Docsity
Docsity

Prepare for your exams
Prepare for your exams

Study with the several resources on Docsity


Earn points to download
Earn points to download

Earn points by helping other students or get them with a premium plan


Guidelines and tips
Guidelines and tips

Bacterial Toxins: Production, Types, and Impact on Health, Study notes of Toxicology

An in-depth exploration of bacterial toxins, their production mechanisms, and the role they play in causing diseases. It covers various types of toxins, including exotoxins and endotoxins, their modes of action, and the impact on human health. The document also discusses the importance of bacterial toxins in foodborne diseases and the detection methods for endotoxins.

Typology: Study notes

2021/2022

Uploaded on 09/27/2022

bartolix
bartolix 🇬🇧

4.8

(17)

304 documents

1 / 11

Toggle sidebar

This page cannot be seen from the preview

Don't miss anything!

bg1
Bacterial Toxigenesis
Food Toxicology
Instructor: Gregory Möller, Ph.D.
University of Idaho
Learning Objectives
Define bacterial toxigenesis.
Explore bacterial toxins, their background and nomenclature.
Differentiate exotoxins and endotoxins.
Explore the toxicity, properties, and mode of action of exotoxins.
Understand the toxicity,
properties, and mode of action
of endotoxins (pyrogens).
Understand the origins of sepsis.
Review the detection of
endotoxins. Bacterial Toxigenesis
The ability to produce toxins:
a mechanism of bacterial disease.
• Cell-associated lipopolysaccharide (LPS) toxins are referred to as
endotoxins.
Extracellular diffusible toxins are referred to as exotoxins.
Bacterial Toxins
[Substances] that are toxic to eukaryotic cells as measured in a
variety of ways, Orndorff 1992
A microbial substance able to induce host damage, Madigan et al.
1997
Any organic microbial product
or substance that is harmful
or lethal to cells, tissue cultures,
or organisms, Atlas 1995
Bacterial Toxins
[Toxins are] a common and series cause of tissue damage,
especially in bacterial infection, Mims et al. 1993;
Disease is frequently determined by production of microbial
toxins, Murray et al. 1994
pf3
pf4
pf5
pf8
pf9
pfa

Partial preview of the text

Download Bacterial Toxins: Production, Types, and Impact on Health and more Study notes Toxicology in PDF only on Docsity!

Bacterial Toxigenesis

Food Toxicology Instructor: Gregory Möller, Ph.D. University of Idaho

Learning Objectives

  • Define bacterial toxigenesis.
  • Explore bacterial toxins, their background and nomenclature.
  • Differentiate exotoxins and endotoxins.
  • Explore the toxicity, properties, and mode of action of exotoxins.
  • Understand the toxicity, properties, and mode of action of endotoxins (pyrogens).
  • Understand the origins of sepsis.
  • Review the detection of endotoxins.

Bacterial Toxigenesis

  • The ability to produce toxins: a mechanism of bacterial disease.
  • Cell-associated lipopolysaccharide (LPS) toxins are referred to as endotoxins.
  • Extracellular diffusible toxins are referred to as exotoxins.

Bacterial Toxins

  • ‘[Substances] that are toxic to eukaryotic cells as measured in a variety of ways’, Orndorff 1992
  • ‘A microbial substance able to induce host damage’, Madigan et al. 1997
  • ‘Any organic microbial product or substance that is harmful or lethal to cells, tissue cultures, or organisms’, Atlas 1995

Bacterial Toxins

  • ‘[Toxins are] a common and series cause of tissue damage, especially in bacterial infection’, Mims et al. 1993;
  • ‘Disease is frequently determined by production of microbial toxins’, Murray et al. 1994
  • ‘Bacterial [toxins] are thus important determinants of bacterial virulence’ Poxton and Arbuthnott 1990

Bacterial Toxins

  • ‘Microbial toxins are components or products of microorganisms which, when extracted and introduced into host animals, can reproduce disease symptoms normally associated with infection without infestation by those microorganisms’, Williams and Clarke, 1998

Bacterial Toxins

  • Terms
    • Exotoxin = extracellular protein toxin
    • Endotoxin = lipid A portion of Gram-neg outer membrane
    • Enterotoxin = toxin that acts on gastrointestinal tract, producing typical food poisoning symptoms
  • Nomenclature
    • Named for host cell attacked: cytotoxin, neurotoxin
    • Named for producer or disease: cholera, Shiga
    • Named for activity: lecithinase, adenylate cyclase
    • Letter designation: exotoxin A

Endotoxins and Exotoxins

  • Endotoxins are cell-associated substances that are structural components of the outer membrane of Gram-negative bacteria. - Released from growing bacterial cells - Released from cells which are lysed from effective host defense (e.g. lysozyme) - Released from activities of certain antibiotics (e.g. penicillins)
  • Exotoxins usually secreted by bacteria but in some cases they are released by lysis of the bacterial cell.

