m m

m m
m m
 M
  
   
   

   
ym Have no true nucleus, nuclear membrane, & mitotic apparatus. 

 
ym Have nucleoid: 1.m planctomycetes: have nucleoid
m package of DNA in e cytoplasm. surrounded by a nuclear envelope
m wonsists of a single continuous circular molecule ranging in size from 0.58 to
 almost 10 million base pairs.
with 2 membranes.
2.mVibrio cholerae & Brucella

  m no. of nucleoid & chromosomes depend on e growth condition. melintesis: have 2 dissimilar
m apidly growing bacteria have > nucleoid per cell than slowly growing one. chromosomes.
m t is Feulgen-positive: indicates e presence of DNA (negati vely charged). 3.mBorrelia burgdorferi & Streptomyces
ym Nuclear region is filled with DNA fibrils. coelicolor: have linear chromosome.
ym ack of autonomous plastids such as mitochondria & chloroplasts.
ym electron transport enzymes r localized in e cytoplasmic membrane.
ym n photosynthetic bacteria: e photosynthetic pigments r located in intracytoplasmic membrane systems such as lamellae,
chromatophores, or chlorosomes.
ym n cynobacteria: have thylakoids & e major accessory pigme nts used for light harvesting r found on e surface of e
thylakoid membranes.
ym torage of reserve material:
m s in the form of insoluble granules which appear as refractile bodies in the cytoplasm
m Function in e storage of energy or as a reservoir of structural building block.
m |ost r bounded by a thin nonunit mebrane consisting lipids.
ym ypes of inclusion bodies of storage materials:
1.m poly--hydroxybutyric acid (pHB):
m A lipid-like compund consisting of chains of -hydroxybutyric acid units connected through ester linkages.
m produced when e source of nitrogen, sulfur, or phosporus is limited while carbon is excess.
2.m Gylogen:
m Formed when carbon is excess.
m sed s carbon source when protein & nucleic aci synthes r resumed same as pHB.
3.m ulfur:


 
 m prokaryotes oxidize reduced sulfur compounds such as hydrogen sulfite & thiosulfate to produce intracellular
granules of elemental sulfur.

 m hen e reduced sulfur is limiting, e granuled sulfur is oxidized to sulfate & e granule will slowly disappear.
4.m phosphate:
m norganic phosphate is accumulated in e form of granules of polyphosphate.
m granules can be used as sources of phosphate for nucleic acid & phospholipid synthesis to suport growth.
m granules r also termed as volutin granules or metachromatic granules because they stain red with a blue dye.
5.m w 2 fixation:
m Autotrophic bacteria contain polyhedral bodies surrounded by a potein shell (carboxysomes).
m bodies contain e key enzyme of w 2 fixation, ribulosebisphosphate carboxylase.
6.m |agnetosomes:
m ntracellular crystal particles of iron magnetite that allow certain aquatic bacteria to exhib it magnetotaxis.
m urrounded by a nonunit membrane containing phospholipids, proteins, & glycoproteins.
7.m Gas vesicles:
m provide buoyancy for aquatic bacteria.
m mebrane is a 2 nm thick layer of protein, impermeable to water & solutes but permeable to gas.
8.m wytoskeletal protein:
m play cytoskeletal roles.
m Actin homologs (|reB & |bl) help to determine ell shape, segregate chromosomes, & localize protein with e cell.
m Nonactin homologs (FtsZ) & bacterial cytoskeletal proteins (ecY & |inD) help to determine cell s hape, regulate
cell division, & segregate chromosomes.

 ym wonsists of capsule, cell wall, cell membrane, flagella, & pili.
ym Function to protect bacteria from hostile environments such as extreme osmolarity, harsh chemical, & antibiotic.


ym Also called cytoplasmic membrane which is visible in electrom micrograph as thin section.
ym womposed of phospholipids & 200 kinds of proteins which make up 70% of the membrane mass.
ym Have no sterols except mycoplasma which incorprate sterols into their membrane when growing in

 the sterol-containing media.
ym At least 50% of e cytoplasmic membrane must be in semifluid state for cell growth to occur. At low
temperature, this is done by incorpration of unsatured fatty acid into e phospholipids of e
membrane.
1.m permeability & ransport:
ym Forma a hydrophobic barrier impermeable to most hydrophilic molecules which can only be
transported through:
a.m passive transport: relies on diffusion, uses no energy, & operates only when e solute is at higher

 concentration outside than inside e cell. he examples are simple diffusion, facilitated diffusion,
M  & channel protein.
b.m Active transport: 2 types depending on e source of enegy employed:
m on-coupled transport: moves a molecule across e c ell membrane across e cell membrane at

  e expense of a previously established ion gradient such as proton -motive or sodium-motive
force. here r 3 basic types i.e. uniport, symport, & antiport.
m ABw transport: employs Ap directly to transport solutes into e cell. t is facilitated by
specific binding proteins.
c.m Group translocation: e substrate is phosphorylated during e process, no concentration gradient
is involved & it allows bacteria to utilize their energy sources by couling transport with
metabolism.
d.m pecial transport process: some bacteria secrete siderophores (compound that chelate Fe &
promotes its transport as a soluble complex) such as hydroxamic acid.
 2.m lectron ransport & xidative phosphorylation:
ym cell membrane has a functional analog with e mitochondrial membrane.
ym t contains cytochromes & other enzymes & e component of respiratory chain for oxidative
phosphorylation.
3.m xcretion of Hydrolytic xoenzymes & pathogenicity protein:
ym cell membrane excretes hydrolytic enzymes that degrade e polymers to subunits small enough
to enetrate e cell membrane.
ym Macteria secrete them directly into e external medium or into e periplasmic space b/w e
peptidoglycan layer & eouter membrane of e cell wall (gram ve).
4.m Miosynthetic Function:
ym cell membrane is e site of e carrier lipid on which e subunits of e cell wall r assembled.
ym enzymes for cell wall biosynthesis & phospholipid synthesis are located here too.
5.m whemotactic ystems:
ym Attractants & repellents bind to specific receptors in e bacterial membrane.
ym A complex polymer consists of:
1.m A backbone of alternating N acetylglucosamine & N acetylmuramic acid.
m ame in all species.
2.m A set of identical tetrapeptide side chain attached to e N acetylmuramic acid.
m Dary from species to species.
m n all specis, there r features in common. |ost hae  -alanine at position 1, D-glutamate at
position 2, & D-alanine at position 4. position 3 is e most variable one Gram ve


 

bacteria have diamnopimelic acid (DAp) while Gram +ve bacteria have -lysine.
  3.m A set of identical peptide cross -bridges.
m Dary from species to species.
m n many Gram ve cell wall, e cross bridge consists of a direct peptide lin kage b/w DAp
amino group of 1 side chain & e carboxyl group of e terminal D -alanine on e 2nd side
chain.
ym n Gram +ve bacteria, there r 40 sheets of peptidoglycan & on 1 or 2 sheets in Gram ve
bacteria.
1.m echoic & eichuronic Acids:
ym wonsists of glycerophosphate or ribitol phospate residues which r connected by phosphodiester
linkages & usually have other sugars & D-alanine attached (to position 2 or 3 of glycerol or
position 3 or 4 of ribitol).
ym vely charged & partially responsible for e ve charge of e cell surface.
ym 2 types i.e. wall techoic acid (A) which is covalently linked to peptidoglycan & membrane
techoic acid or lipotechoic acid (A) whish is associated with lipids.
ym ie b/w e cytoplasmic membrane & e peptidoglycan layer, extending through pores in e
peptidglycan layer.

  ym repeat unit may be glycerols (joined by 1,3 or 1,2-linkages), ribitol (joined by 1,5-linkages),
ym functions:  
 or complex consists of both joined to glucose, galactose, or N-acetylglucosamine.
a.mGives osmotic ym Functions:
protection. M
  a.m |ake up a polyanionic network or matrix with peptidoglycan which provides functions
b.mHelps in cell relating to elsticity, porosity, tensile, strenghth, & electrostatic properties of e envelope.
division. b.m wonstitue e major surface Ag of those Gram +ve species.
c.m erves as a primer c.m Associated (A) with e | protein molecule forming microfibril that facilitate e
for its own attachment of S pyogenes to animal cells.
biosynthesis. ym eichuronic acids: r similar polymer but e repeat units include sugar acids & r synthesized in
d.mite of antigenic place of techoic acids when phosphate is limiting.
determinants. 2.m polysaccharides:
ym xception to ym wonsists of sugars such as mannose, arabinose, rhamnose, & glucosamines as well as acidic
mycoplasma sugars such as glucuronic acid & mannuronic acid.
which r cell wall- ym xist as subunits of polysaccharides in e cell wall.
lacking bacteria
containing no 1.m uter |embrane:
peptidoglycan. ym Milayered in structure with inner leaflet resembles cell membrane while e outer leaflet
contains p.
ym Functions:
a.m protection from deleterious substances such as bile salts by excluding hydrophbic
molecules.
b.m Has channel protein called porins that permit passive diffusion of low | hydrophilic
compounds.
c.m Has a relatively high antibiotic resistance of Gram ve bacteria.
d.m participates in anchoring to peptidoglycan layer through mpA protein.
e.m Acts as sex pilus rece ptor in F-mediated bacterial conjugation through mpA protein.
f.m wontains proteins involved in e transport of specific molecules such as Dit. M12.
   2.m ipopolysaccharide (p):
M
  ym wonsists of a complex glycolipid called ipid A, attached to a polysaccharide & a te rminal
series of repeat units.
ym ipid A consists of phosphorylated glucosamine disaccharide units to which r attached long
chains fatty acids ( -Hydroxymyristic is unique to e lipid).
ym polysaccharide core has p & includes ketodeoxyoctanoic acid (KD ) & a heptose.
ym repeat unit composed of linear trisaccharides & others & known as Ag which functions to
cover e bacterial surface & exclude hydrophobic compounds through its hydrophilic
carbohydrate chain.
ym Functions:
a.m vely charged p molecules r noncovalently cross-bridged by divalent cations to
stabilize e membrane & provide a barrier to hydrophobic molecules.
b.m Has an important virulance factor (lipooligosaccharides- e sialylation of its N-
acetyllactosamine causes molecular mimicry in e host).
 3.m ipoprotein:
ym wross-link e outer membrane & peptidoglycan layers.
ym wontains 57 aa, representing repeats of a 15 -aa-sequence.
ym lipid component consists of diglyceride thiother linked to a terminal cysteine which s
noncovalently inserted in e outer membrane.
ym Function: to stabilize e outer membrane & anchor it to peptidoglycan layers.
4.mperisplasmic pace:
ym pace b/w e inner & outer membrane which contains peptidoglycan layers & a gel -like
solution of proteins.
ym 20-40% of cell volume.
ym periplasmic proteins:
ym wontains a high concentration of D-glucose which r variously substituted with glycerol
phosphate & phosphatidylethanolamine residues or -succinyl esters.
ym hese r called membrane-derived oligosaccharides which control osmoregulation.
ym womposed of:
1.m peptidoglycan.
2.mAn external asymmetrical lipid bilayer.
3.m inner leaflet which contains mycolic acids (contain waxes) lnked to an arabinoglycan.
4.m outer leaflet which contains other extractable lipids.
ym Advantages:

 

1.m highly ordered lipid embeded with proteins allows slow passage of nutrients & drugs by
  forming water-filled pores.
2.m hydrophobic structure renders e bacteria resistant to many harsh chemicals like strong acid.
3.mts permeability to hydrophilic molecules slows e bacterial grothw rate.
ym Known as Acid-Fast because when introduced to a dye by heating or treatment with
detergent, e dye cant be removed by dilute Hw.
ym .g.: mycobacteria.
ym |any both Gram +ve & -ve bacteria posses 2D crystalline subunit -type lyer lattice of protein or
glycoprotein molecules (-layer) as outermost component.
ym -layer:

   1.m womposed of a single kind protein molecule which capable of self-assembly.

  2.mesistant to proteolytic enzymes & prot ein-denaturing agents.
3.mprotects e cell from wall-degrading enzymes & from e invasion of predatory bacerium.
4.m |aintains e cell shape.
5.m|ay involve in cell adhesion to host epidermal surface.
ym Glycocalyx:
1.m Definition: e polysaccharide-containing material lyingoutside e cell.
2.m Formed from sucrose.
3.m Functions: resistant to desiccation & correlates dental carries with sucrose consumption in human.


 ym wapsule:
1.m Definition: a condensed, well-defined layer closely surrounding e cell that excludes particles.


  2.m Functions in e:
-m nvasiveness of pathogeinc bacteria.
-m protection from phagocytosis.
-m Adherence to bacteria to surface of their environment.
ym lime layer: glycocalyx which is loosely associated with e cell & does not exclude particles .
ym hread-like appendages composed of protein, 12 30 nm in diameter.
ym rgan of locomotion, 3 types of arrangement:
1.m peritrichous: flagella all around e body of e bacteria, e.g. Salmonella typhi.
2.m Amphitrichous: a single flagellum at each pole.
3.m ophotrichous: bunch of flagella at one or both ends, e.g. Spirilium minus.
4.m |onotrichous: 1 flagellum at 1 end, e.g. V. cholerae.

 ym Muilding blocks: thousands molecules of a protein subunit called flagellin.
   ym flagellum is formed by e aggregration of s ubunits to form a helical structure
ym highly antigenic: H Ag.
ym tructure for attachment: a complex composed of a hook (short curved structure acts as universal
joint b/w motor & flagellum) & basal body (bears a set of rings, i.e. one pair in Gram +ve & two pairs
in Gram ve).
ym semirigid helical rotors to which e cell imparts a spining movement.
  
ym Flagellar motor & its components r located in e cell envelope.
ym rigid surfce appendages.
ym horter & finer than flagella.
ym womposed of structural protein subunits termed pilins.

 ym |inor protein called adhesins r located at e tips of e pili & r responsible for attachment properties.
ym 2 types:
M  1.m rdinary pili: adherence of symbiotic & pathognic bacteria to host cells.
2.m ex pili: attachment of donor & recipients cell in bacterial conjugation.
ym pilin molecules r arranged helically to form a straight cylinder that does not rotate & lacks a complete basal body.
ym pili grow from inside of e cell outward.


