MCB 2010 Miami Dade College Laboratory Investigation of Unkwown Organism Report

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MCB 2010

Miami Dade College

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This is for the LAB portion of Microbiology 2010. ASSIGNMENT IS DUE APR 20, 2020 @ 2130 EST

-Need to identify an unknown organism, based on info given from slideshows attached

-Need to answer approx 60 yes/no questions from the info above (will aid in Identifying the organism)

-finally, write a small APA-style report on the organism, writing topic and criteria attached

I will upload 5 more files (Studypool only allows 5 files to be uploaded at a time)

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Unknown Project Introduction Unknown Organism #2 Introduction Morphological and Cultural Characteristics Part 1 Unknown Project The exercises in this presentation will allow you to go through the process of identifying an unknown bacterium. As you identify the organism, you will gain an understanding of the morphological, cultural, and physiological characteristics of bacteria. Many of the tests are biochemical in nature and you will gain a thorough understanding of the chemistry of the tests that were performed on the organisms. Please make sure to use your lab manual as reference. You will be required to download a number of files to help you keep track of the information and determine the identification of unknown organism. Watch video https://mediaplayer.pearsoncmg.com/assets/secs-microbio-mlt-unknown-id Step 1: Accumulate Information • You will gather information that pertains to your unknown organism’s morphological, cultural, & physiological characteristics. • Record data carefully and keep notes “Tests List”. • After sufficient information has been recorded, you can consult dichotomous keys and a portion of Bergey’s Manual to identify the bacteria which will help you to narrow down and eventually identify the organism to exact Genus species. • Dichotomous keys are key for the identification of organisms based on a series of choices between alternative characteristics. For bacteria, the first choice will be the Gram stain result! Files to be downloaded and printed • • • • Tests List – print at start of unknown project Dichotomous key #1 Dichotomous key #2 to be printed out Bergey’s Manual pages later in project Evaluations of cultural and morphological characteristics Colony Morphology. Lab Ex. 2-2 (Page 36) • Color, shape, size, and • 5 basic categories of texture of microbial colony morphology growth are determined by – Shape the genetic makeup but – Margin are greatly influenced by – Elevations environmental factors including nutrient – Texture availability, temperature – Pigment production and incubation time. Click here for results Evaluations of cultural and morphological characteristics Gram Stain. Lab Ex. 3-7 (Page 105) Watch video: Click on image or use the following https://www.youtube.com/watch?v=UbGl3CUboPo Evaluations of cultural and morphological characteristics Gram Stain. Lab Ex. 3-7 (Page 105) Click here for results Evaluations of cultural and morphological characteristics Optimum Growth Temperature . Test #5 • Microorganisms cultivated in broth display a variety of growth characteristics, one of those is a preferred temperature. • For this experiment, Unknown Organism was inoculated into nutrient broth (NB). One tube was place at 25°C and another placed at 37°C. • After 48 hours of growth, the tubes will be observed for turbidity. Click here for results Evaluations of cultural and morphological characteristics Growth Pattern on Slants. Lab 2-3 (Page 44) • Agar slants are useful primarily for cultivation and maintenance of stock cultures as well as pigment production • Dense and opaque growth with smooth edges is termed - filiform • Possible growth patterns: A. Flat/dry B. Spreading edge (diffuse edge) C. Microbacterium produce friable (crusty) growth D. Translucent, barely visible • Tryptic soy agar (TSA) slants were inoculated with Unknown Organism using an inoculating needle. A B C D Click here for results Evaluations of cultural and morphological characteristics Growth Pattern in Nutrient Broths. Lab 2-4 (Page 45) • Microorganisms cultivated in broth display a variety of growth characteristics. • Bacterial genera, and frequently individual