Microbiology Flashcards
Why do we culture blood?
to look for bacteraemia - it is normally bacteria free without bacteria flora
How can blood be contaminated?
patient’s skin, surface of the items used to take the culture, fingers of the person taking the blood, laboratory
How long does it take for a blood culture to identify an organism?
1 day to do gram stain and coccsu/bacillus
2-3 days to identify organism and assess whether this is likely to be a contaminant
Why are contaminants a problem?
give wrong treatment, waste of money, side effects for patients, unnecessary equipment, increased length of stay, mislead clinicians, not always able to identify whether it is a contaminant or not, don’t meet contamination targets
What organism is resistant to all B lactams?
MRSA
Disadvantage of using B lactam antibiotics?
can predispose to C.difficile and colonization with resistant bacteria e.g. MRSA, many people report allergies to this group, especially penicillin
What is the most widely used class of antibiotics and how do they work?
B lactams which includes penicillins, cephalosporins, carbapenems and combinations - bind to cell wall and inhibit cells wall sunthesis
Two main types of penicillins?
amoxicillin, flucloxacillin
What antibiotic is used for enterococci and H.influenzae?
amoxcillin - but most enterobacteria are resistant to it
What is flucloxacillin used for?
anti staphylococcal agent as stability to penicillinase
What are B lactamase inhibitors used for?
bacteria resistance is through B lactamase production, so these antibiotics prevent this resistance, so use combinations e.g. co-amoxiclav, tazocin
What are monobactams used for?
e.g. Aztreonam, for gram negative bacilli, including pseudomonas and preferably aerobes, safer for C.diff
What are carbapenems used for?
e.g. imipenem for very broad spectrum hospital acquired infections
Disadvantages of carbapenems e.g. imipenem?
expensive, CI in CNS disorders and can predispose to C.diff
What are cephalosporins used for?
e.g. cefotaxime, ceftazidime for gram negative activity, good CNS penetration and good anti pseudomonal
What are glycopeptides used for?
e.g. vancomycin for gram positive only and reserved for serious, resistant ones e.g. MRSA (Iv only and toxic)
What are macrolides used for?
e.g. erythromysin, clarithromycin for staph and strep activity, used to soft tissue and throat infections if allergic to penicillins (no gram negative cover)
What are lincosamindes used for?
e.g. clindamycin for staph, strep and anaerobes and C.diff and PMC good bone concentrations and well absorbed orally
What are tetracyclines used for?
mild MRSA infections and chlamydias
What are aminoglycosides used for and how do they work?
e.g. gentamicin for enterobacteriaceae, pseudomonas and staphylococci, synergy with cell wall antibiotics against streptococci and enterococci - inhibit ribosomal activity and protein synthesis
Disadvantages of aminoglycosides?
more toxic than glycoproteins so nephro and oto toic levels must be monitored carefullly
What is fusidic acid used for?
serious anti staphylococcal agents e.g. osteomyelitis, only used in combination
What is oxazolidinone used for and how does it work?
grampositive including MRSA and VRE - inhibits bacterial protein synthesis and blocks initiatio complex formation
Disadvantages of oxazolidinone?
only 1 class e.g. linezolid and are expensive and can cause bone marrow suppression if used for >2 weeks
What are nuclei acids/quinolones used for and how do they work?
e.g. ciprofloxacin - UTIs, enterobacteriacease pseudomonas and staph - inhibitt DNA gyrase
What is rifampicin used for and how do they work?
TB and general Staph’s and meningitis prophylaxis - must be used in combination - interfere with nuclei acid synthesis or function
What is metronidazole used for?
anti anaerobic for bacteria and protozoa
What likely organisms are seen in UTIs?
e coli, staph sparophyticus, enterococci
Antibiotics for UTIs?
trimethoprim, amoxicillin, clavulanate, cephalosporin
Likely organisms to cause COPD?
Hinfluenzae, pneumococcus, Moraxella catarrhalis
Treatment of COPD infection?
amoxicillin with clavulanate, tetracycline, trimethoprim
Likely organisms to cause pneumonia?
pneumococus
Treatment of pneumonia?
B lactams, erythromycin
Likely organisms in a wound?
staph aureus, strep pyogenes, anaerobes
Treatment of a wound?
flucloxacillin, erythromycin, co-amoxiclav
How can antibiotics be misused?
use broad spectrum when narrow are just as effective, treat for too long, treating with IV when oral is as effective, using combination when one is a effective, failure to use a dose appropriate for renal and hepatic function, using antibiotics for highly unlikely organisms, failure to step down organism once known
Dangers of antibiotic misuse?
adverse drug reactions which increases with multiple drug prescribing, harmful drug interactions, errors in prescribing monitoring, can cause super infection and antibiotic resistance
What is the difference between bacteriostatic and bactericidal?
