12+-+Micro+Lab+-+Antimicrobial+Drugs_Fillable
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Troy University, Phenix City *
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MICROBIOLO
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Biology
Date
Apr 3, 2024
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Antimicrobial Drugs and Sensitivity LAB#12 Learning Objectives: Student should explain the term minimal inhibitory concentration (MIC) Students will perform and antibiotic sensitivity test for a gram positive and a gram negative bacterium. Students will provide the rationale for the agar diffusion technique. Materials Required: Growing Broth Culture (2) Nutrient Agar Plates (2) Sterile Swab Bunsen Burner Ethanol Forceps Antibiotic Discs Test Tube Holder Ruler or Measurement tool for MM (second day)
Instructions for this assignment: Students will be provided with 4 different antibiotics and two petri plates. Students will test the effectiveness of 4 different antibiotics using a gram positive and a gram negative bacteria. Record the names of the antibiotics in the table in the lab report by name and disc code.
Antibiotic Sensitivity Protocol A.
Please be certain to label plates prior to spreading bacteria. B.
Mix the tube of bacteria well while the top is secure. C.
Immediately after mixing remove a sterile swab from package and place the swab into mixed bacterial sample. D.
Roll the swab gently on the agar plate, making every effort to contact every area of the agar plate. Turn the plate and swab the plate again coming in contact with every are of the plate again. Repeat this until you have turned the plate in a complete circle. Repeat this at least 2 times. E.
Place the plate near the flame from the Bunsen burner and leave the lid slightly open so that the plate will dry. F.
On the writing surface of the plate (bottom) divide the plate into 4 quadrants labeling them 1 through 4. G.
Repeat steps A through F for the second agar plate. H.
Before applying the antibiotic discs to each plate label the table in this lab with the 4 selected antibiotics. Make sure that the antibiotics are the same for each plate. I.
Alcohol flame the forceps making sure that your hand is above the forceps and the alcohol is dripping away from your hand. J.
Remove the antibiotic disc for quadrant 1 and place the disc in the center of quadrant. Then tap the disc slightly to ensure good contact with the media. K.
Repeat this step for quadrant 2 – 4. Please be certain you are placing the proper antibiotic in the proper quadrant. L.
Repeat steps F through K for the second agar plate. M.
Once you are finished with both agar plates place the plates in the incubator media side up.
Lab Report:
Antimicrobial Drugs and Sensitivity
Purpose: ___________________________________________________________________ ___________________________________________________________________ ___________________________________________________________________ Day 2 A.
Remove inoculated plates from the incubator.
B.
Measure in millimeters (mm) and record the diameter of the zone of inhibition (area of
clearing around the disc) in the table below and evaluate each using the key to
determine if your results are Susceptible (S), Intermediate (I) or Resistant (R).
DATA TABLE ANTIBIOTIC NAME DISC CODE Staphylococcus epidermitis Escherichia coli Zone Size S, I or R Zone Size S, I or R 1. 2. 3. 4. Conclusion: State a conclusion of your results for each bacteria and for each antibiotic treatment. __________________________________________________________
__________________________________________________________
__________________________________________________________
__________________________________________________________
__________________________________________________________
__________________________________________________________ Streptomycin S10
Tetracycline
Te30
Penicillin
Chloramphenicol
13mm
23mm
P10
13mm
C30
24mm
34mm
19mm
25mm
14mm
I
I
S
S
R
R
S
S
To determine which antibiotics are susceptible, intermediate, or resistant to certain bacterias. Streptomycin is Intermediate to Staph Epi and E.Coli. Tetracycline is susceptible to both S.Epi and E.Coli. Penicillin is resistant to both S.Epi and E.Coli. Chloramphenicol is susceptible to both S.epi and E.Coli.
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Related Questions
Give the uses/functions and images of each apparatuses.
Basic Laboratory Equipment
Uses/Functions
Picture
1. Microscope
2. Colony Counter
3. Autoclave
4. Microbiology incubator
5. Drying oven
6. Refrigerator (microbiology)
7. Bunsen burner/alcohol lamp
8. Candle jar
9. Anaerobic jar
10. Microhood or Bacteriologic hood/Safety hood/Safety cabinet
11. Bacteriologic filters (Seitz, Chamberlain, Berkfield)
12. Petri dish
13. Culture tubes
14. Hanging drop slide
15. Durham’s tube
16. Staining rack
17. Thermostatically controlled water bath
18. Inoculating loop
19. Inoculating needle
20. Vials
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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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Questions:
1. What happens to the number of bacteria as you streak
from one sector to another?
