Microbiology Flashcards
What is a pathogen?
Organism that causes disease
What is a commensal?
Organism that colonises host but causes no disease normally
What is an opportunist pathogen?
Microbe that only causes disease if host defence is compromised
What is virulence/pathogenicity?
The degree to which an organism is pathogenic
What is the first stage of gram staining?
Fixation of clinical materials to microscope slide (heat/methanol)
What happens in gram staining after fixation?
Application of primary stain: crystal violet (all cells turn purple)
What happens after the primary stain in gram staining?
Application of mordant (iodine): crystal violet-iodine complex formed
What happens after application of mordant in gram staining?
Decolourisation step: distinguishes gram +ve and gram -ve, use acetone or ethanol
What happens after decolourisation in gram staining?
Application of counterstain: safranin to stain gram -ve pink
What do gram positive bacteria look like after gram staining?
Stains purple
What do gram negative bacteria look like after gram staining?
Stains pink
How can you remember the gram staining method?
“Come In and Stain” Crystal violet Iodine Alcohol Safranin
What is the difference between gram positive and gram negative bacteria?
Gram positive: thick layer of peptidoglycan (cell wall)
Gram negative: thin layer of peptidoglycan (high lipid content)
What are the 2 different types of gram positive cocci?
Staphylococcus and streptococcus
What are the characteristics or staphylococcus?
Clusters/catalase +ve
What are the characteristics of streptococcus?
Chains/catalase -ve
Give some examples of gram positive rods.
Corneybacteria, Mycobacteria, Listeria, Bacillus, Nocardi
What test can be done to determine the type of staphylococcus?
Coagulase test
What bacteria is it if it tests positive after a coagulase test?
S.aureus or MRSA
What bacteria is it if it tests negative after a coagulase test?
Coagulase negative staph (S.epidermidis/S.saphrophiticus)
What is coagulase?
Enzyme produced by S.aureus that converts soluble fibrinogen in plasma to insoluble fibrin
What test can be done to determine the type of streptococcus?
Haemolysis on blood agar
What does alpha haemolysis indicate?
Alpha haemolytic strep
What does beta haemolysis indicate?
Lancefield A, B, C + G - S.pyogenes
What does gamma haemolysis indicate?
Lancefield D (S.bovis/enterococcus)
What does alpha haemolysis look like?
Partial lysis (greening)
What does beta haemolysis look like?
Complete lysis (clear)
What does gamma haemolysis look like?
No lysis
What is the haemolysis test?
Uses hydrogen peroxide to test reaction with haemoglobin
Give some examples of alpha haemolytic chains.
S.pneumoniae
Viridans group streptococci
Give some examples of beta haemolytic chains.
Group A strep (S.pyogenes)
Group B strep (S.agalactiae)
Group G strep (S.dysgalactiae)
Give examples of gamma (non) haemolytic chains.
Group D strep (S.bovis, enterococcus)
What infections are associated with S.aureus?
Impetigo, boils, cellulitis, endocarditis, toxic shock syndrome (skin infections)
What infections are associated with S.epidermidis?
Surgical wound infections, septicaemia, endocarditis
What infections are associated with S.saphrophiticus?
Acute cystitis
What infections are associated with S.pneumoniae?
Pneumonia, otitis media, sinusitis, meningitis
What infections are associated with viridans group strep?
Oral strep, deep organ abscesses
What infections are associated with group A strep (S.pyogenes)
Cellulitis, tonsillitis, impetigo, scarlet fever, pharyngitis (throat + skin infections)
What infections are associated with group B strep (S.agalactiae)?
Postpartum infection, neonatal sepsis, neonatal meningitis
What infections are associated with group G strep (S.dysgalactiae)
Severe mastitis, rheumatic fever, throat infections
What infections are associated with S.bovis?
Endocarditis, UTIs, sepsis (colorectal)
What infections are associated with enterococcus?
Endocarditis, UTIs, intra-abdominal + pelvic infections
Where is S.aureus found?
Nasal passages + skin
Where is S.epidermidis found?
