Environmental Mutagenesis Lab Manual - MIC-637
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Questions and Answers

What is the primary objective of LAB NO: 1?

  • To study soil health
  • Degradation of octane
  • Isolation of soil-degrading bacteria (correct)
  • Analysis of UV radiation effects
  • Soil-degrading bacteria are known to worsen soil contamination.

    False

    What type of hydrocarbons are studied in LAB NO: 3?

    Octane

    LAB NO: 4 deals with the degradation of ____ by soil bacteria using spray plate technique.

    <p>diesel</p> Signup and view all the answers

    What were the characteristics of the colonies observed in the negative control?

    <p>Well-defined, smooth-edged colonies of uniform size and pigmentation</p> Signup and view all the answers

    Match the labs with their main focus:

    <p>LAB NO: 1 = Isolation of soil-degrading bacteria LAB NO: 3 = Degradation of octane LAB NO: 4 = Degradation of diesel LAB NO: 5 = UV radiation induces mutagenesis in bacteria</p> Signup and view all the answers

    Which process do soil-degrading bacteria contribute to?

    <p>Nutrient cycling</p> Signup and view all the answers

    The test plate showed significantly larger and well-formed colonies.

    <p>False</p> Signup and view all the answers

    What type of cells were primarily observed in the positive control after 10 minutes of UV exposure?

    <p>Mostly intact pink rods</p> Signup and view all the answers

    Which of the following species is oxidase-positive?

    <p><em>Pseudomonas</em></p> Signup and view all the answers

    UV radiation has no effect on bacteria.

    <p>False</p> Signup and view all the answers

    The methodology used in the experiment involved observing bacteria exposed to ___ radiation.

    <p>UV</p> Signup and view all the answers

    Polycyclic aromatic hydrocarbons (PAHs) are not considered mutagenic or carcinogenic.

    <p>False</p> Signup and view all the answers

    What environmental problem is associated with greasy soil?

    <p>Hydrocarbon contamination</p> Signup and view all the answers

    Match the observation condition with its outcomes:

    <p>Negative Control = Well-defined colonies with abundant growth Positive Control (10 min) = Some clumping and fragmented cells Test Plate (20 min) = Pronounced abnormalities and minimal growth Overall Implication = Induces lethal mutations in bacteria</p> Signup and view all the answers

    What ecological process does the degradation of benzene by microorganisms contribute to?

    <p>Environmental health</p> Signup and view all the answers

    Soil-degrading bacteria can transform pollutants into more or less ____ forms.

    <p>mutagenic</p> Signup and view all the answers

    The cotton-soaked method is used to study the biodegradation of __________ by soil bacteria.

    <p>benzene</p> Signup and view all the answers

    Match the following controls with their descriptions:

    <p>Positive control = A minimal media plate + glucose Negative control = A minimal media plate Test plate = A minimal media plate with soaked benzene cotton balls</p> Signup and view all the answers

    What is the purpose of making wells in the plates during the experiment?

    <p>To place soaked benzene cotton balls</p> Signup and view all the answers

    Incubation of the plates occurs at a temperature of 25°C.

    <p>False</p> Signup and view all the answers

    What method is used to isolate individual bacterial colonies in the experiment?

    <p>Serial dilution</p> Signup and view all the answers

    What type of media was used in the experiment to observe microbial growth?

    <p>Minimal media</p> Signup and view all the answers

    Fungal contamination was observed only on one of the plates during the experiment.

    <p>False</p> Signup and view all the answers

    What are the names of the two bacterial genera mentioned that were viable on nutrient agar?

    <p>Bacillus and Pseudomonas</p> Signup and view all the answers

    During the experiment, fungal contamination on the filter paper suggested that the experimental conditions favored _____ growth.

    <p>fungal</p> Signup and view all the answers

    Match the following microbial aspects with their relevance to the experiment:

    <p>Hydrocarbons = Environmental pollutants Aseptic conditions = Prevent contamination Fungal contamination = Interfered with assessment Minimal media = Differentiated microbial growth</p> Signup and view all the answers

    What could be a potential future direction to reduce contamination in experiments?

    <p>Improving aseptic techniques</p> Signup and view all the answers

    The presence of diesel in the environment does not pose any ecological risks.

    <p>False</p> Signup and view all the answers

    What role did the filter paper serve during the experiment?

    <p>It was used to observe fungal contamination.</p> Signup and view all the answers

    What is the primary objective of the study?

    <p>To evaluate the degradation of hydrocarbons by soil microorganisms</p> Signup and view all the answers

    The study shows that Bacillus and Pseudomonas are effective in degrading benzene.

    <p>False</p> Signup and view all the answers

    What are two environmental sources of hydrocarbons mentioned in the content?

    <p>Industrial activities, oil spills</p> Signup and view all the answers

    The type of media used to grow Bacillus and Pseudomonas was called _____ media.

