Microbiology Flashcards
What is microbiology?
Microbiology is the study of all living organisms that are too small to be visible to the naked eye.
Who is the father of microbiology?
Antonie van Leeuwenhoek
What are the four major groups of Human Pathogens, and what are they?
-Protozoa: Single-celled animals.
-Fungi: Higher plant-like organisms.
-Bacteria: Generally small, single-celled.
-Viruses: Very small obligate parasites, non-living.
Which Human pathogens are eukaryotes and prokaryotes?
- Eukaryotes: Protozoa and Fungi.
-Prokaryotes: Bacteria.
What are the 8 eukaryotes’ characteristics?
- Size 5-50 µms
- Complex (Compartmental)
- Frequently multi-cellular
- Linear chromosome + Histones
- Introns/Exons
- 80S Ribosomes
- No/Flexible Cell Wall (sterols)
- Cell cycle (mitosis/meiosis
What are the 9 prokaryotes’ characteristics?
- Size 0.5-10 µms
- Simple (relatively)
- Often single-celled
- Single Circular Chromosome
- Gene structure (introns rare)
- 70S Ribosomes
Co-transcription/translation - Rigid Cell Walls (PG)
- Rapid Cell cycle
What are the 10 characteristics of eukaryotic’s cell structure?
- Cell membrane
- Nucleus
- Centriole/Centrosome
- Nucleolus/Ribosomes
- Endoplasmic Reticulum (ER)
- Cytosol
- Mitochondria
- Golgi
- Cytoskeleton
- Secretory Vesicles, Lysosomes
What are the 10 characteristics of a prokaryotic’s cell structure?
- Plasma Membrane
- Cell Wall (Peptidoglycan)
- Nucleoid (DNA & associated proteins)
- Ribosomes
- Cytoplasm
- Capsule, Flagellar, Pili
Draw both Eukaryotic cells and Prokaryotic cells’ cell structure.
Check it.
What is the structure of the cytoplasmic membrane and its function?
Lipid bilayer which acts as a selective membrane. It retains the cytoplasm and transports molecules between the extracellular and intracellular environments.
What are bacteria sensitive to?
Disruption of the plasma membrane by physical or chemical methods.
What is a nucleoid?
The nucleoid is a chromatin-dense area within the cytoplasm and contains the bacterial DNA, associated proteins and RNA that are responsible for controlling the bacteria’s activity and reproduction
What is the Prokaryotic Protein Synthesis like? (6 steps)
- Gene (DNA)- Undergoes transcription
- mRNA (DNA-dependent RNA polymerase)
- Ribosome (tRNA) - Undergoes translation
- Protein
- 2nd, 3rd, 4th Structure
- Export/Assembly/Processing
What is the Peptidoglycan Cell Wall?
The peptidoglycan (murein) sacculus is a unique and essential structural element in the cell wall of most bacteria.
What is the structure of the Peptidoglycan Cell Wall like?
- There are continuous sugar chains: NAM-NAG
- To connect those chains, peptide side bridges are built by five amino acids.
What are the cell walls of Gram +ve like, and what is it a target of?
- Rigid layer
- Barrier
- Repeated Polysaccharide
structure - Gram +ve
- Multi-layer & often secondary
polymer - Target of Penicillin
What are the cell walls of Gram -ve like, and what is it a target of?
- Rigid layer
- Barrier
- Repeated Polysaccharide
structure - Gram–ve
- Outer membrane
- Often decorated with LPS
- Periplasmic space
- Target of Penicillin
What are lipopolysaccharides?
The primary function of LPS is to provide structural integrity and a permeability barrier to protect the bacterial cell from entering deleterious molecules such as toxins and bile salts during its inhabitation in the gastrointestinal tract.
What organisms (Gram-ve or Gram+ve) have Flagella & Fimbriae
Both
What are flagella?
Flagellum, plural flagella, hairlike structure that acts primarily as an organelle of locomotion in the cells of many living organisms.
What are fimbriae?
Fimbriae are long filamentous polymeric protein structures located at the surface of bacterial cells. They enable the bacteria to bind to specific receptor structures and colonize specific surfaces.
Draw the flagellar motor
From bottom to top:
-Series of rings
-Inner membrane
-Rod
-Peptidoglycan
-Outer membrane
-Hook
-Filament
What can flagella and fimbriae be for?
Adherence, sex and motility
What five things do prokaryotic cells need for growth?