  
 




  
m
m m
m m

  


   


 ym S. aureus: ubiquitous, ant. nares (50% 70% of healthy person), skin, mucous membrane of & G, hospital.
ym S. epidermis: skin, gut, & .
ym S. saprophyticus: decaying plant & animal, urinary bladder, & lower urinary tract of young female.


 ym Gram-positive spherical cells. ym pathogenic staphylococci:
ym Arranged in grape-like irregular clusters. 1.m Hemolyze blood.
ym Grow readily on many types of media. 2.m woagulate plasma.
ym Active metabolically: fermenting carbohydrates & 3.m produce various extracellular enzymes & toxin.
producting pigments (white to deep yellow) & lactic acids. 4.m apidly develop resistance to antimicrobial
ym woagulase test: S. aureus positive, others negative. agents, drying, & heat.

 
    ym pherical cells about 1 m in diameter arranged in irregular cluster.
ym iquid cultures: single, pairs, tetrads, & chains cocci.
ym Young cocci stain strongly Gram positive, on aging: many become Gram negative.
ym Nonmotile & do not form spores.
ym ysis under influence of drugs like penicillin.

 ym Grow readily on aerobic or microaerophilic condition & most rapidly at 37 ow.
ym wolonies:

   1.m n solid media: round, smooth, raised, & glistening.
2.m S. aureus: gray to deep golden yellow colonies, creamy growth, causes discoloration, & hemolyzes blood.
3.m S. epidermidis: gray to white colonies.
4.m S. saprophyticus: grey or white colonies.
ym pigment production:
1.m Form pigment best at room temperature (20 25 ow).
2.m Develop pigment only prolonged incubation.
3.m No pigment is produced anaerobically or in broth.
  ym esistance to antimicrobial drugs:
1.m esistance to penicillin:
m Due to e production of -lactamase production.
m nder control of plasmids which r transmited by trasduction & perhaps also by conjugation.
2.m esistance to nafcillin:
m ndependent of -lactamase roduction.
m esistance is due to mecA gene which encodes a low affinity penicillin binding proteins.
3.m usceptibility to vancomyocin:
m usceptible if |w is < 2 g/m.
m ntermediate susceptibility if |w is 4 8 g/m.
m esistant if |w is > 16 g/m.
4.m plasmid-mediated resistance to tetracyclines, erythomycins, aminoglycosides, & etc.
5.m olerance:
m mplies that staphylococci r inhibited by a drug but not killed by it.
m wan be attributed to lack of activation of autolytic enzymes in e cell wall.
1.m peptidoglycan:


ym A polysaccharide polymer containing linked subunits providing e rigid exoskeleton of e cell wall.

 ym Destroyed by strong acid or exposure y lysoenzyme.
ym licits e production of -1 & opsonic Abs by monocytes.
ym A chemoattractant for p|N leukocytes.
ym Have endotoxin like activity & activates complement.
2.m eichoic acids: polymers of glycerol or r ibitol phosphate which linked to peptidoglycan & can be antigenic.
3.m protein A: a cell wall component that binds to e Fc portion of gG molecules except gG 3 . his makes it as self.
4.m wapsules:
ym nhibits phagocytosis by p|N leukocytes.
ym Have coagulase or clumping factors that bind to fibrinogen yielding aggregration of e bacteria.
1.m watalase: converts H 2 2 into water & oxygen & differentiates staphylococci from streptococci.
  2.m woagulase:
  ym An enzyme-like protein that clots oxalated or citrated plasma.
ym Minds to prothrombin & e complex initiates fibrin polymerization.
ym Deposits fibrin on e bacterial surface & alters their ingestion by phagocytic cells.
3.m ther enzymes:
4.m wlumping factor: responsible for adherence of e organisms to fibrinogen & fibrin. hen mixed with plasma, it forms
clumps.
5.m xotoxins:
ym -toxin: heterogenous protein that acts on eukoryotic cell membrane. Also acts as a potent hemolysin
ym -toxin: degrades sphingomyelin which toxic to many cells.
ym '-toxin: heterogenous & dissociates into subunits in nonionic detergents. t disrupts biologic membranes & causes
diarrheal disease.
ym >-hemolysin: efficiently lyse Mw by causing pore formation in e cellular membranes that increase cation
permeability.
6.m eukocidin: consists of 2 components w hich act synergistically to kill Mws.
7.m xcofoliate toxins: consists of 2 distinct proteins of same molecular weight:
ym pidermolytic toxin A: chromosomal gene product & heat -stable.
ym pidermolytic toxin M: plasma mediated & heat-labile.
ym Yield e generalized desquamation of e staphylococcal scalded skin syndrome.
ym superantigen.
8.m oxic shock snydrome toxin: a superantigen which promotes e manifestations associated with fever, shock, &
multisystem involvement includng desquamative skin rash.
 U.m nterotoxin:
ym superantigen, heat-stable, & resistant to e action of gut enzymes.
ym emetic effect is e result of wN stimulation after e toxin acts on neural receptors in e gut.
ym on a chromosomal element called a pathogenecity island which interacts with bacteriophages to pr oduce
toxin.

    ym prototype of staphylococcal lesion is furuncle or localized abscess.
ym mechanism: production of coagulase which coagulates fibrin around e lesion & within e lymphatic l formation of
a wall that limit e process l accumulation of inflammatory cells & fibrous tissues at e centre l liquefaction of
necrotic tissue occurs l drainage of e liquid centre l granulation tissue formation l healing.
ym rganisms may spread via e lymphatics & bloodstream results in s uppuration within veins associated with
thrombosis.
ym complications:
1.m steomyelitis: in a terminal blood vessel of e mataphysis of a long bone leading to necrosis & chronic
suppuration.
2.m pneumonia, meningitis, emphyma, endocarditis or sepsis with suppuration in any organs.
3.m kin infection: impetigo, pyoderma, or acne.
4.m Mullous exfoliation: e scalded skin syndrome caused by e production of exfoliative toxin.
5.m oxic shock syndrome caused by -1.
1.m ocalized staphylococcal infection:



ym Appears as pimple, hair follicleinfection, or abscess.
 ym here is an intense, localized, painful inflammatory reaction that undergoes central suppuration & heals quickly
when e pus is drained.
ym wall of fibrin & cells prevent e organism from spreading.
2.m Direct contamination of a wound: e.g. in postoperative staphylococcal wound infection or infection following
trauma.
3.m Macteremia: due to bacterial dissemination results in endocarditis, meningitis, organ dysfunction, & intense focal
suppuration.
4.m Food poisoning: characterized by a short incubation perid (1 8 hours) with violent nausea, vomiting, diarrhea, &
rapid convalescence.
5.m oxic shock syndrome: manifested by an abrupt onset high fever, vomiting, diarrhea, myalgias, scarlatiniform rash,
& hypotension with cardiac & renal failure.
1.m Gram staining: purple clustering cocci. 5.m phosphate test.
  
 2.m pigmentation. 6.m Deoxyribonuclease test.
 3.m woagulation test. 7.m |annitol fermentation.
4.m protein A test. 8.m watalase positive.
1.m reat with penicillinase resistant penicillins: 2.m f e organism is methicillin resistant treat with:
 
ym wloxacillin ym Dancomycin
ym Nafcillin ym wlindamycin
ym Dicloxacillin ym Gentamcin
ym wycloserine
ym Fusidic acid
ym ifampicin
m
m


 


   
ym wlassification into many categories has been based on:
1.m wolony morphology & hemolytic reactions on blood agar.
2.m erologic specificity of e cell wall group-specific substance or capsular Ags.
3.m Miochemical reactions & resistance to Ags.
4.m cologic features.
ym wategories of streptococci:
1.m Hemolysis:
a.m -hemolysis: complete disruption of Mws with clearing of e blood around e bacterial growth.
b.m -hemolysis: incomplete reduction of hemoglobin & e formation of green pigment.
c.m >-hemolysis: non-hemolytic steeptococci.
2.m ancefield classification (group-specific substance):
m Mased on e carbohydrate contained in e cell wall of many streptococci.

 
  m basic serologic grouping is ancefield A-H & K-.
a.m Group A: rhamnose-N acetylglucosamine, Streptococcus pyogenes.
b.m Group M: rhamnose-glucosamine polysaccaharide, Streptococcus agalactiae.
c.m Group w: rhamnose-N acetylgalactosamine, Streptococcus equislimilis.
d.m Group D: glycerol teichoic acid containing D-alanine & glucose, Streptococcus faecalis.
e.m Group F: glucopyranosyl-N acetylgalactosamine.
3.m wapsular polysaccharide: based on e antigenicity of e capsular polysaccharides.
4.m Miochemical reactions:
m sed for species that typically do not react with commonly used Ab preparations for e group -specific
substances.
m Mased on biochemical tests such as sugar fermentation, presence of enzyme, test for susceptibility, & etc.
ym |any streptococci r classified according to e combination of e above features.



  
ym pherical & ovoid cocci arranged in chains which divide in a plane perpendicular to e long axis of e chain (one
 
    plane only).
ym treptococci r gram -positive, however as a culture ages & e bacteria die, they can appear to be gram -negative.
 ym |ost group A strains produce capsules composed of hyaluronic acid which impede phagocytosis.
ym cell wall contains proteins, carbohydrates, peptidoglycans, & hair-like pilli (|-protein) which r covered with
lipoteichoic acids (for attachment to epithelial cells).
ym |ost grow in solid media as discoid colonies of 1 -2 nm in diameter.
ym S. pyogenes r -hemolytic which produces large zones of -hemolysis around colonies greater than 0.5 mm in

 diameter.
ym 2 forms of colonies:

   1.m |atte colonies: strains that produce much | protein & generally r > virulent.
2.m Glossy colonies: strains that produce little | protein & often r not virulent.
ym hey r pY +ve but bacitracin sensitive.
ym nergy is obtained from e utilization of glucose with e production of lactic acid.
ym Growth tends to be poor on solid media or broth unless enriched with blood or tissue fluid.
  ym |ost pathogenic strains grow best at 37 o w.
ym |ost streptococci r facultative anerobes & grow under aerobic & anaerobic conditions while e peptostreptococci
e obligate anaerobes.
1.m | protein:
ym A major virulent factor of group A S. pyogenes.
ym Appears as hair-like projections on e cell wall.
ym presence indicates virulence & can resist phagocytosis, while its absence indicates no virulence.
ym mmunity to infection is related to e presence of type -specific Abs to | protein.
ym | protein molecule has a rod-like coiled structure that separates functional domains. structure allows

 for a large no. of sequence changes while maintaining function.

 ym Has 2 classes, (may be a virulent determinant for rheumatic fever) & .
2.m substance:
ym No r/ship to e virulence of e streptococci.
ym substance is acid & heat labile.
ym btained from streptococci by proteolytic digestion.
ym t permits differentiation of certain types of streptococci by agglutination of specific antisera.
3.m Nucleoproteins: also known as p substances whi ch make up most of e streptococcal cell body.
1.m treptokinase (fibrinolysisn): transforms e plasminogen into plasmin that will digest fibrin & other protein.
2.m treptodornase: depolymerizes DNA.
3.m Hyaluronidase:
ym plits hyaluronic acid which aids in spreading infecting microorganisms.
ym antigenic & specific for each bacterial or tissue source.
4.m pyrogenic exotoxins (erythrogenic toxin):
  ym xist in 3 antigenically distinct streptococcal pyrogenic exotoxins of A, M, & w.
ym xotoxin A (| protein types 1 & 3) has been associated with streptococcal shock syndrome & scarlet fever.
  ym Acts as superantigens which activate polyclonal cells.
5.m Diphosphorydine nucleotidase: ability to kill leukocytes.
6.m Hemolysins:
a.m treptolysin : protein which is hemolytically active in e reduced state & responsible for hemolysis seen
when growth is in cuts deep into e medium of blood agar plate.
b.m treptolysin : responsible for e hemolytic zones around streptococcal colonies on blood agar plate
eventhough it is not antigenic.
1.m Diseases attributable to invasion by S. pyogenes , -hemolytic group A streptococci:
a.m rysipelas: massive brawny edema & a rapidly advancing margin of infection.
b.m wellulitis: acute, rapidly spreading infection of e skin & subcutaneous tissues which is associated with pain,
tenderness, swelling, & erythema. lesion is not raised with indistinct boundries.
c.m Necrotizing fasciitis (streptococcal gangrene): extensive & very rapidly spreading necrosis of e skin &
subcutaneous tissue (flesh-eating bacteria).
d.m puerperal fever: a septicemia originating in e infected woud (endometritis).
e.m Macteremia/sepsis: follows e infection of traumatic or surgical wounds which rapidly can be fatal.
2.m Diseases attributable to local invasion with S. pyogenes & their by-products:
a.m Streptococcal sore throat:
ym D/t e adherence of S. pyogenes to pharyngeal epithelium via lipoteichoic acids.
ym n infant & small children: e sore throat occurs as subacute nasopharyngitis with thin serous discharge,
little fever & cervical lymph node enlargement.
ym n older children & adults: > acute & characterized by intense nasopharyngitis, tonsiltis, & intense
redness & edema of mucus membrane with purulent exudates, high fever, & tender cervical lymph


 nodes.
b.m treptococcal pyoderma:
 ym ocal infection of superficial layers of skin called impetigo.
ym wonsists of superficial vesicles that break down & eroded areas whose denuded surface is covered with
pus & encrusted.
ym t spreads by continuity & higly communicable especially in hot & humid climates.
ym wwidespread infection occurs in eczematous or wounded skin or in burn which progress to cellulitis.
3.m nvasive group A streptococcal infections:
a.m treptococcal toxic shock syndrome:
ym wharacterized by shock, bacteremia, respiratory failure, & multiorgan failure.
ym Death occurs in 30% of patients.
ym nfection tends to follow minor trauma such as necrotizings fasciitis, myositis, & etc.
ym rythema & desquamation may occur.
b.m carlet fever:
ym pyrogenic exotoxins A-w also cause scarlet fever in association with S. pyogenes pharyngitis or with soft
tissue infection.
ym rash appears on e trunk after 24 hours of illness & involve e extremities.
 4.m poststreptococcal diseases:
a.m Acute glomerulonephritis:
ym Develops 3 weeks after S. pyogenes skin infection.
ym |ay be initiated by AgAb complexes on gromerular basement membrane.
ym wlinical features: blood & protein in urine, edema, high blood pressure, & urea nitrogen retention with
low level of serum complement.
ym ome die, some develop chronic form & kidney failure, & majority recovers completely.
b.m heumatic fever:
ym esults in damage to heart muscle & valve d/t cross reaction b/w cell membrane Ags & human heart
tissue Ags.
ym onset is preceded by S. pyogenes infection 1-4 weeks earlier.
ym patients with > severe streptococcal sor e throat have a greater chance of developing rheumatic fever.
ym ymptoms: fever, malaise, a migratory non-suppurative polyarthritis, & inflammation of all parts of
heart.
ym carditis leads to thickened & deformed valves & to small perivascular granuloma in myocardium
which r replaced by fibrous tissues.
ym heumatic fever has a marked tendency to be reactivated by recurrent streptococcal infection
whereas nephritis does not.
1.m Direct detection - the antigen is extracted from a throat swab. he antigen extract will then bind with antibody
specific to the group A streptococcal carbohydrate. his has classically involved agglutination of antibody
coated beads. However, simpler tests have been recently introduced. esults are available within minut es.
2.m ancefield grouping of isolated beta hemolytic colonies (see above).
  
 3.m wolonies are beta hemolytic (figure 5) and their growth is inhibited by bacitracin (pre sumptive diagnosis).
 4.m patient serum shows antibodies to streptolysin or other streptococcal antigens. his is important if delayed
clinical sequelae occur.
ym Meta hemolysis is caused by two hemolysins and
the former is inactive in the presence of oxygen. hus,
stabbing of the plate increases the intensity of the hemolysis reaction.
ym All r susceptible to penicillin G & most r susceptible to erythromycin.
  ym ome r resistant to tetracyclines.
 
ym Gram positive bacteria which r lancet-shaped diplococcic or arranged in chains.
 
    ym eadily lysed by surface active agents which remove or inactivate e inhibitor of cell wall autolysins.
ym pneumococci is inhibited by ptochin.
ym pneumococci forms small round colonies which r at 1 st dome-shaped but later develop a central plateau with
an elevated rim.


ym t is -hemolytic on blood agar form smooth colonies.
  ym |ost energy r obtained from fermentation of glucose leading to e production of lactic acids (limits growth).
ym pneumococci which produce large amounts of capsules produce large mucoid colonies.
1.m womponent structures: pneumococci have cell wall which contains peptidoglycan & teichoic acid which r
 
 immunogically distinct for each > U0 types.

 2.m Quelling reaction: i.e. capsule swelling test for rapid identification & typing of e organisms.
1.m production of disease:
ym Diseases r produced through e ability to multiply in tissues.
ym capsules resist phagocytosis from phagocytosis.
ym No toxins r produced, but gA protease is produced to help in colonization.
2.m oss of natural resistance which predispose to pneumococcal infection:
a.m Diral & other respiratory tract infection that damage cell surface which leads to mucus accumulation,
bronchial obstruction, & injury.

  b.m Alcohol & drugs that depress phagocytic activity, cough reflex, & facilitates aspiration of foreign material.
c.m Abnormal circulatory dynamic.
d.m |alnutrition & general debility.
3.m pathology of pneumococcal infection:
ym pneumococcal causes an outpouring of fibri nous edema fluid, Mws, & Mws into e alveoli which leads to
consolidation of e lungs. However, e alveolar wall remains intact.
ym hey reach e bloodstream through lymphatic drainage & many reside in sinuses & middle ear.
ym ater, mononuclear active cells phagocytose e debris & e lquid phase gradually reabsorbed.
ym onset is usually sudden, with fever, chills, & sharp pleural pain.



ym sputum is similar to alveolar exudates which is bloody or rusty in color.
 ym complications: meningitis, endocarditis, & septic arthritis.
1.m tained smear: gram stain shows many p|Ns & many Mws.
 M     2.m wapsule swelling test: positive.
   3.m wulture: blood culture reveals -hemolytic.
ym D w: penicillin G with high doses.
 
ym pneumococci remain susceptible to vancomycin.

  ym pneumococci r resistant to tetracycline & erythromycin.

 
 w


 


 


ym hey r 0.5-1.0 m in diameter & several micrometers long.
ym hey posses irregular swelling at one end that gives e club-shaped appearance.
ym Granules (staining deeply with aniline dye) r irregularly distributed within e rod which gives a beaded
appearance.
ym ndividual corneybacteria tends to lie parallel or at acute angles to one another.
  ym colonies:
m mall, granular, gray, with irregular edges, & have small zone of hemolysis.
m n potassium tellurite, e colonies r brown to black with a brown -black halo because tellurite is reduced
intracellularly.
ym 4 biotypes of corneybacteria: gravis, mitis, intermedius, & belfanti.
ym hey grow aerobically & tend to pleomorphism in microscopic & colonial morphology.
ym n oefflers serum medium, thay grow > readily & also called as Klebs -oeflerr Macillus.
ym hey r spread by droplets or by contact to susceptible persons, & then they grow on mucous membranes & skin
abrasion.
ym Diphtheria toxin:
m wharacteristics: Heat-labile polypeptides & bipartite molecule which is composed of fragment A & M.
m production:
-m toxin is produced by toxigenic bacteria.
-m oxigenic & lysogenic bacteria r produced when nontoxigenic bacteria r infected by bacteriophage from
toxigenic diphtheria bacilli. trait is subsequently heredirity.
 


