1) Virulence Factors And Their Role

Virulence factors should never be considered independently of the host's defenses; the clinical course of a disease often depends on the interaction of virulence factors with the host's response. An infection begins when the balance between bacterial pathogenicity and host resistance is upset In essence, we live in an environment that favors the microbe, simply because the growth rate of bacteria far exceeds that of most eukaryotic cells. Furthermore, bacteria are much more versatile than eukaryotic cells in substrate utilization and biosynthesis. The high mutation rate of bacteria combined with their short generation time results in rapid selection of the best-adapted strains and species. In general, bacteria are much more resistant to toxic components in the environment than eukaryotes, particularly when the major barriers of eukaryotes (skin and mucous membranes) are breached. The pathogenesis of many bacterial infections cannot be separated from the host immune response, for much of the tissue damage is caused by the host response rather than by bacterial factors Virulence factors help bacteria to (1) invade the host, (2) cause disease, and (3) evade host defenses. The following are types of virulence factors: 1) Adherence Factors: Many pathogenic bacteria colonize mucosal sites by using pili (fimbriae) to adhere to cells. 2) Invasion Factors: Surface components that allow the bacterium to invade host cells can be encoded on plasmids, but more often are on the chromosome. 3) Capsules: Many bacteria are surrounded by capsules that protect them from opsonization and phagocytosis. 4) Endotoxins: The lipopolysaccharide endotoxins on Gram-negative bacteria cause fever, changes in blood pressure, inflammation, lethal shock, and many other toxic events. 5) Exotoxins: Exotoxins include several types of protein toxins and enzymes produced and/or secreted from pathogenic bacteria. Major categories include cytotoxins, neurotoxins, and enterotoxins. 6) Siderophores: Siderophores are iron-binding factors that allow some bacteria to compete with the host for iron, which is bound to hemoglobin, transferrin, and lactoferrin. 7) Superantigens: Microbial antigens that have in common an extremely potent activating effect on T-cells that bear a specific variable region. Superantigens cross-link the variable region with class II MHC proteins regardless of the peptide binding in the T-cell receptor's pocket. The result is a transient expansion and subsequent death and energy of the T-cells with the appropriate variable regions.

2) Bacterial Threshold And Quorum Sensing

Quorum sensing is the regulation of gene expression in response to fluctuations in cellpopulation density. In similar fashion, some social insects use quorum sensing to determine where to nest. In addition to its function in biological systems, quorum sensing has several useful applications for computing and robotics. Quorum sensing can function as a decision-making process in any decentralized system, as long as individual components have: (a) a means of assessing the number of other components they interact with and (b) a standard response once a threshold number of components is detected. Quorum sensing bacteria produce and release chemical signal molecules called auto inducers that increase in concentration as a function of cell density. The detection of a minimal threshold stimulatory concentration of an auto inducer leads to an alteration in gene expression. Gram-positive and Gram-negative bacteria use quorum sensing communication circuits to regulate a diverse array of physiological activities. These processes include symbiosis, virulence, competence, conjugation, antibiotic production, motility, sporulation, and biofilm formation. In general, Gram-negative bacteria use acylated homoserine lactones as auto inducers, and Grampositive bacteria use processed oligo-peptides to communicate.

3) Toxins - Endotoxin And Exotoxin - Their Role In Pathogenesis

Bacterial Toxin Bacterial toxin is characteristics of any bacterium that enhance its pathogenicity, which is the ability to cause disease. There are two general types, which are endotoxin and exotoxin. Endotoxin Endotoxin is the toxin that kept within the bacterial cell and to be released after destruction of the bacterial cell wall or bacterial cell lysis. These are heat-stable, lipopolysaccharide (LPS) components of the outer membranes of gram-negative bacteria only. The key effects of endotoxins on vertebrates are mediated by their interaction with specific receptors on immune cells such as monocytes, macrophages, dendritic cells, and others. Upon challenge with endotoxin, these cells form a broad spectrum of immune mediators such as cytokines, nitric oxide, and eicosanoids. Exotoxin Exotoxin is the toxin protein that are secreted or released by both gram-positive and gramnegative bacteria towards the hosts cell. An exotoxin can cause damage to the host by destroying cells or disrupting normal cellular metabolism. They are highly potent and can cause major damage to the host. Besides, exotoxins are also susceptible to antibodies produced by the immune system, but many exotoxins are so toxic that they may be fatal to the host before the immune system has a chance to mount defences against it.

