (0.5)

2

Dilution and Viable Titer Dilution, dilution factor, CFU and Viable Titer present, and information correct (0.25 each) 1

Week 4/5: Isolation plate present with full colony morphology (7 characteristics: shape, size, color, margin, elevation etc.) (0.5) Comparison of colony morphology to the previous (original) colony morphology (0.5)

1

Isolation and Gram Stains Both gram stains present (40x and 100x objective) – (0.5)

2

With shape, arrangement, color, and gram stain category listed and explained, student states correctly objective/ magnification (0.5) If stains do not match or are mixed, a statement of possible error that occurred is included. If they do match, a statement included about this. (0.5) Comparison of the gram stain results to the previous (original) morphology and explanation on why (0.5)

Week 6: Biochemical Tests

All tests mentioned. 5.5

(11 tests) In results column, students must mention for each test: positive/negative, color and pH (0.25 for each test)

In description column, students must mention for each test: overall reaction that occurred or enzyme that is present

and any other characteristics about the bacteria (eg. aerobic). Be as specific and detailed as possible (0.25 for each test)

Week 7: All tests mentioned

2

Selective and Differential (4 tests)

In results column, students must mention for each test: positive/negative, color, pH. (0.25 for each)

In the description column, students must mention for each test: overall reaction that occurred or enzyme that is

present. Be as specific and detailed as possible. (0.25 for each)

Week 7/8: API Test Kit picture shown, all tests described as either positive or negative (1)

1.5 API Test Kits API Reading sheet shown with results written and final 7-digit code (0.25)

API Web results included with mention of final identification of %ID. (0.25)

Week 8: Bergey’s manual steps used, and final identification shown (0.75) along with a screenshot of the flowchart used for identification (0.25).

1 Bergey’s Manual

Students include a table comparing observed results to expected results for their API ID of their unknown

2

Week 9: Table contains all biochemical tests and API tests listed with observed results obtained throughout the semester (0.5)

Comparison of ID Table contains expected results of each of the biochemical and API tests listed. Students can use the Bergey’s Manual on Canvas (0.5)

Students highlight areas of the table in which observed and expected results do not match (0.5) Table contains citations for all information obtained from outside resources (0.5)

Week 10/11: Picture of Kirby Bauer Assay included (0.5) 1.5

Kirby Bauer All 6 antibiotic measurements match the classifications (R/I/S) based on standards for known bacteria (1)

Results Write-up Paragraph

Results write-up paragraph is present and includes all portion of the results in a summary format

4

Students are not repeating what is already stated in the results table but rather summarizing the important results from each section.

Week 3: Dilution pattern comparison among three plates and viable titer (0.75)

Week 4/5: Does isolation plate morphology match dilution previous step? Is it a pure culture? (0.75)

Week 6/7: Which tests positive and which enzymes are present. (1)

Week 8: API identification stated (0.25)

Week 8: Bergey’s identification stated (0.25)

Week 10-11: Antibiotic classifications stated (0.5)

No discussion takes place within the paragraph (0.5)

Discussion (10 points)

5 – 8

paragraphs

Student addresses colony and cellular morphology and if it matches expectation (through support from citation)

1 If no citation, -0.5 point

Students address biochemical tests and if it matches expectations. Which did/did not? (through support from citation) 1

If no citation, -0.5 point

Comparison of API Kit to Bergey’s manual identification. Which was more effective? 1

Student addresses susceptible antibiotics and if it matches expectation (through support from citation)

1 If no citation, -0.5 point

Student addresses if research questions proposed in introduction were answered or not. 2

Possible sources of error stated AND why the error caused an issue within the experiment. Minimum two errors discussed. If there are no errors, the student addresses potential areas for error and why they would cause an issue.

2

Discussion of all potential environments your identified bacteria can be found. (1) Citations present to support (0.5) Does this match your sampled environment? (0.5)

1

Discussion of the importance/relevance of identified bacterial sample 1

Future Directions

1-2

paragraphs (2 points)

Student proposes a new follow-up experiment and proposes methodology which makes sense and builds on the information obtained in this lab report Note: Do not point out errors in current experiment or suggest the same methodology used in this experiment

1

Student mentions the application of both current and future results in a real-world setting 1

References (4 points)

References APA format used in references section: inclusion of hanging indentations and alphabetical order 1

In-text citations

In-text citations done correctly (Tomasetti et al. 2021) and is used to support information that is not original in introduction section

0.5

In-text citations done correctly (Tomasetti et al. 2021) and is used to support information that is not original in discussion section