2. What happens to the colonies when the bacteria are
separated well?
3. How bacteria does each colony come from?
many
4. Why do you make sure that the inoculating loop is red
hot?
5. What happens when the streaking is not correct?
6. When do
you
flame the mouth of the cultures tubes?
7. Why you should not use the inoculating loop when it
is red hot?
arrow_forward
Question:-
Describe control strains used in the clinical microbiology laboratory and explain their maintenance in the laboratory. ( write BY WORD and all steps I need).
Introduction
Discussion
References
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Give the uses/functions and images of each apparatuses.
Basic Laboratory Equipment
Uses/Functions
Picture
1. Microscope
2. Colony Counter
3. Autoclave
4. Microbiology incubator
5. Drying oven
6. Refrigerator (microbiology)
7. Bunsen burner/alcohol lamp
8. Candle jar
9. Anaerobic jar
10. Microhood or Bacteriologic hood/Safety hood/Safety cabinet
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Activity 2. Media Used in Isolating Coliforms
You are a group of microbiologists tasked to test the presence of coliforms in the newly built water station. You are
asked to isolate E. coli, Salmonella, and Shigella. Enumerate the media used for each step of isolating these
bacteria. Include pictures and references.
Bacteria
Enrichment
(Broth/Agar)
Presumptive test
(Broth/Agar)
Isolation Media
(Broth/Agar)
E. coli
Salmonella
Shigella
1. How can you confirm that E. coli, Salmonella, and Shigella are present in the newly built water
station? Explain.
2. What is the difference between nutrient broth and nutrient agar?
3. What is the importance of these steps (Enrichment, Isolation, and presumptive test) in isolating bacteria?
Conclusion about the process of isolating bacteria
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Subject : Environmental Microbiology
Can u use the information given below to answer these 2 question
1.Provide an aim for this lab
2. Provide objectives
DISCUSSION QUESTIONS
What is the relationship between the resolution power and the useful magnification that may be obtained with the light microscope?
What determines the resolving power of the lens system?
What is the limit of resolution obtainable with the light microscope?
How you will distinguish between bright field and dark-field microscopy and provide a specific example where each would be method of choice for observing a culture of bacteria?
What advantages does electron microscopy have over light microscopy?
What are disadvantages of electron microscopy over light microscopy?
#Compare the use and the methodology of TEM with SEM? Provide at least one example where each would be the method of choic
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Cause and Effect Analysis describe the effect on the test procedure and explain briefly
The zones of adjacent antibiotic disks overlap.
Antibiotic disk cartridge was left open for more than 7 days.
Antibiotic disk was placed closely at the edge of the plate.
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INSTRUCTION: Give the principle or significance of the following practices:
1. Flaming the mouth or lid of culture tubes or plates after opening and before closing them.
2. Flaming the inoculating needle or loop before and after inoculation.
3. Holding caps, lids, or cotton plugs rather than putting them on the table while inoculating.
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Instruction: answers must be in numbers.
You are culturing bacteria using a petri dish, the bacteria grow very well on this plate you have. However, your microbiology instructor wants to know how many bacteria per milliliter (bacteria/mL) are on your plate. And as you have remembered during the first day of inoculating this bacteria you used a imL aliquots sample of a 1:10,000 dilution, and the total colony you counted on this plate is 170. How many bacterial per milliliters would that be?
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I need help with microbiology/2010
1. Describe where the following items should be discarded:
a) gloves
b) petri dishes
c) test tubes
d) microscope slides
2. Describe the safety procedures for the following hypothetical situations:
a. You spilled a full test tube of bacterial culture on the bench top.
b. You notice flames coming from the tubing of your Bunsen burner
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Guide Questions:
Differentiate the types of culture media based on its physical state and uses.
What makes agar good support for microbial growth?
Define and contrast the terms “sterile” and “clean”.
List 5 other methods of sterilization and describe the principle/s and when each method is used.
What are the requirements to support microbial growth?
