Microbiology π¦ Flashcards
What is microbiology?
Microbiology is the study of living organisms (βmicroorganismsβ or βmicrobesβ); simple in structure and usually small in size (cannot be seen with the naked eye).
What does the study of microbiology include?
They include bacteria, fungi, protozoa, and viruses.
What is the importance of microbiology?
Microorganisms cause many diseases and some microorganisms have been used in the manufacture of antibiotics and foodstuffs.
What are creatures classified into?
- Animalia (Eukaryotic): helminths
- Plantae (Eukaryotic)
- Protista (Eukaryotic): Protozoa
- Fungi (Eukaryotic)
- Monera (Prokaryotic): cellulars like bacteria and acellular like a virus
What is another term for prokaryotic?
Pre-mature
What is the difference between cellular prokaryotic and eukaryotic?
Nuclear membrane: absent - present Chromosomal number: haploid - diploid Histones: absent - present Ribosome: 70s - 80s Peptidoglycan: present - absent Mitochondria: absent - present Mitosis: absent - present Cell wall sterols: absent - present Membrane-bounded organelles: absent - present
What is the size of bacterial cells?
Measured by micron (micron= 1/ 1000 mm).
What is the shape of bacterial cells?
- Cocci or spherical: e.g. Staphylococci.
- Bacilli or cylindrical: e.g. Diphtheria bacilli.
- Spiral:
ο· One curve: e.g. Vibrio.
ο· More than one: e.g. Spirochetes, Spirillum.
What is the habitat of bacterial cells?
a. Parasitic (need host): bacterial flora (commensally) and pathogenic bacteria.
b. Saprophytic: free-living in soil, air, and water.
What is the structure of bacterial cells?
- Surface Structures (bacterial envelope):
β Cell wall.
β Cytoplasmic membrane.
β Capsule or slime layer. - Internal structures:
- Nuclear body
- Flagellae
- Inclusion bodies
- Ribosomes
- Fimbria
- Mesosomes
What are the characters of the cell wall?
It is the rigid layer outside the cytoplasmic membrane.
What is the chemical structure of the cell wall?
Composed of peptidoglycan.
What are the Differences between Gram-positive and Gram-negative cell walls?
Peptydoglycan:
ο· Thick (~ 40 sheets)
ο· Comprising up to 50% of the cell wall.
ο· Thin (One or two sheets)
ο· Comprising only 5β10% of the cell wall.
Special Components:
ο· Teichoicacid:ribitol or glycerol.
ο· Polysaccharides.
- Outer membrane (thick).
ο· Lipoprotein.
ο· Lipopolysaccharide: o Lipid A (the endotoxin). o Polysaccharide (somatic antigen).
2- Periplasmic space
ο· Between cytoplasmic membrane and outer
membrane and contains hydrolytic enzymes and penicillinase.
What is the function of the cell wall?
SPA GTG
- Preservation of the shape of the cell.
- Protectiveagainst high internal osmotic pressure.
- Antigenic character:
ο§ In Gram-positive: Teichoic acid.
ο§ In Gram-negative: somatic βOβ antigen(Polysaccharide). - Toxicity: The lipid A endotoxin of Gram-negative cell wall.
- Cell wall is responsible for Gram staining reaction (Gram-positive bacteria stain violet while Gram-negative bacteria stain pink)
- Cell wall is the target for the action of some antibiotics: penicillin and cephalosporins and vancomycin.
What are the enzymes that attack the cell wall?
The peptidoglycan is hydrolyzed by lysozyme found in tears, saliva, and nasal secretions.
What are the characters of the cell membrane?
It is a very thin elastic membrane that lies immediately under the cell wall.
What is the chemical structure of the cell membrane?
- composed of biphospholipids and proteins.
- prokaryotes have no sterols in the cytoplasmic membrane except for Mycoplasma.
What is the function of the cell membrane?
- Permeability and transport: transport nutrients into and waste products out of the cell.
- Electron transport and oxidative phosphorylation: for energy production (ATP).
- Excretion of hydrolytic enzymes.
- Biosynthetic function: carries enzymes and molecules for the biosynthesis of the cell wall, DNA, and membrane lipids.
