Microbiology for dentists theme 1 Flashcards
What are examples of microorgansims ?
bacteria viruses archaea fungi protozoa
What are the origins of life ?
4 bn years ago cellular life came into being
2 bn years ago modern eukaryotes existed
0.5 bn years ago multicellular life evolved
What is evolution ?
the process of change over time which results in new species
What is phylogeny ?
evolutionary relationships between organisms
How is phylogeny determined ?
genetic relationhips- looking at the rRNA gene
Why is the rRNA gene used ?
present in all organisns
shows a high degree of variability
What is the process for determining phylogeny ?
isolate DNA
Make copies of the rRNA gene
sequence the DNA
analyse and compare
Which microorganisms dominate the tree of life ?
bacteria
arachaea
eukarya
What are example of eukarya ?
protozoa
animals
fungi
plants
What are archaea ?
they diverged from bacteria
they have some similarities to eukarya - similar DNA replication
correlation with periodontal disease
What are protozoa ?
single celled eukaryotes
found in aquatic environments- dental unit waterlines
graze on bacteria
What is an example of protozoa ?
plasmodium- malaria
What are fungi ?
simple eukaryotes
make multicellular structures
grow as branched tubes
What is the fungi cell wall made of ?
chitin
What are viruses ?
obligate intracellular parasites
acellular
nucleic acid in a protein coat
evovle rapidly
What are prions ?
proteinaceous infectious agents
proteins that have changed conformation from an alpha helix to a beta pleated sheet
they are autocatalytic- can cause further conformational changes
leads to degeneration in brain function
extremely resistant to heat
How can prions be transmitted ?
contamianted food
hormone treatments
surgical instruments
How is a prion formed ?
normal protein adopts a misfolded protein shape
prion binds to normal proteins
leads to the conversion of more protesins
What do prions aggregate to form ?
amyloid fibrils
How are amyloid fibrils made ?
from the stacking of beta pleated sheets
What do amyloid fibrils do ?
disrupt cell function
cause neuro-degenerative disorders
What type of neuro-degenerative disorders do prions cause ?
transmissable spongiform encephalopathies
What are prions resistant to ?
heat resistant
protease resistant
What are the 3 categories of prion disease ?
spontaneous
acquired
inherited
What is an example of a spontaneous prion disease ?
sporadic CJD
What is an example of inherited prion disease ?
familial CJD
What is an example of acquired prion disease ?
Kuru
Why do prions have long incubation periods ?
they replicate slowly
What property of prions allows them to form plaques ?
theyre insoluble
How is neuronal tissue death via prions visible ?
spongiform pathology
What is the relevance of prions to dentistry ?
dental tissues are considered low risk
could be iatrogenic transmission
difficult to destroy- essential to autoclave instruments and use single-use instruments
What is a major problem with prions ?
they cant be detected as they have no assay
What is the basic structure of a virus ?
a nucleic acid and a protein capsid coat
What are the 2 types of proteins ?
enveloped and non-enveloped
What are enveloped viruses ?
they consist of the nucleic acid and the protein capsid and the lipid layer which is embedded with glycoproteins
What are the 3 types of viral genome shapes ?
linear
circular
segmented
What are the 2 types of nucleic acid ?
DNA and RNA
single stranded or double stranded
What is the purpose of the viral genome ?
to encode for proteins- structural, proteases and enzymes
Which enzymes do viruses need ?
reverse transcriptase
DNA polymerase
RNA polymerase
What is the mode of action for a virus ?
- genome codes for proagation proteins
- Infection of the host
- Genome replication
- Invasion of immune system
Which types of virues are most likely to cause disease ?
Single stranded RNA
Double stranded DNA
What is the life cycle of a DNA virus ?
phagocyte brings virus virus infects epithelial cell harnesses replication machinery trasncribe into RNA RNA is translated into viral proteins lysis - pathology
What is the life cycle of an RNA virus ?
viruses contain reverse transcriptase- convert viral RNA into DNA
Replicate DNA
Integration into the genome
transcribed and translated
more RT Made, capsid proteins and envelope proteins to assemble new viruses
What is persistence ?
some proviruses will insert into the viral chromosome and remain there- evade the immune system - latent
What is an example of an RNA enveloped virus ?