    <p>minimal</p> Signup and view all the answers

    Match each microorganism to its expected result in the study:

    <p>Bacillus = Negative for benzene degradation Pseudomonas = Negative for benzene degradation</p> Signup and view all the answers

    Which of the following represents a potential risk of hydrocarbons in the environment?

    <p>Their potential to bioaccumulate</p> Signup and view all the answers

    The tests conducted showed positive results for both bacterial species at all experimental stages.

    <p>False</p> Signup and view all the answers

    What method was used on Day 1 to prepare a master plate?

    <p>Stabbing on nutrient agar</p> Signup and view all the answers

    What is bioremediation primarily used for?

    <p>To break down hydrocarbons</p> Signup and view all the answers

    The positive control plate had diesel sprayed on it.

    <p>False</p> Signup and view all the answers

    Which two genera of bacteria were tested for diesel degradation?

    <p><em>Bacillus</em> and <em>Pseudomonas</em></p> Signup and view all the answers

    The agar plate marked as the __________ control confirmed bacterial viability on the minimal media.

    <p>positive</p> Signup and view all the answers

    Match the bacterial growth levels with their corresponding observations.

    <p>Positive Control = +++ (growth) Negative Control = ++ (less growth) Test Plate = ++++ (more growth)</p> Signup and view all the answers

    What method was used to evaluate the bacteria's ability to metabolize diesel?

    <p>Spray plate technique</p> Signup and view all the answers

    The study indicated that these bacteria can effectively degrade diesel.

    <p>False</p> Signup and view all the answers

    How long were the plates incubated during the experiment?

    <p>3 days</p> Signup and view all the answers

    Study Notes

    Environmental Mutagenisis Lab Manual - MIC-637

    • Lab course in charge: Ms. IQra
    • Student name: Kainaat
    • Seat no: EH2121042

    Index (Lab Experiments)

    • Lab No. 1 (15-10-2024): Isolation of soil-degrading bacteria
    • Lab No. 2 (22-10-2024): Degradation of benzene from soil microorganisms using the cotton soak method.
    • Lab No. 3 (22-10-2024): Degradation of hydrocarbons (octane) by soil microorganisms.
    • Lab No. 4 (4-12-2024): Degradation of hydrocarbon (diesel) by soil bacteria using spray plate technique.
    • Lab No. 5 (11-12-24): UV radiation induces mutagenesis in bacteria.

    Lab No. 1: Isolation of Soil-Degrading Bacteria

    • Objective: Isolate soil-degrading bacteria
    • Introduction: Soil bacteria break down organic matter, crucial for soil health and nutrient cycling. Some bacteria can transform pollutants into mutagenic compounds.
    • Soil Bacteria Types: Pseudomonas and Bacillus are effective in hydrocarbon degradation.
    • Sample Collection: 1 gram of greasy soil from a petrol pump collected.
    • Methods:
      • Serial dilution of soil sample (diluted to 10-¹ to 10-⁶)
      • Plating: 0.1mL of dilutions spread on nutrient agar plates.
      • Incubation: Plates incubated at 37°C for 24 hours.

    Lab No. 2: Degradation of Benzene by Soil Microorganisms

    • Objective: Observe benzene degradation by soil microorganisms
    • Introduction: Benzene, a common pollutant, is degraded by some soil microorganisms through various processes, often involving breaking down the compound into less harmful substances.

    Lab No. 3: Degradation of Hydrocarbons (Octane)

    • Objective: Observe degradation of octane by soil microorganisms.
    • Introduction: Octane is a hydrocarbon commonly found in petroleum, and soil microorganisms play a vital role in its degradation.
    • Methodology: Use a minimal media to observe growth patterns, as well as replicate of cultures using a sterile replicator.

    Lab No. 4: Degradation of Hydrocarbons (Diesel)

    • Objective: Observe degradation of diesel by soil bacteria using a spray plate technique.
    • Introduction: Diesel is a complex mixture of hydrocarbons and degrading it is an environmental problem.
    • Methodology: Spray diesel onto minimal media plates and observe bacterial growth.

    Lab No. 5: UV Radiation and Mutagenesis in Bacteria

    • Objective: Determine the effects of UV radiation on bacterial mutagenesis.
    • Introduction: UV radiation can cause DNA mutations in bacteria, potentially impacting their survival and growth.
    • Methodology: Expose bacterial cultures (Pseudomonas) to different UV radiation durations (10 minutes, 20 minutes) and observe the results on minimal media plates.

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    Description

    This lab manual focuses on environmental mutagenesis, detailing experiments aimed at isolating soil-degrading bacteria and understanding their role in pollutant degradation. Students will conduct a series of experiments analyzing the effectiveness of various soil microorganisms in breaking down hydrocarbons and the impact of UV radiation on mutagenesis. Each lab is designed to enhance practical skills in microbiology and environmental science.

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