species within a genus, demonstrate characteristic growth patterns in broth that provide useful information when attempting to identify an organism. • Location and how organisms grow help with identification and gives us clues to if they are motile or non-motile. Evaluations of cultural and morphological characteristics • • Growth Pattern in Nutrient Broths. Lab 2-4 (Page 45) A = flocculence B and C = uniform fine turbidity (UFT) • Hint that it may be motile, though not a true motility test • D and E = sediment • Hint that it may non-motile, not a true motility test • F and G = pellicle • Organisms float on top because they produce a waxy cell wall (remember Mycobactierium species are acid fast and have mycolic acid in their cells walls A B C D E F G Evaluations of cultural and morphological characteristics Growth Pattern in Nutrient Broths. Lab 2-4 (Page 45) • For this experiment, Unknown Organism was inoculated into nutrient broth (NB). After 48 hours of growth, the tubes will be observed for turbidity. Click here for results Evaluations of cultural and morphological characteristics Fluid Thioglycollate Medium. Lab 2-7 (Page 50) • Fluid thioglycolate medium (FTM) is a liquid medium designed to promote growth of a wide variety of fastidious microorganisms. • This media is used to determine oxygen requirements • During making of FTM tubes, a gradient of oxygen concentration occurs – higher O2 at top, lower O2 at bottom • Growth patterns: L to R (top image from A B C D E lab book) A. B. C. D. E. Aerotolerant anaerobe Facultative anaerobe Obligate/strict anaerobe Obligate/strict aerobe Microaerophile Watch video: https://youtu.be/AJG18sQd8mU In these tubes dots represent bacterial growth/turbidity Evaluations of cultural and morphological characteristics Fluid Thioglycollate Medium. Lab 2-7 (Page 50) • Microorganisms cultivated in FTM display growth representing their level of oxygen requirement. • For this experiment, Unknown Organism was inoculated into fluid thioglycollate medium (FTM) and incubated at 37°C. • After 48 hours of growth, the tubes will be observed for location of growth or turbidity. Click here for results Result slides follow View isolated colonies below. Choose 2-3 descriptive words to describe colonies. Fill in Test List #1 Click to return to previous slide View isolated bacteria below. Fill in Test List #2 and Test List #3 Click to return to previous slide View and interpret nutrient broth tubes below. Tube C is un-inoculated control. Fill in Test List #5 37°C 25°C Click here to return to previous slide A B Control C View and interpret TSA slant below. Fill in Test List #7 Possible choices: • Friable growth • Flat/dry growth • Spreading edge growth • Translucent growth Click to return to previous slide View and interpret FTM tube below. Fill in Test List #4 Possible choices: • Facultative anaerobe • Obligate/strict aerobe Click to return to previous slide Examine Tube B and compare with the un-inoculated control (Tube A). Choose the growth pattern and record in Test #8 Possible choices: a. Uniform fine turbidity b. Sediment c. Pellicle d. Flocculence A Click here to return to previous slide B Unknown Project Introduction Unknown Organism #2 Fermentation and Respiration Tests Part 2 Differential Tests • Bacteria have been differentiated based on their enormous biochemical diversity. • These are called differential or biochemical tests. • These tests usually provide a single KEY substrate ( ex: particular sugar or amino acid) whereas the organism can carry out a specified metabolic reaction or series of reactions. • We will use individual tests to learn about metabolic pathways and mechanisms both shared and unique to various bacteria. Fermentation Tests • Fermentation tests are to determine whether your unknown is capable of fermenting various sugars. • Fermentation is the partial break down of organic molecules mainly sugars. • Microbial fermentation of sugars produces three major products: acid, alcohol and gas • Detection of fermentation is done with tests using differential media such as Phenol Red broth or general media MR-VP medium. • Not all bacteria ferment all the sugars. • The main product of bacterial fermentation of sugar is variety of organic acids. • Some bacteria may produce gases such as CO2 or hydrogen in addition to acids. • pH change is an excellent