HIV
What is an example of an RNA non-enveloped virus ?
Rhino
What is an example of a DNA enveloped virus ?
Herpes
What is an example of a DNA non-enveloped virus ?
HPV
What are some viruses that are systematic but cna have oral effects ?
MMR
What are the oral signs of measles ?
rash-kopliks spots
What are the oral signs of mumps ?
salivary gland inflammation
What are the oral signs of rubella ?
Rash
What are some viral infections of the mouth ?
hand, foot and mouth disease
Herpes
HIV
What is hand, foot and mouth disease ?
enterovirus that causes oral blisters
What is Herpes ?
Large class of structurally similar viruses
What is HPV ?
linked to oral cancer
causes papillomas of the oral mucosa and the genitals
Which HPV are associated with HPV ?
16 and 18
Which HPV are restricted to the oral cavity ?
13 and 32
What does HIV cause ?
AIDS
What is HIV Associated with ?
oral candidosis
NUG
Which cells does HSV 1 infect ?
mucosal epithelial cells
neuronal cells
How is HSV 1 latent ?
hides in the immune system until reactivated
What does reactivated HSV1 cause ?
stomatitis
How is HSV1 transmitted ?
through saliva
What are the factors that lead to NUG ?
stress
smoking
HPV
What plaque control methods are used for NUG ?
ultrasonic
chlorohexidine
metronidiazole
OHI
What are the symptoms of HSV1 ?
cold sores
What are the symptoms of Varicella-zoster ?
child- chicken pox
adult-shingles
What are the symptoms of epstein barr ?
Glandular fever
What are the symptoms of cytomegalovirus ?
congential defects
What are the symptoms of HHV8?
Kaposis sarcoma-affects AIDS patients
How is varicella - zoster virus trasnmitted ?
aerosols and direct contact
How do shingles appear ?
in the facial nerve tract
Why arent kids vaccinated against chicken pox ?
because its mild
also vaccinating kids reduced the boosting of immunity ex-chicken pox adults get from being exposed to chicken pox kids
How is Hepatitis B transmitted ?
blood saliva STI
What are the severe effects of Hepatitis B ?
hepatocellular carcinoma
Is Hep B present in the mouth ?
it is present in the mouth but does not cause oral disease
How can you look for viral infection ?
clinical apperance
lab tests- ELISA and sero conversion to look for antibodies
viral culture
histology from a biopsy
How can we prevent viral infections ?
cross infection control
vaccines
Which drugs are used in anti-viral healthcare ?
acyclovis
What are the characteristics of prokaryotes ?
no membrane bound organelles
haploid
no fusion of gametes
Why are fungi described as chemoorganotrophs ?
secrete extracellular enzymes which digest organic material
What are hyphae ?
network of filaments of fungi
What are mycelium ?
they grow over surfaces in a network
What is the fungi cell wall made of ?
chitin
What is different about fungi cell membranes ?
they contain ergestrol rather than cholesterol
What colour is the gram stain in GP bacteria and why ?
Purple
thick peptidoglycan layer able to retain the crystal-iodide complex
What are flagella ?
rotating protein motors that allow motility
made of flagellin sub units
What is rotation in flagella couple to ?
flow of protons across the membrane
What is the method for the gram stain ?
create a heat fixed smear flood with crystal violet add iodide complex decolourise with alcohol counterstain with safarnin
What are acid-fast bacterial cell walls ?
very high lipid/wax content
contain mycolic acids
found in mycobacterium TB
What is the method for detecting acid fast bacterial cell walls ?
detected with carbol fuschin
driven in with heat
destain with 3% alcohol
What are the cytoplasmic membranes of gram positive and gram negative cell walls ?
phospholipid bilayer
What is the structure of gram negative outer membranes ?