Exotoxins

  • Most well-characterized family of toxins.
  • Secreted by bacterium as soluble proteins.
  • Most act catalytically
  • Highly specific in the substrate utilized
    • Tissue cells, organs, or body fluid
  • Highly specific mode of action.
  • Site of damage caused by the toxin indicates the location of the substrate. - Enterotoxin, neurotoxin, leukocidin, or hemolysin.

Protein Toxins: Cytotoxic Activity

  • Certain protein toxins have very specific cytotoxic activity
    • Attack specific types of cells.
    • e.g. tetanus or botulinum toxins attack only neurons.
  • Some toxins have fairly broad cytotoxic activity and cause nonspecific death of all sorts of cells and tissues, eventually resulting in necrosis. - Staphylococci, streptococci, clostridia, etc.
  • Some broadly lethal but with unknown specifics - e.g. anthrax toxin LF.

Protein Toxins: Strongly Antigenic

  • In vivo antibody (antitoxin) neutralizes the toxicity of bacterial proteins.
  • Protein toxins are inherently unstable
    • In time they lose their toxic properties but retain their antigenic properties.
  • Toxoids are detoxified toxins which retain their antigenicity and their immunizing capacity.

Protein Toxins: Toxoids

  • The formation of toxoids can be accelerated by treating toxins with a variety of reagents - Formalin, iodine, pepsin, ascorbic acid, ketones, etc. - 37ºC, pH 6-9 for several weeks.
  • Resulting toxoids can be use for artificial immunization where the primary determinant of bacterial virulence is toxin production. - e.g. diphtheria and tetanus.
  • Toxoids can be genetically engineered.

Gram-Negative Bacteria: Endotoxins

  • An important part of the toxicity of these organisms is conferred through the release of endotoxins, as occurs in septicemia, toxic shock syndrome, and sometimes in food poisoning.
  • Endotoxins = lipopolysaccharides.
    • A part of cell membrane envelopes.
  • Killed bacteria can release endotoxins as they decay. - E. coli , Salmonella , Shigella , Pseudomonas , Neisseria , Haemophilus , and other pathogens.

Endotoxins (Pyrogens)

  • Cause a wide variety of serious reactions such as fever, shock, changes in blood pressure, and in other circulatory functions.

Endotoxins

  • Endotoxins are toxic to most mammals
  • Regardless of the bacterial source, all endotoxins produce the same range of biological effects in the animal host.
  • Most of our knowledge of the biological activities of endotoxins derives not from the study of natural disease but by challenge of experimental animals.

Endotoxins

  • The injection of living or killed Gram-negative cells, or purified LPS, into experimental animals causes a wide spectrum of nonspecific pathophysiological reactions.
  • Includes: fever, changes in white blood cell counts, disseminated intravascular coagulation, hypotension, shock and death.

Endotoxins

  • Injection of fairly small doses of endotoxin results in death in most mammals.
  • The sequence of events follows a regular pattern: (1) latent period;

Agents of Foodborne Infections

That Also Produce Toxins

  • Shigella sp.
    • Shiga toxin
  • Listeria monocytogenes
    • Listeriolysin
  • Salmonella sp.
    • Enterotoxin, cytotoxin
  • enterohemorrhagic E. coli
    • Shiga-like toxin
  • Vibrio parahaemolyticus
    • Hemolysin
  • Yersinia enterocolitica
    • Enterotoxin

Bacterial Cell Structures

Bacterial Cell Wall

Bacterial Cell Wall

Gram-Positive Cell Wall

Gram-Negative Cell Wall

Lipoplysaccharide (LPS) GNB

  • Major constituent of outer surface of the outer membrane
  • Covers ~75% of the OM
  • Gives colonies a smooth appearance
  • ~3-10% of the total dry cell weight
  • 3-4 million LPS molecules per cell

Lipopolysaccharide Structure

LPS Lipid A

  • Lipid A is a powerful biological response modifier that can stimulate the mammalian immune system. - During infectious disease caused by Gram - bacteria, endotoxins released from, or part of, multiplying cells have similar effects on animals and significantly contribute to the symptoms and pathology of the disease encountered.