- Chemotactic function: contain receptors of binding and repellents.
What are the characters of the capsule?
-Some bacteria can produce a gelatinous layer surrounding the cell outside the cell wall.
Capsule: condensed well-fined layer closely surrounds the cell.
Glycocalyx: polysaccharide-containing material lying outside the cell.
Slim layer: if glycocalyx is loosely surrounding the cell.
What is the chemical structure of the capsule?
Consists of polysaccharides, except in bacillus anthracis(protein polymers).
What is the function of the capsule?
- Virulence factor as it protects the bacterial cell from phagocytosis.
- Protects the cell wall against attack by bacteriophages, complement, and lysozymes.
- Antigenic (K-antigen): used in serodiagnosis or vaccine preparation.
What is the definition of flagella?
Long thread-like, helical filaments.
What are the types of flagellate?
- Monotrichous (single polar flagellum).
- Lophotrichous (multiple polar flagellae).
- Amphitrichous (One flagellum in each pole of the
cell) . - Peritrichous (flagella distributed over the entire
cell) e.g. E.coli.
What is the structure of flagella?
made up of a contractile protein called flagellin.
What are the functions of flagella?
- It is the organ of Motility:
> Movement toward the optimal nutrients.
Movement toward optimal oxygen concentration in aerobic bacteria.
Choosing the locality suitable for colonization.
Assist pathogenic bacteria in penetration through a viscid mucous secretion.
- Highly Antigenic (H-antigens).
What are viruses?
- Viruses are not cells, i.e. they do not have a nucleus or organelles.
- The smallest infectious agents (20 to 300 nm in diameter).
- They are obligate intracellular parasites (they do not have ribosomes and canβt be planted on agar)
- contain one kind of nucleic acid (RNA or DNA)
What is the difference between prokaryotes and acellular organisms?
- Prokaryotes: they do not have a membrane-bound nucleus
- Acellular: do not have a cell membrane
What is a virus particle (virion) composed of?
- Protein coat (capsid)
- Nucleic acid core
- Envelope (in some viruses)
What are the characters of viral capsid?
o It surrounds viral nucleic acid.
o made up of subunits called capsomers.
o Each capsomer is consisting of one or several proteins.
o The capsid with the enclosed nucleic acid is called the nucleocapsid.
What are the functions of the capsid?
a) It protects the viral genome against inactivation by nuclease enzymes.
b) The arrangement of capsomers (symmetry) is either icosahedral, helical, or complex.
c) Participates in adsorption of virions to susceptible cells. It determines the antigenicity
D) has a role in attachment.
What is viral symmetry?
1- Icosahedral (enveloped and non enveloped)
2- Helical (enveloped and non enveloped)
3- Complex (rare)
What are the characters of viral nucleic acid?
ο DNA (single molecule) or RNA (single or segmented).
ο Single or double-stranded (but one molecule)
ο Linear or circular.
ο It is the infectious part of the virus and codes for viral structure and non-structural proteins.
What are the characters of the viral envelopes?
ο Lipoprotein, the lipid from the host cell membranes and protein is virus-specific.
ο Glycoproteins in the form of spike-like projections on the surface attach to the host cell receptors.
ο It determines viral antigenicity and specificity
What determines the antigenicity of the virus?
The envelope and the capsid
What are the spikes of HIV and the receptor it works on respectively?
Gp120 and cd4
What are the steps of the viral replication cycle?
1-Attachment (adsorption by glycoprotein spikes)
2-Penetration
3-Uncoating
4-Viral gene expression (transcription) and protein synthesis (translation for the synthesis of coat)
5-Viral nucleic acid synthesis (replication for the synthesis of more versions)
6. Assembly (reunion)
7. Release
Attachment step of virus
ο Attachment of the virus to the host cell
ο It is receptor-specific
Penetration step of the virus
ο In non enveloped viruses penetration occurs by crossing the plasma membrane directly or by receptor-mediated endocytosis.
ο In enveloped viruses penetration occurs by fusion of viral envelope with cell membrane or with the membrane of endosome at the cell surface
Uncoating step of the virus
ο It is the release of viral nucleic acid by cellular enzymes.