 -m actual production of toxin occurs when e prophage of e lysogenic diphtheria become induced & l yses e
cells.
-m toxigenicty is under control of e phage gene & e invasiveness is under control of bacterial gene.
m |echanism of action:
-m Fragment A: inhibits polypeptide chain elongation by inactivating e elongation factor F-2 with e
presence of NAD.
-m Fragment M: no independent activity but required for e transport of fragment A into e cell.
-m oxin fragment A inactivates F-2 by catalyzing a reaction that yields free nicotinamide and inactive
adenosine diphosphate-ribose-F-2 complex.
-m esult: abrupt arrest of protein synthesis which leads to necrotizing & neurotoxic effects.
1.m ffects of bacilli themselves:
ym Formation of a false membrane which adhere to underlying tissue.
ym Diphteria produce toxin which is absorbed into e mucus membranes l destruction of epithelium l
superficial inflammatory response l e necrotic epithelium becomes embedded in exuding fibrin & red &
white cells l grayish pseudomembrane is formed (over tonsil, pharynx, or larynx).
ym t is a grayish white, wrinkle, & tough which is difficult to strip off.
ym f removed, it will leave behind minute bleeding spot. regional lymph nodes in e neck enlarged & there
might be marked edema of e entire neck.
  ym n fauceal & laryngeal diphtheria e false membrane may cause mechanical obstructi on of e passage.
ym ounded or skin diphtheria occurs chiefly in tropics: e membrane may form an infected wound which fails to
heal.
ym However, diphtheria does not actively invade deep tissues & practically never enters bloodstream.
2.m ffects of exotoxin:
ym diphtheria within e membrane produce toxin actively.
ym oxin is absorbed & causes distant toxic damage: n e heart, muscle, liver, kidneys, adrenal, &
a.m parenchymatous degeneration. sometimes nerve damage (results in paralysis
b.m Fatty infiltration. of soft palate, eye muscles, & extremities.
c.m Necrosis with gross hemorrhage.
1.m nitial features: sore throat & fever.
2.m ater development d/t to membrane formation & e toxin :
a.m prostration & dyspnea which leads to suffocation.


 b.m rregularities in cardiac rhythm d/t toxic myocarditis.


 c.m paralysis effects on vasomotor control of e blood vessels.
d.m oxic & interstitial nephritis in kidneys.
e.m Fatty changes & hemorrhage in adrenal glands.
f.m Nerves: neuritis.
1.m Gram stain:
ym My using e Dcaron swabs from nose, throat, or other suspected lesion.
ym |ust be done before e registration of antibiotic.
2.m wulture: by using a oeflerr slant (12-18 hours) or a tellurite plate (36-48 hours).
 3.m oxigenicity test:


 a.m lek test: immersed a filter paper in diphtheria toxin l set it on serum agar l streak e strain of w.
diphtheria on e agar at e right angle to e strip paper l incubate at 37 ow precipitation (toxin-antitoxin
interaction).
b.m An immunochromographic strip assay: a highly sensitive assay.
c.m pw & A.
1.m reatment of diphtheria antitoxin is mandatory for strong clinical suspicion of diphtheria.


  2.m Antimicrobial drugs such as penicillin & erythromycin.

!

 3.m Active immunization with diphtheria toxoid (vaccine) to yield adequate antitoxin level. sually in combination
with tetanus toxoid or pertussis vaccine.



 

"




 


1.m ypical organisms:
ym pores r wider than e diameter of e rods.
ym pores r located centrally, subterminally, or terminally.
ym |oat species r motile & possess peritrichous bacteria.
2.m wulture & colony:
ym wlostridia r anaerobes which grow under anaerobic condition.
  ym ome r aerotolerant & will also grow in ambient air.
ym hey grow well in blood-enriched media used to grow other bacteria.
ym colony:
m |ultiple sizes (large-raised & small) colonies which can be spread on agar surface.
m |any produce zones of hemolysis on blood agar.
3.m Growth: clostridia can ferment a variety of sugars, digest protein, turn milk into acids & undergo stormy
fermentation.
w
 

ym wauses tetanus & is worldwide in distribution in e soil & in e feces of horses .
ym wan be distinguished by specific flagellar Ags.
!
!

#
ym All share a common Ags which may be masked & produce neurotoxin, tetanospasmin.
ym spores r centrally located.
ym toxin is tetanospasmin which is cleaved by a bacterial protease into 2 peptides linked by disulphide bond.
ym |echanism: toxin binds to receptors on e presynaptic membranes of motor neurons l it migrates by e retrograde
axonal transport system to e cell bodies to spinal cord & brain stem l it diffuses to terminals of inhibitory cells l
$

it degrades synaptobrevin (protein for docking of neurotransmitter vesicles) l release of inhibitory glycine &
GAMA is blocked l hyperreflexia, muscle spasm, & spastic paralysis results.
ym xtremely small amount of toxin can be lethal to human.
ym w. tetani is not an invasive organism. hus, e infection remains localized in e area of devitalized tissue into which e
spores have been introduced.
  ym Germination of e spores & development of vegetative organisms r aided by:
1.m Necrotic tissue. Aid e establishment of low
2.m walcium salts. redox potential.
3.m Associated pyogenic infection.
ym incubation period: 4-5 days to as many weeks.
ym disease is characterized by tonic contraction of voluntary muscles:
m |uscular spasm of e area of injury & infection l muscle of e jaw (trismus & lock jaw) which contract so that


 mouth cant be opened l spasm of other voluntary muscle l tonic spasm l by e presence of external
stimuli, generalized muscle spasm can occur.


 m n newborn infant, risus sardonicus occur d/t a peculiar grin caused by acute spasm of facial muscles.
ym patient is fully conscious & pain may be intense.
ym Death usually results from interference with e mechanics of respiration.
ym |ortality rate in generalized tetanus is very high.
 ym clinical pictures & history of injury is of diagnostic test.


 ym proof of isolation of w. tetani must rest on e production of toxin & its neutralization by specific antitoxin.

1.m Active immunization with toxoids (vaccine).

ym produced by detoxifying w toxin with formalin & concentrating it.
ym 3 injections should be given for e initial course with a booster injection given upon entry to school.
2.m proper care of wound contaminated with soils.
3.m prophylactic use of antitoxin.


  ym | administration which neutralizes e toxin that has not been fixed to nerv ous tissues.

!

 4.m Administration of penicillin.
ym trongly inhibits e growth & stops further toxin production.
ym |ay also control associated pyogenic infection.
5.m urgical debridement for removing e necrotic tissues.
6.m patients who develop symptoms of tetanus should be given muscle relaxant, sedation, & D assisted ventilatoxin to
neutralize e toxin.
w



ym toxin produced causes a spreading infection which has lethal , necrotizing, & hemolytic properties.
ym -toxin of w. perfringens type A is a lecithinase & its lethal action is proportionate to e rate at which it splits
lecithin to phosphorylcholine & diglyceride.
$
 ym theta toxin has similar hemolytic & necrotizing effects. t is not a lecithinase but a DNAase & hyaluronidase, a
collagenase that digests collagen is subcutaneous tissue & muscle.
ym hey also produce enterotoxin when grown in meat dishes which induces intense diarrhea in 6 -18 hours.
ym spores r terminally located.
1.m nvasive clostridial infection:
ym pores reach tissue by contamination or traumatized area or from intestinal tract l spores germinate,
multiply, & ferment carbohydrate in tissue l form gases l tissue distention & blood supply interference l
secretion of necrotizing toxin & hyaluronidase l spread of infection l extension of tissue necrosis l
 


 opportunity for increased bacteria growth, hemolytic anemia, & severe toxemia death.
2.m Gas gangrene or clostridial myonecrosis:
ym nvolves mixed infection of toxigenic clostridia, proteolytic clostridia, & various cocci & gram-negative
organisms.
 ym rganism from fecal flora may contaminate a wound & may grow & multiply in poorly perfused
tissue.ifection spread rapidly & gas can be felt in tissue.
ym From a contaminated wound, e infection spreads in 1-3 days to produce precipitation in e subcutaneous tissue &
muscle, results in foul-smelling discharge, rapidly progressing necrosis, fever, hemolysis, toxemia, shock, & death.


 ym disease: anaerobic fasciitis or cellulitis.


 ym nterotoxin:
m oxin forms when e organisms sporulate in e gut.
m onset of diarrhea (w/o vomiting & fever) is 6 -18 hours, but e illness lasts only 1 -2 days.
1.m Gram stain of specimens obtained from wound, pus, & tissue with e presence of large gram positive rods
 suggestion for gas gangrene clostridia.
2.m wulture in chopped meat-glucose medium & thioglycolate medium & onto blood agar plates incubated


 anaerobically.
3.m Final identification rests on toxin production & neutralization by specific antitoxin.
ym reatment:
1.m prompt & extensive surgical debridement of e involved area & excision of all devitalized tissues.


  2.m Administration of antimicrobial drugs such as penicillin at e same time.

!

 3.m Antitoxin in e form of concentrated gs & polyvalent antitoxin.
ym prevention: early & adequate cleansing of contaminated wounds & surgical debridement together with e
administration of antimicrobial drugs.
w
 
 
ym wauses botulism which is worldwide in distribution & found in soil & occasionally in animal feces.
!
!

# ym pores of e organism r highly resistant to heat, withstanding 100 o w for several hours.
ym However, heat resistance is diminished at acid pH or high salt concentration.
ym oxin is liberated during e growth & during e autolysis of e bacteria.
ym even antigenic varieties r known from A-G, but only ype A, M, & cause human illnesses.
ym ype A & M r associated with a variety of foods & type with fish products.
$
 ym Motulinum toxin is absorbed from e gut & binds to receptors of presynaptic membranes of motor neurons of e
peripheral nervous system & cranial nerve.
ym proteolysis of e target protein in e neuron inhibits e release of Ach at e synapse, resulting in lack of muscle
contraction & paralysis.
ym toxin is destroyed by heating for 20 minutes at 100 o w.
ym illness is d/t intoxication resulting from e ingestion of food with e bacteria.
ym most common food: spiced, smoked, vacuum -packed, or canned alkaline foods that r eaten w/o cooking.
 



ym n e food, e spores germinate & under anaerobic condition, vegetative forms grow & produce toxin.
ym toxin blocks Ach at N| results in faccid paralysis.
ym ymptoms begin 18-24 hours after ingestion with visual disturbances, inability to swallow, & speech difficulties,
signs of bulbar paralysis r progressive & death d/t respiratory paralysis or cardiac arrest.


 ym No fever is associated, e patient remains fully conscious shortly before death.


 ym mortality rate is high.
ym ecovered persons do not develop antitoxin in e blood.
ym Newborn infants: poor feeding, weakness, & sign of paralysis which leads to sudden infant deah syndrome.
 ym toxin can be demonstrated in e serum & leftover of e food by using A or passive hemagglutination.


 ym antigenic type of toxin is identified by neutralization of specific antitoxin in mice.

1.m prompt D administration of antitoxin with customary precautions.



  2.m Adequate ventilation must be maintained by mechanical respirator.

!

 3.m risk from home-canned food can be reduced by boiling e food > 20 minutes before consumption.
4.m oxoid for active immunization.




 

 

 


ym prevalent in soil, water, air, & on vegetation.
ym typical cells:
m arge aerobic gram positive rods occurring in chains.
m |easure 1x3-4 m, have square ends, non-motile, & spore r located at e center of e rods.
m se simple sources of nitrogen & carbon for energy & growth.
m ts capsule contains poly-D-glutamic acid which is antiphagocytic where its gene in plasmid.
m B. anthracis which is non-capsulated is not virulent & does not induce anthrax in test animal.
 
ym colonies:
m round & have a cut glass appearance in transmitted light.
m Hemolysis is uncommon but gelatin is liquefied & growth in gelatin stab resembles as inverted fir tree.
ym spores:
m resistant to environmental changes, dry heat, & certain chemical disinfectant for moderate periods.
m persist for years in dry earth.
m wan be sterilized by autoclaving.
ym A zoonosis where human is accidently infected by contact with infected animals or their products.
 


 ym infection is acquired by e entry of e spores through:
1.m njured skin (cutaneous anthrax): U5%.
 2.m arely e mucus membrane (G anthrax): very rare.
3.m My inhalation of spores into e lungs (inhalation anthrax): 5%.
ym mechanism: e spores germinate in e tissue at e site of entry l vegetative organisms grow l formation of a
gelatinous edema & congestion l spread via lymphatic to e bloodstream l multiply freely in e blood & tissues
l death.
ym Anthrax toxin:
m |ade up of 3 proteins:
1.m protective Ag (pA): binds to specific cell receptors & following proteolytic activation, it forms a
membrane channel that mediates entry of F & F into e cell.
2.m dema factor (F): an adenylyl cyclase, with pA it forms a toxin known as edema toxin.
3.m ethal factor (F): with pA, it forms lethal toxin which is e major virulence factor & cause death.
m anthrax toxin genes r on another plasmid.
ym nhalation anthrax or woolsorters disease: spores r inhaled & phagocytosed in e lungs l transported by lymphatic
drainage to e mediastinal lymph nodes l germination occurs l toxin production l development of
hemorrhagic mediastinitis & sepsis l death.
1.m n susceptible animal:
ym organisms proliferate at e site of entry.
ym capsules remain intact & e organisms r surrounded by a large amount of proteinaceous fluid containing few
leukocytes.
  ym leukocytes help them in rapid dissemination & in reaching e bloods tream.
2.m n resistant animal:
ym organisms proliferate for a few hours in e present of massive accumulation of leukocytes.
ym wapsules gradually disintegrate & disappear.
ym organisms remain localized.
1.m wutaneous anthrax:
ym ccurs on e exposed surface of e arms or hands, followed by face & neck.
ym A prurutic papule develops 1-7 days after e entry of spores or e organisms through scratch.
ym papule rapidly changes into a vesicle or small ring vesicles that coalesce & a necrotic ulcer develops.
ym lesion is 1-3 cm in diameter & has a characteristic of central black eschar.
ym After 7-10 days, eschar is fully developed & it dries, loosens, separates, & heals by granulation & leaves scar.
ym |arked edema occurs with lymphangitis & lymphadenopathy.
ym ystemic signs & symptoms: fever, malaise, & headache.


 2.m nhalation anthrax:


 ym incubation period: as long as 6 weeks.
ym arly clinical manifestations: marked hemorrhagic necrosis & edema of e mediastinum.
ym ther complications: substernal pain, pronounced mediastinal widening, hemorrhagic pleural effusion, cough,
sepsis, & hematogenous spread to G causing bowel ulceration, or to meninges causing hemorrhagic
meningitis.
ym mortality rate is high.
3.m G anthrax:
ym G anthrax is acquired through ingestion of spores or spread of organisms from e intestinal tract.
ym wlinical signs: abdominal pain, vomiting, & bloody diarrhea.
1.m pecimens examination:
ym pecimens r obtained from fluid or pus from a local lesion, blood, & sputum.
ym wan be examined using:
a.m F staining techniques.
b.m Gram stain: large gram positive rods.
2.m wulture:
 a.m Mlood agar plate:


 ym Non-hemolytic gray to white colonies with a rough texture & a ground -glass appearance.
ym womma-shaped outgrowths (|edusa Head) may project from e colonies.
b.m emisolid medium: non-motile organisms r present.
c.m |ice or guinea pigs: killed upon intraperitoneal injection.
3.m erological test:
ym A is used to measure Abs against edema & lethal toxins.
ym A positive results is a fourfold change on a single titer of greater than 1:32.
1.m Antibiotics r effective & must be started early: ciprofloxacin & penicillin G with gentamycin or streptomycin.
2.m propghylaxis with ciprofloxacin & doxycycline for potential exposure.
3.m wontrol measures:


  a.m Disposal of animal carcasses by burning or b y deep burial in lime pits.