bacteriostatic - prevents bacteria growth by inhibiting protein synthesis, DNA replication or metabolism, reduces toxin production
bactericidal - kills bacteria by inhibiting cell wall synthesis, good if poor penetration
Why do antimicrobials want a large MIC?
minimum inhibitory conc - to attach to more binding sites to inhibit bacteria
What factors must be considered in drug administration choice?
will it penetrate, pH of the site, is antimicrobial lipid soluble
How do bacteria resist antimicrobials?
change antimicrobial site by changing binding site configuration, destroy or inactivate the antimicrobal, prevent antimicrobial etery by modifying bacterial membrane porin channel site, numbers or selectivity, remove antimicrobial from bacterium using export pumps
How does bacterial resistance develop?
intrinsically naturally resistant so subpopulations can develop it too
acquired - not all subpopulations are equally resistant
Gram positive antimicrobial resistant bacteria?
MRSA, VRE
Gram negative antimicrobial resistant bacteria?
B lactamases, ESBL, carbapenenases, AmpC B lactamase resistant
What methods can be used to test antimicrobial resistance?
antimicrobial sensitivity testing, chromogenic plates, mechanism specific tests, genotypic methods, breakpoint plates
What is a pathogen?
an organism that is capable of causing disease
What is a commensal?
an organism which colonises the host but causes disease in normal circumsatnces
What is an opportunist pathogen?
only causes disease if host defences are compromiseed
What is pathogenicity?
the degree to which an organism is pathogenic
What is asymptomatic carnage?
when a pathogen is carried harmlessly at a tissue site where it causes no disease
Components to a bacteria?
capsule, cell wall, outer and inner membrane, pili, chromosome, may also have plasmids
What is the function of a capsule on bacteria?
super coated so difficult for phagocytes to ingest
What is used to stain acid fast bacilli?
Ziehl Neelsen stain
What colour is a gram positive stain?
purple/blue
What colour is a gram negative stain?
pink
What is the difference in gram positive and negative cell wall composition?
n = more lipopolysaccharide, with lipoprotein, less peptidoglycan with inner and out membrane
p = more peptidoglucan with lipoteichoic acid and cytoplasmic membrane
What is the idea bacterial environment?
temperature
What are the 4 stages of bacterial life?
lag, exponential (log), stationary, death
What is endotoxin?
component of outer membrane of bacteria produced by lipopolysaccharide gram negative, has non specific action, stable effect of heat, weak antigenicity and not convertable to toxoid
What is exotoxin?
secreted by gram positive (and negative), can be converted to toxoid, strong antigenicity, liable effect of heat and specific action
What is involved in a gram stain?
apply crystal violet stain to fix bacteria, add iodide to bind to crystal violet and help fix it to cell wall, decolourize with ethanol and counter stain with safranin (pink)
in gram negative, the decolourizze interacts with lipids and cells lose their lipopolysaccharide membrane and the crystal violet complex so appears pink
in positive, the decolourize dehydrate the cell wall and crustal violet is trapped in the multilayed peptidoglycan
Methods for infection control?
decontaminate medical equipment, single used items, manage peripheral and central IV lines, manage short term catheters, outbreak control plan, specific antimicrobial prescribing policies, personal protective equipment worn by all staff, dispose sharps into sharps bin, never res heath or bend needles or overfill sharps bin, hand hygeiene
What is an endogenous infection?
infection of a patient by their own flora, especially in hospitalised patients with invasive devices and surgical aptients
Prevention of endogenous infection?
good nutrition and hydration, antispepsis/skin prep where indicated, control underlying disease, remove linses and catheters as soon as clinically possible, reduce antibitotic pressue as much as clinically possible
What are protozoa?
eukaryotic organisms that consume bacteria, algae and microfungi
What are the 5 main protozoa?
flagellates - allows movement, reproduce by binary fision
amoebae - flow through cytoplasma, produced by pseudopodia
sporozoans - no locomotory extensions, most intracellular parasites and reproduce by multiple fission
ciliates - cilia beat rhythmically with macro and micro nuclei
microsporida - makes resistant spores, unique polar filament, coiled inside spore
What has malaria incidence increased?
parasite resistance, climate changes, increased travel to area
How do viruses cause disease?
virus causes invagination of the cell membrane and attaches to cell receptors and released into cell cytoplasm, carry nuclei acids and replicate using cells proteins and forms a defense mechanism
virus moves in vacuole and acidification causes a change in surface proteins of virus to release products, this replicates until the cell lysis or exocytosis so it can spread to other cells, budding causes parts of the virus to be pinched off
How does a virus cause damage?
direct destruction of host cells e.g. polio, modification of host cell structure or function e.g. rotavirus and HIV, damage through overreactivity of host in response to infection e.g. hepatitis B and C, damage through cell proliferation and immortalisation e.g. HPV, virus evasion of the host defences
What makes up a cell wall of fungi?
polysaccharides and chitin, most don’t have a capsule, and the cell wall is the sterol rich cytoplasmic membrane
What are the different forms of fungi?
moulds, yeasts, dimorphic
Who are most at risk of getting a fungal infection?
immunocompromised, especially neutrophil deficient (those who are healthy only get superficial infections e.g. athletes foot)
What does the fungal cell wall stain with?