LPS
asymmetrical
What are fimbriae ?
protein filamentous structures
enable cells to adhere to surfaces
aid in biofilm production
What are pili ?
used in conjugation- DNA transfer
used in adhesion
found in streptococus pyogenes
type 4 have a twitching motility
What are capsules made of ?
polysaccharides and the proteins
What is the purpose of the capsule ?
aid adherence - biofilm production
evade phagocytosis
stop desiccation (protect against drying out )
What are cell inclusions ?
packets of energy resources
What is the purpose of cell inclusions ?
safe way to store energy and avoid osmotic stress
What are some examples of cell inclusions ?
glycogen
poly-B-hydroxybutyric acid
What are endospores ?
certain bacteria can only form them
highly differentiated cells that are very resistant
How are endospores destroyed ?
by autoclaving
What is an examples of endospores ?
clostridium dificil
arent killed by antibiotics
What do bacteria require to grow ?
they consume and produce energy-
they require macronutrients and micronutrients
water
trace elements
What are heterotrophs ?
obtain carbon from organic material
What are autotrophs ?
obtain carbon from C02
What are auxotrophs ?
cant synthesis organic compounds like amino acids
What is fermentation ?
an anaerobic process
organic compound used as the electron acceptor and donor
What is respiration ?
aerobic process
electron donor is oxidised with oxygen
Where do fermentation and respiration happen ?
in both humans and bacteria
What is homolactic fermentation ?
fermentation of hexose sugars leads to 2 lactate and 2 H+
What is the relevance of lactic acid ?
major product found in dental caries
Which bacteria in plaque ferment to produce lactic acid ?
streptococus mutans
What is the first step in both respiration and fermentation ?
glycolysis
How is ATP produced in fermentation ?
substrate level phosphorylation
How is ATP produced in respiration ?
oxidative phosphorylation
electron transport chain
How much ATP is made in aerobic respiration ?
38 ATP
included glycolysis and the citric acid cycle
What is the purpose of the citric acid cycle ?
to produce reducing agents
What protein is needed for ATP synthesis in respiration ?
FOFI ATPase
What is FOFI ATPase ?
A highly abundant proten growing in bacterial cell walls
How does FOFI ATPase generate ATP ?
flow of protons into cells
How do Oral bacteria use FOFI ATPase ?
streptococus mutans uses ATPase to pump protons out of cells when conditions are too acidic to reduce the acidity
What is the definition of bacterial growth ?
irreversible increase in biomass and usually numbers of bacteria
How can we measure bacterial growth ?
microscopy
colony forming units
absorbance
growth curves
What is the process of bacterial cell division ?
binary fission DNA replication cell elongation septum formation distinct cells cell separation distinct cells
How is bacterial growth described ?
exponential
number of cells doubles during a constant time
What is the risk of such a high increase in a short time ?
increases the risk of pathogenic bacteria
What are the phases of bacterial growth ?
exponential
ststionary
death
What are the 3 methods used for measuring bacterial growth ?
microscopic counts
viable and serial dilutions
tubidimetric methods
Why is e.coli used as a lab model organism ?
more is known about it than any organism
grows rapidly and has simple nutritional requirements
established genetics and genome
What is the e.coli genome like ?
circular chromosome made of 4.6 million base pairs
encodes around 4400 base pairs
What is an operon ?
genes encoding enzymes in a single biochemical pathway
What are plasmids ?
small circular pieces of DNA found in varying numbers in the cell
What types of genes do chromosomes carry ?
genes that are useful but arent essential for survival
eg. antibiotic resistance
How can genetic information be exchanged by plasmids ?
bacteria die and release plasmids that can be absorbed by bacteria
What are the 3 types of horizontal gene transfer ?
conjugation
transformation
transduction
What does horizontal gene transfer have the ability to do ?
ability to change the genotype and the phenotype of an organism
make the bacteria “fitter”- multiply faster or give resistance
What is DNA transformation ?
DNA uptake freely from the environment
What is transduction ?
bacteriophage mediated
bacteriophage will infect a bacterial cell and take up the host DNA
it will then inject the DNA into another cell and then integrate it into the chromosome - donor DNA will take up new functions
What is conjugation ?
pilus mediated
pilus extends from the donor to the recipient and adheres
plasmids are transported down the pillus
How is DNA incorporated into the chromosome of the recipient ?