!

 b.m Decontaminating by autoclaving of animal products.
c.m protective clothing & gloves for handling potentially infected materials.
d.m Active immunization of domestic animals with live attenuated vaccines.
e.m mmunization of people with high occupational risk.
m
m
m
m
m
m
m
m
m m
  

"

 


ym Also known as fungus bacterium. ym properties:
ym wlass 1: Actinobacteria which consists of: m Acid (alcohol) fastness: resist decolorization by acidified
!
!

# 1.m Actinomyces. alcohol after stained with basis fuchsin.
2.m Nocardia. m presence of mycolic acids in e cell wall.
3.m rynebacteria. m A G + w content of DNA are 61-71%.
4.m hodococcus.
ym At least 100 species.
ym wausative agents of tuberculosis, leprosy, & opportunistic infection (saprophytes).
ym ome r slow grower (> 7 days) & some r rapid grower (< 7 days).
ym |orphology:
m od-shaped, aerobic, & do not form spores.
m ome r coccobacillary, filamentous, or branched.
ym taining:
1.m Ziehl-Neelsen stain: hot staining (result: red in color).
 

 2.m Kinyoun: cold staining.
3.m Auramine or fluorescence: yellow-orange fluorescence.
ym wulture:
1.m |. leprae cant be grown in culture, only in mouse foot pad & U banded armadillos.
2.m thers: owenstein-ensen () medium (inspissed egg media), |iddlebrook 7HU (liquid), |iddlebrook 7H10 &
7H11 which r agar based. (wultures color is green).
ym pigment production (yellow to orange):
1.m cotochromogens: form pigment in e dark.
2.m photochromogens: form pigment after exposure to light.
3.m Non-chromogens: no pigmentation at all.
|



 



ym hin straight uncapsulated rods measuring about 0.4 x 0.3 m.
ym AFM, obligate aerobe which r non-motile & non-spore forming.
  ym urvive in milk, organic matters, pasture land, phenolic, & hypochlorides.
ym ensitive to alcohol, formaldehyde, glutaraldehyde, heat, & D light.
ym esistant to acids, alkali, & quaternary ammonium compounds.
ym Derive energy from e oxidation of many simple carbon compounds.
ym Found mainly in cell wall which can induce hypersensitivity type 4 in previousl y infected animal.
ym constituents of tubercle bacilli:
1.m ipids:
"
 
 m nclude mycolic acids, waxes, & phosphatides which r largely bound to protein & polysaccharide.


 m |uramyl peptide complexed with lipid can cause granuloma formation & caseous necrosis.
2.m proteins: elicit e tuberculin reaction & formation of a variety of Abs.
3.m polysaccharides: can induce immediate type of hypersensitivity & can serve as Ags in reaction with sera of
infected persons.
ym Dirulence is d/t bacterial invasiveness & ability to survive in macrophages.
ym No toxin r produced.
ym mmune response is mediated by cellular immunity which leads to hypersensitivity type 4.
ym 2 types of tuberculosis:
1.m primary tuberculosis:
m ite of infection:
a.m sually lungs:
-m port of entry is t hrough respiratory tract by inhalation of bacilli.
-m primary complex or Ghon focus: enlargement of hilar lymph nodes.
-m wan be spread to other parts of body through hematogenous or lymphatic spread.
b.m ccasionally G: primary complex consists of tonsil & cervical lymph nodes en largement (which results
in scrofula or purplish in e region) as well as enlargement of ileocaecal & messentric lymph nodes
enlargement.
c.m kin: primary complex consists of regional lymph nodes enlargement (prosectors wart).
m mechanism (1-2 weeks time): cell clones (Ag specific) l cytokines l macrophage activation l
granuloma formation l central caseation (usually limit e 1 st infection) l quiescent l fibroblast scar &
calcification l recovery or incomplete destruction of my cobacterium l dormant l immunocompromised
 


 condition leads to reactivation l post-primary tuberculosis.
m 2 principal lesions:
a.m xudative type:
-m wonsists of an acute inflammation reaction with edema fluid, p|Ns, & monocytes around e
tubercle bacilli.
-m een in lungs that may heal by resolution & leads to massive necrosis of tissue.
-m During this phase, tuberculin test is positive.
b.m productive type or chronic granuloma:
-m wonsists of 3 zones:
i.m A central area of large, multinucleated giant cells containing tubercle bacilli. ater, e area will
undergo casaetion.
ii.m A mid zone of pale epitheliod cells arranged radially.
iii.m A peripheral zone of fibroblasts, lymphocytes, & monocytes.
-m A caseous tubercle may break into a bronchus, empty its contenst & form a cavity which then
heals by fibrosis or calcification.
2.m post-primary tuberculosis:
m sually caused by tubercle bacilli that have survived in primary lesions.
m wharacterized by longer local lesions, minimal lymphatic involvement, frequent cavitation, & usually
 involved e apical region of lungs which is e highest oxygen concentration.
m eactivation can be spontaneously when e is low.
m process of granuloma formation is e same but > extensive with large area of caseation, a condition known
as tuberculomata.
m protease & cytokines levels r high.
m mechanism: tubercle bacilli l erode wall of bronchus l liquefied content l expectorated l aerobic l
bacilli growth increases.
m n immunocompromised patients, e cavitation is rare
however, lymphatic & hematogenous spread is
common: cryptic disseminated tuberculosis.
1.m uberculin test: old tuberculin & purified protein derivative (ppD).
2.m |antoux test: intracutaneously.
3.m Heaf test: guns with 6 prongs.
4.m ine test:
 ym ndemic areas: > 10 mm.
ym Non-endemic areas: < 5 mm.



ym whildren & immunocompromised.
5.m aboaratory specimen: sputum, bronchial washing, biopsy, gastric aspirates, wF, & pleural fluids.
6.m wulture:  medium or middlebrook media.
7.m Drug susceptibility testing: in  medium containing doubling dilutions of drugs.
8.m X-ray.


  1.m terilizing: rifampicin & pyrazinamide.
2.m Macteriocidal: isoniazid, streptomycin, & ethambutol.

!

 3.m Macteriostatic: ethionamide, prothionamide, cycloserine, thiacetazone, & D-aminosalicyclic acids.
|






ym eprosy: a chronic mycobacterial disease primarily affecting e peripheral nervous system & secondarily involving skin
& other tissues.
ym tiology: mycobacterium leprae:
m unculturable in e lab, but can be grown in mouse footpad & U banded armadillos.
!
!

# m |an is e only significant reservoir.
m ransmission is via nasal secretion by droplets & by skin.
m Acid-fast bacilli are singly & arranged in parallel bundles or in globular masses.
m ften found in scraping from skin or mucous membrane & within endothelial cells of blood vessels or in e
mononuclear cells.
ym principle target cells r chwann cells which leads to nerve damage & results in anesthesia & muscle paralysis.
ym kin:
m Non-specific lesions with pigmentary changes (hypopigmented): anesthesia or hypoaesthesia.
m ften heals spontaneously.
ym wlinical menifestations depends on immune status of patients, ranging from tuberculoid to lepromatous.
ym Morderline tuberculoid < mid-borderline < borderline lepromatous.

Hyperactive Anergy
ym Few localized skin lesions with few bacilli ym Numerous skin lesions: often confluent.
(paucibacillary). ym Macilli are many (multibacillary) with clustering.
 


 ym ntense granulomatous response: damage in major ym lowered: anergy.
nerve (hardening). ym ar lobes: bacillary +++++

ym ther signs & symptoms:

1.m yes:
m veitis. eads to blindness.
m worneal infection to eyelid paralysis.
2.m Mone resorbtion:
m wollapse of nasal bone.
m hortening of fingers & toes (autoamputation).
3.m Neuritis, orchitis, & immune-complex nephritis.
1.m History & physical examination.
2.m plit skin smear & purich biopsy from ear lobes, nasal secretion, & nasal mucosa.
3.m Histology: granuloma & caseation.
4.m AFM:
ym Diable if stained strongly & evenly.
 ym Dead when stained unevenly & weakly.


 ym |orphological index (|):
m High |: active disease.
m ow |: with chemotherapy.
5.m kin test:
a.m epronin test (only guide).
b.m Fernandez (early): reaction in 1-2 days.
c.m |itsude (late): reaction in 7 days (usually ve in ).
1.m Dapsone: diaminophenyl suphone-DD.
2.m DD: resistant, thus |D with rifampicin & clofazimine.


  3.m ther drugs: prothionamide, ofloxacin, & minocycline.

!

 4.m H :
a.m paucibacillary: rifampicin & dapsone for 6 days.
b.m |ultibacillary: rifampicin, dapsone, & clofazimine for 2 years or more.

m
m
%&' ()(*)*
ime from onset wharacteristics
1 o complex develops
3-6 weeks
uberculin conversion is +ve
progressive healing of 1 o complex
2-6 months
possibility of pleural effusion
6-12 months possibility of military or meningeal tuberculosis
1-3 years possibility of bone or joint tuberculosis
3-5 years possibility of gastrourinary or chronic tuberculosis
m
m
(+, -*
 *%( 
Macillary index (record bacteria in high power)
1+ 1-10 per 100 fields
2+ 1-10 per 10 fields
3+ 1-10 per field
4+ 10-100 per field
5+ 100-1000 per field
6+ > 1000 per field

m




m

 


!
!

# 

 


  
 


 


 


 
 

 
 
 

!


   
1.m egressive theory: Diruses may be degenerate form of intracellular parasites.
2.m progressive theory: Diruses r derived from cellular NA & DNA components.
ym Normal cellular nucleic acids that gained e ability to replicate autonomously & therefore evolve.
 ym DNA viruses came from mitochondrial DNA, plasmid, or transposable elements. hey then evolve coat
proteins & transimissibility.
ym NA viruses from mNA.
3.m wombination theory: Diruses evolved through a combination of e above mechanisms.
ym bligate intracellular parasites.
ym mall (20 300 nm in diameter) filterable through bacterial-tight filters.
ym wontain:
  a.m A single type of nucleic acid, either DNA or NA.

 

 b.m A protein coat (capsid) consisting of individual protein units (capsomeres).
c.m A host derived lipid membrane (envelope) through which may be inserted viral protein (soikes or
peplomers).
ym |ultiply inside living cells by using e biosynthetic machinery of e host cell.
1.m Dirion morphology: size, shape, types of symmetry, peplomers, or membranes.
2.m Dirus genome properties: types of nucleic acid, size of genome, strandedness, & etc.

 
   
 3.m physicochemical properties of virion: molecular mass, buoyant density, pH stability, & etc.
a.m Disease: encephalitis & 
 4.m Genome organization & replication: gene order, number, & position of open reading frames.
hepatitis. 5.m Antigenic properties.
b.m |odes of transmission: 6.m Miologic properties: natural host range, mode of transmission, vector relationship, & etc.
arbovirus. 1.m Double stranded DNA: Adenoviruses, Herpesviruses, poxviruses, & etc.
c.m Host cells: animal, M   2.m ingle stranded +ve sense DNA: parvoviruses.
plant, & bacteria.  !"#$%&!' 3.m Double stranded NA: eoviruses.
d.m issue or organ: 4.m ingle stranded +ve strand NA: picornaviruses, ogaviruses, & w.
&"(&'%)!*
adenoviruses & 5.m ingle stranded ve sense NA: rthomyxoviruses, habdoviruses, & etc.
enteroviruses. + 
6.m egmented: rthomyxoviruses.
&"($ "&!$ 7.m Non-segmented: habdoviruses.
1.m ypes of symmetry:
a.m wubic symmetry:
ym cosahedral pattern: e most efficient arrangement for subunits in a closed shell.
ym Have 20 equilateral triangle faces, 12 vertices, & 5 or 3 or 2 fold axes of rotational symmetry.
ym vertex units have 5 neighbors (pentavalent) & all others have 6 (hexavalent).
ym Have 60 identical subunits on e surface of icosahedrons.
ym viral nucleic acid is condensed within e isometric particles (virus -encoded core proteins) or cellular
histones.
ym .g.: poliovirus.
b.m Helical symmetry:
 ym protein subunits r bound in periodic way to e viral nucleic acid, winding it into a helix.
 
    ym viral nucleic acid protein complex (nucleocapsid) is coiled inside lipid-containing envelope.
ym All known examples of animal viruses with helical symmetry contain NA genome (except
rhabdoviruses), have flexible nucleocapsids that r wound into a ball inside envelopes.
c.m womplex structures: > complicated in structure such as poxviruses (brick shaped with ridges in e xternal surf.).
2.m nvelopes:
a.m nveloped:
ym polyhedral capsid: Herpes simplex virus.
ym Helical capsid: nfluenza virus. All animal viruses with helical nucleocapsids r
b.m Non-enveloped: enveloped & have NA genome.
ym polyhedral capsid: poliovirus.
ym Helical capsid: Macteriophage virus.
1.m Functions of structural proteins:
ym Facilitate transfer of e viral nucleic acid from one cell to another.
ym protect e viral genome against inactivation by nucleases.
ym participate in e attachment of e virus particle to a susceptible cell.
ym provide e structural symmetry to e virus particles.

 
ym Determine e antigenic characteristics & activities of e virus.
2.m nzymes:
ym present in very small amounts.
ym ssential for e initiation of e viral replicative cycle when e virion enters a host cell.
ym .g.: NA polymerase & reverse transcriptase.


 ym ncodes e genetic information necessary for replication of e virus.


  ym size:
a.m Diral DNA genome: 3.2 kbp to 375 kbp.
b.m Diral NA genome: 7 kbp to 30 kbp.
ym Diral NA genome:
m |ay be single linear molecule.



 m |ay be loosely associated with e virion.
m isolated NA of viruses with +ve sense genomes is infectious & functions as mNA
within e infected cell.
m Dirions carry an NA polymerase that in e cell transcribes e genomic NA molecules
into several complementary NA molecules which serve as mNA.
ym |any viral genomes have been sequenced which reveal e genetic r/ship among isolates.
ym |ay be characterizaed by its G + w content.
 ym lipid is acquired when e viral nucleocapsid buds through a cellular membrane in e
course of maturation.
ym Mudding occurs only at sites where virs -specific proteins have been inserted into e host cell
membranes.
ym specific phospholipid composition of e virion envelope is determined by e specific type of


cell membrane involved in e budding process.

 ym here r viral glycosated proteins protruding from e envelope & exposed on e external surf.
of e virus particle.
ym here r unglycosated proteins of viral origin underneath e envelope that anchor e particle.
ym ipid-containing viruses r sensitive to ether or other organic solvents which leads to loss of
infectivity.
ym Diral envelopes contain glycoprotein which r virus-encoded (e sugar often reflects host cell)
ym Functions:
a.m Attaches e virus particle to a target cell by interacting with cellular receptor.


 
b.m ften involved in e membrane fusion step of infection.
c.m mportant viral antigen.
d.m Frequently involved in e interaction of e virus particle with neutralization Ab.
1.m Heat & cold:
ym cosahedral virus: stable, losing little infectivity after several hours at 37 o w.
ym nveloped virus: heat-labile, rapidly dropping in titre at 37ow, lose infectivity after prolonged storage at -
U0ow, & sensitive to repeated freezing & thawing.
ym Diral infectivity generally destroyed by heating at 50 60 ow for 30 minutes.
ym Dirus can be preserved by storage at subfreezing temperatures.
ym ome may withstand lyophilization & be preserved at room temperature (> resistant when heated in dry
state).
2.m alt:
ym |any viruses can be stabilized by salts in concentration of 1 mol/, e.g.: |gwl 2 & |g 4 (important in vaccine)

   ym alt stabilization can maintain e viral potency for weeks at ambient temperature.
 3.m pH:
ym sually stable b/w pH of 5.0 to U.0.
ym ome r resistant to acidic condit ion but all r destroyed in alkaline condition.
4.m adiation: D light, X-ray, & high energy particles inactivate viruses & lose its infectivity.
5.m photodynamic inactivation: viruses become susceptible to inactivation by visible light when stained with vital
dyes because the dyes bind to e nucleic acid. hus may inactivate progeny virus & cease plaque development.
6.m ther susceptibility: enveloped viruses r e most susceptible.
7.m Detergent:
ym Non-ionic detergents solubilize lipid constituents of viral membranes.
ym Anionic detergents solubilize viral envelopes & disrupt capsids into several polypeptides.
8.m Formaldehyde: destrots virus infectivity by reacting with nucleic acids (single stranded genomes r > affected).




!


   
1.m lectron microscopy (|):
    a.mransmission |: -ve staining of whole virus particles, staining of ultrathin section, & metal shadowing.
b.mcanning |.
 