4) Superantigen With Example

An antigen is defined as any substance, not only infectious agents, but also environmental toxins, drugs, food molecules, and altered membrane molecules, that induce a state of sensitivity and/or immune responsiveness after a latent period of days to weeks, and which reacts with antibody proteins and/or other immune mediators, of a sensitized subject. Superantigens are a class of antigens that cause non-specific T-cell activation, and massive release of immune cytokines. Superantigens are able to activate up to 20 percent of the body's T-cells, which would explain the link between respiratory and skin allergies, and probably intestinal sensitivity and allergic reaction. Superantigens would disrupt the strength of our adaptive immune system, which is the learned ability to target antigens with a high specificity. Scientists have discovered that superantigens can activate auto-reactive T cells and induce severe relapses in multiple sclerosis, where the body goes through periods where the immune system attacks its own nerve sheaths, or myelin. In animal studies, the superantigens were able to activate auto-reactive T cells not involved in the initial bout of the multiple sclerosis. Superantigens have also been implicated in acute diseases, such as food poisoning and toxic shock syndrome. We see that superantigens present quite a health threat. Some examples of superantigens are proteins found on the membranes, or secreted from, microbes such as Staphylococcus and Streptococcus bacteria, which are called exotoxins. These bacteria are quite common in the environment, and exist naturally on the skin and in the intestinal tract of humans. The bacteria are able to evolve genetic traits that protect them from danger.

5) Slime And Biofilm

Slime A slime layer in bacteria is an easily removed, diffuse, unorganized layer of extracellular material that surrounds bacteria cells. Specifically, this consists mostly of exopolysaccharides, glycoproteins, and glycolipids. The slime layer is not to be confused with the S-layer, a separate and highly organised glycoprotein layer surrounding many bacterial cells. The function of the slime layer is to protect the bacteria cells from environmental dangers such as antibiotics and desiccation. The slime layer also allows bacteria to adhere to smooth surfaces such as prosthetic medical devices and catheters. Biofilm A biofilm is any group of microorganisms in which cells stick to each other on a surface. These adherent cells are frequently embedded within a self-produced matrix of extracellular polymeric substance (EPS). Biofilm EPS is a polymeric conglomeration generally composed of extracellular DNA, proteins, and polysaccharides. Biofilms may form on living or non-living

surfaces and can be prevalent in natural, industrial and hospital settings. The microbial cells growing in a biofilm are physiologically distinct from planktonic cells of the same organism, which, by contrast, are single-cells that may float or swim in a liquid medium. Microbes form a biofilm in response to many factors, which may include cellular recognition of specific or non-specific attachment sites on a surface, nutritional cues, or in some cases, by exposure of planktonic cells to sub-inhibitory concentrations of antibiotics.[4][5] When a cell switches to the biofilm mode of growth, it undergoes a phenotypic shift in behaviour in which large suites of genes are differentially regulated.

1) Cardinal Features Of Inflammation

There are five: 1) Redness (Rubor) and Heat (Calor) Due to vasodilatation and increase rate of blood flow to the micro circulation 2) Swelling (Tumor) due to vascular permeability that is increase and plasma fluids that will leak into the injured tissue 3) Pain (Dolor) brought about by the pressure of fluids or swelling through the nerve endings. 4) Loss of function/ movement brought by the swelling and pain.

5) Microbial Mimicry
Mimicking our own cells (molecular mimicry), killing or hiding in white blood cells and suppression of our immune response by secretion of toxins. Pathogenic mimics can possess a structural architecture that differs markedly from that of the functional homologues. Therefore, proteins of dissimilar sequence may have a common structure which elicits an autoimmune response. It has been hypothesized that these virulent proteins display their mimicry through molecular surfaces that mimic host protein surfaces. Bacteria not only mimic our own cells to avoid our immune system, but can also mimic a virus, inducing the wrong immune response. Auto immunity: The failure of an organism in recognizing its own constituent parts as self, which allows an immune response against its own cells and tissues.

6) Selective And Differential Media

Selective Media 1) Selective media contain one or more agents that are inhibitory to all organisms except those being sought. In other words, this media select for the growth of certain bacteria to the disadvantage of others. 2) Inhibitory agents used for this purpose include dyes, bile salts, alcohols, acids and antibiotics. 3) An example of selective media is phenylethyl alcohol agar, which inhibits the growth of aerobic and facultatively anaerobic gram-negative rods and allows gram-positive cocci to grow.

