Exercise 23 Lab Report
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May 9, 2024
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Laboratory Report
Student: Date: Section: 177
23
23 The Staphylococci: Isolation and Identification
A. Results
1. Tabulation
After examining your mannitol salt agar plate, record the presence (+) or absence (−) of staphylococcus colonies in the appropriate columns. After performing coagulase tests on the various isolates, record the results as (+) or (−) in the appropriate columns. Collect data from your classmates to complete the chart.
Item
FOMITE
Colonies on
MSA
Colonies on
MSA
Coagulase
Coagulase
NOSE
CULTURE
Fomite
Nose
TOTAL TESTED
TOTAL COAGULASE
POSITIVE
TOTAL COAGULASE
NEGATIVE
PERCENTAGE
COAGULASE
POSITIVE
PERCENTAGE
COAGULASE
NEGATIVE
178
2. Microscopy
Provide drawings here of your various isolates as seen under oil immersion (Gram staining).
The Staphylococci: Isolation and Identification (continued)
3. Record of Culture and Test Results
Record your observations for all tests in the table below. Using Table 23.1, determine the likely identity of each isolate.
Unknown
GROWTH
ON MSA
(+/–)
FERMENTATION
OF
MANNITOL (+/–)
COAGULATION
OF
PLASMA (+/–)
DNASE
(+/–)
NOVOBIOCIN
(S/R)
SPECIES
HEMOLYSIS ON
BAP (α/β/γ)
Nose
Fomite
B. Short-Answer Questions
1. Describe the selective and differential properties of mannitol salt agar (MSA) for the isolation and iden-
tification of staphylococci.
2. Describe the differential property of blood agar for the isolation and identification of staphylococci.
3. Why is the coagulase test considered to be the definitive test for S. aureus
?
4. What is the role of coagulase in the pathogenesis of S. aureus
?
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Related Questions
ISOLATE #1
Results for "Isolate #1"
***Please fill out the table****
1. Blood agar:
2. Nitrate Test:
Test
Results
Interpretation
Blood agar (type of hemolysis)
Nitrate test
3. Gelatinase Activity:
Gelatinase activity
Resistant or Susceptible?
Novobiocin Resistance
Zone Diameter:
4. Novobiocin Resistance:
5. Coagulase Test:
Coagulase (Sure-vue test)
1. Based on your results, what is the possible identification (Genus and species) of “Isolate #1"?
I 9 10 11 12 3 14
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ISOLATE #2
Results for "Isolate #2":
***Please fill out the table****
1. Blood agar:
2. Nitrate Test:
Test
Results
Interpretation
Blood agar (type of hemolysis)
Nitrate test
3. Gelatinase Activity:
Gelatinase activity
Resistant or Susceptible?
Novobiocin Resistance
Zone Diameter:
4. Novobiocin Resistance:
5. Coagulase Test:
Coagulase (Sure-vue test)
2. Based on your results, what is the possible identification (Genus and species) of "Isolate #2"?
nhes
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General Instructions:Choose 1 bacteria/diseases and fill out the details. An example of an answered template is also provided for your reference.
Causative Agent and Disease Profile for S. aureus
Template and Example
ITEM
MSM
PROFILE
MICROBIAL PROFILE
I
MICROORGANISM/CAUSATIVE AGENT
Staphylococcus aureus
A
GRAM REACTION
(+)
B
OXYGEN REQUIREMENT
Facultative Aerobes
C
SIZE
1.5 µm
D
SHAPE
Cocci in clusters
E
HABITAT
Normal flora of skin/anterior nares/pharynx
F
DISCOVERY
G
MICROSCOPIC IMAGE
II
DISEASE PROFILE
Scalded skin syndrome
A
DISEASE/S
Skin and Wound Infections
Scalded Skin Syndrome
Toxic Shock Syndrome
Food Poisoning Pneumonia
B
SYMPTOMS OF THE DISEASE
A high fever · Nausea and vomiting · A rash on your palms and soles that resembles a sunburn
C
INCUBATION PERIOD
2 and 4 hours (range 30 minutes to 8 hours)
D
MODE OF…
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BONUS (15 points)
The fallowing series of dilution was prepared from a specimen to determine the number of bacteria.