Skin
Where is S.saphrophiticus found?
Female genital tract + perineum
Where is S.pneumoniae found?
Nasopharynx (+ rest of upper resp tract)
Where is viridans group strep found?
Oral cavity mostly, also upper resp tract, female genital tract + GI tract
Where is S.pyogenes found?
Resp tract
Where is S.agalactiae found?
Lower GI tract + female genital tract
Where is S.dysgalactiae found?
GI tract + genital tract
Where is S.bovis found?
GI tract
Where is enterococcus found?
GI tract
What antibiotics can be used to treat S.aureus?
Flucloxacillin, Co-Amoxiclav, Macrolides (susceptible to most)
What antibiotics can be used to treat MRSA?
Gentamicin, Vancomycin
What antibiotics can be used to treat S.epidermidis?
Co-Amoxiclav
What antibiotics can be used to treat S.saphrophiticus?
Nitrofurantoin, trimethoprim
What antibiotics can be used to treat S.pneumoniae?
Oral Amoxicillin, IV Benzylpenicillin
What antibiotics can be used to treat viridans group strep?
Amoxicillin
What antibiotics can be used to treat S.pyogenes?
Amoxicillin (any penicillins)
What antibiotics can be used to treat S.agalactiae?
Amoxicillin
What antibiotics can be used to treat S.dysgalactiae?
Amoxicillin
What antibiotics can be used to treat S.bovis?
Amoxicillin
What antibiotics can be used to treat enterococcus?
Amoxicillin
What antibiotics do beta haemolytic strep respond to?
Any penicillin (amox, co-amox, fluclox etc)
Who is most vulnerable to S.epidermidis?
Immunocompromised patients
What percentage of cystitis infections are caused by S.saphrophiticus?
5-8%, most commonly in young women
What is the most virulent viridans group strep?
S.milleri
What is the most commone cause of bacterial infections in newborn babies?
S.agalactiae
What is group A strep?
S.pyogenes
What is group B strep?
S.agalactiae
What is group G strep?
S.dysgalactiae
What is group D strep?
S.bovis, enterococcus
What test is used to determine the type of gram negative bacillus?
Appearance on MacConkey agar
What does a pink appearance on a MacConkey agar suggest?
Lactose fermenting: E.coli, klebsiella
What does a white appearance on a MacConkey agar suggest?
Non-lactose fermenting: shigella, salmonella, pseudomonas, proteus
How is the type of lactose fermenting bacteria determined?
Biochemical identification (API strip) + sensitivity tests
How is the type of non-lactose fermenting bacteria determined?
Oxidase test
What does a positive oxidase test indicate?
Pseudomonas
What does a negative oxidase test indicate?
Shigella, salmonella, proteus
What does a positive oxidase test look like?
Purple
What does a negative oxidase test look like?
Cream coloured
What is an oxidase test used for?
To determine if a bacterium produces certain cytochrome c oxidases
Give some examples of gram negative cocci.
Neisseria (gonorrhoea), Moraxella, anaerobic cocci - Veillonella
Give some examples of lactose fermenting bacteria.
E.coli
Klebsiella
Give some examples of non-lactose fermenting bacteria.
Pseudomonas
Shigella
Salmonella
Proteus
What are the 2 types on non-lactose fermenting bacteria?
+ve and -ve oxidase
What infections are associated with E.coli?
UTIs, traveller’s diarrhoea, cholecystitis, cholangitis
What infections are associated with klebsiella?
Pneumonia, meningitis, surgical wound infections
What infections are associated with pseudomonas?
Pneumonia
What infections are associated with shigella?
Shigellosis (diarrhoea, fever)
What infections are associated with salmonella?
Salmonellosis (diarrhoea)
What infections are associated with proteus?
UTIs, related to stone formation
What infections are associated with neisseria?
Gonorrhoea, meningitis
Where is E.coli found?
GI tract
Where is klebsiella found?
GI tract
Where is pseudomonas found?
GI tract
Where is shigella found?
