Microbiology Study on Bacterial Genes in Water

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Questions and Answers

What is the purpose of the study?

To search for enterotoxin-producing bacteria in home tap water, RO drinking water, and home storage tanks of Basrah governorate.

What percentage of bacteria in the study were found to be positive for the sea gene (enterotoxin encoding gene)?

46%

What genetic element carries the sea gene, which encodes enterotoxin A?

Temperate phages.

What are the three major stages of biofilm formation?

<p>Attachment, Maturation, and Dispersion.</p> Signup and view all the answers

What does the ica operon encode for?

<p>Intercellular adhesion.</p> Signup and view all the answers

Which genes were tested in the study using Polymerase Chain Reaction (PCR)?

<p><em>sea</em>, <em>icaA</em>, and <em>icaD</em></p> Signup and view all the answers

What is the name of the protein encoded by icaA?

<p>N-acetylglucosaminyl-transferase.</p> Signup and view all the answers

What is the role of icaR?

<p>It is a regulator that controls the transcription of the <em>ica</em> operon.</p> Signup and view all the answers

What is the significance of the ica operon in the context of enterotoxins?

<p>The <em>ica</em> operon helps bacteria adhere to surfaces, enhancing their ability to secrete enterotoxins.</p> Signup and view all the answers

The sea gene was found in all the bacterial species tested in the study.

<p>False (B)</p> Signup and view all the answers

The presence of ica genes in bacteria enhances their ability to cause infections.

<p>True (A)</p> Signup and view all the answers

The presence of ica genes was only detected in bacteria isolated from home storage tanks.

<p>False (B)</p> Signup and view all the answers

The study concluded that RO drinking water is completely free of pathogenic bacteria.

<p>False (B)</p> Signup and view all the answers

What conclusion can be drawn from the findings of the study regarding the presence of ica and sea genes in bacteria?

<p>The presence of both <em>ica</em> and <em>sea</em> genes in bacteria isolated from various water sources suggests that these bacteria have the potential to cause infections and secrete high concentrations of enterotoxins.</p> Signup and view all the answers

How many bacterial species tested were found positive for both sea and ica genes?

<p>4</p> Signup and view all the answers

Why is the presence of enterotoxin-producing bacteria in drinking water a concern?

<p>All of the above (D)</p> Signup and view all the answers

What is the primary mechanism by which ica genes enhance bacterial morbidity?

<p>They enhance bacterial adhesion to surfaces. (D)</p> Signup and view all the answers

Flashcards

Exotoxins

Soluble proteins secreted by bacteria, often consisting of two parts: one for binding to host cells and another for causing harm.

Enterotoxins

A specific type of exotoxin that affects the intestines, causing watery diarrhea by stimulating fluid secretion.

sea gene

A gene encoding for Enterotoxin A, often carried by phages, which can transfer horizontally to other bacteria.

Temperate phages

A family of viruses that infect bacteria, carrying genes that can alter bacterial properties.

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Horizontal Gene Transfer

The transfer of genetic material between bacteria via mechanisms other than direct inheritance, like through phages.

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ica operon

A group of genes involved in the production of polysaccharide intercellular adhesin (PIA), which helps bacteria stick together and form biofilms.

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Biofilm

A complex community of microorganisms attached to a surface and encased in a slimy matrix.

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Polysaccharide intercellular adhesin (PIA)

A substance produced by bacteria that helps them adhere to surfaces and form biofilms.

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Biofilm Formation

The process by which bacteria attach to a surface and begin to form a biofilm.

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Planktonic bacteria

Individual bacteria moving freely in a liquid environment.

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Attachment

The stage of biofilm formation where individual bacteria initially attach to a surface.

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Maturation

The stage of biofilm formation where attached bacteria start to grow and multiply, forming a complex structure.

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Dispersion

The stage of biofilm formation where bacteria detach from the biofilm and disperse into the surrounding environment.

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Flagellar motility

A type of bacterial motility that involves rotating flagella to propel the bacteria through liquid.

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Type IV pili-mediated motility

A type of bacterial motility that involves extensions called pili that help the bacteria move across surfaces.