There were 62 colonies on the agar plate prepared by transferring 0.3 ml from the tube number 4.
Calculate the dilution factors for each tube.
What is the cell concentration in the original specimen?
Calculate the total number of cells in tube number 2.
étv
A) F12
F9
F10
F7
%
&
%3D
delete
5
{
T
Y
J
K
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LABORATORY REPORT: Aseptic Technique & Inoculation of Bacteria
Guide Questions.
1. Why is direct flaming preferred when disinfecting loops and needles?
2. Why is it important to flame the entirety of the loop and not just the tip? What consequences can be seen when this process is not done correctly?
3. What is the difference between quadrant streak method A from method B?
4. Why do we use a pencil instead of a pen when labelling culture tubes or plates? Are there other alternatives in labelling?
5. Why is an inoculating needle used in Subculturing Techniques? Can a loop be used instead?
NOTE: Please try to answer all of the question asked, i promised to give you a good ratings
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Case #2
Clinical History: This 29-year-old male's illness began 10 weeks prior to death, with an
episode of "flu". Two weeks later his urine became "smoky". He was found to have
hematuria, albuminuria and elevated BUN (180 mg/dl). He died from a pulmonary
embolus.
The throat culture obtained exhibited gram positive cocci in chains. It also showed beta-
hemolysis on sheep blood agar (SBA) and was catalase negative.
1. What was the most likely organism?
A. Streptococcus pyogenes
B. Streptococcus viridans
C. Staphylococcus aureus
D. Streptococcus pneumonia
2. What is the BEST diagnosis at the time of death?
3. What are the other possible sequela of this infection?
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Analysis of the pathological constituents
GIVE THE FOLLOWING:
- Name of Test/Test Reagent
- Positive Result
1) Protein (Note: Presence of protein in urine is termed as proteinuria or albuminuria)
A. Heat and Acetic acid test
Fill a test about ¾ full of urine.
Heat the upper portion gently to boiling for 1 -2 minutes being careful not to shake the tube.
Rotate the tube to prevent overheating. A turbidity may be due to albumin, phosphates or carbonates.
Add 3 drops of acetic acid drop by drop while boiling between each drop. If turbidity disappears, it is due to carbonates and phosphates.
B. Heller’s nitric acid test
Place 1 ml of urine sample in a test tube.
Hold the test tube in an inclined position and carefully pour 1 drops of conc HNO3 down the side of the test tube.
Note formation of white ring at the zone of contact of the two solutions which is an indication of the presence of protein.
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Analysis of the pathological constituents
GIVE THE FOLLOWING:
a. Name of Test/Test Reagent
b. Positive Result
Blood (Note: Presence of blood in urine is termed as hemolobinuria)
To 1 ml of freshly prepared Benzidine solution (sat’d sol’n of Benzidine in glacial Acetic acid) and 10 drops of urine sample.
Add 5 drops of 3% H2O2. A positive result is indicated by a blue green color.
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A 1.5-year-old child developed vomiting, diarrhea, and fever. Stool sample were inoculated into the Endo media. After 18 hours on the surface of the medium grew medium-sized, round, slightly convex red colonies with a metallic luster. The doctor suspected Escherichia coli.
1. Name the composition of the Endo agar media.
2. Describe the properties of bacterial colonies on the Endo media.
3. What purpose differential diagnostic Endo media used for?
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amylasw test
test for?
medium+ - reagents
+ Test/why?
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Helping tags: Biology, bacterial count, dilution, serial dilution
WILL UPVOTE, just pls help me answer the following questions and explain them clearly. Pls
show complete solutions also for the computation part. Thank you.
1. A bacterial culture was grown for 9 hours. At 3-hour interval, the culture was sampled to
determine the population of the culture, by transferring 25 ml of the suspension to 225
ml 0.85% NaCl. Three consecutive dilutions were further made by using 1 ml aliquot in 9
ml of 0.85% NaCI. One ml from each dilution was plated in each of duplicate plates. The
following table shows the results of the plating method.