 2.m X-ray diffraction pattern of crystallized viruses: Has a resolution power down to e atom level.
3.m N| nuclear magnetic resonance.
1.m Direct observation in e electron microscope: Diruses can be visualized in preparations from tissue extracts & in
ultrathin sections of infected cells.
   2.m edimentation in e ultracentrifuge:
ym f particles r suspended in a liquid, they will settle to e bottom at e rate that is proportionate to their size.

ym r/ship b/w e size & shape of a particle & its sedimentation rate permits determination of particle size.
3.m womparative measurement: M/w viruses or b/w bacte ria & viruses.
1.m Detection of virus-infected cells (multiplication of viruses):
a.mDevelopment of wp.
b.mAppearance of a virus-encoded protein such as hemagglutinin.
c.mAdsorption of Mw to infected cells: hemadsorption (d/t virus -encoded hemagluitinin).

    d.mDetection of virus-specific nucleic acid.
 e.mDiral growth in an embryonated chick egg: may result in e death of e embryo.
2.m nclusion body formation:
ym site of development of e virions.
ym |ay be considered as diagnostic aid.
1.m physical method:
ym Quantitative assays such as pw can determine e no. of viral genome copies in a sample.
ym Dirus sequence variation may reduce virus detection & quantification by this method.
    ym physical wounting under |: by comparison with a standard suspension of small latex particles.
 ym Hemagglutination Assay.
2.m Miological method:
ym Depend on measurement of animal death, animal infection, or wp in tissue culture.
ym |ultiple cycles of replication & cell killing produce a small action of infection or plaque.
 ym plaque Assay: A single plaque can arise from a single infectious virus particle called plaque forming unit (pF) .
ym nd point Dilution Assays: to estimate e 50% infectious dose (semi -quantitative).
ym starting material: large volumes of tissue culture medium, body fluids, or infected cells.
ym t is a multistep procedures:
1.m woncentration of e virus particles by:
a.m precipitation with ammonium sulphate, ethanol, or polyethylene glycol, or by ultrafiltration.
b.m Hemagglutination & elution for orthomyxoviruses.
2.m nce concentrated, virus then can be separated from host materials by:
a.m A preliminary purification: Differential centrifugation which removes most nonviral material.
b.m final purification method:
i.m Density gradient centrifugation: n rat e zonal centrifugation, e concentrated virus is layered onto a


  preformed linear density gradient of sucrose or glycerol. During e centrifugation, e viruses sediment
 as a band at a rate determined 1 o by e size & weight of e virus particle.
ii.m High speed centrifugation: nvolves e density gradient of cesium chloride, potassium citrate, or


 sucrose (one which is least toxic to e virus is to be chosen). Dirus particles migrate to an equilibrium
position where e density of e solution = buoyant density & form a visib le band.
ym ther methods:
1.m wolumn chromatography: virus is bound to a substance such as diethylaminoethyl or phosphocellulose &
eluted by changes in pH or salt concentration.
2.m Zone electrophoresis: permits e separation of virus particles from contaminants n e basis of charge.
3.m pecific antisera: remove virus particles from host.
ym cosahedral viruses r easier to purify than envelope viruses b/cause e latter contain variable amounts of envelope
per particle, thus e virus particles e heterogenous in both size & density.
1.m particle can be obtained only from infected cells or tissues.
2.m particles obtained from various sources r identical.
3.m degree of infective activity of e preparation varies directly with e no. of particles present.

  4.m Destruction of e physical particle is associated with a loss of viral activity.
5.m wertain properties of e particles & infectivity must be shown to be identical.
 6.m absorption spectrum of e purified particles in e D range should coincide wi th e D inactivation spectrum of e


 virus.
7.m Antisera prepared should react with e infective particles & vice versa.
8.m particles should be able to induce characteristic disease in vivo.
U.m passage of e virus in tissue culture should result in e production of e p rogeny.
ym easons to inactivation:
1.m o sterilize laboratory supplies & equipment.
2.m o disinfect surfaces or skin.
   3.m o make drinking water safe.
4.m o produce inactivated virus vaccines.
  ym |ethods of inactivation:

   1.m terilization accompanied by steam under pressure, dry heat, ethylene oxide, & gamma irradication.
2.m urface disinfectants include sodium hypochlorite, glutaraldehyde, formaldehyde, & peracetic acid.
3.m kin disinfectants include chlorhexidine, 70% ethanol, & iodophores.
4.m Daccine production may involve D or detergents to inactivate e viruses.
ym Generally, viruses r more sensitive than bacteria & fungi to inactivation by physical & chemical agents.
ym nveloped viruses r more sensitive than naked ones.

   ym Half life of viruses at certain temperatures:
1.m 60o w: measured in seconds.
  2.m 37 o w: measured in minutes.

  3.m 20 ow: measured in hours.
4.m 4 ow: measured in days.
5.m -70 o w or lower: measured in years.
1.m Freezing of small aliquots of virus suspended in medium containing protective protein such as albumin followed
by storage at:
  a.m emperature of w 2 ice: -70 ow.
    b.m emperature of liquid nitrogen: -1U6o w.
2.m Freeze-Drying (lyophilization): dehydration of a frozen viral suspension under vacuum followed by storage at
 4 ow or -20 ow.
3.m tabilization in one molar concentration of certain salts.


  m

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 ym For virus to multiply it must obviously infect a cell.
ym All must make proteins with 3 sets of functions:
1.m nsure e replication of e genome.
2.m package e genome into virus particles.
3.m Alter e metabolism of e infected cell so that viruses r produced.

  

 

,
  ,

1.m Attachment:
ym .e. interaction of a virion (viral attachment protein) with a specific receptor (mostly glycoprotein) site on e surface of a cell.
ym eceptor binding is believed to reflect fortuitous configurational homologies b/w a virion & a cell.
ym absence or presence of receptors determines e cel l tropism & pathogenesis of a virus.
2.m penetration:
ym .e. virus particle is taken up or engulfed inside e cell.
ym |ay involve 3 mechanisms:
a.m eceptor-mediated endocytosis by using endosome & also called as viropexis.
b.m ranslocation or direct penetration oe e enti re virion across e plasma membrane.
c.m Fusion of e viral envelope with e plasma membrane of e cell: involve e interaction of a viral fusion protein with a 2 nd cellular receptor
or coreceptor.
3.m ncoating:
ym .e. e physical separation of e viral nucleic acid from e outer structural components of e virion so that it can function.
ym genome may be released as a free nucleic acid or as a nucleocapsid (usually contain polymerases).
ym ccurs concomitantly with or shortly after penetration.
ym |ay require acidic pH in e endosome & e infectivity of e parental virus is lost at uncoating stage.

       


ym specific mNAs must be transcribed from e viral nucleic acid for successful expression & duplication of genetic information.
ym hen, viruses use cell component to translate e mNA. However, virus -specific proteins may regulate e extent of transcription of e genome or
e translation of viral mNA.
ym Diral protein is synthesized in e cytoplasm on polyribosome composed of v irus specific mNA & host cell ribosome. larger e virus, e >
independent of cellular function & is > susceptible to antiviral chemotherapy (> virus -specific processes r available for drug action).
ym eplication strategy:
1.m NA containing viruses:
m -ve sense viruses: carry NA polymerase to synthesize mNA.
m |ost genetic information is expressed at e same time.
m idest variation in strategies of gene expression:
a.m ome virions carry polymerases.
b.m ome systems utilize subgenomic messages which sometimes generated by splicing.
c.m ome viruses synthesize large polyprotein precursors that r processed & cleaved to generate final gene products.
m Diral genomic NA is generally duplicated in cell cytoplasm.
2.m DNA containing viruses:
m arly viral protein r synthesized soon after inf ection & late protein r made only late in infection, i.e. after viral DNA synthesis.
m DNA viruses that replicate in e nucleus use host cell DNA & NA polymerases & processing enzymes.
 
  -
1.m |orphogenesis or assembly:
ym nvolve e assembly of all e components necessary for e formation of e mature virion at a particular site of e cell.
ym During this stage, e basic structure of virus is formed & n ewly synthesized viral genomes & capsid polypeptide assemble together to form
progeny viruses.
ym cosahedral capsid can condense in e absence of nucleic acid, whereas nucleocapsids of viruses with helical symmetry cannot f orm w/o
viral NA.
ym Non-enveloped viruses accumulate in infected cells & e cell eventually lyse & release e virus particles.
ym site of assembly varies for different viruses.
2.m |aturation:
ym .e. e stage of e life-cycle at which e virus becomes infectious.
ym sually involves structural changes in e particle, often resulting from specific cleavage of polyproteins to form e mature in dividual protein
products.
ym nveloped viruses may mature by a budding process. o that they r not infectious until they have acquired e envelopes. heref ore,
infectious progeny do not accumulate in e cell.
ym However, excess amounts of viral components may accumulate & lead e formation of inclusion body as wp which cause cell death.
3.m elease:
ym ytic viruses (most non -enveloped): e cell breaks open & release e virus.
ym nveloped viruses: acquire e lipid membrane as e virus buds out through e cell membranes. Dirion envelope proteins r picked up during
this process as e virus is extruded.
ym Mudding may or may not kill e cell (controlled by e virus): e physical interaction of e capsid proteins on e inner surf. of e cell membrane
forces e particle out though e membrane.
1.m |ammalian cells can only make DNA & NA in e nuclear compartment i.e. viruses replicating in e cytoplasm must
make their own polymerase.
2.m here is no mammalian cell enzyme able to use NA as a template for NA or DNA synthesis. Diruse s with an NA


  genome have to make their own polymerase.
  3.m serve |any NA viruses with a +ve sense strand do not need to contain a virion -associated polymerase. genomic NA can
as mNA, to make e various viral proteins including polymerase enzymes, upon cel l entry.
4.m NA viruses with a ve sense strand must contain a virion associated polymerase to make a +ve sense strand NA i.e.
mNA upon cell entry.
 !
"+ 




   

  ym ypes of viral infection:
1.m productive infection: occurs in permissive cells & results in e production of infectious virus (few to thousand).

  2.m Abortive infection: fails to produce infectious progeny, either because e cells r non -permissive or unable to
support e expression of all viral genes or because e i nfecting virus may be defective.
3.m atent infection: may ensue with e persistence of viral genomes, e expression of no or a few viral genes, & e
survival of infected cells.

 

 


   
 ym hen host cells caused to be lysed when virus progeny virions released. ym .g.:
ym wan be seen in cell culture which is known as cytophatic effect (wp). 1.m poliovirus: cell lysis.
ym wp: virus-induced damage to e cells that alters its microscopic 2.m mallpox & influenza
appearance. viruses: cell rounding.
ym effect: cells r killed. 3.m Adenovirus: cell clumping.

 
 ym Host cells r not killed (lysed) but undergo neoplastic transformation. ym .g.: NA tumor (oncogenic) virus
ym ntire genome integrates into that of host cells & expresses which r:
     oncogenes. 1.m Human cell leukemia virus.
ym Host cells r transformed to a state of uncontrolled cell division. 2.m arcoma virus.
ym wontinuous release of infectious viral progeny is usually by budding. 3.m |ammary tumor virus.


 ym nfected cells & virus coexist over a long period of not.
ym exact mechanism is not known.
ym Defective interfering particles temperature sensitive mutants & infection (all implicated).
ym .g.: lymphocytis chonemeningitis.
 
 

 
ym initial stage: same as for productive viral infection.
ym hen, non-productive infection can have different courses: apoptosis, non -productive transformation, & latency.
ym |ay be due to:
1.m A block on one stage of replication cycle
2.m Diral genome may be defective. hus, replication cant be completed
3.m Diral genome mat be too small: parvoviridae (need 2 nd helper virus like adenovirus).
4.m |utation.
ym arly viral replication steps: trigger host cell apoptosis response.
ym Host & virus both have no propagation.


  ym .g.: vaceinra virus in whinese hamster ovary cells results in cell rounding, condensation of chromatin, &
fragmentation of DNA.
ym Dirus infects e cell. hen, virus oncogenes integrate with cell DNA leading to cell transformation.
 
 
 ym ransformed cells do not yield infectious progeny virus: non-productive.
     ym .g.: papilloma virus (ware-viruses) of 77 types, some r causally involed in cancer of servix & cancer of
anogenital organs.
ym n 1 oinfection, viral infection produces progeny (productive).
ym hen, 2 o latent infection affects neural cells (for life).
ym Diral genome remains intact during latency in nucleus of host cell.
ym wan reemerge t/out life: especially during immunosuppresed.
 

ym .g.:
1.m HD & H2D: cause chickenpox & shingles (appear in accordance to dermatosome).
2.m pstein-Marr virus (MD): cause nasopharyngeal carcinoma, Murkitts lymphoma, & infectious
mononucleosis.
ym A type of virus-cell interaction. ym wharacteristics of transformed cells:
ym properties of all change dramatically. 1.m oss of contact inhibition of growth.
ym ransform cells have similar properties to tumor cells. 2.m Fewer requirements for serum factors.
     ym .g.: Herpes virus, adenovirus, hepadna virus, 3.m ndefinite no. of cell divisions.
papovavirus, & poxvirus. 4.m Diral Ag exposed.
5.m Fibroretinas absent.
6.m Fetal Ag present.
1.m kin:
ym Mreached through transcutaneous infection & D route.
ym nfection via vectors, usually insects. .g.: togavirus, flavivirus, bunyavirus, reovirus, & rhabdovirus (bite).
   ym Few r from skin lesion: HD, varicella virus, mollusoum contagiosum, & ebola virus.
ym atrogenic infection: Hepatitis M, w, & D while D: papilloma virus, HD, Hepatitis A virus, & HD.
2.m |ucosa: oropharyngeal, G, respiratory tract, & eye.
ym nce infection is established, virus spread:
1.m At e site of entry:
a.m well to cell: viral particles r released at apical end of epithelium.
b.m Disseminated: viral particles r released at basal end of epithelium via hematogenous (viremia) &
 
  lymphatic spread.
2.m Neural spread:
ym |echanism is similar to entry into other cells.
ym ncoated nucleocapsid is carried passively along axons & dendrites by axoplasmic particles.
ym .g.: rabbit virus, poliovirus, reovirus, & herpes virus.

 
 



 

 

!
"


   
ym |utation is a heritable change in e genotype & virus which subjected to mutation causes minor or subtle
genetic changes.
ym |echanisms of mutation:
1.m My e effects of chemical or physical mutagens such as D light & x -ray on nucleic acids.
2.m My e natural behavior of e bases that make up nucleic acids (resonance from keto to same in all
enol & from amino to imino form). viruses.
3.m hrough e fallibility (lack of fedility) of e enzymes that replicate e viral nucleic acids (viruses with high-
fidelity transcriptases have relatively low mutation rates & vice versa).
ym |utation rates:
1.m DNA viruses: similar to those of eukaryotic cells because their replicatory enzymes have proofreading
functions. 10 -8 to 10-11 errors per incorporated nucleotide.
2.m NA viruses: lack of proofreading function in their replicatory enzymes. 10 -3 to 10-4 errors per incorporated
nucleotide.
ym ypes of mutations:
1.m ethal mutation: mutations that interfere with e essential functions of attachment, penetration, uncoating
replication, assembly, & release. apidly lost from e population.
2.m Neutral mutation: d/t redundancy of e genetic code which results in either no change in e viral protein or in
   replacement of an amino acid by a functionally similar amino acid.
3.m Favorable mutation: mutations that do not impair essential viral functions which persist or become fixed in
a virus population.
ym phenotypic Dariations caused by mutation (e mutants):
1.m wonditional-lethal mutants: mutants that r lethal under non-permissive condition but that yields normal
infectious progeny in permissive con dition.
2.m emperature-sensitive mutants: grow at low temperatures (cold-adaptation mutants) but not at high
temperature.
3.m Host-range mutants: able to grow in permissive cells but r abortive in non -permissive cell.
4.m Antigenic drift virus (minor antigenic variation): antigenically altered viruses which r able to cause disease
in previously resistant or immune hosts. .g.: antigenic drift in e epitope of e hemagglutinin spike of
influenza A virus.
5.m Attenuated Daccine trains from mutation: occur in e mutations result in (1) - (3) mutants but with intact
antigenicity & reduced pathogenicity. .g.: e abin vaccine strain of polioviruse.
6.m Mack mutations (reversion): vaccine strain which has fewer mutations & thus less stable & is subjected to
restore e neural virulence. his vaccine strain causes disease in vaccinated individuals.
7.m thers: drug resistant mutants & suppressor mutation (intragenic & extragenic).