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IcaA

A protein that contributes to the formation of PIA by transferring N-acetylglucosamine molecules to the growing polysaccharide chain.

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IcaD

A protein that ensures the correct folding and insertion of IcaA into the bacterial membrane.

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IcaB

A bacterial enzyme that removes acetyl groups from mature PIA.

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IcaC

A transmembrane protein that extends the growing polysaccharide chain during PIA synthesis.

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IcaR

A regulatory protein that binds to the icaA promoter region and can suppress the expression of the ica operon.

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Polymerase Chain Reaction (PCR)

The process of amplifying a specific DNA sequence using PCR.

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Agarose gel electrophoresis

The separation of DNA fragments based on their size by applying an electric current to a gel.

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sea gene

The gene encoding for enterotoxin A, which can be used as a marker for the presence of enterotoxin-producing bacteria.

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icaA and icaD genes

The genes encoding for the polysaccharide intercellular adhesin (PIA), which are associated with biofilm formation.

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Frequency of gene detection

The percentage of bacteria in a sample that were found to carry a specific gene.

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Drinking water (RO)

Water that is treated to remove impurities and contaminants.

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Home storage tanks

Water that is stored in tanks or pipes for later use.

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Home tap water

Water that comes directly from your tap.

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Adhesion

The ability of bacteria to stick to surfaces and form biofilms.

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Morbidity

The increased ability of bacteria to cause disease due to factors like toxin production or biofilm formation.

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Study Notes

Title Page

  • Study on frequency of sea and ica genes in bacteria from watery sources of Basrah governorate.
  • Research completed for a Bachelor's degree in Microbiology.
  • Author: Wafiq Yasin Ismael
  • Supervisor: Prof. Dr. Munaff Jawdat Abd-Al Abbas
  • Date: 2023/2024

Abstract

  • Study investigated enterotoxin-producing bacteria in home tap water, treated water (RO), and storage tanks in Basrah Governorate.
  • 46% of bacteria in all water sources were positive for the sea gene (enterotoxin encoding gene).
    • Home tap water: 55.8%
    • RO drinking water: 39.3%
    • Home storage tanks: 40.9%
  • Ica operon (intercellular adhesion) was detected in 14.6% of bacteria.
    • Home tap water: 5.8%
    • RO drinking water: 15.1%
    • Home storage tanks: 27.2%
  • Four samples showed positive results for both sea and ica genes.
    • 32-67A and E. coli from home tap water.
    • 51-Acinetobacter junii from RO water
    • 17-Kluyvera georgiana from home storage tanks.

Introduction

  • Exotoxins are soluble proteins secreted by bacteria, responsible for effects on host cells.
  • Enterotoxins affect intestinal epithelium, causing watery diarrhea and potentially toxic shock syndrome.
  • Enterotoxins are often carried by temperate phages, potentially transferring between bacterial species.
  • Enterotoxins are stable to digestion and extreme conditions.
  • Staphylococcus aureus is a common source of enterotoxins.
  • Enterotoxin genes can contribute to pathogenicity in non-infectious diseases.

Biofilms

  • Biofilms are communities of microorganisms adhering to a surface.
  • Biofilms exhibit survival strategies to evade the host defense system, altering metabolic processes.
  • Biofilms often resist antimicrobial treatments.
  • Bacteria in biofilms may cause persistent infections that are difficult to resolve.
  • The initial stages of biofilm formation involve attachment and microcolony formation.

Methods and Materials

  • Study involved 89 water samples from different sources in Basrah Governorate analyzed via PCR to detect the presence of particular bacterial genes.
  • Collected DNA samples were previously identified.
  • Results analyzed using gel electrophoresis.
  • Primer sequences and PCR conditions are included.

### Results (Summary)

  • Sea gene detected in 46% of bacteria in different water sources.
  • Ica operon detected in 14.6% of bacteria.
  • Genetic overlap (sea and ica genes) in some bacterial types.

Discussion

  • Study's findings highlight the presence of enterotoxin-producing bacteria in water sources.
  • The presence of adhesion genes (icaA and icaD) in bacteria suggests high concentrations of toxins.
  • The study's practical importance: water used for human consumption should be free of bacteria and their toxins.

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