- Food
- Temperature
- Hydrogen Ion Conc
- Osmotic protection
- Oxygen
What foods are required for Prokaryotic Growth?
-C source organic, e.g. proteins/sugars
inorganic, e.g. fix CO2
-O & H
-N source, eg. Amino acid Ammonia
-Inorganic salts P, S, K, Mg, Ca, Fe
-Trace Elements Zn, Cu, Mn, Ni, Mo,
-Vitamins (small organic cofactors)
e.g. Biotin Folic Acid, Niacin
What temperatures are required for Prokaryotic Growth? Depending on the three organisms?
- Psychrophiles (low temp)
- Thermophiles (high temp)
- Mesophiles (body temp)
What is the human commensals’ pH?
6.8-7.2
Regarding osmotic protection, what are the human commensals?
0.85% NaCl
What five prokaryotes need oxygen for growth?
Aerobes
Micro-aerophiles
Facultative anaerobes
Obligate Anaerobes
Capnophilic
Draw the Bacterial growth curve
- Lag Phase
- Exponential Phase
- Stationary Phase
- Decline Phase
What are the four classifications of Microorganisms?
- Appearance
(Shape; Size; Arrangement; Cell Wall; i.e. Gram-positive/Gram-negative) - Growth Requirements
(Aerobic/Anaerobic (e.g., strict anaerobes)
Requirement for Blood products (e.g. serum proteins)
Sensitivity to agents (e.g. NaCl, Bile, K tellurite)) - Enzyme/metabolic tests
(Coagulase test, Catalase test.
Hemolysis (Streptococci ONLY),
Biochemical profiling (e.g. carbohydrates metabolized)) - Molecular tests
(Immunological tests, e.g., cell surface antigens
DNA sequencing, e.g., qPCR or 16sRNA
Protein profiling e.g., Mass-spec analysis)
How are prokaryotic pathogens identified?
Direct observation under the microscope from a sample or cultured colony, usually fixed & stained.
What are the five shapes of bacteria?
- Cocci
- Bacilli
- Vibrio
- Spirilla
- Spirochaetes
Division in one plane for cocci produces…
Chains of 4-20 cocci. E.g. Streptococcus
Division in three planes for cocci produces…
Clumps. E.g., Staphylococcus
What are the two divisions of Bacillus/Bacilli?
-Rod-shaped bacteria
-Chains of bacilli
Where are chains of bacilli more common?
More common with Gram +ve bacilli
What is fusiform?
Long slender rods & species include emerging pathogens in the GI tract, mouth, head & neck.
Slightly curved rod: Gram+ve human pathogen, true or false?
FAAAAAAAAAAALSE.
Slightly curved rod: Gm –ve human pathogens
What are the two kinds of Spiral-shaped bacteria?
- Rigid spiral bacterium
- Flexible spiral bacterium
Example of Rigid spiral bacterium
Spirillum
3 Examples of Flexible spiral bacterium
Spirochaetes
Treponema & Borrelia
What are spores?
A spore is a cell produced by certain fungi, plants (moss, ferns), and bacteria. Spores are involved in reproduction. Certain bacteria make spores as a way to defend themselves. Spores have thick walls. They can resist high temperatures, humidity, and other environmental conditions.
What are toxins?
It is a poisonous substance that is a specific product of the metabolic activities of a living organism and is usually very unstable, notably toxic when introduced into the tissues, and typically capable of inducing antibody formation.
What do Gram+ stains like?
Crystal violet/iodine complex
What do Gram- stains like?
Pink
What are the limitations of Gram stain?
Not all organisms stain well with Gram stain
What two organisms do not stain well with Gram stain?
- Mycobacterium tuberculosis
- the organism that causes TB
- has a lipid-rich/waxy cell wall that does not take up the stain
- Treponema pallidum
- a spirochaete organism that causes syphilis, a sexually transmitted infection/disease
Meaning of aerobic
The organism grows in oxygen/air
Meaning of Obligate Aerobes
Require oxygen (in the laboratory)
Meaning of Obligate Anaerobes
Tolerate oxygen
Meaning of Capnophillic
Prefer high CO2 levels.
Microaerophillic low oxygen levels required.
What are the two selective media for the growth of specific prokaryotes?
- Mannitol Salt Agar (MSS)
-7.5% Salt allows preferential isolation of Staphylococci
-Mannitol fermentation produces acid & turns agar/colony yellow - Salmonella-Shigella (SS)
-bile salts inhibit coliforms
Why is selective media important?