 M. 

ym ecombination results in e production of progeny virus that carries trait not found together in either of e
parents.
ym Generally occurs b/w members of e same virus type & yields genetically stable progeny.
ym 2 mechanisms of recombination:
1.m ndependent reassortm ent:
m ccurs when related viruses that have segmented genomes exchange segments during replication.
m hese genes r unliked & assorted at random.
m nly NA viruses have segmented genomes such as orthomyxoviridae, reoviridae, arenaviridae, & etc.
m .g.: Antigenic shift (immediate & major antigenic change) in nfluenza A virus (b/w animal & human
strain) which gives rise to pandemic infection.
2.m ntramolecular recombination:
m Known to occurs in viruses with non-segmented genomes either by:
a.m DNA viruses: occur by annealing, breakage, crossing over, & reunion.
b.m NA viruses: occurs by strand strand switching or strand jumping where no breakage in viral NA
molecules.

M   m esults in e production of:
   a.m New viral serotypes (antigenic types).
b.m Diruses with altered virulence.
c.m eactivation of genetically inactivated viruses.
ym Genetic eactivation:
1.m |arker rescue:
m ccurs b/w e genome of active virion & e genome of a virus particle that has been inactivated in some
way.
m esults in e rescue of certain markers (active gene) of inactivated parents which r present in viable
progeny.
m None of e progeny produced r identical to e inactivated parent, but r genetically stable.
2.m |ultiplicity reactivation:
m production of infectious virus by a cell infected with 2 or > virus particles of e same strain, each of
which has suffered a lethal mutation in a different gene.
m ccurs b/w e damaged nucleic acids of e parents, producing viable genome that can replicate.
m greater e damage to e parental genomes, e larger e no. of inactive particles required per cell to
ensure e formation of viable genome.
ym .e. e inhibition of multiplication of one of 2 viruses infecting e same cell cultures or e whole animals.
ym Does not occur with all viral combinations, 2 viruses may infect & multiply within e same cell as efficiently as in
single infection.

 ym wauses of interference:
1.m ne virus may inhibit e ability of e second to adsorb to e cell, by blocking or destroying its receptors.
2.m ne virus may compete with e second for components of e replication apparatus.
3.m first virus may cause e infected cell to produce an inhibitor that prevents replication of e second virus.

 


 M. 
 
 
ym een in enveloped viruses. 2 types:


   1.m polyploidy: virus particles having multiple copies (nucleocapsid) of same genome.
2.m Heteroploidy: virus particles having multiple copies of different genomes.
ym 3 types:

 

 1.m ranscapsidation.
 2.m |ixed capsidation.
3.m |ixed envelopes.
ym interaction of viral gene products in cell infected with 2 viruses, one or both of which may be defective.
ym esults in e replication of one or both under condition in which replication will not ordinarily occur.


   ym ne virus provides a gene product in which e second is defective, allow ing e second to grow.
 ym genotypes of e 2 viruses remain unchanged.
ym However, if both mutants r defective in e same gene product, they will not able to complement each others
growth.
ym Gene transfer to mammalian cells through transformation, transfection, & microinjection.

 ym ecombinant viruses r developed to carry normal human gene that could be useful as vectors for gene therapy.
ym arget diseases: diabetes, cystic fibrosis, wD, & etc.



!
!



   
ym purpose: to utilize e immune response of e host to prevent viral disease.
 ym mmunity to viral infection is based on e development of an immune response to specific Ags located on e surface
of virus particles (for enveloped virus r e surface glycoprotein) or virus -infected cells.

 


ym nactivated vaccines r made by purifying viral preparations to a certain extent & then inactivating viral infectivity
in a way that does minimal damage to e viral structural proteins.
ym Generally stimulate e development of circulating Ab against e coat proteins of e virus, conferring some degree of
resistance but e replicative function should be destroyed.
ym nactivation may be mediated by heat or chemicals such as formaldehyde or -propiolactone.
ym Advantages:
1.m No reversion to virulence by e vaccine virus.
2.m Gives sufficient humoral immunity if boosters r given.
 3.m wan be used in immunodeficient patients.
4.m ometimes better withstands adverse environmental conditions in e tropics.


 ym Disadvantages:
1.m xtreme care is required in their manufacture to make certain that no residual live virulent virus is present in e
vaccines.
2.m immunity conferred is often brief & must be boosted which involved e concern about e possible effects (H
reactions) of repeated administratio n of foreign proteins.
3.m parenteral administration of this vaccine sometimes gives limited protection because local resistance (gA) is
not induced adequately at e natural portal of entry.
4.m cell-mediated immune response to inactivated vaccine is generally p oor.
5.m ome killed-virus vaccines have induced H to subsequent infection.
ym prepared from attenuated strains that r almost or completely devoid of pathogenicity but r capable of inducing a
protective immune response.
ym |ethods of attenuation:
1.m Administration of pathogenic or partially attenuated virus by an unnatural route: used in e immunization of
military recruits against AD using enterically coated live adenovirus types 4, 7, & 21.
2.m passage of e virus in an unnatural host or h ost cells to induce host-range mutation:
m sed in e most vaccines for men & animals.
m xamples of human vaccines attenuated by inducing host -range mutation:
a.m 17D vaccine strain of yellow fever passed in mice & chick embryos.
b.m ive-attenuated poliovaccine were passaged in monkey kidney cells.
c.m |easles passaged in e chick embryo fibroblast.
d.m ubella & mumps.
3.m Development of temperature-sensitive mutants: used in e conjunction with e above method.
 
ym Advantages:


 1.m Act like e natural infection with regard to their effect on immunity.
2.m hey multiply in e host & tend to stimulate longer-lasting Ab production, to induce a good cellular immunity,
& to induce Ab production & resistance at e portal of entry.
ym Disadvantages:
1.m risk of reversion to greater virulence during multiplication within e vaccine & underattenuation.
2.m nrecognized adventitious agents latently infecting e culture substrate may enter e vaccine stocks.
3.m storage & limited shelf life of attenuated vaccines present problems, but can be overcome by e use of viral
stabilizers.
4.m nterference by coinfection with a naturally occurring, wild-type virus may inhibit e replication of e vaccine
virus & decrease its effectiveness.
5.m preparation instability & heat lability.
6.m wannot be given to immunocompromised or pregnant patients.
7.m pread to contacts of vaccine who have not consented to be vaccinated.
 ym Daccines with purifying viruses & viral Ag.
ym xamples:
M 1.m HA vaccines for influenza A & M.


 2.m HMs Ag vaccine: in e past derived from e plasma of chronic carriers.
ym problem: increasing purification may lead to loss of immunogenicity & this may necessitate coupling to an
immunogenic carrier protein or adjuvants such as aluminium salt.
ym produced by recombinant DNA technology.
ym 2 types:
1.m ecombinant viral proteins:
m Dirus proteins expressed in bacteria, yeasts, & mammalian cells.
m xamples:

M  a.m . coli r 1 st to be used, but e expressed proteins r not glycosylated & caused drawback.


 b.m ecombinant Hep M vaccine is e only recombinant vaccine licensed at present.
2.m Hybrid virus vaccine:
m An alternative application of recombinant DNA technology.
m xample: polyvalent live hybrid virus vaccine e gene of several viruses or even other organism can be
inserted into virus (e.g.: Daccinia virus) to generate a hybrid virus v accine that can express several
exogenous immunizing Ags.
ym vaccination depends on e identification of immunogenic sites. .g.: food & mouth disease.
ym Advantages of defined viral Ags or peptides:
1.m production & quality control simpler.
2.m No NA or other viral or external protein, therefore < toxic.
 
 3.m afer in cases where viruses r oncogenic or establish a persistent infection.
4.m Feasible even if virus cannot be cultivated.


 ym Disadvantages of defined viral Ags or peptides:
1.m |ay be less immunogenic than conventional inactivated whole-virus vaccines.
2.m equires adjuvant or carrier protein.
3.m equires 1 o curse of injection followed by boosters.
4.m Fails to elicit w|.
ym Has been developed d/t e ability of anti-idiotype Abs to mimic foreign Ags & induce against e viruses, bacteria,
& protozoa.
 
 ym Advantages of anti-idiotype antibodies:
M  1.m Have many potential uses as viral vaccines, particularly when e Ag is difficult to grow or hazardous.
2.m Have been used to induce immunity against a wide range of viruses, including HMD, rabies, Newcastle, disease
virus & FeD, reoviruses, & polioviruses.
ym DNA coding for e foreign Ag is directly injected into a relatively no -dividing tissue such as muscle of an animal so
that e foreign Ag is directly produced by e host cells.
ym Advantages:
1.m DNA is relatively inexpensive & easier to produce than conventional vaccines.
2.m time for development is relatively short which may enable timely immunization against emerging infectious
diseases.
ym precautions:
 

 1.m Delivery of e DNA to cells is still not optimal.
2.m possibility that e vaccines DNA will be integrated into host chromosomes & turn on e oncogene or turn off e
tumor suppressor gene.
3.m xtended immunostimulation by e foreign Ag may provoke chronic inflammati on or autoAM production.
ym xamples:
1.m 1st clinical trial: HD in 1UU5 & subsequent trial in 1UU6. echnique used: direct injection of plasmids which is
2.m Daccines for influenza virus, HD, & cell lymphoma. taken up by host cells for expression.
ym Advantages: ym Disadvantages: non-proteinous Ags like polysaccharides
1.m Fast access to virtually every Ag. & glycolipid cannot be used.
 2.m No-cultivable can be approached.


    3.m Non-abundant Ags can be identified.
4.m Ags not expressed in vitro can be identified.
5.m No-structural proteins can be used.
ym proteins separated from virus particles r generally much less immunogenic than e intact particles.
ym his different is d/t e change in configuration of a protein when it is released from e structural requirements of e
virus particle.
ym Attempts to enhance e immunogenic activity of separated proteins by:
1.m Adjuvants:
m potentiate e with modes of action:

   a.m Functions to localize & slowly release Ag at or near e site of administration.
 b.m Functions to activate Apws to achieve effective Ag processing or presentation.
m |aterials used as adjuvants:
 
 a.m Aluminium salt: e only used for human vaccines.

  b.m |ineral oils: vaccines administrated as a water-in-oil emulsion which is slow in breakdown &
gradually release e Ag. t can be potentially carcinogenic.


 c.m Macterial cell wall: mycobacterial product (Freunds adjuvants).
2.m iposomes:
m .e. artificial lipid membrane spheres into which immunizing proteins (& adjuvants) r incorporated.
m wan stimulate w|.
m immunostimulating complexes have activities equivalent to those of e virus particles from which e
proteins r derived.
1.m Different types of virus may cause similar disease.

 M 2.m Antigenic drift & shift, especially true of NA viruses & those with segmented genomes.
3.m Animal reservoirs which can lead to reinfection after elimination from e human population.


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ym Arbovirus: arthropod borne virus (those infecting human r zoonotic).
 ym |ultiply in tissue of arthropods & r passed into new vertebrates when arthropods take a subsequent blood meal after
a period of extrinsic incubation.
ym No taxonomic significance with 535 heterogenous group of virus.
ym nder 7 taxonomic families & many r zoonotic which do not require arthropod vectors.

Family Genus hape Nucleic acid nvelope

1.m Alpha virus
ogaviridae wube +ve ssNA +
2.m ungroup
Flaviviridae Flaviviparous wubic +ve ssNA +
1.m Munyavirus
2.m phlebovirus
     Munyaviridae 3.m Nairovirus Helical -ve ssNA +
4.m ukuvirus
5.m Hantavirus
1.m rbivirus
eoviridae 2.m wotivirus wubic dsNA -
3.m nassigned
1.m Desicuvirus
habdoviridae 2.m yssavirus Mullet-shape -ve ssNA +
3.m nassigned
ym 3 clinical syndromes:
1.m Fever of an undifferentiated type with or w/o a moculopapular rash & usually benign.
  2.m ncephalitis often with high fatality rate.
3.m Hemorrhagic fever which is frequently severe & fatal.
ym degree of viral multiplication & its predominant site of localization in tissue determine e clinical syndromes.

ym Dengue (breakbone fever) is a mosquito -borne infection caused by a flavivirus that is characterized by fever, severe
headache, muscle & joint pain, nausea & vomiting, eye pain, & rash.

    ym 4 serotypes of dengue virus transmitted in tropical area b/w 35 oN + 35 o.
ym Dectors: A. aegypti (principle), A. albofictus, A. polynesiesi, A. cochi, & A. scuttelaris bebrideus.
ym Definitive & reservoir host: only human, thus not a zoonosis.
ym principle vector which is A. aegypti: m n endemic area: 1/20 mosquitoes is infected.
m Mread around human, but aviposits in stagnant water. m ransmission in tropical area is t/out e year, high

  m Adults take shelter indoor. during raining season, high humidity, long lifespan,
m Mite during 1-3 hours, in e morning & late afternoon. high temperature, & low extrinsic incubation
m Fly as long as 0.8 to 2.0 km. period.
1.m Dengue fever:
ym sually is a self-limited infection.
ym wlinical disease begins 4-7 days after infection.
ym onset of fever may be sudden with malaise (d/t cytokine response) , chills, & headache.
ym pain develops especially in e back, joints, muscles, & eyeball: d/t HD perivascular mononuclear infiltrate & lipid
accumulation (high creatinine phosphokinase & hepatic transaminase).
ym Diremia at e onset of fever & may persist for 3 -5 days but e temperature returns to normal after 5 -6 days &
subside on e 3 rd day & rise again about 5-8 days after onset (saddleback pattern).
ym ash (lymphatic dermal vasculitis) may appear on e 3 rd or 4 th day & lasts for 1-5 days.
ym Dirus infects bone marrow & causes peripheral cytopenia & hemophagocytosis.
2.m Dengue hemorrhagic fever (DHF):
ym |ay occur in individuals with passively acquired or preexisting nonneutralizing heterologous dengue Ab d/t

  previous infection with a different serotype of virus.

ym nitial symptoms stimulate normal dengue, but e patients condition is abruptly worsen.
3.m Dengue shock syndrome (D):
ym wharacterized by shock & hemoconcentration.
ym Fatal cases d/t kuppfer cells hyperthropy & focal ballooning.
ym hock is d/t sudden extravasation of plasma into extravascular compartment including pleura & abdominal
cavity usually when fever has declined.
ym D is also d/t high NF, s, FN->, & complement activation.
ym esults: loss of fluid, hypoproteinemia, & albuminemia.
ym eversal usually within 48 hours of DHF.
ym pathogenesis: 2ndary infection l heterologous or enhancing, non-neutralizing Abs increase uptake of e virus l
increase replication in Fc-bearing cells (Abs mediated immune enhanced) l high NF-, wD8, & 2 l cross
reactivity l wD4 & wD8 increase l high FN-> & -2 l D.
ym Family: flaviviridae.
ym Dirions:
    m wonsists of | (membrane) glycoprotein & (envelope) glycoprotein.
m Dimmers packed horizontally head to tail on virion surface.
m nstable in heat, D light, disinfectant (alcohol & iodine), acid, & ether.
1.m Dengue fever: acute fibrile disease, headache, musculoskeletal pain, rash, fever, malaise, rash, irritability, & .



2.m DHF & D:


 ym ffusion of pleura (80%), ascites, gallbladder, pararenal, liver, spleen, & pericardial.
 ym |yocardial dysfunction, metabolic acidosis, respiratory distress, encephalitis, liver & renal failure.
ym Fatality rate: up to 50%.
3.m Adults:
ym > severe & acute.
ym whills, severe frontal headache, retroorbital pain, severe musculoskeletal & lumbar back pain.
ym kin flushed & rashes.
1.m pw: for rapid identification & serotyping of dengue virus in acute phase serum during e period of fever.
2.m Diral isolation: done 1 week after onset by inoculation of a mosquito cell line with patients serum, coupled with NA
assays.
3.m wulture: specimen obtained from biopsy or autopsy & cultured in suckling mice.
 M   4.m erological test:
ym My cross-reactivity of gG to heterologous flavivirus Ags.
ym pecimen obtained from blood & wF.
ym .g.: A & hemagglutinin inhibition test.
  ym reatment is symptomatic & supportive (fluid replacement therapy).
ym |ainstay:

  1.m public health: insectidal fogging & vaccine (under development).
2.m personal hygiene at home.