indicator of metabolic activity. • pH indicators such as phenol red or methyl red can be used to show pH changes. Fermentation Tests Phenol Red Broth. Lab 5-3 (Page 158) • Phenol Red (PR) is a differential test medium used to determine fermentation and gas production. It can be used to differentiate members of Enterobacteriaceae and to distinguish them from other Gram-negative rods. • All ingrediants are the same except for the sugar • pH indicator is phenol red – Neutral is red – Acidic is yellow – Basic is pink • Gas indicator is bubble or clear zone inside the Durham tube Watch video: https://www.youtube.com/watch?v=vJcL9f52Zuk https://www.youtube.com/watch?v=3os3odC_UFU Keep in mind that we use four sugars, glucose, lactose, mannitol and arabinose BUT the inoculation and interpretation of results are the same Fermentation Tests Phenol Red Broth. Lab 5-3 (Page 158) • Some bacteria can grow in phenol red broth but do not use the carbohydrate provided. For energy, these grow using only the protein supplied. • As the bacteria grow, they remove amine groups from amino acids to grow and in the process produced ammonia, NH3. Ammonia is basic and raises the pH. Phenol red is pink at high pH. – See last tube in Figure 5-7. • Often, the letter K is used to demonstrate degradation of proteins BUT this result is still a negative test result for the fermentation of that sugar. Fermentation Tests Phenol Red Broth. Lab 5-3 (Page 158) • For this experiment, Unknown Organism was inoculated into four different phenol red carbohydrate broths and incubated at 37°C. • The sugars used are – – – – Glucose Lactose Mannitol Arabinose • After 48 hours of growth, the tubes will be observed for color change and production of gas. • Use Table 5-2 and pictures in Figure 57 to interpret your Unknown Organisms fermentation of sugars results. Click here for results Fermentation Tests Methyl Red and Voges-Proskauer Tests (MRVP). Lab 5-4 (Page 161) • MRVP broth is a combination medium used for two tests. • MR test is designed to detect organisms capable of performing a mixed fermentation. • VP is designed for organisms that are able to ferment glucose, but quickly convert their acid products to acetoin and 2,3-butanediol. Watch video: https://www.youtube.com/watch?v=LS5x0d9XPCA Fermentation Tests Methyl Red and Voges-Proskauer Tests (MRVP). Lab 5-4 (Page 161) • For this experiment, Unknown Organism was inoculated into MRVP broth and incubated at 37°C. • After incubation, aliquots were removed and reagents added according to Procedural Diagram 5-14. Click here for results Tests to Identify Microbial Ability to Respire (Catalase, Oxidase and Nitrate Reduction Tests) • Respiration is the complete breakdown of organic compounds, like sugar, to release energy. • Respiration involves in 3 stages; Glycolysis, Krebs Cycle and Electron Transport Chain (ETC). • The final electron acceptor at the end of ETC could be oxygen (aerobic respiration) or inorganic compound such as nitrate or sulfate(anaerobic respiration) • Respiration tests directly or indirectly demonstrate the presence of ETC. Respiration Tests Catalase Test. Lab 5-5 (Page 165) • Aerobic respiring bacteria use oxygen as the final electron acceptor. HOWEVER, not all electrons end up in the appropriate place! • As oxygen enters into cellular reactions, it may be transformed into several toxic products: – O2·- (superoxide ion free radical) – H2O2 (hydrogen peroxide) • Various types of oxygen free radicals (unpaired electrons) are collectively called reactive oxygen species (ROS) – Disrupt cell membranes; destroys lipids, proteins, and DNA • Organisms that produce these toxins also produce enzymes capable of breaking them down. Respiration Tests Catalase Test. Lab 5-5 (Page 165) • The enzymes catalase and superoxide dismutase are necessary to neutralize toxic forms of O2 • Superoxide dismutase converts superoxide ion to hydrogen peroxide • Catalase converts hydrogen peroxide to water and gaseous O2 • When catalase and superoxide dismutase enzymes are present, they indicate O2 can be used by bacteria • Obligate aerobes, facultative anaerobes, aerotolerant anaerobes, and microaerophiles contain these enzymes to breakdown these ROS – Obligate anaerobes lack these enzymes Respiration Tests Catalase Test. Lab 5-5 (Page 165) • For this experiment, Unknown Organism was inoculated into a TSA slant and incubated at 37°C. • After incubation, a large amount of growth was added to a microscope slide. • Aseptically, two drops of hydrogen peroxide was added directly onto the bacteria and immediately observed for the production of bubbles. • Use Table 5-5 to interpret results. Click here to see results Respiration Tests Oxidase Test. Lab 5-6 (Page 168) • Many aerobes, microaerobes, facultative anaerobes, and even some anaerobes have electron transport chains (ETC). • The functions of the ETC are: 1. 