Medical biology, microbiology, virology, immunology department Microbial Mechanisms of Pathogenicity by As.Prof. O.V. Pokryshko Main Features of Pathogenic Microorganisms. Pathogenicity. This is the potential capacity of certain species of microbes to cause an infectious process. Virulence signifies the degree of pathogenicity of the given culture (strain). Virulence, therefore, is an index of the qualitative individual nature of the pathogenic microorganism. Virulence in pathogenic microbes changes under the influence of natural conditions. The virulence of pathogenic microorganisms is associated with adherence, invasiveness, capsule production, toxin production, aggressiveness and other factors. The adherence Adherence factor Filamentous hemagglutinin Fimbriae Glycocalyx or capsule Pili Slime Description Causes adherence to erythrocytes Help attach to solid bacteria to solid surfaces Inhibits phagocytosis and aids in adherence Bind bacteria together for transfer of genetic material Tenacious bacterial film that is less compact than a capsule Teichoic and lipoteichoic Cell wall components in gram acid positive bacteria that aid in adhesion Adherence bacteria to cell surfaces Adherence of vibrio cholera on the mucose Capsule production Capsule production makes the microbes resistant to phagocytosis and antibodies, and increases their invasive properties. Thus, for example, capsular anthrax bacilli are not subject to phagocytosis, while noncapsular variants are easily phagocytized. The role of capsular material in bacterial virulence. Some pathogenic microorganisms (B. anthracis, C. perfringens, S. pneumoniae, causative agents of plague and tularaemia) are capable of producing a capsule in animal and human bodies. Certain microorganisms produce capsules in the organism as well as in nutrient media (causative agents of rhinoscleroma, ozaena, pneumonia). Invasive properties of pathogenic bacteria Virulent microbes are characterized by the ability to penetrate tissues of the infected organism (invasive properties). collagenase and hyaluronidase immunoglobulin A protease leukocidins M-protein protein A Collagenase and hyaluronidase degrade collagen and hyaluronic acid, respectively, thereby allowing the bacteria to spread through subcutaneus tissue (Streptococci, Staphylococci, Clostridium ). Immunoglobulin A protease degrades IgA, allowing the organism to adhere to mucous membranes, and is produseed chiefly by N. gonorrhoeae, Haemophilus influenzae, and S. pneumoniae. Leukocidins neutrophilic macrophages. can destroy leukocytes M-protein of antiphagocytic. S. pyogenes both and is Protein A of S. aureus binds to IgG and prevents the activation of complement. Coagulase, which is produced by S. aureus and accelerate the formation of a fibrin clot from its precursor, fibrinogen (this clot may protect the bacteria from phagocytosis by walling off the infected area and by coating the organisms with a layer of fibrin) The invasion of cells by bacteria Toxin production According to the nature of production, microbial toxins are subdivided into exotoxins and endotoxins. More than 50 protein exotoxins of bacteria are known to date. Exotoxins easily diffuse from the cell into the surrounding nutrient medium. They are characterized by a markedly distinct toxicity, and act on the susceptible organism in very small doses. Exotoxins have the properties of enzymes hydrolysing vitally important components of the cells of tissues and organs. Exotoxins exert their effects in a variety of ways – by inhibition of protein synthesis, inhibition of nerve synapse function, disruption of membrane transport, damage to plasma membranes. Exotoxins may be devided into fifth categories on the basis of the site affected: neurotoxins (tetanotoxin, botulotoxin) C. tetani, C. botulinum, B. cereus, S. aureus; cytotoxins (enterotoxins, dermatonecrotoxin) E. coli, Salmonella spp., Klebsiella spp., V. cholerae, C. perfringens; functional blocators (cholerogen), V. cholerae; membranotoxins (hemolysins, leucocidin), S. aureus; exfoliatin S. aureus. Action of the hemolysin on red blood cells MICROORGANISM TOXIN DISEASE ACTION Clostridium botulinum Several neurotoxins Botulism Paralysis; blocks neural transmission Clostridium tetani Neurotoxin Tetanus Spastic paralysis; interferes with motor neurons Corynebacterium diphtheriae Cytotoxin Diphtheria Bordetella pertussis Pertussis toxin Whooping cough Blocks G proteins that are involved in regulation of cell pathways Streptococcus pyogenes Hemolysin Scarlet fever Food Lysis of blood cells Staphylococcus aureus Enterotoxin Poisoning Aspergillus flavus Cytotoxin Aflatoxicosis Amanita phalloides Cytotoxin Mushroom food poisoning Blocks protein synthesis Intestinal inflammation Blocks transcription of DNA, thereby stopping protein synthesis Blocks transcription of DNA,thereby stopping protein synthesis Endotoxins are more firmly bound with the body of the bacterial cell, are less toxic and act on the organism in large doses; their latent period is usually estimated in hours, the selective action is poorly expressed. According to chemical structure, endotoxins are related to glucoside-lipid and polysaccharide compounds or phospholipid-protein complexes. They are thermostable. Some endotoxins withstand boiling and autoclaving at 120°C for 30 minutes. Action of the endotoxin Endotoxin in the bloodstream Differences between exotoxins and endotoxins exotoxins endotoxins Proteins Heat labile Lipopolysaccharides Heat stable Actively secreted by cells, diffuse into surrounding medium form part of cell wall,do not diffuse into surrounding medium Readily separable from cultures by physical means such as filtration Action often enzymic Obtained only by cell lysing Specific pharmacological effect for each exotoxin Non-specific action of all endotoxins No enzymic action Specific tissue affinities Active in very minute doses Highly antigenic No specific tissue affinity Active only in very large doses Weakly antigenic Stimulate formation of Do not stimulate formation antitoxin which neutralizes of antitoxin toxin Converted into toxoid by Can not be toxoided formaldehyde Produced by both grampositive bacteria and gram-negative bacteria Frequently controlled by extrachromosomal genes (e.g. plasmids) Produced by gramnegative bacteria only Synthesis directed by chromosomal genes genes In characterizing pathogenic microbes a unit of virulence has been established. Dlm (Dosis letalis minima), representing the minimum amount of live microbes which in a certain period of time bring about 95-97 % death of the corresponding laboratory animals. the absolute lethal dose of pathogenic microbe Dcl (Dosis certa letalis) which will kill 100 % of the experimental animals has been established. At present LD50 (the dose which is lethal to one half of the infected animals) is considered to be the most suitable, and may serve as an objective criterion for comparison with other units of virulence.