Gomorra methernamine silver of periodic acid schiff reagent as it is too weak for gram stain, KOH
How do yeasts and moulds repoduce?
yeasts - asexual budding
moulds - spore formation
Shape of yeasts and moulds?
yeasts - round/oval
moulds - tubular
How do yeasts and moulds grow?
yeasts - grow in tissue and bud as tubular forms as a pseudohyphae
moulds - grow by longitudinal extension and branch to form interwoven mycelium
What are the 3 types of fungal infection?
superficial mycoses - in skin, hair, nails, mucus membranes with yeast and ringworm
subcutaneous mycoses - pigmented fungi with septate dark brown cells singly or in clusters e.g. madura foot
systemic mycoses - primary or opportunist pathogens
Treatment of fungal infections?
drugs to target sterols in cell membrane, topical or systemic
antifungal therapy binds to serols in cell walls, destabalising it and inhibits RNA and DNA synthesis and inhibits ergosterol biosynthesis and mircotubule assembly
Examples of fungal infections?
candidasis - on mucous membranes of mouth and vagina, a commensal yeast that causes disease when microflora disturbed
ringworm - direct or indirect transfer of infection keratin e.g. athelets foot
aspergillois - spores in soils and ducts can be invasive, allergic or asperfillomia
cryptococcosis - capsulate yeast in immunosuppressed
Is microbacteria gram positive or negative?
weak gram positive with a high lipid ontent
How can mycobacteria cause infection
survive a long time in macrophages and are slow growing, so immunodeficient are susceptible
How does leprosy cause damage?
has granulomas formed as a response to try and contain mycobacteria, these can cause damage to nerves which means risk of tumour and burns and the nose can collapse
How do mycobacteria appear in a Ziehl Neelson stain?
turns red then destained to be blue
How long does it take to culture TB?
solid = 3-8weeks liquid = 1-3 weeks - then can used mycobacteria growth indicator tube using flurometric detection
What does PCR do in TB culturing?
amplifies nucleic acids, purifies and concetratesTB, sonicates to release henomic material and performs PCR, rapid result and detects rifampicin resistance using fluorensence
What test is used to detect re-activity of T cells for TB
tuberculin skin test looks for cell mediated immune defense
Difference between lepromatous and tuberculoid leprosy?
LL - lesions full of bacilli, poorly formed granulomatus, extensive skin lesions, Th2 response
TL - tissue hypersensitivity and granulomatas, causing tissue damage, immune reaction, Th1 response and CD4+ T cell response, producing IFN-Y and TNF-a
4 drugs for TB treatment?
rifampicin, isoniazid, pyrazinamid, ethambutol for 2 months followed by INH and RIF for 4 months
Examples of gram positive cocci?
staphylococci, streptococci, enterococci
Examples of gram negative cocci?
neisseria and moraxella
Examples of gram positive bacilli?
actinomyces, bacillus, clostridia, diptheria, listeria monocytogenes
Examples of gram negative bacilli?
E coli, campylobacter, pseudomonas, salmonella, shigella, proteus
What is the catalase test?
to see if the microbe has catalase which destroys H2O2 and produces O2, if O2 is produced it is positive and is staphlococcus and it has catalase, it no O2 it is negative and streptococcus
Purpose of catalase test?
distinguishes between staphylococcus and streptococcus
What test is used on gram positive cocci?
catalase test
What test is used on staphylococci?
coagulase test
What is the coagulase test?
coagulase converts fibrinogen to fibrin and differentiates between staphylococcus aureus and staphylococcus epidemidis (positive is s.aureus and appears cream/yellow, negative is white)
How does s.aureus appear?
pigmented clusters, coagulated (solid), had DNAase
How does s.aureus cause disease?
high disease potential, pore forming toxin proteases. toxic shock syndrome toxin, spread by aerosal and touch and can colonize skin wounds
Treatment of s.aureus?
flucloxacillin (MRSA is resistant to B lactams though so needs vancomycin)
How does s.epidemidis appear?
non pigmented clusters, non coagulated, no DNAase
How does s.epidimidis causes disease?
low disease potential and s opportunistic and forms persistent biofilms
What test is done to distinguish streptococci?
haemolysis on blood agar
What are the 3 outcomes of haemolysis on blood agar and how do they appear?
beta haemolytic strep - clearing of agar around colonies due to strepolysin O and S production
alpha haemolytic strep (viridans strep) - greening of agar around the colonies due to H2O2 production
gamma haemolysis - no lysis