2. Transport electrons down a chain of molecules/enzymes to the terminal electron acceptor (oxygen in aerobic respiration) Generate a proton motor force that drives the production of ATP The last enzyme in the ETC found in some bacteria is called cytochrome C oxidase. • Application of this test is to identify bacteria containing cytochrome C oxidase • Respiration Tests Oxidase Test. Lab 5-6 (Page 168) • For this experiment, Unknown Organism was inoculated into a TSA slant and incubated at 37°C. • After incubation, a large amount of growth was transferred onto the reagent slide, BBLTMDrySlideTM. This slide contains the substrate for the enzyme cytochrome C oxidase. • Immediately observe for color change within 20 seconds. • Use Table 5-6 to interpret results. Click here for results Respiration Tests Nitrate Reduction Test. Lab 5-7 (Page 171) • Anaerobic respiration involves the reduction (or transfer of electrons) to an inorganic molecule (not oxygen!). • Virtually all members of Enterobacteriaceae perform a one-step reduction of nitrate to nitrite. • This media contains the inorganic molecule nitrate. • This test is used to determine if the bacteria contain the enzyme nitrate reductase. Respiration Tests Nitrate Reduction Test. Lab 5-7 (Page 171) A B • Reagents are testing for the presence to nitrite in the broth. • If the test is positive, this means nitrate → nitrite • A=presence of nitrite = positive result nitrate reductase is present • B=control tube Respiration Tests Nitrate Reduction Test. Lab 5-7 (Page 171) • For this experiment, Unknown Organism was inoculated into nitrate broth and incubated at 37°C. • After incubation, aliquots were removed and reagents added according to Procedural Diagram 5-28. • Reagents are testing for the presence of nitrite in the broth. nitrate → nitrite • If the test is positive, nitrate reductase is present Click to view results Result slides follow Use Table 5-2 and pictures in Figure 5-7 to interpret your Unknown Organisms fermentation of sugars results. Record results for Tests # 9, 10, 11,and 12 Click here to return to previous slide Use Figure 5-10 and Table 5-3 to interpret and record results for MR. Record in Test #13. Click here to return to previous slide Use Figure 5-13 and Table 5-4 to interpret and record results for MR. Record in Test #14. View results below. Fill in Test List #15 Click here to return to previous slide View results below. Fill in Test List #16 Click here to return to previous slide Tube B is the control tube. Interpret the result in tube A. Fill in Test list #17 Click here to return to previous slide Unknown Project Introduction Unknown Organism #2 Other Biochemical Tests Conclusion of Unknown Project Part 4 Other Biochemical Tests Citrate as a Sole Carbon Source. Lab 5-8 (Page 175) • Citrate-permease enzyme breaks down sodium citrate • Sodium citrate is the only carbon-containing compound in the medium. – Most bacteria would prefer to use glucose or an easier compound to break down – Defined medium • Ammonium phosphate provides the sole nitrogen source in the medium. • Citrate-positive bacteria produce ammonia (NH3) and ammonium hydroxide (NH4OH), both of which alkalinize the medium and turn it blue. Other Biochemical Tests Citrate as a Sole Carbon Source. Lab 5-8 (Page 175) • Bromthymol Blue is the pH indicator in this medium. – Green at pH 6.9, Blue at pH 7.6 • For this test an inoculating needle was used to add a light inoculum on the Simmons citrate agar slant with the Unknown Organism. • Tube was incubate at 37 °C for 48 h. Click here to view results Other Biochemical Tests Phenylalanine Deaminase Test. Lab 5-11 (Page 182) • Organisms that produce phenylalanine deaminase can be identified by their ability to remove the amine group (NH2) from the amino acid phenylalanine. • To interpret the