Brock Biology of Microorganisms (15th Edition)
Brock Biology of Microorganisms (15th Edition)
15th Edition
ISBN: 9780134261928
Author: Michael T. Madigan, Kelly S. Bender, Daniel H. Buckley, W. Matthew Sattley, David A. Stahl
Publisher: PEARSON
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Chapter 19, Problem 1AQ

Design an experiment for measuring the activity of sulfuroxidizing bacteria in soil. If only certain species of the sulfur oxidizers present were metabolically active, how could you tell this? How would you prove that your activity measurement was due to biological activity?

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Summary Introduction

To discuss:

Design an experiment to measure the activity of sulfur-oxidizing bacteria in the soil. How to detect, if only certain sulfur oxidizing species present were metabolically active. How to prove the activity measurement was because of biological activity.

Introduction:

Sulfur oxidizing bacteria (for example, Thiobacillus, Thermothrix,Sulfolobus, and Thiothrix) can convert hydrogen sulfide (H2S) into sulfate (SO42) or other forms of sulfur. They can grow in various environments, for example, soil and water. MAR-FISH is a fluorescence in situ hybridization (FISH) combined with microautoradiography (MAR). MAR-FISH can simultaneously identify microorganisms and their metabolic property.

Explanation of Solution

An experiment to measure sulfur-oxidizing bacteria from soil:

  • Isolate Thiobacillus species (gram-negative bacteria) from sulfide rich soil (for example, paddy field) followed by serial dilution in water.
  • Add 5 ml water containing sample (bacteria) to 50 ml thiosulfate mineral salts medium (thiosulfate MSM).
  • Incubate the medium at 30°C for 7 days. The formation of turbidity and reduced pH of the thiosulfate MSM medium indicates the bacterial growth.
  • Further streak plate method (onto the solid medium of thiosulfate MSM) can be used to isolate the pure culture of Thiobacilli.
  • Incubate the petri plate at 30°C for 7 days.
  • Screen the purified culture of Thiobacilli in the MSM consisting of Na2S but not of Na2S2O3.
  • The sulfide medium used for the growth of bacteria is the sole source of energy.
  • Further, Thiobacillus can be confirmed by gram-staining.
  • Sulfur-oxidizing bacteria, such as Thiobacillus species can be used to study.

To measure the activity of sulfur oxidizing bacteria in the soil, the radioisotope H235S should be used. The soil should be incubated with radioisotope (H235S) to know whether the sulfide oxidation occurred or not. After incubation, the presence of 35SO42- should be analyzed to prove the activity of sulfur-oxidizing bacteria in the soil. The presence of 35SO42- proves that the sulfide oxidation has occurred in the test. To prove that the oxidation occurred due to biological activity, sterile the soil (medium) before the addition of labeled H2S. This is used as a control in this experiment. If the oxidation activity observed in the experiment is biological, it should be absent in the control. The MAR-FISH can be used to identify the presence of metabolically active bacteria in the soil.

MAR-FISH (microautoradiography- fluorescence in situ hybridization) technique is used to evaluate the microbial identification (phylogeny) with measurement of metabolic activities. MAR-FISH is used to assess the metabolism of a specific radiolabeled substance by microorganisms in the natural sample and also help to identify those particular microorganisms. Therefore, MAR-FISH can be used to identify the metabolism of radioisotope H235S by microorganisms in this experiment.

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Chapter 19 Solutions

Brock Biology of Microorganisms (15th Edition)

Ch. 19.4 - How does viability staining differ from stains...Ch. 19.4 - What types of environments limit the application...Ch. 19.4 - Why is it incorrect to say that the GFP is a...Ch. 19.4 - Prob. 1CRCh. 19.5 - What structure in the cell is the target for...Ch. 19.5 - FISH and CARD-FISH can be used to reveal different...Ch. 19.5 - Why is CARD-FISH more suitable than FISH for...Ch. 19.6 - What could you conclude from PCR/DGGE analysis of...Ch. 19.6 - What surprising finding has come out of many...Ch. 19.6 - How has next-generation sequencing technology...Ch. 19.6 - QWhich method, ARISA or T-RFLP, would provide more...Ch. 19.7 - Prob. 1MQCh. 19.7 - What are the advantages and disadvantages of...Ch. 19.7 - Why might a microarray be superior to using...Ch. 19.8 - Prob. 1MQCh. 19.8 - How do environmental genomic approaches differ...Ch. 19.8 - Prob. 3MQCh. 19.8 - Prob. 1CRCh. 19.9 - Prob. 1MQCh. 19.9 - If a large pulse of organic matter entered the...Ch. 19.9 - Q What are the major advantages of radioisotopic...Ch. 19.10 - What is the simplest explanation for why lunar...Ch. 19.10 - What is the expected isotopic composition of...Ch. 19.10 - How might exchange of metabolites among members of...Ch. 19.10 - Will autotrophic organisms contain more or less...Ch. 19.11 - How could NanoSIMS be used to identify a...Ch. 19.11 - Prob. 2MQCh. 19.11 - How does MAR-FISH link microbial diversity and...Ch. 19.11 - Q What can MAR-FISH tell you that FISH alone...Ch. 19.12 - How can stable isotope probing reveal the identity...Ch. 19.12 - What key method is required to do genomics on a...Ch. 19.12 - Prob. 3MQCh. 19.12 - How would you use cytometric cell sorting to...Ch. 19 - Design an experiment for measuring the activity of...Ch. 19 - You wish to know whether Archaea exist in a lake...Ch. 19 - Design an experiment to solve the following...Ch. 19 - Design a SIP experiment that would allow you to...
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