Lipopolysaccharide Lipid A

LPS Mode of Action

  • Bound by plasma proteins LPS-binding proteins (LBP).
  • LBP interacts with receptors on monocytes and macrophages and other types of receptors on endothelial cells.
  • In monocytes and macrophages three types of events are triggered during their interaction with LPS:
  1. Production of cytokines, (IL, TNF, etc.) which stimulate production of prostaglandins and leukotrienes (powerful mediators of inflammation and septic shock).
  2. Activation of the complement cascade. (cause histamine release leading to vasodilation) and effect neutrophil chemotaxis and accumulation. The result is inflammation.
  3. Activation of the coagulation cascade:
    • a. Coagulation → fibrinolysis and hemorrhaging → blood clotting cascade → bradykinins and other vasoactive peptides → coagulation, thrombosis → acute disseminated intravascular coagulation → depleted platelets and clotting factors → internal bleeding.
    • b. Inflammation
    • c. hypotension.

Inflammation → Sepsis

  • The LPS net effect is to induce inflammation, intravascular coagulation, hemorrhage, and shock.
  • LPS also acts as a immune system B cell mitogen stimulating the polyclonal differentiation and multiplication of B-cells and the secretion of immunoglobulins (e.g. IgG).

Infection/Intoxication: Inflammation Cycle

  • Macrophage migration inhibitory factor (MIF) is a pro-inflammatory pituitary and immune cell cytokine and a critical mediator of septic shock.

Inflammation Cycle: Toll Receptor

  • Indoor and outdoor detection of air, water or dust contamination
  • Medicine:
  • Detection of Gram-negative bacterial infection, diagnosis of sepsis

Pyrogen Limits: Pharmacological Products

  • Amoxicillinum natricum 0.25EU/mg
  • Clindamycini hydrochloridium 0.58EU/mg
  • Water for intravenous infusion 0.25EU/ml
  • Therapeutic devices for cerebrospinal contact 0.06EU/ml
    • 1EU = 0.2 ng LPS

Detection of Endotoxin

  • Biological tests:
    • Rabbit Pyrogen test**
    • Limulus Amebocyte Lysate test**
    • Neutrophil Chemiluminescence test
  • Non-biological endotoxin detection:
    • Chemical markers (3-OH fatty acids, Kdo)
    • Detection by molecules specifically recognizing LPS

Detection of Endotoxin – Biological Tests

  • For most of the 20th Century, the Rabbit Pyrogen Test was the standard method of testing for pyrogenicity. This test, which took approximately 4 hrs, is accomplished by injecting the substance being analyzed into a rabbit’s ear.
  • If the animal developed a fever, it confirmed the presence of pyrogens.

Detection of Endotoxin – Biological Tests

  • L imulus A mebocyte L ysate test (LAL)
  • The Atlantic horseshoe crab Limulus polyphemus

Detection of Endotoxin – Biological Tests

  • The LAL Test was commercially introduced during the 1970s.
    • In 1977, the FDA described conditions for the use of LAL as an end-product test for endotoxin in human biological products and medical devices.
  • To obtain the lysate required for the LAL test, a small amount of horseshoe crabs’ blood is drawn.
  • Next, blood cells (amebocytes) are separated and lysed to obtain the cellular proteins.

Limulus Amebocyte Lysate test (LAL)

  • Gel clot LAL provides a simple +/- result
  • Chromogenic end-point LAL offers a quantitative result and exhibits less product interference than LAL methods utilizing the clotting protein.
  • Kinetic turbidimetric LAL gives quantitative results but its use of the clotting protein limits its sample compatibility.
  • Kinetic chromogenic LAL provides automation and greater sensitivity to 0.005 EU/ml (1pg of LPS)

Limits of Rabbit Pyrogen Test and LAL

  • Cannot be used for:
    • Diagnostic testing of blood and other body fluid for endotoxin content
    • Testing of concentrated salts solutions
    • Testing of chemicals
    • Solutions of various proteins
  • Non-biological endotoxin detection used.

Endotoxin Levels in Ground Beef (LAL)