ο Uncoating renders viral nucleic acid accessible for transcription and replication.
Viral nucleic acid synthesis (replication) step of the virus
by using a strand of the parental nucleic acid as a template for the production of progeny DNA or RNA molecules.
Assembly step of the virus
Assembly of viral nucleic acid and protein coats to form mature virus particles.
Release step of the virus
Virus particles are released from the cell either by:
ο Budding through the outer cell membrane as in enveloped viruses (taking a part of the bilayer)
ο Rupture of the cell membrane and release of the mature particles in unenveloped viruses.
How are viruses laboratorally detected?
Direct or indirect (virus isolation) and serologic detection of antiviral antibodies
How are viruses directly detected in the laboratory?
1) Detection of Virus particles by (electron microscope), inclusion bodies by (LM) (in cytoplasm and nucleus)
2) Detection of viral antigens by (EIA, RIAβ¦)
3) Detection of the viral nucleic acid by (PCR) and other
molecular techniques.
How are viruses indirectly detected in the laboratory?
-Viruses are obligate intracellular parasites and can be cultivated on:
a) Tissue cultures:
ο Pieces of animal or human tissues are trypsinized to get separate cells.
ο Cells are grown in media containing amino acids, vitamins, calf serum, and antibiotics.
ο A monolayer or sheet of cells is formed on the flat side of the container within a few days. Viruses are inoculated on the monolayer (we notice the change on the cells done by the virus)
b) Embryonated egg.
c) Animal inoculation.
How are viruses serological detected in the lab?
By serological methods (ELISA, RIAβ¦).
How is the prevention of viruses?
Vaccination and public health measures.
How are viruses treated?
ο Antiviral drugs are medicines that cure or control virus infections.
ο antiviral agents tend to be narrow in the spectrum and have limited efficacy. (Unlike antibiotics)
What are the types of antiviral drugs?
1) Inhibitors of Herpesviruses
2) Inhibitors of Retroviruses
3) Inhibitors of other viruses
4) Interferon
What are examples of Inhibitors of Herpes viruses?
Nucleoside analogue : inhibits virus-specific DNA polymerase e.g. Acyclovir, Ganciclovir, and rhindisevir (for covid)
What are examples of Inhibitors of Retroviruses?
- Reverse transcriptase inhibitors
- protease inhibitors
- fuzeon
- integrate inhibitors
What is the function of reverse transcriptase inhibitors?
- inhibit reverse transcriptase of HIV
- e.g. Azidothymidine (AZT), dideoxyinosine and bcavar sulfate
What is the function of protease inhibitors
- saquinavir and indinavir for treatment of HIV
What is the action of fuzeon?
- blocks the viral and cellular membrane fusion step involved in the entry of HIV into the cell
What is the action of integrase inhibitors?
- inhibit integrase enzyme of the virus
Give examples for inhibitors of other viruses
- amantadine
- zanamivir and oseltamivir
- ribavirin
What is the function of amantadine?
inhibits Influenza A virus uncoating
What is the function of zanamivir and oseltamivir?
inhibit viral neuraminidase of influenza A and B viruses (inhibit viral release from an infected cell to other cells
What is the function of ribavirin?
inhibits both DNA and RNA polymerase enzymes. Used for treatment of HBV, HCV infections, and RSV pneumonitis
How are interferons produced?
Infection of cells with viruses induces the production of proteins that are known as interferons because they were found to interfere with viral replication in previously uninfected tissue culture cells.
What are the types of interferons?
There are 3 types of interferons:
- interferon-Ξ± (IFN-Ξ±)
- interferon -Ξ² (IFN-Ξ²),
- interferon-Ξ³ (IFN-Ξ³), which is induced by activated T cells.
What is the mechanism of action of interferons?
- IFN-Ξ± and IFN-Ξ² are secreted by the infected cell and then bind to a common cell-surface receptor, known as the interferon receptor, on both the infected cell and nearby cells.
- Interferon induces the synthesis of several host cell proteins e.g. RNA dependent protein kinase (PKR) that contributes to the inhibition of viral replication
What is the definition of the genome?
