Microbiology: Generalities, Infection biology, Diagnostic microbiology, Parasitology Flashcards
Gram Staining Procedure
P = V M = I D = A C = S
Primary Stain = crystal Violet Mordant = Iodine Decolorizing Agent = Acetone Counterstain = Safranin
Gram Staining Procedure
Primary Stain = crystal Violet Mordant = Iodine Decolorizing Agent = Acetone Counterstain = Safranin
Bacteria NOT seen in Gram stain? Alternative approach?
These Rascals May Microscopically Lack Color
These Rascals May Microscopically Lack Color
Treponema - Spirochetes: Darkfield microscopy
Rickettsia - Giemsa/Tissue stains
Mycobacteria - Acid-Fast Stain
Mycoplasma - Serology
Legionella - Silver stain
Chlamydia - Giemsa stain (inclusion bodies)
Bacteria NOT seen in Gram stain? Alternative approach?
These Rascals May Microscopically Lack Color
Treponema - Spirochetes: Darkfield microscopy
Rickettsia - Giemsa/Tissue stains
Mycobacteria - Acid-Fast Stain
Mycoplasma - Serology
Legionella - Silver stain
Chlamydia - Giemsa stain (inclusion bodies)
Obligate Aerobes
Nosy and Nagging Pests Must Breathe Lots of oxygen
Nocardia Neisseria Pseudomonas Mycobacteria Bordetella/ Brucella/ Bacillus cereus Legionella
Obligate Aerobes
Nocardia Neisseria Pseudomonas Mycobacteria Bordetella/ Brucella/ Bacillus cereus Legionella
Obligate Anaerobes
ABC of Anaerobes
Actinomyces
Bacteroides
Clostridium
Obligate Anaerobes
Actinomyces
Bacteroides
Clostridium
Prophage Coded Bacteria
(Lysogenized strains) of ABCDE
(Lysogenized strains) of ABCDE: Non-essential, for virulence
shigA-like toxin: EHEC Botulinum Cholera Diphtheria Erythrogenic strain (of S. pyogenes/ GABHS/ scarlet fever)
Prophage Coded Bacteria
(Lysogenized strains) of ABCDE: Non-essential, for virulence
shigA-like toxin: EHEC Botulinum Cholera Diphtheria Erythrogenic strain (of S. pyogenes/ GABHS/ scarlet fever)
Normal Flora
Skin Nose Mouth Dental plaque Colon Vagina
Normal Flora
Skin: Staphylococcus epidermidis Nose: Staphylococcus aureus Mouth: Viridans streptococci Dental plaque: Streptococcus mutans Colon: Bacteroides, Escherichia coli Vagina: Lactobacillus vaginalis, Escherichia coli, Streptococcus agalactiae
Salma Hayek and her curli hair
Salmonella has surface proteins called ‘curli’ which mediate binding to endothelium and to extracellular proteins such as fibronectin
Enzyme in bacterial invasion, a.k.a. “spreading factor”
Hyaluronidase
What does IgA protease do?
Allows adherence to mucous membranes
Bacteria with IgA protease
SHine My Gong
Bacteria with IgA protease: SHiNe My Gong
Streptococcus pneumoniae
Haemophilus influenzae
Neiserria meningitidis
Neirserria gonorrhea
Streptococcus pyogenes anti-phagocytic factor
M protein
Staphylococcus aureus protein A function
Prevents complement activation
Which bacteria have “ADP-ribosylation” as the MOA of their exotoxins?
Diphtheria
Cholera
E.coli (heat-labile)
B. pertus
ADP-ribosylation is the addition of one or more ADP-ribose moieties to a protein. It is a reversible post-translational modification that is involved in many cellular processes, including cell signaling, DNA repair, gene regulation and apoptosis
Which bacteria have “superantigen” as the MOA of their exotoxins?
TSST (S. aureus)
Staphylococcal enterotoxin
Erythrogenic toxin (S. pyogenes)
Which bacteria have “protease” as the MOA of their exotoxins?
Tetanus (tetanospasmin, C. tetani)
C. botulinum
Lethal factor of B. anthracis toxin
Scalded skin toxin (S. aureus)
Which bacteria have “lecithinase” as the MOA of their exotoxins?
Clostridium perfringens alpha toxin
MOA of shiga toxin
Kills cells by cleaving 60S ribosomal subunit
Which bacteria have endotoxin?
eNdotoxin is an integral part of gram Negative cell walls
LPS toxic component: lipid A (overproduction of cytokines, complement cascade, coagulation cascade)
All bacteria have cell walls composed of peptidoglycan except
Mycoplasma pneumoniae
All gram-positive have NO endotoxin except
Listeria monocytogenes
All bacterial capsules are composed of polysaccharide except
Bacillus anthracis
All exotoxins are heat-labile except
Staphylococcal enterotoxin
Bacteria : Agar
Clostridium perfringens
Egg yolk (McClung-Toabe)
Bacteria : Agar
Corynebacterium diphtheriae
Tellurite (Loeffler’s medium, will produce black colonies)
Bacteria : Agar
Group D Streptococci
Bile esculin
Bacteria : Agar
Staphylococci
Mannitol salts
Bacteria : Agar
N. meningitidis and N. gonorrhea from sterile sites
Chocolate
Bacteria : Agar
N. gonorrhea from non-sterile sites
Modified Thayer-Martin
Antibiotic content of Modified Thayer-Martin agar
Vancomycin
Colistin
Nystatin
Trimethoprim
Bacteria : Agar
Haemophilus influenzae
Chocolate + Factors V and X
Agar : Bacteria
Lowenstein-Jensen
Mycobacterium tuberculosis (yellowish brown, “buff, rough, tough”, 4 weeks!)