Homologous recombination
Give an example of transformation using tryptophan ?
e.coli strains that lacka a tryptophan pathway grow on an a tryptophan lacking agar- no growth
take DNA from tryptophan from tryptophan positive cells - cells will uptake DNA
grow on tryptophan agar- see visible growth
What are the goals of genetic manipulation ?
knock out genes
change individual amino acids in a protein
FISH
produce recombinant proteins like insulin
What are the steps in Recombinant DNA technology ?
Creation of the recombinant DNA
Cloning of the DNA in an organism
Using the DNA - express the gene
How can we clone recombinant DNA ?
amplify the DNA by introducing it into a living cell which will produce many identical copies
What are the tools for Recombinant DNA technology and why ?
enzymes- to cut DNA and RNA
vectors - act as a vehicle to transport the DNA into host cell for replication
cells- Amplify and express the DNA
DNA and RNA - Raw genetic material
Which enzymes are needed in recombinant DNA technology ?
Restriction enzymes
SNA Ligase
Taq polymerase
Reverse transcriptase
What do restriction enzymes do and give an example ?
cleave and cut DNA at very specific sequences- molecular scissors
eg. Restriction endonuclease
What does DNA ligase do ?
stick DNA fragments together
What does Taq polymerase do ?
used in PCR- creates multiple copies of the DNA fragment
What does reverse transcriptase do ?
convert RNA into DNA
How are restriction enzymes produced and whyn ?
naturally produced by bacteria to cleave bacteriophage DNA
Which sequences do restriction enzymes recognise and what do they produce ?
recognise 4-8 base pairs in a palindromic sequence
produce either sticky or blunt ends
What are sticky ends ?
they are overhanging at either the 5’ or 3’ end
compatible overhangings are annealed with DNA ligase
What do vectorrs contain that allows the insertion of the new recombinant DNA ?
restriction sites so DNA can be efficiently introduced into host DNA
How can we check which cells have taken up the DNA ?
the plasmid will have an antibiotic resistance gene - if the cell has taken up the plasmid the gene will be expressed and the cells will be able to be grown on agar containing penicillin
What do vectors contain to allow for expression of the inserted gene ?
regulatory sequences
How do plasmids replicate ?
independent of the bacterial chromosome
What do plasmids have that allows recognition by the host replication machinery ?
origin of repication
What is the polylinker ?
a piece of synthetic DNA that allows the insertion of new gene
What does the polylinker contain ?
lacZ gene which codes for the B-galactosidase enzyme
which turns a dye from blue to white
If the polylinker is disrupted - gene is successfully added- blue colonies
What are other example of vectors beside plasmids ?
phagemids
cosmids
bacteriophages
What are cosmids and phagemids ?
genetically engineered hybrids which replicate as a plasmid but can be packaged as a bacteriophage
How is the recombinant plasmid inserted into the cells ?
via transformation- either by electroporation or using Calcium chloride heat shock, 42 degrees for 3 mins
Where does each cell in a colony arise from ?
a single e.coli cell
each cell in the colony will contain copies of the same plasmid - clones
What were the disadvantages of using amimal insulin ?
side effects
difficult to purify
autoimmune reactions
How is recombinant insulin made ?
the gene for proinsulin has 2 introns - DNA is first converted to mRNA
then reverse transcribed into DNA to get rid of introns
the gene is inserted into a plasmid
plasmid inserted into cell and cloned in E.coli
Examples of other proteins made using recombinant technology ?
Erythropoietin
Tpa
How can recombinant DNA be applied dentally ?
recombinant DNA can be used to manipulate genes
create a genetically altered strain of S.mutans that doesnt produce lactic acid
What is a planktonic solution ?
a solution of cells that are homogenous , single species and nutrient rich
What is a surface associated collection ?
they have limited hetrogeneity
single species
nutrient rich
What are natural collections of bacteria ?
in the from of a structured community with organic matter and active microbes
heterogenous
mixed species
nutrient limited (gradient of nutrients)