0.5

All citations mentioned in references section are cited as in-text references throughout the text 0.5

A minimum of 8 references are used throughout the report. 0.5

Formatting (4 points)

Introduction Proper grammar and sentence structure flows. Sentences do not contain pronouns: I, our, me, us, we etc. Section does not cross minimum or maximum guidelines (minus 0.5)

0.5

Results

Proper grammar and sentence structure flows. Sentences do not contain pronouns: I, our, me, us, we etc. (0.5) Results write up section does not cross minimum or maximum guidelines (minus 0.5)

0.5

Each table has a numbered title and caption at its heading and information in the section flows properly (0.5) 1

Pictures included in results section are all cropped, formatted, and labelled (where applicable) consistently and its aspect ratio is consistent with each image type

0.5

Discussion Proper grammar and sentence structure flows. Sentences do not contain pronouns: I, our, me, us, we etc. Section does not cross minimum or maximum guidelines (minus 0.5)

0.5

Future Directions Proper grammar and sentence structure flows. Sentences do not contain pronouns: I, our, me, us, we etc. Section does not cross minimum or maximum guidelines (minus 0.5)

0.5

Overall

All bacteria names are properly written throughout the report. Italicization with no improper capitalization 0.5

All tables and text are formatted similarly throughout: sizing is the same, font and font size throughout report are the same

0.5

Total point assignment 50

Assignment Percentage in final grade 20%

Helpful link to APA formatting guidelines: https://owl.purdue.edu/owl/research_and_citation/apa_style/apa_formatting_and_style_guide/general_format.html

https://owl.purdue.edu/owl/research_and_citation/apa_style/apa_formatting_and_style_guide/general_format.html

,

Activity #1 Section: U01

Lab Title: Introduction to Staining. Student Name: Thalia Ramos Gigato

Date: 01/25/2022 Student ID: 6141550

Staining Technique

Species

Objective

Picture

Description

Cellular Morphology

Simple Stain

Yeast- crystal violet

40x

Shape: cocci Arrangement: Clustered Color: Purple Staining characteristics: Crystal Violet Gram positive

Simple Stain

Yeast- Safranin

40x

Shape: cocci Arrangement: clustered Color: pink Staining characteristics: Safranin Gram negative

Gram Stain

Escherichia coli

40x

Shape: rods Arrangement: mostly in pairs Color: pink Staining characteristics: Gram negative

Gram Stain

Staphylococcus aureus

40x

Shape: rods Arrangement: clustered Color: purple Staining characteristics: Crystal violet, Safranin

Gram positive

Gram Stain

Bacillus megaterium (24h)

40x

Shape: rods Arrangement: clustered Color: purple Staining characteristics: Crystal violet, safranin Gram positive

Gram Stain

Bacillus megaterium (48h)

40x

Shape: rods Arrangement: clumped Color: purple Staining characteristics: Crystal violet, safranin Gram positive

Gram Stain

Mycobacterium sp.

40x

Shape: cocci Arrangement: clustered Color: pink Staining characteristics: Gram negative

Acid fast stain

Staphylococcus aureus

40x

Shape: spherical Arrangement: clumped Color: purple/blue Staining characteristics:

Gram positive

Acid fast stain

Mycobacterium sp.

40x

Shape: rod Arrangement: in chains Color: pink Staining characteristics: Carbol fuchsin, acid alcohol, methylene blue. Gram positive Note: The results aren’t consistent since the color should be purple, this might be caused by a cross-contamination.

Spores Stain

Bacillus megaterium

(24h)

40x

Shape: rods

Arrangement: clumped

Color: purple

Staining characteristics: Gram positive

Spores Stain

Bacillus megaterium

(48h)

40x

Shape: rods

Arrangement: clustered

Color: purple

Staining characteristics: Gram positive

,

Activity #2

Section: U01

Lab Title: Isolation and Identification of Bacteria from Wild Samples.

Date: 02/08/2022

Student Name: Thalia Ramos Gigato

Student ID: 6141550

Table 1: Population survey of environmental sample

Environment

Picture

Description

Dilution Factor

Viable Titer

Plant

Population Survey

Unique colonies: 22

Colony Morphology:

Size: small

Shape: punctiform

Color: creamy white

Margins: even

Elevation: flat

Surface: smooth

Opacity: opaque

Other: spread out, more in the right side.

102

CFU: 22

Viable Titer: 2.2×10^4 CFU/mL

Calculations.