Agar : Bacteria
Thiosulfate citrate bile salts (TCBS)
Vibrio
cholera - yellow
parahemolyticus - bluish green
Agar : Bacteria
Bordet-Gengou or Regan-Lowe charcoal medium
Bordetella pertussis
Agar : Bacteria
Charcoal-yeast extract
Legionella pneumophila
Agar : Bacteria
Skirrows
Campylobacter
Helicobacter
Agar : Bacteria
Barbour-Stoenner-Kelly (BSK)
Borrelia burgdorferi (Lyme disease)
Agar : Bacteria
Eaton
Mycoplasma pneumoniae
Agar : Bacteria
Cetrimide
Pseudomonas aeruginosa
Agar : Bacteria
Xylose-Lysine-Deoxycholate (XLD)
Salmonella
Shigella
Agar : Bacteria
Ellinghausen-McCullough-Johnson-Harris (EMJH)/ Fletcher’s
Leptospira interrogans
Difficult to culture, hence other tests are used for diagnosis
Chlamydia (intracellular)
Gonorrhea (NAAT is used)
M. tuberculosis (AFB, PCR)
Antigen detection tests
IF (B. pertussis, L. pneumophila, Rabies)
EIA like ELISA
Other agglutination tests
Antibody detection tests
Western blot
Lepto MAT
COPT
Gold standard for diagnosis of VIRAL diseases
PCR
usually, serology first, then nucleic acid amplification
Serologic Tests for Syphilis
Nontreponemal (screening only)
VDRL
RPR
Antigen (cardiolipin, cholesterol, purified lecithin)
USR, TRUST
Serologic Tests for Syphilis
Treponemal Antibody Tests (for confirmation)
TPEIA (if nontreponemal test is positive)
EIA/ELISA vs. RIA
enzyme-labelled antibodies, measured by spectrophotometer vs. radio-labelled antibodies, measured by gamma counter
Latex agglutination tests are used for
Streptococcal A pharyngitis
Bacterial meningitis
Cryptococcus (CALAS / Cryptococcal Antigen Latex Agglutination System)
Diagnosis for HIV:
Screening
Confirmatory
Gold standard
ELISA
Western Blot
PCR
Drugs: Folate synthesis inhibitors
Sulfonamides
Tripmethoprim
Drugs: RNA polymerase inhibitor
Rifampicin
Drugs: Cell membrane inhibitors
Amphotericin
Ketoconazole
Polymyxin
Drugs: DNA gyrase inhbitors
Fluoroquinolones
Drugs: Cell wall synthesis inhibitors
Beta-lactam antibiotics (Carbapenems, Cephalosporins, Monobactams, Penicillins)
Others: Bacitracin, Fosfomycin, Vancomycin
Drugs: Protein synthesis inhibitors
Aminoglycosides Chloramphenicol Clindamycin Macrolides Mupirocin Streptogramins Tetracyclines
BBB Penetrability: Excellent with or without inflammation (8 drugs/drug classes)
Sulfonamides Chloramphenicol Trimethoprim Metronidazole Rifampicin Isoniazid Fluconazole Flucytosine
BBB Penetrability: Good only with inflammation
Penicillins Cephalosporins: Cefuroxime (2nd gen), 3rd gen parenteral (except Cefoperazone), 4th gen Imipinem + Cilastatin Meropenem Aztreonam Ciprofloxacin Vancomycin
BBB Penetrability: Minimal or not good even if with inflammation (4 drug classes)
Aminoglycosides
Tetracyclines
Lincosamides
Macrolides
BBB Penetrability: No passage even if with inflammation (3 drug classes)
Polymyxins
1st and 2nd gen Cephalosporins
Amphotericin B
Cephalosporins are not active against 3 organisms
MRSA
L. monocytogenes
Group B Strep (enterococci)
Resistance mechanisms: Beta lactams
Hydrolysis
Mutant PBP
Resistance mechanisms: Tetracycline
Active efflux from cell (gram pos and neg)
Resistance mechanisms: Aminoglycosides
Inactivation by enzymes
Impermeability (strict anaerobes and streptococci)
Resistance mechanisms: Sulfonamides
Overproduction of target (more PABA)
Resistance mechanisms: Fluoroquinolones
Mutant DNA gyrase
Resistance mechanisms: Chloramphenicol
Reduced uptake into cell
Resistance mechanisms: Vancomycin
Reprogramming of D-ala D-ala (becomes Lac-Lac)
S. aureus (GISA: glycopeptide intermediate S. aureus)
Resistance mechanisms: Quinupristin/Dalfopristin
Ribosomal methylation
Resistance mechanisms: Macrolides/Erythromycin
RNA methylation
Drug efflux
Resistance mechanisms: Fluconazole
Active efflux (Candida spp.)
Resistance mechanisms: Pseudomonas aeruginosa
Impenetrability (multiple antibiotic classes)
Resistance mechanisms: Modification of antimicrobial agent
PBP alteration in MRSA
Penicillin resistance of S. pneumo and N. meningitidis
Altered peptidoglycan in enterococci
RNA polymerase mutation (Rif resistance in MTB)
DNA gyrase mutation in enterobactericeae (Cipro resistance)
Ribosomal gene mutations (resistance to protein synthesis inhibitors)
Dihydrofolate reductase mutations (resistance to trimethoprim)
Resistance mechanisms: Absence of antimicrobial target
Echinocandin (-fungin) resistance in Cryptococcus spp.