Sampling
COUNTS
2nd dilution
3rd dilution
4th dilution
0,0
30: 35
250;245
1st dilution
1st
2nd
3rd
0; 0
240; 235
TNTC
55; 60
5; 6
TNTC
TNCT
TNTC
TNTC
a) Illustrate the dilution series used and label the final dilution of each dilution.
b) Determine the bacterial count (CFU/ml) every 3 hours of incubation for 9 hours.
Show all computations.
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A. A
Aa A
三,三,行,E
AaBbCcDdE Aa
AaBbCcDdF
AaBbCcDdF
AaBbCcDd
A
Normal
No Spacing
Heading 1
Heading 2
=y updates, fixes, and improvements, choose Check for Updates.
1. You have found a putative virus which is able to infect a bacterium causing increased
mortality and resistance to all antibacterial agents. You have systematically purified the
sample but have an unknown concentration of viruses. You perform the following serial
dilutions.
0.5ml
1ml
5ml
0.1ml
2.5ml
1ml
15ml
1
3
6
7
Phage Buffer
100.5ml
9ml
14.5ml
99.9ml 25ml
5ml 985ml
Tube Dilution in each Test Tube (Show Tube Dilution in each Test Tube (Show your
your work)
#
#
work)
4
1
6.
O Focus
glish (United States)
W
30
DII
DD
000
000
F4
80
F7
F8
F9
F10
F3
F5
F6
$4
&
4
5
6
8.
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Matching Type
Match the the condition/description to the correspondingtest/specimen to be conducted or evaluated. Choose
the correct answer.
Polymerase Chain Reaction Test
Rapid Antibody Test
Thyroxine Stimulating Hormone
Blood typing
Blood Chemistry
Microscopic Stool Examination
Complete Blood Count
Physical Stool Examination
Urinalysis
Alanine Aminotransterase Level
Susceptibility test
Human Chorionic Gonadotropin
Glycated Hemoglobin
1. Fasting Blood Sugar
2. Helminth Infestation
3. Presence of COVID-19 virus as antigen
4. Organ damage by Acetaminophen *
5. Antibiotic Resistance
6. Effectiveness of Metformin
7. Pregnancy
8. Ilicit drug use by bus drivers
9. Hypothyroidism
10.Identification of IgG and IgM against COVID-19 virus
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B. Microbial Processes
Study the diagrams and answer the given questions.
Pepartery
For items no. 1 to 6, refer to the diagram at
Glucose
the left corner.
1. What process is shown in the diagram?
2. In what part of a bacterium does the
process occur?
3. What are the raw materials of the process?
Dnydryacetore
phoghee (DHAP
Enerr
eenserving
stage
2NAD
NADH
4. What stage involves the consumption of
two ATP molecules?
2000-0 synopen
5. What stage involves the production of four
ATP molecules?
Phoy
PEP
ZADP
Pyuv ae
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Case Study: You are asked to inoculate lactose media (broth) with E. coli that was growing exponentially in glucose media (broth). You
monitor the growth of these cells in the lactose broth using a viable plate count technique and plot the Growth Curve for E. coli growing c
lactose. After a day of monitoring the cells, you note that the culture entered into lag phase growth initially and is now currently growing
exponentially. In addition, you find that the cultures doubling time is 30 minutes. You know that the doubling time of E. coli in glucose
media is 20 minutes. You did not change the oxygen content or temperature at Which you were growing the cells, E. coli is still aerobically
respiring at 37°C.
Question: If the lac operon is 'on' what does this imply about the cell?
O The cell is dying and the lac operon is going to induce apoptosis. Apoptosis is truly cell death and the cells will enter into the death phase of the Growth
Curve.
O Lactose is being catabolized and glucose is no…
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Please answer and explain
TOPIC: STAINING FECAL SMEARS
1.) What are the advantages of staining fecal smears with permanent stains (namely: Iron hematoxylin stain, Wheatley's trichrome stain, and modified acid-fast stain)? Enumerate more than two and explain.