Differential Media 1) Differential media employ some factor that allows colonies of one bacterial species or type to exhibit certain metabolic or culture characteristics that can be used to distinguish them from other bacteria growing on the same agar plate. 2) One commonly used differential media is MacConkey agar, which differentiates between gram-negative bacteria that can and cannot ferment the sugar lactose. Note that those not able to ferment lactose appear light pink or relatively colourless while those capable to ferment appear deep purple colour. 3) Another example is blood agar ( eg : sheep) and chocolate agar.

7) Provisional Diagnosis
A provisional diagnosis is one that is initially determined to be the diagnosis, except for the fact that all test results have not been received or analysed. When the full medical work-up has been completed, the provisional diagnosis is usually changed to a definite diagnosis. Nevertheless, the provisional diagnosis sets in motion the first stage of treatment and illness management.

8) Siderophore
Siderophores or iron carriers are defined as relatively low molecular weight, ferric ion specific chelating agents elaborated by bacteria and fungi growing under low iron stress. The role of these compounds is to scavenge iron from the environment and to make the mineral, which is almost always essential, available to the microbial cell. Interest in it has accrued with the realization that most aerobic and facultative anaerobic microorganisms synthesize at least one siderophore. Siderophores have been related to virulence mechanisms in microorganisms

pathogenic to both animals and plants. In addition, they have clinical applications and are possibly important in agriculture. Siderophores have a very high affinity for iron and are able to solubilize and transport ferric iron (Fe3+) in the environment and also to compete for iron with mammalian proteins such as transferrin and lactoferrin. The majority of bacteria and fungi use siderophores to solubilize and transport iron. Microorganisms can use either siderophores produced by themselves or siderophores produced by other microorganisms. The many different types of siderophores can generally be classified into two structural groups, hydroxamates and catecholate compounds. Despite their structural differences, all form an octahedral complex with six binding coordinates for Fe3+. Siderophores have potential applications in the treatment of some human diseases and infections. Some siderophores are used therapeutically to treat chronic or acute iron overload conditions in order to prevent iron toxicity in humans. Individuals who have defects in blood cell production or who receive multiple transfusions can sometimes have too much free iron in the body. However, in order to prevent infection during treatment for iron overload, it is important to use siderophores that cannot be used by bacterial pathogens. A second clinical application of siderophores is in antibiotic delivery to bacteria. Some gram-negative bacteria are resistant to antibiotics because they are too big to diffuse through the outer-membrane porins. However, siderophore-antibiotic combination compounds have been synthesized that can be transported into the cell using the siderophore receptor.

11) Selective Toxicity Of Antibiotics And Example

The selectively toxicity of antibiotic means that they must be highly effective against the microbe but have minimal or no toxicity to humans. In practice, this is expressed by a drug's therapeutic index (TI) - the ratio of the toxic dose (to the patient) to the therapeutic dose (to eliminate the infection). The larger the index, the safer is the drug (antibiotic) for human use. The selective toxicity of antibiotics is brought about by finding vulnerable targets for the drug in the microbe that do not exist in the human that is given the drug. Most antibiotics in clinical usage are directed against bacterial cell wall synthesis, bacterial protein synthesis, or bacterial nucleic acid synthesis, which are unique in some ways to bacteria. For example, the beta lactam antibiotics (penicillin and its relatives) inhibit peptidoglycan synthesis in the cell wall. Humans have neither a cell wall nor peptidoglycan and so are unaffected by the action of the drug.

12) Acid Fast Staining

The acid-fast stain is a differential stain used to identify acid-fast organisms such as members of the genus Mycobacterium . Acid-fast organisms are characterized by wax-like, nearly impermeable cell walls.They contain mycolic acid and large amounts of fatty acids, waxes, and complex lipids. Acid-fast organisms are highly resistant to disinfectants and dry conditions. Because the cell wall is so resistant to most compounds, acid-fast organisms require a special staining technique. The primary stain used in acid-fast staining, carbolfuchsin, is lipidsoluble and contains phenol, which helps the stain penetrate the cell wall. This is further assisted by the addition of heat. The smear is then rinsed with a very strong decolorizer, which strips the stain from all non-acid-fast cells but does not permeate the cell wall of acid-fast organisms. The decolorized non-acid-fast cells then take up the counterstain.