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INSTRUCTION: Give the principle or significance of the following practices:
NOTE: *please add references*
1. Flaming the mouth or lid of culture tubes or plates after opening and before closing them.
2. Flaming the inoculating needle or loop before and after inoculation.
3. Holding caps, lids, or cotton plugs rather than putting them on the table while inoculating.
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Instructions:
Give at least five different microbial control methods you have on your home as listed on the pre-task activity. Identify its microbial control method (Physical or chemical), Specific component (heat, phenol, etc), Mode of action (denatures protein, disrupt cell membrane, etc), Use (sterilize, disinfect, antiseptic), Level of activity (Sterilant, high, intermediate, low disinfectant)
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Direction: Read and analyze the following laboratory experiment and answer the
following question.
PART 1: SURFACE AREA AND CELL SIZE
Materials: Agar containing NaOH, and the pH-indicator dye phenolphthalein
cured into cubes of various size, 3 plastic cups, HCl, metric ruler, paper towels.
Methodology:
1. Safety: Wear goggles and nitrile gloves while completing this lab.
2. Obtain three different size blocks of pink or blue agar. Using a ruler,
measure the length, width, and height of the three blocks given below. Cut
the agar according to the given dimension.
Small = 1 cm x 1 cm x 1 cm
Medium = 2 cm x 2 cm x 2 cm
•
• Large = 1 cm x 1 cm x 6 cm
3. Record your data.
4.
Pour HCl or vinegar into two small cups. Place the one larger "cell" into one
cup and the two smaller cells in the other cup. Start timing 30 minutes.
5. After 30 minutes, remove the cells and blot them dry with a paper towel.
6. Using your ruler, measure the distance the HCl has diffused into the blocks
as shown on the…
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Serial dilution
QUESTION 2
What type of agar is TSA? (select all that apply)
Nutrient
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Procedures/ Spccific test sample
used
Tests
Indication/Purpose
Draize Acute Skin Irritation Test
Draize Eye Irritation Test
Coagulation and Liquefaction of Proteins
Chorioallontoic Membrane (CAM)
Assay
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LAB QUESTIONS – ASSESSMENT AND CRITICAL
THINKING
1. When is a simple negative stain used?
2. Why do microorganisms remain unstained in the simple
negative staining procedure?
3. What information can be obtained from a simple positive
staining procedure?
4. What is the difference between a simple and differential
stain?
5. What is the reagent and purpose for each of the following
gram stain steps?
a. Primary stain
b. Mordant
c. Decolorizer
d. Counterstain
6. Which step is the most crucial or most likely to cause poor
results in the Gram stain? Why?
7. What part of the bacterial cell is most involved with Gram
staining, and why?
8. What cell wall component is responsible for the acid-fast
property of mycobacteria?
9. Is a gram stain an adequate substitute for an acid-fast stain?
Why or why not?
10. Which area on a streak plate will contain the greatest
amount of growth? The least amount of growth? Explain
your answers.
11. What is a colony (as viewed on an agar plate)? How can a
pure…
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TOPIC: KATO-THICK & KATO-KATZ TECHNIQUES
What are the differences between Kato-Thick smear and Kato-Katz technique? Briefly explain each.
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6
2
4
3
5
LABORATORY REPORT FORM
EXERCISE 25
ASSAY OF ANTIMICROBIAL AGENTS: DISK-DIFFUSION METHODS
What is the purpose of this exercise?
Evaluate
1.
For each of the disinfectants or antiseptics tested, determine the active ingredient. What class of inhibitory
agent does it belong to and what is its mode of action?
CHEMICAL AGENT
Lysol
Sanitizer
Foaming
Handwash
Detergent.
Desinfecting
Hand
Sanitizer
ACTIVE INGREDIENT(S)
Alhyl
Dinethylaenzy/a.
chloroxylenol 0.3%
Alkyl, Dimethyl benzyl
ammonium chlorides
ethy benzyl A. C.
Ethyl Alcohol 70%
1/Cleansing
s Pray
Hydrogen
Peroxide
Benzalkonium
Lidocaine HC)
An Septic
CLASS OF
AGENT
MODE OF ACTION
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Cause-and-Effect Analysis: Given the conditions below, describe the effect on the staining procedure and explain briefly:
1. Excessive heat was applied during fixation
2. Low concentration of crystal violet used during gram staining
3. Excessive washing between steps
4. Insufficient decolorization
5. Excessive counterstaining
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Question:-
How to perform a bactericidal test bactericidal test of Dental Pulp Stem Cell (DPSC)?