(22CFU/0.1mL) x10^2 = 22,000 = 2.2×10^4 CFU/mL

Population Survey

Unique colonies: N/A

Colony Morphology:

Size: N/A

Shape: N/A

Color: N/A

Margins: N/A

Elevation: N/A

Surface: N/A

Opacity: N/A

Other: No colonies presented in the plate.

104

CFU: No colonies

viable Titer: Cannot be calculated

Calculations

N/A

Population Survey

Unique colonies: 33

Colony Morphology

Size: small

Shape: punctiform

Color: creamy white

Margins: even

Elevation: flat

Surface: smooth

Opacity: opaque

Other: The colonies appear smaller than the colonies on the plate 10^2

106

CFU: 33

Viable Titer 3.3×10^8 CFU/mL

Calculations

(33 CFU/0.1mL) x10^6= 330,000,000= 3.3×10^8 CFU/mL

The first plate has many colonies but not as much as the third plate, while the second plate doesn’t have any colonies present, just appear sweaty. Also, the colonies on the 10^6 plate are smaller than the ones presented in the 10^2 plate.

Table 2: Gram Stains characterization of chosen colonies

Isolated Colony

Dilution

Picture

Description

Picture

Description

Section: U01 Group: 5

Colony #1

10^-2

Colony Morphology:

Size: small

Shape: circular

Color: creamy white

Margins: even

Elevation: flat

Surface: smooth

Opacity: opaque

40x objective

Cellular Morphology:

Size: small

Shape: rods

Arrangement: clustered, bacillus

Color: pink

Staining characteristics: The stain was pink indicating being Gram negative.

100x objective

Colony #2

10^2

Colony Morphology:

Size: small

Shape: circular

Color: creamy white

Margins: even

Elevation: flat

Surface: smooth

Opacity: opaque

40x objective

Cellular Morphology:

Size: small

Shape: cocci

Arrangement: clumped, streptococcus

Color: pink

Staining characteristics: Since the stain turned pink indicates being Gram negative.

100x objective

Table 3: Other characterized colonies

Isolated Colony

Dilution

Picture

Description

Picture

Description

Colony #3

10^2

Colony Morphology:

Size: small

Shape: circular

Color: creamy white

Margins: even

Elevation: flat

Surface: smooth

Opacity: opaque

40x objective

Cellular Morphology:

Size: small

Shape: rods

Arrangement: clustered, bacillus

Color: pink

Staining Characteristics: Since the stain turned pink indicates that it might be Gram negative.

Other: After staining the bacteria turned pink which indicates possible being Gram negative but there are always the possibility of some cross contamination altering the results.

100x objective

Colony #4

10^2

Colony Morphology:

Size: small

Shape: circular

Color: creamy white

Margins: even

Elevation: flat

Surface: smooth

Opacity: opaque

40x objective

Cellular Morphology:

Size: small

Shape: rods

Arrangement: clustered, streptobacillus

Color: purple

Staining Characteristics: After staining the bacteria turned purple which indicates being Gram positive.

Other: Since the rest of the bacteria have stained pink and this is the only one turning purple it could mean there was some cross contamination in the process.

100x objective

Colony #5

10^2

Colony Morphology:

Size: small

Shape: circular

Color: creamy white

Margins: even

Elevation: flat

Surface: smooth

Opacity: opaque

40x objective

Cellular Morphology:

Size: small

Shape: rods

Arrangement: clustered, bacillus

Color: pink

Staining Characteristics: Since the bacteria appears pink it could indicate being Gram negative.

Other: The bacteria appear to be Gram negative but there could be some cross contamination altering the results.

100x objective

,

MCB3020L General Microbiology Laboratory

Activity #3

Section: U01

Lab Title: Further Isolation of Bacterial Sample: Characterization, Gram Staining and Re-streaking.

Date: 02/15/20222

Student Name: Thalia Ramos Gigato.

Student ID: 6141550

CLASS ACTIVITY WORKSHEET

Table 1: Isolation of chosen colony #1

Note: Some of this information is obtained from your previous class activity.

Sample

Picture

Description

Isolated streak on nutrient agar plate #1

Colony Morphology:

Size: small

Shape: punctiform

Color: creamy white

Margins: even

Elevation: flat

Surface: smooth

Opacity: opaque

Other: They aren’t as much spread out as the plates from last week.

Does this match the previous colony chosen from serial dilution plates? Why or why not?

Yes, they have the same colony morphology.

Is it pure? Why or why not?

They look about the same.

Gram stains of tertiary streak

40x objective

Cellular Morphology:

Size: small

Shape: rods

Arrangement: cluster

Color: pink

Staining characteristics: Gram negative, thin peptidoglycan.