Because the presence of specific substances permits the growth of one organism over another.
What is differential media?
The incorporation of chemicals produces visible changes in colonies that facilitate
identification (differentiation)
What are the two differential media?
- MacConkay Agar
- bile salts & crystal violet
- Lactose only C source & neutral red indicates fermentation.
- facilitate identification of Enterobacteriaceae.
-Eosin & Methylene Blue (EMB) - lactose salts & two dyes
- allows identification of lactose fermenters, e.g., E.coli
What are the three colony characteristics?
- Form
- Elevatiom
- Margin
What are the 6 characteristics in form?
- Punctiform
- Circular
- Filamentous
- Irregular
- Rhizoid
- Spindle
What are the five characteristics of elevation?
- Flat
- Raised
- Convex
- Pulvinate
- Umbonate
What are the 6 characteristics of margin?
- Entire
- Undulate
- Lobate
- Erose
- Filamentous
- Curled
What are the three categories of the identification of Streptococcus spp?
Alpha (partial), Beta (complete) & Gamma (no hemolysis)
What are the Lancefield Groupings?
A, B, C, F, G
Lancefield grouping is a system of classification that classifies catalase-negative Gram-positive cocci based on the carbohydrate composition of bacterial antigens found on their cell walls.
What do you do after Lancefield Grouping?
You add the size:
- Pyogenic (Large)
- S. milleri (Small)
What 2 branches does the biochemical characterization divide?
- Metabolic Profiling
* Utilisation of Carbon sources; (acid production)
* Utilisation of amino acids; - Exo-Enzyme production
* Catalase (2H2O2 to 2H2O & O2)
* Coagulase (clot plasma)
S. aureus + ve
* Hydrolysis of Lipid Clostridia species
* Urease (Urea converted to ammonia & CO2)
How do you carry out the identification
- Inoculate/Resuspend a Single colony
- Transfer 100 µls into automated biochemical profiling.
What are serological tests for?
This section analyses blood samples for the presence of viral antigens and antibodies (IgG and IgM) to help diagnose diseases and check immune status.
What is the process of identification in serological tests? (6 steps)
- Host immune response to the antigen by raising antibodies.
- Check if the antibody is specific to microbe/virus (Polyclonal) or single component (monoclonal)
- Detect presence of specific IgM antibody to virus/microbe
- Demonstrate IN VITRO by agglutination reaction
- It has a rapid detection of viruses (24 hours)
- It can identify specific serotypes of bacteria.
What is the key concept in serology?
Antibody/Antigen interaction
What is the detection of viruses in serological tests like?
It is rapid, within 24 hours.
How can it be possible to identify the stage of infection?
Using two different tests.
What is agglutination, and why is it important?
This is the immune process where antibodies bind to multiple pathogens forming clumps. Enhancing phagocytosis and preventing the pathogens from infecting other host cells.
What is genome sequencing, and what is its function?
Scientists use a process called genomic sequencing to decipher the genetic material found in an organism or virus. Sequences from specimens can be compared to help scientists track the spread of a virus, how it is changing, and how those changes may affect public health.
What are the other three pieces of DNA technology and their definitions?
-qPCR: qPCR stands for quantitative polymerase chain reaction and is a technology used for measuring DNA using PCR.
-Next-generation sequencing: It’s a massively parallel sequencing technology that offers ultra-high throughput, scalability, and speed. The technology determines the order of nucleotides in entire genomes or targeted regions of DNA or RNA.
-Nanopore sequencing: is a unique, scalable technology that enables direct, real-time analysis of long DNA or RNA fragments. It works by monitoring changes to an electrical current as nucleic acids are passed through a protein nanopore. The resulting signal is decoded to provide the specific DNA or RNA sequence.
What can successful amplification of DNA
target indicate?
The presence of an organism or even specific
virulence factor
What is 16sRNA?
The 16S rRNA gene encodes the small subunit ribosomal RNA molecules of ribosomes, responsible for the essential process of converting genetic messages to functional cell components via the translation of mRNA to proteins.
What is MALDI-TOF?
Matrix-assisted laser desorption/ionization time of flight.
Mass spectrometry (MS) has become a widely used technique for rapidly and accurately identifying bacteria, mycobacteria, and specific fungal pathogens in the clinical microbiology laboratory.