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Case Study: You are asked to inoculate lactose media (broth) with E. coli that was growing exponentially in glucose media (broth). You
monitor the growth of these cells in the lactose broth using a viable plate count technique and plot the Growth Curve for E. coli growing on
lactose. After a day of monitoring the cells, you note that the culture entered into lag phase growth initially and is now currently growing
exponentially. In addition, you find that the cultures doubling time is 30 minutes. You know that the doubling time of E. coli in glucose
media is 20 minutes. You did not change the oxygen content or temperature at which you were growing the cells, E. coli is still aerobically
respiring at 37°C.
Question: If you want to grow E. coli as quick as possible would you grow it on lactose or glucose?
O Mixing the two carbon sources would provide the most optimal growth conditions.
O Lactose because of the generation time being faster.
O Lactose and an anaerobic environment.
O Glucose…
arrow_forward
Case Study: You are asked to inoculate lactose media (broth) with E. coli that was growing exponentially in glucose media (broth). You
monitor the growth of these cells in the lactose broth using a viable plate count technique and plot the Growth Curve for E. coli growing on
lactose. After a day of monitoring the cells, you note that the culture entered into lag phase growth initially and is now currently growing
exponentially. In addition, you find that the cultures doubling time is 30 minutes. You know that the doubling time of E. coli in glucose
media is 20 minutes. You did not change the oxygen content or temperature at which you were growing the cells, E. coli is still aerobically
respiring at 37°C.
Question: In order to determine the doubling time of E. coli a viable plate countris performed. This means that you are truly counting
O Dead cells only via a microscopic count
O Live cells only via membrane filtration or serial dilution
O All cells via membrane filtration or serial…
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Identify whether the statements are TRUE or FALSE
1. You should use P-20 micropipettes to transfer volumes for spread plating.
2. You should use P-200 micropipettes to transfer volumes for pour plating.
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Esc
F1
F2
F3
F4
F5
F6
F7
F8
F9
F10
F11
40
%23
5. What is the physical appearance (solid, liquid, semi-solid) of the following media: broth tube, agar slant
tube, agar deep tube, and agar plate. What are you testing when you introduce an inoculum with a needle on
an agar deep tube?
at
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Procedure:
1. Prepare 5 test tubes. Place 1 ml of 1% starch and add 10 drops of saliva to
each tube. Mix thoroughly.
2. Place the first tube in ice water, the 2nd
tube leave at room temperature, the 3rd
tube in 40°C , the 4th
tube at 60°Cwater bath and the 5th
tube boil for 2
minutes..
3. Leave the 4 tubes in their respective temperatures for 30 minutes. The 4th
tube
allows to stand for 30 minutes after heating for 2 minutes.
4. Test the contents of each tube with iodine and benedict’s tests.
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MATCHING TYPE. Match the phlebotomy materials found in Column A to the Collection Method found in Column B.
Column A: Materials 1. Cotton 2. Plain glass tube3. Red top glass tube4. Gauge 23 syringe5. Two-way needle6. Adapter7. Lancet8. Lavender top plastic tube9. Tourniquet10. Micropore
Column B: Collection MethodA. Syringe MethodB. Evacuated SystemC. Winged CollectionD. Capillary CollectionE. All of the aboveF. Syringe and Evacuated onlyG. Evacuated and Winged
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No SIM ?
10:33 PM
60%
السؤال
Staphalococcus is differed from
Stretpococcus by:
Catalse test
O Coagulase test
O Gram negative
O Phosphatease
حفظ الإجابة
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Wil completed a full plate titer assay and there were 25 plaques on his 10^-7 plate. What is the titer of his high titer lysate? 10uL of each dilution were used for the plaque assays.
O 2.5X10^11 pfu/mL
O 2.5X10^7pfu/mL
O 2.5X10^9 pfu/mL
O 2.5X10^8 pfu/mL
O 2.5X10^10 pfu/mL
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Area of no growth around an antibiotic disk indicates_________________.