!
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ym nique property: ability to establish lifelong persistent infections in their hosts & to undergo periodic reactivation.
However, e reactivated infection may be clinically quite different from e disease caused by 1 o infection, but is >
serious in immunocompromised patients.
ym tructure & composition:
1.m Dirion: large & spherical virus with diameter of 150 200 nm (enveloped) & of 125 nm (naked).
 2.m Genome: double-stranded, linear, 125-240 kbp DNA in e form of toroid & reiterated sequences, surrounded
by a protein coat that exhibits icosahedral symmetry & has 162 capsomeres.
3.m Nucleocapsid: surrounded by an envelope that is derived from hosts nuclear membrane & contains viral
glycoprotein spikes about 8 nm long & Fc receptors.
4.m proteins: > than 35 proteins in e virion.
5.m egument: an amorphous, asymmetric structure b/w capsid & envelope.
1.m -herpesviruses:
ym Fast-growing, cytolytic viruses that tend to establish latent infection in neurons.
ym |embers: HD 1&2 & Daricella-zoster virus.
2.m -herpesviruses:

 
  ym low-growing & cytomegalic viruses which become latent in secretory glands & kidneys.
ym |embers: w|D, HHD-6, &HHD-7.
3.m >-herpesviruses:
ym nfect & become latent in lymphoid cells.
ym |embers: MD & HHD-8.
ym Dirus binds to receptor on envelope glycoprotein (glycosaminoglycan) l enters e cell by fusion with e cell
membrane l e capsid is transported trough e cytoplasm to a nuclear pore l capsid uncoating l DNA
associated with e nucleus l viral DNA becomes circle immediately upon release from e capsid l Dp16, a
tegument protein, complexes with several cellular protein & activat e initial viral gene expression l immediate-


  early genes r expressed l -proteins r produced l e proteins permit expression of e early set of genes l
translated into -proteins l viral DNA replication begins l late transcripts r produced & produce >-proteins l
maturation occurs by budding of nucleocapsids through e altered inner nuclear membrane l enveloped viral
particles r transported by vesicular movement to e cell surface.
ym ength of replication is 18 hours (HD) 70 hours (w|D).



ym 2 serotypes of HD-1 & HD-2 which grow rapidly & r highly cytolytic.
ym genomes:
m arge ( 150 kbp) & can encode at least 70 polypeptides.
m imilar in organization & exhibit substantial sequence homology.
m wan be distinguished by sequence analysis or by restriction enzyme analysis of viral DNA.
ym hey share several cross -reactive epitopes with each other & also with DZD.


 ym |ode of transmission:
1.m HD-1: spread by contact usually involving infected saliva or tears. |an is e only
2.m HD-2: transmitted sexually or from a maternal genital infection to a newborn. natural host.
ym ypes of viral glycoproteins:
1.m ne (gD): e most important inducer of 3.m g: an Fc receptor.
neutralizing Abs. 4.m Glycoprotein G: type-specific & allow for
2.m Glycoprotein w: w3b-binding protein. antigenic discrimination b/w HD-1 & HD-2.
1.m HD-1:
m 1 o infection occurs early in life (6 months to 3 years) & usually asymptomatic with development of Abs, but e virus
is not eliminated.
m A carrier state is established t/out life & punctuated by transient recurrent attacks of HD.

    m isk factors: lower socio -economic population with crowded living condition & poorer hygiene.
m Generally causes infection above e belt.
m frequency of recurrenc es varies widely among individuals.
2.m HD-2:
m eroprevalence is higher among women than men & higher among blacks than white.
 m Abs r seldom found before puberty d/t D.
m ecurrent genital infections may be symptomatic or asymptomatic & tend to recur > often than HD -1.
m |aternal genital HD infection poses risks both to mother & fetus.
m Genital HD infection increases in HD type 1 patients.
1.m pathology:
ym wytolytic infection d/t necrosis of infected cells with inflammatory response.
ym esion in e skin & mucous membrane is e same in HD -1 & HD-2.
ym wytophatic effects:
a.m well ballooning.
b.m productions of wowdry type A intranuclear inclusion bodies.
c.m |argination of chromatin.
d.m Formation of multinucleated giant cells.

  e.m well fusion: provides method for cell to cell spread.
o
2.m 1 infection: Dirus enters through mucosal surface or broken skin l replication at e site of infection l invasion of local
nerve ending l transported to dorsal ganglia by retrograde axonal flow l further replication l latency.
3.m atent infection:
ym stablish latency in: trigeminal ganglia (HD-1) & sacral ganglia (HD-2).
ym atency: non-replicating state which uses microNA to prevent cell death & maintaining e latency. atency lasts
for e lifetime & no virus can be recovered b/w recurrences.
ym provocative stimuli can reactivate e virus such as axonal injury, stress, or exposure to D light.
ym eactivation: virus follows axons back to e peripheral site & replication proceeds at e skin or mucous membrane.
1.m ropharyngeal disease (HD-1):
ym |ost frequently in small children (1-5 years) & involves e buccal & gingival mucosa of e mouth.
ym ncubation period: 3- 5 days with clinical illness lasts 2-3 weeks.
ym ymptoms: fever, sore throat, vesicular & ulcerative lesions, gingivostomatitis, pharyngitis, tosillitis & malaise.
ym ecurrent disease: cluster of vesicles localized at e border of e lip with intense pain which fades over 4 -5 days.
ym Desicles l pustules l crusting stage l heal w/o scarring after 8 -10 days.
2.m Keratoconjunctivitis (HD-1):
ym ecurrent lesion: dendritic keratitis, corneal ulcers, or vesicles on e eyelids.
ym here may be progressive involvement of corneal stroma with permanent opacification & blindness.
3.m Genital herpes (HD-2):
ym 1 0 infections:
m asts about 3 weeks & characterized by vesiculoulcerative lesion of e penis or cervix, vulva, vagina, &
perineum which is painful & present with fever, malaise, dysuria, & inguinal lymphadenopathy.
m womplications: extragenital lesions, aseptic meningitis, & viral excretion ( 3 weeks).
ym ecurrences: common & mild with appearance of vesicle about 10 days.
4.m kin infections:
ym ccur in abrasion that becomes contaminated with e virus.


 ym esions r seen on e fingers of dentists & hospital personnel & e body of wrestlers.
 ym Mecome severe & life threatening in individuals with skin disorders that permits extensive local viral replication &
spread:
a.m czema herpeticum: a potentially serious disease that occurs in patients with eczema.
b.m Herpetic withlow which arise from implantation of e virus into e skin & typically affect e fingers.
5.m ncephalitis:
ym A necrotizing encephalitis which is localized & usually at hippocampus.
ym Has a high mortality rate & often survive with neurological defects.
6.m Neonatal herpes:
ym Acquired in utero, during birth (through birth canal with herpetic lesion: avoid by using cesarean) o r after birth
(from family members or hospital personnel).
ym 3 categories of disease:
a.m esions localized to e skin, eyes, & mouth. Diral pneumonitis or
b.m ncephalitis with or w/o localized skin involvement. intravascular
c.m Disseminated disease involving multiple organs such as wN. coagulopathy.
ym |ortality rate of untreated disease is 50% & e prognosis can be worst.
7.m nfections in immunocompromised patient s:
ym nvolved patients who r HD patients, organ transplantation, malignancies, malnutrition, & immunosuppressed d/t
diseases or therapy.
ym |any newborns acquire passively transferred maternal Abs which lost after 1 st 6 months of life which is e period of
greatest susceptibility until 2 years old.
ym HD-1 Abs begins to appear at e early childhood, & by adolescence, they appear in most persons.
  ym HD-2 Abs rise during e age of adolescence & sexual activity.
ym n 1o infections, g| appears 1 st, then followed by gG & gA Abs that persist for long periods.
ym Abs do not prevent reinfection or re activation of latent virus but may be modify subsequent disease.
1.m wytopathology: specimen obtained from e base of e vesicles & e presence of multinucleated giant cells indicates HD is
present.
2.m solation & identification: virus may be isolated from herpetic lesions, throat washing, wF, & stool both during 1 o &
asymptomatic periods, but is not sufficient evidence for indication of causative agent.
3.m erology: Abs appear in 4-7 days after infection & reach a peak in 2 -4 weeks which is useful to document to recent
 M   infection.
4.m Direct detection:
a.m lectron microscopy of vesicle fluid: ra[ but cant distinguish b/w HD & DZD.
b.m pw: now is used routinely for e diagnosis of herpes simplex encephalitis (from wF).
c.m mmunofluoresence of skin wrappings: can distinguish b/w HD & DZD.
1.m Drug therapy:
  ym D w: acyclovir, valacyclovir, & vidarabine which r e inhibitors of viral DNA synthesis.

  ym drugs may suppress clinical manifestation, shorten time to healing, & reduce recurrences of genital herpes.
2.m Daccination: purified glycoprotein Ags found in e viral envelope.
 3.m prevention: susceptible patients should be protected from e virus.
 
  
ym properties of e virus:
m t has no animal reservoir, but virus propagates in e cultures of human embryonic tissue & produce typical
intranuclear inclusion bodies.
m wp r > focal & spread much > slowly than HD.
m nfectious virus remains strongly cell -associated & serial propagation is > easily accomplished by passage of infected
cells than of tissue culture fluids.


 m ame virus produces both chickenpox & shingle, & children who have recovered from zoster virus -induced infection
r resistant to varicella.
ym verview of e diseases:
1.m Daricella (chickenpox): mild, highly contagious disease (children) characterized by a generalized vesicular eruption
of e skin & mucous membranes. disease is severe in adults & immunocompromised patients.
2.m Zoster (shingle): a sporadic, incapacitating disease of adults & immunocompromised individuals that is
characterized by a rash on e skin innervated by a single sensory ganglion.
ym Daricella & zoster occur worldwide.
ym Daricella is highly communicable & is a common epidemic disease of childhood, but rare in adulthood (common in
winter & spring).

    ym A live attenuated vaccine is available.
ym Daricella spreads readily by airborne droplets & by direct contact but r less common in zoster because e virus is absent
from e upper respiratory tract.
ym A varicella patient is infectious shortly before e appearance of rash to e 1 st few days of rash.
1.m Daricella:
ym oute of infection: e mucosa of e upper respiratory tract or e conjunctiva.
ym nitial replication in regional lymph nodes l 1o viremia spreads virus l replication in liver & spleen l 2o viremia
involving infected mononuclear cells transports virus to skin l rash develops l swelling of epithelial cells, ballooning
degeneration, & accumulation of tissue fluid l vesicle formation.
ym Dirus replication & spread r limited by host humoral & cellular .

  2.m Zoster:

ym kin lesions r histopathologically identical to varicella.
ym ensory neurons & ganglia: acute inflammation & only single ganglion may be involved.
ym distribution of lesions in skin corresponds closely to areas on innervation from an individual dorsal root ganglion.
ym aning immunity allows viral replication to occur in a ganglion, causing intense inflammation & pain. Dirus travels
down e nerve & induces vesicle formation.
ym wellular immunity is e most important host defense & reactivations r sporadic & recurs infrequently.
1.m Daricella:
ym ncubation period of typical disease is 10 -21 days.
ym ymptoms: malaise & fever l rash on e trunk l rash on e face, limbs, buccal & pharyngeal mucosa in e mouth l
successive fresh vesicles appear in crops l all stages of macules, papules, vesicles, & crusts may be seen at one time (e
rash lasts about 5 days).
ym womplications: rare in normal children but encephalitis can occur & varicella pneumonia is most common com plication
& mortality rate is very low.
ym Neonatal varicella:
m infection is contracted from e mother just before or after but w/o sufficient to modify e disease.


 m Dirus is often widely disseminated & may prove fatal.
 m Daricella according to e week of gestation:
a.m 1st 20 weeks of pregnancy: scarring of skin & hypoplasia of limbs (3% chance of transmission).
b.m After 20 week gestation: DZD can cross e placenta in e late stages of pregnancy to infect e fetus congenitally &
e disease very fom mild to a fatal disseminated infection.
ym mmunocompromised patients: increased risk of complications & Dw may occur & rapidly fatal.
2.m Zoster:
ym tarts with severe pain in e area of skin & mucosa supplied by one or > group of sensory nerves & ganglia.
ym Few days after onset, a crop of vesicles appears along e dermatotomes & e trunk, head, & neck r most commonly
affected.
ym complication in elderly is postherpetic neuralgia which is protracted pain that may continue for months.
ym previous infection with varicella is believed to confer lifelong immunity to varicella.
ym Abs induced by varicella vaccine persist for at least 20 years.
  ym Zoster occurs in e presence of neutralizing Ab to varicella.
ym ncreases in varicella Ab titer may occur in persons with HD infection.
ym wellular immunity is important host defense for recovery.
1.m Direct detection: electron microscopy may be used for vesicle fluids but cant distinguish b/w HD & DZD, but F on skin
scrapping can distinguish b/w e two.
2.m Dirus isolation: rarely carried out as it requires 2 -3 weeks for result.
 M   3.m erology: e presence of DZD gG is indicative for past infection & immunity, while e presence g| is indicative of recent
1o infection.
4.m zank smear: multinucleated giant cells r seen which r absent i n non-herpetic lesions.
1.m reatment:
ym Daricella in normal children is mild & self-limiting.
ym Acyclovir is shown to accelerate e resolution of e disease & should be given promptly immunocompromised
individuals with varicella infection & normal individuals with serious complications such as pneumonia &
 
encephalitis.

  2.m prevention:

ym A live attenuated varicella vaccine is generally used & is highly effective at inducing protection from varicella in
children, but less in adults & U5% effective in preventing severe disease.
ym hingle vaccine is effective in older adults at reducing both e frequency of outbreaks of zoster & e severity of
 disease that occur.

   
ym biquitous herpesviruses belong to -herpesvirus subfamily.
ym Has e largest genetic content of human herpesviruses with DNA genome of 240 kbp.
ym Nucleocapsid is 105 nm in diameter with 162 capsomers.
ym cell surface glycoprotein acts as an Fc receptor that can non -specifically bind e Fc region of Abs which may help in
immune system invasion.



ym w|D is very species-specific & cell-type specific.
ym w|D replicates very slowly in cultured cells & infection is spread 1 o cell to cell.
ym wp: perinuclear cytoplasmic inclusion, intranuclear inclusions typical of herpesvirus, multinucleated giant cells,
enlargement of many cells, & inclusion-bearing cytomegalic cells.
ym nfection sources: breast milk, urine, saliva, semen, & cervical secretion.
ym w|D is endemic in all over e world causingcytomegalic inclusion disease.
ym prevalence of infection varies with socioeconomic status, living conditions, & hygienic practices.
ym Abs prevalence may be moderate in adults of high economic status.
ym Human r e only one known host for w|D & once infected, e person carries e virus for life which may be activated from
time to time.
ym ransmission requires close person to person contact & virus may be shed in urine, saliva, semen, breast milk, & white

   
blood cells.
ym dominant routes r oral & respiratory spread but can also be spread transplancentally, by blood t ransfusion, by organ
transplantation, & by sexual contact.
ym eactivation of latent maternal infections can lead to vertical transmission.
ym w|D infections r markedly increased in immunosuppressed populations such as HD patients & organ transplant
recipients.
1.m Normal host:
ym |ay be transmitted person to person requiring close contact with virus -bearing material.
ym ncubation period: 4-8 weeks in normal older children & adult.
ym Dirus causes systemic infection & r isolated from lung, liver, esophagus, colon, kidneys, monocytes, & M cells.
ym disease is known as infectious mononucleosis -like syndrome.
ym w|D establishes lifelong latent infections & can be shed intermittently from e pharynx & urine.
ym w| is depressed during 1 o infection which contributes to e persistence of viral infection & may take several months
to recover.
2.m mmunosuppressed host:
ym 1 o infection is > severe in immunosuppressed persons such as organ transplantation, malignancy, chemotherapy, &