N/A (from water)
Where is salmonella found?
Intestines (from raw meats, poultry, eggs)
Where is proteus found?
Intestines
What antibiotics are used to treat E.coli?
Co-amoxiclav, Trimethoprim, Nitrofurantoin
What antibiotics are used to treat pseudomonas?
Tazocin, Gentamicin, Quinolones
How is klebsiella treated?
Do sensitivity testing
What antibiotics are used to treat shigella?
Quinolones, Azithromycin
What antibiotics are used to treat salmonella?
Amoxicillin, Quinolones, Macrolides
What antibiotics are used to treat proteus?
Penicillins, cephalosporins
What antibiotics are used to treat neisseria?
Cephalosporins
What bacterium causes the majority of UTIs?
E.coli
Why is klebsiella hard to treat?
Some strains highly resistant
How does pseudomonas present?
Do not usually cause infections in healthy people - if so infection is mild
What is the pathology behind meningitis?
Inflammation of the pia + arachnoid mater - microorganisms infect the CSF
Not always an infective cause - can have chemical/ post-surgical etc
What are the symptoms of meningitis?
Stiffness of the neck, photophobia + severe headache
Infective: fever, malaise
Petechial rash associated with meningococcal meningitis
What causes bacterial meningitis in neonates?
E.coli (-ve), Group B Strep (+ve), Listeria monocytogenes (+ve)
What causes bacterial meningitis in infants?
Neisseria meningitidis (-ve), Haemophilus influenzae (-ve), S.pneumoniae (+ve)
What causes bacterial meningitis in young adults?
Neisseria meningitidis (-ve), S.pneumoniae (+ve)
What causes bacterial meningitis in the elderly?
Neisseria meningitidis (-ve), S.pneumoniae (+ve), Listeria monocytogenes (+ve)
What are the 2 most common causes of bacterial meningitis?
N.meningitidis (-ve) + S.pneumoniae (+ve) = diplococci
What viruses can cause meningitis?
Mumps virus, echo virus, coxsackie virus, herpes simplex virus, poliovirus
What drugs can induce meningitis?
NSAIDs, trimethoprim
How is meningitis diagnosed?
Blood culture
CSF sample (lumbar puncture at L4) for microscopy + sensitivity testing
Nose + throat swabs for viral
What does a CSF sample look like that indicates bacterial meningitis?
Turbid yellow colour, neutrophil polymorphs, raised protein, low glucose
What does a CSF sample look like that indicates viral meningitis?
Lymphocytes, normal protein, normal glucose
What does a CSF sample look like that indicates tuberculous meningitis?
Lymphocytes, raised protein, low/normal glucose
How is bacterial meningitis treated?
Start antibiotics before tests come back if suspected
Cephalosporins: IV cefotaxime/ IV ceftriaxone
If over 50/immunocompromised add IV amoxicillin to cover listeria
One dose oral ciprofloxacin - prophylaxis for contacts
How is meningococcal septicaemia treated?
Immediate IM benzylpenicillin in community/ IV cefotaxime in hospital
How is viral meningitis treated?
Supportive treatment, self-limiting in 4-10 days, acyclovir for HSV meningitis
What antibiotics inhibit cell wall synthesis? What bacteria are they used to treat?
Glycopeptides and beta lactams - gram positive bacteria (have thick cell walls)
Give some examples of glycopeptide antibiotics.
Vancomycin and teicoplanin
What are the different categories of beta lactams?
Penicillins
Cephalosporins
Carbapenems
Give some examples of penicillins.
Benzylpenicillin
Amoxicillin
Flucloxacillin
Give some examples of cephalosporins.
Cephalexin
Cefotaxime
Ceftriaxone
Give some examples of carbapenems.
Imipenem
Estapenem
Which antibiotics inhibit protein synthesis?
Cholaramphenicol
Macrolides
Tetracyclines
Aminoglycosides
Give some examples of macrolides.
Clarithromycin
Erythromycin
Give some examples of tetracyclines.
Doxycycline
Give some examples of aminoglycosides.