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Plasma and clot
e)
Serum and clot
14. If small clots are noted when making a blood smear, one should:
a)
Make the smear and note on the slide that clots are present
Mix the blood specimen for one hour then make the smear
Do nothing; small clots will not affect a blood smear
Request a new specimen before doing the blood smear
Add more anticoagulant to the blood tube, mix, then make the blood smear
15. What is the purpose of doing a differential?
a)
b)
To determine the proportion of RBCS in the whole blood
To count the number of WBC's in whole blood
To determine the proportion of WBC's in whole blood
To microscopically examine RBCS and platelets
To diagnose anaemia
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Related Questions
- ISOLATE #1 Results for "Isolate #1" ***Please fill out the table**** 1. Blood agar: 2. Nitrate Test: Test Results Interpretation Blood agar (type of hemolysis) Nitrate test 3. Gelatinase Activity: Gelatinase activity Resistant or Susceptible? Novobiocin Resistance Zone Diameter: 4. Novobiocin Resistance: 5. Coagulase Test: Coagulase (Sure-vue test) 1. Based on your results, what is the possible identification (Genus and species) of “Isolate #1"? I 9 10 11 12 3 14arrow_forwardISOLATE #2 Results for "Isolate #2": ***Please fill out the table**** 1. Blood agar: 2. Nitrate Test: Test Results Interpretation Blood agar (type of hemolysis) Nitrate test 3. Gelatinase Activity: Gelatinase activity Resistant or Susceptible? Novobiocin Resistance Zone Diameter: 4. Novobiocin Resistance: 5. Coagulase Test: Coagulase (Sure-vue test) 2. Based on your results, what is the possible identification (Genus and species) of "Isolate #2"? nhesarrow_forwardGeneral Instructions:Choose 1 bacteria/diseases and fill out the details. An example of an answered template is also provided for your reference. Causative Agent and Disease Profile for S. aureus Template and Example ITEM MSM PROFILE MICROBIAL PROFILE I MICROORGANISM/CAUSATIVE AGENT Staphylococcus aureus A GRAM REACTION (+) B OXYGEN REQUIREMENT Facultative Aerobes C SIZE 1.5 µm D SHAPE Cocci in clusters E HABITAT Normal flora of skin/anterior nares/pharynx F DISCOVERY G MICROSCOPIC IMAGE II DISEASE PROFILE Scalded skin syndrome A DISEASE/S Skin and Wound Infections Scalded Skin Syndrome Toxic Shock Syndrome Food Poisoning Pneumonia B SYMPTOMS OF THE DISEASE A high fever · Nausea and vomiting · A rash on your palms and soles that resembles a sunburn C INCUBATION PERIOD 2 and 4 hours (range 30 minutes to 8 hours) D MODE OF…arrow_forward
- BONUS (15 points) The fallowing series of dilution was prepared from a specimen to determine the number of bacteria. There were 62 colonies on the agar plate prepared by transferring 0.3 ml from the tube number 4. Calculate the dilution factors for each tube. What is the cell concentration in the original specimen? Calculate the total number of cells in tube number 2. étv A) F12 F9 F10 F7 % & %3D delete 5 { T Y J Karrow_forwardLABORATORY REPORT: Aseptic Technique & Inoculation of Bacteria Guide Questions. 1. Why is direct flaming preferred when disinfecting loops and needles? 2. Why is it important to flame the entirety of the loop and not just the tip? What consequences can be seen when this process is not done correctly? 3. What is the difference between quadrant streak method A from method B? 4. Why do we use a pencil instead of a pen when labelling culture tubes or plates? Are there other alternatives in labelling? 5. Why is an inoculating needle used in Subculturing Techniques? Can a loop be used instead? NOTE: Please try to answer all of the question asked, i promised to give you a good ratingsarrow_forwardCase #2 Clinical History: This 29-year-old male's illness began 10 weeks prior to death, with an episode of "flu". Two weeks later his urine became "smoky". He was found to have hematuria, albuminuria and elevated BUN (180 mg/dl). He died from a pulmonary embolus. The throat culture obtained exhibited gram positive cocci in chains. It also showed beta- hemolysis on sheep blood agar (SBA) and was catalase negative. 1. What was the most likely organism? A. Streptococcus pyogenes B. Streptococcus viridans C. Staphylococcus aureus D. Streptococcus pneumonia 2. What is the BEST diagnosis at the time of death? 3. What are the other possible sequela of this infection?arrow_forward