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Procedure:
1. Prepare 6 test tubes and place 1 ml saliva in each.
2. Add Iml water to tube 1,2 ml to tube 2, 4 ml to tube 3,6 ml to tube 4, 8 ml to
tube 5 and 10 ml to tube 6. Mix thoroughly.
3. Transfer 1 ml of each into 6 separate test tubes (discard excess solution) and
add 1 ml of 1% starch paste. Mix well and heat in a water bath with
temperature maintained at 40°C for 30 minutes.
4. Divide each of the contents of each tube into 2 and test with:
a. Iodine Test – to half of the content of each tube, add 1 drop of iodine in KI
and note the color produced. Compare the intensity of color in each of the
6 tubes.
b. Benedict's – to 1 ml benedict's reagent, add 5 drops of the other half of
each tube and heat in a boiling water bath for 3 minutes. Note color of
precipitate.
5. Rank each tube according to decreasing reaction rate one (1) being the fastest
to hydrolyze and 6 being the slowest.
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Give the uses/functions and images of each apparatuses.
Basic Laboratory Equipment
Uses/Functions
Picture
6. Refrigerator (microbiology)
7. Bunsen burner/alcohol lamp
8. Candle jar
9. Anaerobic jar
10. Microhood or Bacteriologic hood/Safety hood/Safety cabinet
11. Bacteriologic filters (Seitz, Chamberlain, Berkfield)
12. Petri dish
13. Culture tubes
14. Hanging drop slide
15. Durham’s tube
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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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Please match the test with its description to assess your understanding of the kinds of data collected during information gathering and
your understanding of the processes involved in identifying microbes from samples.
1. using macroscopic and microscopic traits for identification appearance
2. tests that determine chemical characteristics including enzyme production and nutritional requirements of the microbe
biochemical tests
3. analyzing the genotype of an organism DNA profiles
4. tests the organism against known antibodies to determine if there is a reaction between the organism and the antibody
immunologic testing v
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困
Topic: laboratory Instrument
Which of the following are TRUE regarding
Which of the following is/are TRUE
the use of laboratory instruments and
regarding the use laboratory instruments
equipment in regards to making of culture
media? "
and equipment? *
Air bubbles on the surface of agar plates
Hot plates can be used to sterilize the
inoculating loop and needle.
can be removed by fanning them with the
luminous flame of the bunsen burner.
If the agar surface is wet, it can be dried by
A drying oven is used to sterilize nutrient
agar, a solid culture medium.
heating at 30-40 deg C in the drying oven
before inoculation is done.
The refrigerator is used to grow
psychrophiles.
All media taken from the refrigerator should
be warmed to room temperature before
use,
P Type here to search
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Please help me answer correctly the table this is my assignment in biochem lab
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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Topic: Preparation of Microbial Suspension
5. Over-dilution with sterile water.
Effect:
Rationale:
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True or False
1. Two challenges regarding IFU are adhering with the IFU's and interpreting complex IFU.
2. Mechanical disinfection is typically more effective than manual techniques.
3. Adding a UV disinfector to a pass-through window is NOT offer a higher level a safety for sterile processing personnel.
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Topic: Pour-Plate Method
2. Molten agar mixed with bacterial suspension has a temperature higher than 45˚CEffect:
Rationale:
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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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- Give the uses/functions and images of each apparatuses. Basic Laboratory Equipment Uses/Functions Picture 1. Microscope 2. Colony Counter 3. Autoclave 4. Microbiology incubator 5. Drying oven 6. Refrigerator (microbiology) 7. Bunsen burner/alcohol lamp 8. Candle jar 9. Anaerobic jar 10. Microhood or Bacteriologic hood/Safety hood/Safety cabinet 11. Bacteriologic filters (Seitz, Chamberlain, Berkfield) 12. Petri dish 13. Culture tubes 14. Hanging drop slide 15. Durham’s tube 16. Staining rack 17. Thermostatically controlled water bath 18. Inoculating loop 19. Inoculating needle 20. Vialsarrow_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_forwardQuestions: 1. What happens to the number of bacteria as you streak from one sector to another? 2. What happens to the colonies when the bacteria are separated well? 3. How bacteria does each colony come from? many 4. Why do you make sure that the inoculating loop is red hot? 5. What happens when the streaking is not correct? 6. When do you flame the mouth of the cultures tubes? 7. Why you should not use the inoculating loop when it is red hot?arrow_forward
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