results, the reagent, ferric chloride (FeCl3) must be added. • The normally colorless phenylpyruvic acid reacts with the ferric chloride and turns a dark green color almost immediately. • Phenylpyruvic acid + FeCl3 → green color Other Biochemical Tests Phenylalanine Deaminase Test. Lab 5-11 (Page 182) • For this test an inoculating loop was used to add a heavy inoculum on a phenylalanine agar slant with the Unknown Organism. • Tube was incubate at 37 °C for 48 h. • After incubation, ferric chloride was added to tube. Click here to view results Other Biochemical Tests SIM Medium Test. Lab 5-20 (Page 202) • SIM is a combination differential media that combines several compatible tests in to one medium. • SIM medium is used for determination of three bacterial activities: 1. Motility 2. Sulfur reduction 3. Indole production from the amino acid tryptophan Other Biochemical Tests SIM Medium Test. Lab 5-20 (Page 202) • Motility is an important differential characteristic of Enterobacteriaceae. • After incubation, the first result to view is motility. • If the growth stays within the stab line, the organism is non-motile, tube B . • If growth radiates outward from the stab, often all the way to the edge of the tube, it is motile, tube A. A B Other Biochemical Tests A B SIM Medium Test. Lab 5-20 (Page 202) • Sulfur reduction is the next result to view. • Hydrogen sulfide (H2S), a colorless gas can be detected when it reacts with ferrous sulfate in the medium to form a black precipitate –ferric sulfide. • Any blacking is a positive result. • Tube A is H2S-negative • Tube B is H2S-positive Other Biochemical Tests SIM Medium Test. Lab 5-20 (Page 202) • The last result to view is indole production. • Indole production occurs in bacteria that contain an enzyme called tryptophanase that hydrolyses the amino acid tryptophan. • This hydrolysis can be viewed by the addition of the reagent Kovac’s solution. • Formation of a red/pink color is a positive result. • Tube A is indole-negative • Tube B is indole-positive A B Other Biochemical Tests SIM Medium Test. Lab 5-20 (Page 202) • For this test a SIM tube was single stabinoculated using an inoculating needle. • Tube was incubate at 37 °C for 48 h. Click here to view results Conclusion of Data Collection • Now that all tests are complete, print out dichotomous keys #1 and #2. • Use these keys to determine the genus of your organism. • To determine the species, use the tables and descriptions found in Bergey’s Manual. Compare your results against the published material in Bergey’s Manual. • Fill out Test List # 27 entry. Conclusion of Data Collection • Your Unknown Organism is one of the following: 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. Bacillus subtilis Bacillus cereus Proteus vulgaris Proteus mirabilis Serratia marcescens Shigella flexineri Klebsiella pneumoniae Pseudomonas aeruginosa Escherichia coli Citrobacter freundii Providencia stuartii Staphylococcus aureus Result slides follow Tube A is the un-inoculated control tube. Interpret the result in tube B. Fill in Test list #23 Click here to return to previous slide View phenylalanine slants after the addition of FeCl3. View and interpret Tube B. Tube A is the un-inoculated control. Fill in Test List #24 Click here to return to previous slide View and interpret the SIM tube below. Kovac’s reagent has been added. Fill in Tests List #6, #25 and #26. Click here to return to previous slide Unknown Project Introduction Unknown Organism #2 Hydrolysis Tests Part 3 Hydrolysis Tests • Reactions that use water (H2O) to split complex molecules are called hydrolysis or hydrolytic reactions. • Enzymes required for these reactions are called hydrolytic enzymes. • Enzymes secreted from the organism to catalyze reactions outside the cell are called extracellular or exoenzymes. • The following tests are designed to identify the actions of both intracellular and extracellular enzymes. Hydrolysis Tests Starch Hydrolysis. Lab 5-12 (Page 184) • Starch is a polysaccharide made up of repeating subunits of a-D-glucose. • Starch is too large to pass through bacterial cell membrane. • Organisms produce extracellular enzymes to hydrolyze starch in sugar subunits. • Secreted enzymes that hydrolyze starch are αamylase and oligo-1,6-glucosidase. Hydrolysis Tests Starch Hydrolysis. Lab 5-12 (Page 184) • Iodine stains starch