Potential errors?

None

Do the 40x and 100x pictures match? Why or why not?

Yes, because the bacteria look the same.

Does this Gram stain result match the previous colony (obtained from serial dilution)? Why or why not?

Yes, because the still have rod shape and are gram negative.

Is it pure? Why or why not?

Yes, because they have the same characteristics.

Gram stains of tertiary streak

100x objective

Gram stains of tertiary streak

40x objective

Cellular Morphology:

Size: small

Shape: rods

Arrangement: cluster

Color: pink

Staining characteristics: gram negative, thin peptidoglycan

Potential errors?

None

Do the 40x and 100x pictures match? Why or why not?

Yes, because they have the same chape, color, and arrangement.

Does this Gram stain result match the previous colony (obtained from serial dilution)? Why or why not?

Yes, because the bacteria is negative and present rod shape.

Is it pure? Why or why not?

Yes, because they look the same.

Gram stains of tertiary streak

100x objective

Gram stains of the Tail Region

40x objective

Cellular Morphology:

Size: small

Shape: rods

Arrangement: cluster

Color: pink

Staining characteristics: gram negative, thin peptidoglycan

Potential errors?

None

Do the 40x and 100x pictures match? Why or why not?

Yes, because the bacteria have the same shape, color, and arrangement.

Does this Gram stain result match the previous colony (obtained from serial dilution)? Why or why not?

Yes, because it shows the same results.

Is it pure? Why or why not?

Yes, because they share the same characteristics.

Gram stains of the Tail Region

100x objective

Gram stains of the Tail Region

40x objective

Cellular Morphology:

Size: small

Shape: rods

Arrangement: cluster

Color: pink

Staining characteristics: gram negative, thin peptidoglycan

Potential errors?

None

Do the 40x and 100x pictures match? Why or why not?

Yes, they look the same.

Does this Gram stain result match the previous colony (obtained from serial dilution)? Why or why not?

Yes, because the bacteria have the same characteristics.

Is it pure? Why or why not?

Yes, because they have same color, arrangement, and shape so it’s considered a pure colony.

Gram stains of the Tail Region

100x objective

Table 2: Isolation of chosen colony #2

Note: Use the guidelines above to help replicate the table for nutrient agar plate #2

Sample

Picture

Description

Isolated streak on nutrient agar plate #2

Colony Morphology:

Size: small

Shape: punctiform

Color: creamy white

Margins: even

Elevation: flat

Surface: smooth

Opacity: opaque

Other: The colonies are close

Does this match the previous colony chosen from serial dilution plates? Why or why not?

Yes, they have the same colony morphology.

Is it pure? Why or why not?

Yes, because they have the same characteristics.

Gram stains of tertiary streak

40x objective

Cellular Morphology:

Size: small

Shape: rods

Arrangement: cluster

Color: pink

Staining characteristics: gram negative, thin peptidoglycan.

Potential errors?

None

Do the 40x and 100x pictures match? Why or why not?

Yes, because they show the same characteristics.

Does this Gram stain result match the previous colony (obtained from serial dilution)? Why or why not?

Yes, because the colony has the same characteristics.

Is it pure? Why or why not?

Yes, because the bacteria have the same shape, arrangement, and color.

Gram stains of tertiary streak

100x objective

Gram stains of tertiary streak

40x objective

Cellular Morphology:

Size: small

Shape: rods

Arrangement: cluster

Color: pink

Staining characteristics: gram negative, thin peptidoglycan

Potential errors?

None

Do the 40x and 100x pictures match? Why or why not?

Yes, because the bacteria still have the same characteristics.

Does this Gram stain result match the previous colony (obtained from serial dilution)? Why or why not?

Yes, because by the characteristics we can assume is the exact same bacteria.

Is it pure? Why or why not?

Yes, because it presents the same shape, arrangement, and color.

Gram stains of tertiary streak

100x objective

Gram stains of the Tail Region

40x objective

Cellular Morphology:

Size: small

Shape: rods

Arrangement: cluster

Color: pink

Staining characteristics: gram negative, thin peptidoglycan.

Potential errors?

None

Do the 40x and 100x pictures match? Why or why not?

Yes, because they have the same colony morphology.

Does this Gram stain result match the previous colony (obtained from serial dilution)? Why or why not?

Yes, because by the characteristics of the bacteria they are the same.

Is it pure? Why or why not?

Yes, because the bacteria show the same shape, arrangement, and color.

Gram stains of the Tail Region

100x objective

Gram stains of the Tail Region

40x objective

Cellular Morphology:

Size: sma

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