Gentamicin
Streptomycin
When are macrolides used?
We generally use macrolides instead of penicillin for those with a penicillin allergy
Which antibiotics inhibit folate synthesis?
Trimethoprim
Sulphonamides - Sulphamethoxazole
Which antibiotics inhibit DNA gyrase?
Fluroquinolones - Ciprofloxacin
Which antibiotics bind to RNA polymerase?
Rifampicin
Which antibiotics cause DNA strand breaks?
Metronidazole
Why shouldn’t you give trimethoprim to a pregnant woman?
Folate is important during pregnancy to prevent spina bifida - inhibits folate synthesis
How common are mycobacteria in the UK?
Rare
Describe the characteristics of mycobacteria.
Aerobic, non-spore forming, non-mobile bacilli
How does a mycobacteria infection present?
Slow-growing causes gradual onset of disease
How is a mycobacteria infection treated?
Requires multi-antibiotic treatment for a prolonged period
What are the 2 main examples of mycobacteria?
TB and leprosy
What staining technique is used to identify mycobacteria?
Ziehl-Neelsen stain used instead of gram staining
What colour do acid-fast bacteria stain in Ziehl-Neelsen staining?
Red
What colour do non-acid fast bacteria stain in Ziehl-Neelsen staining?
Blue
How do viruses survive?
Dependent on host cells for living - have no organelles or cell wall
How do viruses cause disease?
Can cause disease via direct destruction (polio), modification (rotavirus), over-reactivity (hepatitis B), damage through cell proliferation (HPV)
Why are viruses difficult to identify?
Not visible in light microscopy, can’t culture as only reproduce inside live cells
How are viruses identified?
PCR + nucleic acid amplification tests (NAAT) used, can also do serology (look for antibodies in response to virus)
What will be the results of a serology test if there is a virus present?
IgM within 1 week of onset, IgG appears later
How are viral infections treated?
Viral infections are often self-resolving, in severe cases antivirals can be given ‘-vir’, eg acyclovir
What are protozoa?
Microscopic unicellular eukaryotes
How do protozoa survive?
Can be free living or parasitic
How are protozoa classified?
Ameoboids, ciliates, sporozoan, flagellates
Give some common examples of protozoa.
Malaria, giardia lamblia, toxoplasmiosis, trichomonas vaginalis
What type of organism are fungi?
Eukaryotes
Describe the structure of fungi.
Have a cell wall - chitin + glucans (polysaccharides)
How do fungi move?
Move by growing across or through structures or by dispersion in air/ water
What are the 2 forms that fungi exist in?
Yeast and moulds
What is a yeast?
Single cell that divide via budding
What is a mould?
Form multicellular hyphae or spores
How do fungi cause infections?
Only a few can actually cause infection in humans - eg candida, athlete’s foot, nappy rash
How are fungal infections treated?
Antifungals target cell wall/ plasma membrane - ‘-azole’
How common are worm infections?
Common worldwide but rare in UK
What are the 3 groups of worms?
Nematodes (roundworms), trematodes (flatworms), ceratodes (tapeworms)
How do worms reproduce?
Adult worms cannot replicate inside body without a period of development outside the body
What is the pre-patent period?
Interval between infection + appearance of worm eggs/larvae in stool
Describe the immune system’s response to worms.
Poor
Which antibodies are involved in worm infections?
Mainly IgG + IgE mediated
Give some examples of common worm infections.
Hookworm, schistosomiasis
What will be the result of the optochin test if the bacteria are sensitive to it?
Streptococci pneumoniae - clear zone of no growth around disc
What will be the result of the optochin test if the bacteria are resistant to it?
Viridans streptococci and other alpha haemolytic streptococci - there will be growth around the disc
What is the result of the oxidase test if oxidase positive?
Blue - bacteria is AEROBIC e.g. V. cholerae
What is the result of the oxidase test if oxidase negative?
No colour change - bacteria may be aerobic or anaerobic
What does XLD agar look like with salmonella?
Red/pink colonies some with black spots
What does an XLD agar look like with shigella?