brown. After incubation, a plate is flooded with iodine. A brown color represents starch is still present. • Figure 5-43 shows two organisms streaked on a starch plate. • Streak A is a negative result since there is starch next to the streak. • Streak B is positive for starch hydrolysis because the yellow halo surrounding the bacterial growth. • For this test a starch plate was singleline inoculated with the Unknown Organism and incubated at 37 °C for 48 h. A B Use loop Click to view results Hydrolysis Tests Urea Hydrolysis. Lab 5-13 (Page 187) • Urea is a product of decarboxylation of certain amino acids. • Many enteric bacteria posses the ability to metabolize urea. • Application: quickly differentiates urinary tract pathogens Ex. Proteus. • Urea broth contains phenol red. – Neutral is red – Acidic is yellow – Basic is pink Hydrolysis Tests Urea Hydrolysis. Lab 5-13 (Page 187) • Urease hydrolyses urea and produces ammonia (NH3) which raises pH. A B C • Tube B is the un-inoculated control. A positive urease test is shown in Tube A. Hydrolysis Tests Urea Hydrolysis. Lab 5-13 (Page 187) • For this test, the Unknown Organism was inoculated in to urea broth and incubated at 37°C. • Tubes should be examined daily for color change. • It is important to compare to un-inoculated control to determine if color change has occurred. Color change can occur after 24 hours or may take several days in weak positive organisms. Click here for results Hydrolysis Tests Casein Hydrolysis. Lab 5-14 (Page 190) • Casease breaks down the milk protein, casein, into amino acids. • Media contains powdered milk. • Casein hydrolysis test is used for the cultivation and differentiation of bacteria that produce the enzyme casease. • Steak A is casease-negative since the area around the streak is not cleared. • Streak B is casease-positive since the area around the streak has been cleared. • For this test a skim milk plate was single-line inoculated with the Unknown Organism and incubated at 37 °C for 48 h. A B Click here for results Hydrolysis Tests Gelatin Hydrolysis Test. Lab 5-15 (Page 192) • Gelatin is a protein derived from collagen which is a component of vertebrate connective tissue. • Gelatinases hydrolyze gelatin to individual amino acids that organism can then take up and use for metabolic purposes. Hydrolysis Tests Gelatin Hydrolysis Test. Lab 5-15 (Page 192) • Gelatin is solid at 25°C. • Gelatinase-positive organisms secrete gelatinase that will liquefy the medium (Tube A). • Nutrient gelatin liquefies (melts) at 28°C. These tubes need to be incubated at 25°C since incubation at 37°C will lead to false-positives. A B Hydrolysis Tests Gelatin Hydrolysis Test. Lab 5-15 (Page 192) • For this test nutrient gelatin tubes are single stabinoculated using a inoculating needle and then incubated at 25°C. • Gelatinase activity can be very slow in some organism so tubes may need to be incubated for up to 7 days. • It is important to include a non-inoculated control tube to make sure liquidfaction is due to enzyme activity and not temperature related. Click here to view results Hydrolysis Tests Lipid Hydrolysis Test. Lab 5-17 (Page 196) • Lipid is the work generally used to describe all types of fats. • Lipids are hydrolyzed by lipases. • Clear zones appear around streak in a lipase-positive organism. • For this test, a Tributyrin agar plate was single-line inoculated with the Unknown Organism and incubated at 37 °C for 48 h. Click here to view results Result slides follow View starch plate below after iodine has been added. Fill in Test List #18 Steak of Unknown Organism Click here to return to previous slide Compare tubes below. Tube A is the un-inoculated control. Tube B contains the Unknown organism Fill in Test List #19 Click here to return to previous slide View skim milk plate below. Interpret results and fill in Test List #20 Steak of Unknown Organism Click here to return to previous slide View and interpret nutrient gelatin tubes below. Tube B is un-inoculated control treated the same as the Unknown Organism inoculated in tube A. Fill in Test List #21 Click here to return to previous slide View tributyrin agar plate below. Interpret results and fill in Test List #22 Steak of Unknown Organism Click here to return to previous slide
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Explanation & Answer

