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General Microbiology

Types of microscopy used in bacteriology

Light microscopy
Phase contrast microscopy
Fluorescence microscopy
Darkfield microscopy
Transmission electron microscopy
Scanning electron microscopy

Fluorescent microscopy in which ultraviolet rays are used to examine cells after treatment with fluorescent days.

Phase contrast microscope enhances the refractive index differences of the cell components. This microscopy can be used to reveal details of the internal structures as well as capsules, endospores and motility

Electron microscope The resolving power is more than 200 times that of light microscope.
 

The cell cycle

1) Labile cells (GI tract, blood cells)
- Described as parenchymal cells that are normally found in the G0 phase that can be stimulated to enter the G1
- Undergo continuous replication, and the interval between two consecutive mitoses is designated as the cell cycle
- After division, the cells enter a gap phase (G1), in which they pursue their own specialized activities
•    If they continue in the cycle, after passing the restriction point (R), they are committed to a new round of division
•    The G1 phase is followed by a period of nuclear DNA synthesis (S) in which all chromosomes are replicated
•    The S phase is followed by a short gap phase (G2) and then by mitosis
•    After each cycle, one daughter cell will become committed to differentiation, and the other will continue cycling

2) Stable cells (Hepatocytes, Kidney)

- After mitosis, the cells take up their specialized functions (G0). 
- They do not re-enter the cycle unless stimulated by the loss of other cells

3) Permanent cells (neurons)

- Become terminally differentiated after mitosis and cannot re-enter the cell cycle
- Which cells do not have the ability to differentiate ->  Cardiac myocytes

Autoantibodies

Anti-nuclear antibodies (ANA)    Systemic Lupus
Anti-dsDNA, anti-Smith               Specific for Systemic Lupus
Anti-histone                                 Drug-induced Lupus
Anti-IgG                                       Rheumatoid arthritis
Anti-neutrophil                             Vasculitis
Anti-centromere                           Scleroderma (CREST)
Anti-Scl-70                                   Sclerderma (diffuse)
Anti-mitochondria                         1oary biliary cirrhosis
Anti-gliadin                                   Celiac disease
Anti-basement membrane            Goodpasture’s syndrome
Anti-epithelial cell                          Pemphigus vulgaris
Anti-microsomal                            Hashimoto’s thryoiditis

CELLS ORGANELLES

Cell parts:

Mitochondrion – double MB structure responsible for cellular metabolism – powerhouse of the cell

Nucleus – controls synthetic activities and stores genetic information

Ribosome – site of mRNA attachment and amino acid assembly, protein synthesis

Endoplasmic reticulum – functions in intracellular transportation

Gogli apparatus/complex – composed of membranous sacs – involved in production of large CHO molecules & lysosomes

Lysosome – organelle contains hydrolytic enzymes necessary for intracellular digestion

Membrane bag containing digestive enzymes

Cellular food digestion – lysosome MB fuses w/ MB of food vacuole & squirts the enzymes inside. Digested food diffuses through the vacuole MB to enter the cell to be used for energy or growth. Lysosome MB keeps the cell iself from being digested 

-Involved mostly in cells that like to phagocytose
-Involved in autolytic and digestive processes
-Formed when the Golgi complex packages up an especially large vesicle of digestive enzyme proteins

Phagosome 
– vesicle that forms around a particle (bacterial or other) w/in the phagocyte that engulfed it
- Then separates from the cell membrane bag & fuses w/ lysozome to receive contents
- This coupling forms phagolysosomes in which digestion of the engulfed particle occurs

Microbodies:
- Contain catalase
- Bounded by a single membrane bag
-  Compartments specialized for specific metabolic pathways
-  Similar in function to lysosomes, but are smaller & isolate metabolic reactions involving H2O2

-  Two general families:
·        Peroxisomes: transfer H2 to O2, producing H2O2 – generally not found in plants
·        Glyoxysomes: common in fat-storing tissues of the germinating seeds of plants
¨      Contain enzymes that convert fats to sugar to make the energy stored in the oils of the seed available

 Inclusions

– transitory, non-living metabolic byproducts found in the cytoplasm of the cell
- May appear as fat droplets, CHO accumulations, or engulfed foreign matter.

NORMAL MICROBIAL FLORA 

A. Properties. Normal microbial flora describes the population of microorganisms that usually reside in the body. The microbiological flora can be defined as either 
1) Resident flora - A relatively fixed population that will repopulate if disturbed, 

2) Transient flora - that are derived from the local environment. These microbes usually reside in the body without invasion and can
even prevent infection by more pathogenic organisms, a phenomenon known as bacterial interference. 
The flora have commensal functions such as vitamin K synthesis. However, they may cause invasive disease in immunocompromised hosts or if displaced from their normal area. 

B. Location. Microbial flora differ in composition depending on their anatomical locations and microenvironments. The distribution of normal microbial flora.

ANTIGEN-ANTIBODY REACTIONS

Affinity of the antigen-antibody reaction refers to the intensity of the attraction between antigen and antibody molecule.
Antigen-antibody reactions

Reaction test            Modified test

Precipitation  -> Immunoelectrophoresis, Immunoprecipitation
Agglutination -> Latex agglutination, Indirect, Haemagglutination , Coagglutination ,Coombs test

Neutralization  -> Measurement of LD, Plaque assays

Complement fixation  -> Conglutination

Immunofluorescence ->  Indirect immunofiuorescence, Immunoofluoremetric Assay

Enzyme immunoassay -> Enzyme linked, Immunosorbent assay

Radioimmunoassay -> Immunoradiometric assay

Avidity is the strength of the bond after the formation of antigen-antibody complex.

Sensitivity refers to the ability of the test to detect even very minute quantities of antigen or antibody. A test shall be called as highly sensitive if false negative results are absent or minimal.

Specificity refers to the ability of the test to detect reactions between homologous antigens and antibodies only, and with no other. In a highly specific test, false positive reactions will be minimal or absent.

DISINFECTION AND STERILIZATION

•    Sterilization is the best destruction or com removal_of all forms of micro organisms.
•    Disinfection is the destruction of many microorganisms but usually the b spores.
•    Antisepsis is the destruction or inhibition of microorganisms in living tissues thereby limiting or preventing the harmful effect of infection.
•    Astatic Agent  would only inhibit the growth of microorganisms (bacteriostatic, fungistatic, sporostatic).
•    Acidal agent would kill the microorganism (bactericidal. virucidal, fungicidal)
•    Sterilants are the chemicals which under controlled conditions can kill sporinQ bacteria.
 

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