- Analysis of the pathological constituents GIVE THE FOLLOWING: - Name of Test/Test Reagent - Positive Result 1) Protein (Note: Presence of protein in urine is termed as proteinuria or albuminuria) A. Heat and Acetic acid test Fill a test about ¾ full of urine. Heat the upper portion gently to boiling for 1 -2 minutes being careful not to shake the tube. Rotate the tube to prevent overheating. A turbidity may be due to albumin, phosphates or carbonates. Add 3 drops of acetic acid drop by drop while boiling between each drop. If turbidity disappears, it is due to carbonates and phosphates. B. Heller’s nitric acid test Place 1 ml of urine sample in a test tube. Hold the test tube in an inclined position and carefully pour 1 drops of conc HNO3 down the side of the test tube. Note formation of white ring at the zone of contact of the two solutions which is an indication of the presence of protein.arrow_forwardAnalysis of the pathological constituents GIVE THE FOLLOWING: a. Name of Test/Test Reagent b. Positive Result Blood (Note: Presence of blood in urine is termed as hemolobinuria) To 1 ml of freshly prepared Benzidine solution (sat’d sol’n of Benzidine in glacial Acetic acid) and 10 drops of urine sample. Add 5 drops of 3% H2O2. A positive result is indicated by a blue green color.arrow_forwardA 1.5-year-old child developed vomiting, diarrhea, and fever. Stool sample were inoculated into the Endo media. After 18 hours on the surface of the medium grew medium-sized, round, slightly convex red colonies with a metallic luster. The doctor suspected Escherichia coli. 1. Name the composition of the Endo agar media. 2. Describe the properties of bacterial colonies on the Endo media. 3. What purpose differential diagnostic Endo media used for?arrow_forward
- amylasw test test for? medium+ - reagents + Test/why?arrow_forwardHelping tags: Biology, bacterial count, dilution, serial dilution WILL UPVOTE, just pls help me answer the following questions and explain them clearly. Pls show complete solutions also for the computation part. Thank you. 1. A bacterial culture was grown for 9 hours. At 3-hour interval, the culture was sampled to determine the population of the culture, by transferring 25 ml of the suspension to 225 ml 0.85% NaCl. Three consecutive dilutions were further made by using 1 ml aliquot in 9 ml of 0.85% NaCI. One ml from each dilution was plated in each of duplicate plates. The following table shows the results of the plating method. Sampling COUNTS 2nd dilution 3rd dilution 4th dilution 0,0 30: 35 250;245 1st dilution 1st 2nd 3rd 0; 0 240; 235 TNTC 55; 60 5; 6 TNTC TNCT TNTC TNTC a) Illustrate the dilution series used and label the final dilution of each dilution. b) Determine the bacterial count (CFU/ml) every 3 hours of incubation for 9 hours. Show all computations.arrow_forwardA. A Aa A 三,三,行,E AaBbCcDdE Aa AaBbCcDdF AaBbCcDdF AaBbCcDd A Normal No Spacing Heading 1 Heading 2 =y updates, fixes, and improvements, choose Check for Updates. 1. You have found a putative virus which is able to infect a bacterium causing increased mortality and resistance to all antibacterial agents. You have systematically purified the sample but have an unknown concentration of viruses. You perform the following serial dilutions. 0.5ml 1ml 5ml 0.1ml 2.5ml 1ml 15ml 1 3 6 7 Phage Buffer 100.5ml 9ml 14.5ml 99.9ml 25ml 5ml 985ml Tube Dilution in each Test Tube (Show Tube Dilution in each Test Tube (Show your your work) # # work) 4 1 6. O Focus glish (United States) W 30 DII DD 000 000 F4 80 F7 F8 F9 F10 F3 F5 F6 $4 & 4 5 6 8.arrow_forward
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