This is the final one. If you need any clarification don't hesitate to ask. Let me know how i can be of help. Thank you.

Lab record: Unknown identification (Part 1)
**Each members Unknown Data sheet must be included in this report. Make sure your
name is clearly listed at the top of each report.

Mini-report of Unknown Organism (Part 2) (See page 6)
Honor Pledge for Group Projects
Name of Team Members: By including your name on this document
“I accept responsibility for my role in ensuring the integrity of the work submitted
by the group in which I participated.”
1.
2
3
4.

PART 1: Unknown identification- Test Records (50 points)
Unknown Number #________________________

Cell morphology and Gram Stain
Gram Stain Gram negative

Cell
Morphology

Arrangement

Color

Interpretation

Round

White

Gram negative rod

Rod

Ex 2-2: Colony Morphology
Colony Description (Record the observations for the basic categories terms of colony morphology)

Category

Description term

Colony Shape

Round

Margin

Smooth entire

Elevation

Growth into medium

Pigment
production(color)

White

Unknown Cultural Characteristics
Ex 2-4: Growth Pattern in Broth (Record the pattern for your organism and its description)
Uniform fine turbidity
1

Lab record: Unknown identification
Mini-report of Unknown Organism

This hints the organism may be motile though it is not a true motility test.
Ex 2-3: Growth Pattern on Slant (Record the pattern for your organism and its description)

Flat/Dry
Optimum Growth temperature for Culture #
Presumed Optimum temperature for your bacteria is ---------------------250 C--------------.

EX: 2-7: Oxygen Requirement (Growth in Fluid Thioglycollate Medium
(FTM)
Location of growth in FYM

Category based on growth
Facultative anaerobe

Located at the mostly at the top and
distributed througout.

Respiration Tests
Ex 5-5: Catalase Test
Result (observation)

Interpretation (meaning of
result)
Catalase is present

Bubbles

Symbol
(+/-)
+

EX 5-6: Oxidase Test
Result

Interpretation

No color change to blue within
twenty seconds

Symbol
(+/-)

Cytochrome c oxidase is not present

_

Ex 5-7: Nitrate Reduction Test
Gas (Y/N)

N

Reagents
added
(chemical
names)
Inorganic
molecule
nitrate

Result
Color after
reagents

Not red

Interpretation
Zinc
Added
(Yes/NO)

Color
after
zinc

Yes

Red

Nitrates reductase is not
present in the organism

Fermentation Tests
Ex 5-3 Phenol Red Sugars
PR Sugar

Results

Interpretation

Symbol
(+/-)
2

Lab record: Unknown identification
Mini-report of Unknown Organism
Yellow broth, bubble in tube

Fermentation with acid end produc...


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