Red/pink colonies
What are the 3 ways of classifying streptococci?
- Haemolysis
- Lancefield typing
- Biochemical properties
How are streptococci differentiated with haemolysis?
Alpha haemolysis - greenish/brownish e.g Strep. intermedius Beta haemolysis - clear/colourless e.g. Strep. pyogenes Gamma haemolysis - no lysis e.g. Strep. mutans
What are the 4 species of malaria that cause human disease?
- P. falciparum (most common)
- P. ovale
- P. vivax
- P. malariae
How is malaria transmitted?
Transmitted by the bite of the FEMALE ANOPHELES MOSQUITO
How do you treat C. difficile?
In general ANY antibiotic that begins with the letter C can result in clostridium difficile: • Ciprofloxacin • Clindamycin • Cephalosporins • Co-amoxiclav (augmentin) • Carbapanems e.g. meropenem
How do you treat staph aureus?
Flucloxacillin
How do you remember gram positive bacteria?
Sexy - Streptococcus spp. Students - Staphylococcus spp. Can - Corynebacterium spp. Look - Listeria spp. Bad - Bacillus spp. Come morning - Clostridium spp.
How do you remember gram negative bacteria?
Huge - Helicobacter Vaginas - Vibrio cholera Can - Coliforms Never - Neisseria Provide - Parvobacteria Pleasure - Pseudomonas
How do you remember DNA viruses?
H - Herpes viridae A - Adenovirus P - Parvovridae P - Papopviridae H - Hepaviridae Mneumonic - HAPPH
How do you treat gram positive bacteria?
Penicillins
Are staphylococci catalase positive or negative?
Positive
Are streptococci catalase positive or negative?
Negative
Which type of bacteria are most catalase positive?
Many GRAM NEGATIVE bacteria e.g. E.coli and fungi (aspergillus spp.) are catalase POSITIVE
Which bacteria are coagulase positive?
Staphylococcus. aureus is coagulase POSITIVE - clumping
Which bacteria are coagulase negative?
Other staphylococci are coagulase NEGATIVE - no clumping
What is the structure of gram positive bacteria?
- Have a single cytoplasmic membrane
- Have a large amount of peptidoglycan on outer surface
- DO NOT have endotoxin (lipopolysaccharide)
What is the structure of gram negative bacteria?
- Have two membranes; an inner
and outer - Have a smaller amount of peptidoglycan between the membranes
- Antibiotics target the peptidoglycan
- The outer membrane has lipopolysaccharide (ENDOTOXIN) which the immune system can react to - endotoxic shock
What is the general structure of bacteria?
- Capsule:
• A polymer of sugar that protects bacteria from host immune system - can inhibit parts of the innate immune system
• Pneumonia bacteria has a capsule - Cell wall:
• Made up of phospholipid membrane - Bacteria are prokaryotes i.e. they do not have a nuclear membrane (eukaryotes do)
- Bacteria usually have one circular chromosome
- RNA polymerase is present in bacterial cytoplasm
- Cell envelope
Where in the body is open to bacterial colonisation?
Mucosal surfaces are areas open to bacterial colonisation:
- Nasal cavity
- Larynx
- Stomach
- Colon
- Need to keep the lungs, gall-bladder, kidneys & eyes STERILE
What shape are cocci?
Round and spherical
What shape are bacilli?
Rod-like and straight
What can bacteria look like?
Diplococcus (2) Chain of cocci Cluster of cocci Chain of rods Curved rod (vibrio) Spiral rod (spirochaete)
What are the clinical features of malaria?
- Chills & sweats
- Headache
- Myalgia
- Fatigue
- Nausea & vomiting - Diarrhoea
Give some examples of resistant bacteria.
- Aerobic bacteria are unable to reduce metronidazole to its active form thus the antibiotic is harmless to them
- Anaerobic bacteria lack oxidative metabolism required to uptake aminoglycosides
- Vancomycin is not taken up by gram negative bacteria - it cannot penetrate their outer membrane since its too large