It is the total genetic information in an organism.
What are the characters of the prokaryotic genome?
ο Consists of a single copy (Haploid) circular DNA molecule.
ο Range from 580-4600 Kbp
ο Many bacteria contain extrachromosomal DNA materials as a part of the genome called plasmids and transposons.
What is the definition of bacterial extrachromosomal elements?
- These are the DNA material present in a cell other than chromosomal DNA
- The most famous extrachromosomal DNA are:
A. Plasmid.
B. Transposons.
C. Bacteriophage (virus infecting bacteria)
What is the definition of plasmids?
Plasmids are pieces of DNA that exist separate from the chromosome, they contain an origin of replication so they replicate independently.
What are plasmids classified according to?
1- According to the size of the plasmid 2- According to copy number 3- Shape of plasmid 4- Moving plasmid from cell to cell 5- Artificial and natural plasmids 6- according to host range 7- according to compatibility
What are plasmids classified according to size?
Starting from a few hundred base pairs up to 3000 Kbp.
What are plasmids classified into according to copy number per cell?
1- Stringent plasmids
ο 1-2 copies / cell
ο like F- plasmid and phage β plasmid
hybrid (P1)
2- Low copy number plasmids
ο 10-15/copies/cell
ο such as pSC 101
3- High copy number plasmid
ο up to 50 copies/cell
ο like ColE, plasmid
4- Extremely high copy number plasmid
ο these are specifically engineered to be up to 100-200 copies/cell
What are plasmids classified into according to the compatibility of plasmids?
Compatible plasmids:
β’ a cell can maintain more than one plasmid in the same cell (if they carry
different origins of replication).
Incompatible plasmids:
β’ The inability of two plasmids to be maintained in the same cell (if they carry the same origin of replication
What are plasmids classified into according to the shape?
The shape of plasmids can be classified into 3 groups
1- Covalently closed circular (CCC) form:
ο The most common form
ο Present as ds completely closed circular forms (as in E.coli).
2- Semicircular form:
ο Transient form
ο Present as one strand is completely closed, the other strand is opened.
3- Linear:
ο unstable because it is attacked by exonucleases.
What are plasmids classified into according to host range?
1- Broad host range plasmids:
β’ Can replicate in a wide range of bacteria.
2- Narrow host range plasmids:
β’ only replicate in one or a few closely related bacteria.
What are plasmids classified into according to moving from one cell to another?
Conjugative plasmids: which have the tra genes that can mobilize the plasmid from one cell to another by conjugation
Shuttle vectors: plasmids that propagate in 2 diff. Hosts species (yeast and bacteria)
Nonβconjugative plasmids: Cannot be mobilized under any known conditions
What are plasmids classified into according to naturallity or artificiality?
1- natural plasmids:
β’ All the above plasmids are present naturally in bacterial and some yeast cells.
2- Artificial plasmids :
β’ are naturally present plasmid but designed artificially
β’ to be used in genetic cloning as vectors
β’ by adding antibiotic-resistant markers or DNA sequences to be the target of
restriction endonucleases.
What is the importance of plasmids?
- Resistance:
ο Antibiotic resistance.
ο Heavy metals (metal reductase).
ο U/V (DNA repair enzymes). - Conjugation.
- Production: Toxins & enzymes and bacteriocin.
- Biochemical reactions: Sugar fermentation.
- Molecular biology: As a vector.
ο Cloning vectors
ο Gene therapy: These are plasmids used for the insertion of therapeutic genes to express the protein that is lacking in the cells
What are transposons?
Extra-chromosomal small pieces of DNA those are capable of moving themselves from one location in DNA to another, (movable elements or jumping genes).
What are the 3 forms of transposons?
(a) Insertion sequence (IS)
(b) Composite transposons (Tn)
(C) Non β composite transposons
What are the characters of the insertion sequence?
ο The simplest form.
ο They encode only proteins needed for their own transposition
ο Carry repeated nucleotides at their ends (direct repeats or inverted repeats ~ (15-25).
ο Examples: IS 1, 3, and 10.