   
AD patients.
ym Diral excretion is increased & prolonged & e infection is > apt to become disseminated & pneumonia is e most
common complication.
ym n seropositive individuals, e host immune response presumably maintains w|D in a latent state.
3.m wongenital & perinatal infection:
ym w|D can be transmitted in utero with both 1 o & reactivated maternal infection can also be acquired from e
exposure to virus in mothers genital tract during delivery & from maternal breast milk.
ym Fetal & newborn infections with w|D may be severe.
ym Generalized cytomegalic inclusion disease results from 1 o maternal infection.
ym No evidence supports that gestational age at e time of maternal infection affects expression of disease in fetus
1.m Normal host:
ym sually asymptomatic but occasionally causes a spontaneous mononucleosis syndrome (heterophil -negative)
which is mild & rarely results in complications.
ym wharacteristics: malaise, myalgia, protracted fever, liver function abnormalities, & lymphocytosis.
ym ubclinical hepatitis is common, but hepatosplenomegaly in younger children is frequently observed.
2.m mmunocompromised host:
ym womplications: pneumonia, interstitial pneumonitis (M| transplantation), virus-associated leucopenia (solid organ


 transplantation), obliterative boncholitis in lung transplants, graft artherosclerosis (heart transplantation), & w|D -
 related rejection of renal allograft.
ym n AD patients: disseminated disease, gastroenteritis, chorioretinitis, & progressive blindness.
3.m wongenital & perinatal infection:
ym wharacterized by involvement of e wN & D system: may result in death (rate: 20%).
ym wlinical features: intrauterine growth retardation, jaundice, hepatosplenomegaly, thrombocytopenia,
microcephaly, & retinitis.
ym urvivors will develop wN defect, severe hearing loss (10%: deaf), ocular abnormalities, & mental retardation.
ym Acquired infection: virus is shed in e saliva & urine of infected individuals for weeks & months.
ym Abs to w|D occurs in most human sera.
  ym presence of Abs in breast milk does not prevent transmission of infection to breast -feeding infants.
ym |aternal Abs protect > against development of serious disease in e infant than viral transmission.
1.m Direct detection:
a.m Miopsy specimens may be examined histologically for w|D inclusion Abs or for e presence of w|D Ags. However,
sensitivity is low.
b.m pp65 w|D antigenaemia test is now routinely used for e rapid diagnosis of w|D infection in
immunocompromised patients.
 M   c.m pw for w|D-DNA is used in some centers but there may be problems with interpretation.
2.m Dirus isolation:
a.m wonventional cell culture regarded as gold standard but requires up to 4 weeks for result.
b.m > useful & rapid culture method as DAFF test which can provide result in 24 -48 hours.
3.m erology: e presence of g| indicates 1 o infection while e presence gG indicates 2 o infection.
  1.m reatment:

  ym wongenital infection: perhaps an abortion.
ym perinatal & postnatal infection: no treatment.
ym mmunocompromised patients: make e diagnosis early & give prompt antiviral therapy.
ym D w: ganciclovir which reduces e symptoms.
2.m prevention:
ym No licensed vaccine is available but owne strain (live attenuated) has been developed w/o used yet concerns to
its latency & reactivation.
ym |easures taken to avoi d w|D infection in organ transplantation:
a.m creening & matching e w|D status of e donor & recipients.
b.m se of w|D ve blood for transfusion.
c.m Administration of w|D g to seronegative recipients prior to transplantation.
d.m Gadministration of antiviral prophylaxis such as acyclovir & ganciclovir.


M 
ym A ubiquitous >-herpesvirus that causes acute infectious mononucleosis (a polyclonal stimulation lymphocytes).
ym genome:
m 172 kbp, has a G + w content of 5U% & encodes about 100 genes.
m Does not normally integrate into e cellular DNA but forms circular episomes which reside in e nucleus & amplified
during phase in a cell cycle.
ym arget cell is M-lymphocytes:
m nfection leads to e establishment of continuous cell lines indicating e immortaliz ed M-cells.
m |echanism: MD binds to M cell by binding to w3d receptor l MD directly enters latent state in e cells w/o
undergoing complete viral replication l MD enters cell cycle l a limited repertoire of MD genes r expressed l


 indefinite cell proliferation l release virus particles.
m MD-immortalized M cells express different function such as secretion of gs, M cell activation products expression, &
expression of at least 10 viral gene products.
ym atency: viral persistence, restricted virus expression, & e potential of reactivation & lytic replication.
ym eplication: replicates in vivo in epithelial cells of e oropharynx, parotid gland, uterine cervix, & nasopharyngeal
carcinoma.
ym Diral Ags:
a.m atent phase Ags: synthesized by latentl y infected cells which include MNAs & |ps.
b.m arly Ags: nonstructural proteins whose synthesis is not dependent on viral DNA replication.
c.m ate Ags: structural component of e viral capsid.
ym ransmission is by contact with saliva, particularly by kissing.
ym 2 epidemiological patterns:

    1.m n developed countries: 2 peaks of infection i.e. b/w age 1 -6 years old & b/w age 14 -20 years old. ventually, 80-
U0% adults r infected.
2.m n developing countries: infection occurs at 2 years old, & U0% of children r s eropositive.
1.m primary infection:
ym nfection initiates in e oropharynx where viral replication occurs in epithelial cells of e pharynx & salivary gland as
well as M lymphocytes l infected M cell spread e infection t/out e body.
ym Normal individuals: most virus-infected cells r eliminated, but small no. of latently infected lymphocytes persist for
e life time in e host.
ym Autoantibodies r typical of e disease with heterophil Abs that react with Ags on sheep Mw.

    2.m eactivation from latency:
ym ccurs as evidence by increased levels of virus in saliva & of DNA in blood cells.
ym sually clinically silent or subclinical.
3.m wancer:
ym MD is a cause of Murkitts lymphoma, nasopharyngeal carcinoma, Hodgkins disease, & others.
ym patients will have elevated level of Abs to virus specific Ags & e tumor tissues contain MD DNA.
1.m nfectious mononucleosis:
ym ncubation period: 30-50 days.
ym ymptoms: headache, fever & malaise (persists for months after acute illness), fatigue, sore throat, enlarged lymph
nodes & spleen.
ym ypical illness is self limited & lasts about 2-4 weeks.
ym wlinical features: increase in e no. of circulating Mw, mainly large & atypical lymphocytes.
2.m Murkitts lymphoma:
ym ccurs endemically in parts of Africa & papua New Guinea usually in children aged 3-14 years.
ym t restricted to areas with holoendemic malaria which becomes e cofactor of e disease.
ym t responds favorably to chemotherapy.
ym |ultiple copies of MD genome & Ags can be found in M with high titre s of Abs against MD Ags.
ym M shows reciprocal translocation b/w e long arm of chromosome 8 & chromosomes 14, 2, or 22 which results in e


 deregulation of e c-myc gene.
 ym poradic cases occur especially in AD patients which may or may not be associated with MD.
ym n theory, it can be controlled by e eradication of malaria or vaccination against MD.
3.m Nasopharyngeal carcinoma:
ym .e. a malignant tumour of e squamous epithelium of nasopharynx which is very prevalence in . whina but rare in
other parts of e world.
ym |ultiple copies of MD genome & MNA-1 Ags can be found in cells of undifferentiated Npw.
ym High titres of Abs against MD Ags r present & usually e prognosis is poor.
4.m Disease in immunodeficient hosts:
ym After infection, MD maintains a steady low-grade infection in e body, & if in immunocompromised patients, e
virus will be reactivated.
ym n transplant recipient: lymphoproliperative disease may be developed & r fatal with e lesions tend to be
extranodal & in unusual sites such as at G or wN.
ym n AD patients: oral leucopenia & non-Hodgkins lymphoma.
 ym Ducan X-linked lymphoproliperative syndrome which occurs exclusively in males with a defective gene in e X -
chromosomes which counts for half of e fatal cases of immunocompromised patient.
  ym wellular immunity & cytotoxic cells r important in limiting primary infections & controlling chronic infections.
1.m Acute MD infection is usually made by e heterophil Ab test or detection of anti -MD DwA g|.
2.m Murkitts lymphoma: diagnosed by histology or e tumor can be stained with Abs to lambda light chain which will
 M   reveal a monoclonal tumor of M cell origin.
3.m Npw: diagnosed histologically or screening of early lesion of Npw (also for treatment) by determination of e titre of
anti-MD DwA gA.
ym here is no MD vaccine available.
 
ym Acyclovir reduces MD shedding from e oropharynx during period of drug administration but it does not affect e no. of

  MD-immortarlized M cells.


!






   
ym Hepatitis: inflammation of e liver. ym Acute viral hepatitis r characterized by fulminant &
ym |icroscopic characteristics: massive hepatocellular necrosis with jaundice.
1.m potty parenchymal cell degeneration. ym whronic hepatitis:
2.m Necrosis of hepatocytes. 1.m whronic carriers:
3.m A diffuse inflammatory reaction. a.m poradically abnormal aminotranseferase values
4.m Disruption of liver cell cords. & hepatomegaly.

    5.m Kupffer cell hyperplasia. b.m portal inflammation & swollen & pale
6.m periportal infiltration by mononuclear cells. hepatocytes.
7.m Accumulation of macrophages near 2.m whronic active hepatitis: inflammation, necrosis, &
degenerating hepatocytes. collapse of e normal reticulum framework with
ym However, preservation of e reticulum framework bridging b/w e portal triads or terminal hepatic veins.
allows hepatocytes regeneration & liver architecture ym None of e viruses r cytophatogenic & is belived that e
can be maintained. his takes place in 8-12 weeks. cellular damages seen r immune-mediated.
aundice Acute case- womplication/ |ortality from
Dirus ncubation period
(years) fatality rate chronicity chronicity
No chronicity
< 6: < 10%
Average: 30 days. Fulminant,
A 6-14: 40 - 50% None None
(15 - 50 days) cholestatic, &
> 14: 70 - 80%
relapsing hepatitis.
Average: 60-U0 days < 5: < 10% < 5: 30 - U0%
M 1.5m 1.0% 15 25%
(45 - 180 days) > 5: 30 - 50% > 5: 2 10%



Average: 6-7 weeks 60 - 85%
  w < 20% ow 3%
(2 - 26 weeks) wirrhosis: 5 - 20%
verall: 1-3%
Average: 40 days ncreased with age, but
- pregnancy: 15- None
(15 - 60 days) no chronicity
25%

ym Asymptomatic > ymptomatic > Fulminant liver failure > Death.

ym General symptoms: nausea, vomiting, upper abdominal pain, loss of appetite, fever, diarrhea, light colored stool, dark
urine, jaundice, & extrahepatic manifestations.
ym Diral hepatitis: supportive & directed treatment at allowing e hepatocellular damage to resolve % repair itself.
ym whronic liver disease:
1.m HMD: amivudine, a reverse transcriptase inhibitor reduces HMD DNA level, but once stopped, viral replication
resumes & resistant virus mutants r selected.
  2.m HwD: combination therapy of FN- & ribavirin with response rate of up to 50%.
3.m Moth HMD & HwD:
a.m herapy: recombinant interferon-alfa therapy (HMD: 35% remission & HwD: 25% of sustained response).
b.m ith end stage liver damage: orthotopic liver transplantation (problem: risk of reinfection of e graft).


 
ym From family of picornaviridae & genus hepatovirus.
ym size is b/w 27 32 nm spherical particle with cubic symmetry.
ym genome: a linear ssNA genome of 7.5 kb.
ym tability:
m table to treatment with 20% ether, acid (pH 1 for 2 hours), & heat (60 ow for 1 hour).


 m nfectivity can be preserved for at least 1 month after being dried & stored at 25 ow & 42% relative humidity at -
20ow.
m destroyed by: 5.m D radiation (1 minute at 1.1 watts).
1.m Autoclaving (121o w for 20 minutes). 6.m reatment with formalin (for 3 days at 37 ow).
2.m Moiling water (for 5 minutes). 7.m reatment with chlorine (10-15 ppm for 30
3.m Dry heat (180 o w for 1 hour). minutes).
ym utbreaks:
m wommon in families, institutions, day care centres, or military cops.

  
 m D/t fecal-oral route, close contacts with others , & blood exposure.
ym ecurrent explosive epidemic usually result from fecal contamination of food or water.
 ym isk factors: 4.m sers of injection & noninjection drugs.
1.m iving in crowded & poor sanitation. 5.m persons with clotting disorders & chronic liver disease.
2.m ravelers to developing countries. 6.m persons working with nonhuman primates.
3.m |en having sex with men.
ym HAD can be detected in e liver, stool, bile, & blood by A, pw, & NA hybridization assays.
ym HAD is detected in e stool about 2 week prior to onset of jaundice up to 2 weeks after.
ym Anti-HAD in g| fraction: ym Anti-HAD in gG fraction:
 M   m Appears during acute phase. m Appears soon after e onset of disease.
m peaks about 2 weeks after elevation of liver enzymes. m persists for decades.
m Declines to non-detectable levels within 3-6 months. ym Anti-HAD g| confirms e diagnosis by A.
1.m Daccination strategy (licensed for age 1 & above):
a.m arget groups: persons with increased risks of infection & community with historically high risk of HAD.
b.m ide effects: sore at injection site, headache, & malaise with no severe reactions & safe for pregnant ladies.
c.m sage for infants: safe & immunogenic for infants w/o maternal Abs because passively -acquired maternal Abs
blunts e towards e vaccine.

  2.m mmunoglobulin therapy:
a.m pre-exposure: for travelers to intermediate & high HAD-endemic regions.
b.m post-exposure (within 14 days): routine (household & other intimate contacts) & selected situations (institutions &
food prepared by infected food handlers).
3.m Good hygiene practices.
4.m wlean water & sewage system to avoid food & water contamination.

M
ym 3 forms:
1.m pherical particles of 22 nm in diameter made up of HMsAg.
2.m ubular or filamentous form with same diameter but over 200 nm long results from HMsAg overproduction
3.m arger 42 nm spherical virions.
ym outer surface or envelope contains HMsAg & surrounds a 27 nm inner nucleocapsid core that contains HMcAg.
ym viral genome consists of partially double-stranded circular DNA, 3200 bp in length.


 ym 4 phenotypes: adw, ayw, adr, & ayr.
ym HMD is stable:
ym HMD is sensitive to:
1.m At 20 o w for over 20 years.
1.m Higher temperature (100 ow for 1 minute).
2.m o repeated freezing & thawing.
o 2.m onger incubation period (60 ow for 10 hours).
3.m At 37 w for 60 minutes.
o 3.m odium hypochlorite, 0.5% (for 3 minutes).
4.m emains viable after being dried & stored at 25 w.
5.m At pH 2.4 for 6 hours.
6.m o D radiation of plasma & blood.
ym HMD is a worldwide distribution with no seasonal trend or predilection for any age of group.
ym hose who r infected at infant develop chronic disease & at high risk of developing hepatocellular carcinoma.
ym isk factors:
1.m parenteral drug abusers. ym |odes of transmissio ns:
2.m nstitutionalized persons. 1.m HMsAg is found in saliva, nasopharyngeal washing,
3.m Health care personnel. semen, menstrual period, & vaginal secretion.
4.m |ultiply transfused, organ transplantation, 2.m My close contact & sexual intercourse.

  
 3.m My improperly sterilized syringes, needles, scalpels, or
& hemodialysis patients.
5.m Highly promiscuous persons. by tattooing & ear pricing.
6.m Newborns born to mothers with HMD.
ym Global patterns of chronic HMD infection:
1.m High (> 8%), 45% of global population: lifetime risk of infection is > 60% & early childhood infections r common.
2.m ntermediate (2-7%), 43% of global population: lifetime risk of infection is 20-60% & infections occur in all ages.
3.m ow (< 2%), 12% of global population: lifetime risk of infection is < 20% & most infections occur in adult risk goups.
ym DNA polymerase activity, HMD DNA, & HMeAg present early in e incubation period, after e 1 st appearance of HMsAg.
ym During initial phase of infection, high level of HMD particles is detected in e blood.
ym HMsAg is detectable 2-6 weeks in advance of clinical evidence & persists t/out e clinical course of e dise ase but
 M   disappears after 6 months of exposure.
ym Ab to HMsAg is detected after e of HMsAg in low concentration. Detectable by A for HMD Ag, but
ym Anti-HMc g| is detected in acute disease but is low in chronic HMsAg carriers. Abs & pw for viral DNA.
1.m prevention of perinatal HMD transmission. c.m For adolescents (<18 years old).
2.m Daccination strategies: d.m For adults in high-risk groups.

  a.m For all infants. ym ide effects of vaccination r rare, but anaphylaxis appears in
b.m For children in high-risk group. 1:600000 doses given.



ym From family of Flaviviridae & genus Hepacivirus with 6 clades & > 100 subtypes.
ym genome is +ve ssNA virus of U.4 kbp in size which encodes a core protein, 2 envelope glycoproteins, & several
nonstructural proteins.


 ym virus undergoes sequence variation during chronic infection which is called quasi -species (not related to difference in
clinical disease).
ym Genotypes 1 & 4 r > resistant & require > prolonged treatment, but genotype 2 & 3 show a better response to treatment.
ym n |alaysia, 3a > 1 > 2 > 4.
ym HwD infections r extensive t/out e world. ym isk factors:
ym HwD is transmitted by: 1.m llegal injection drug users.
1.m hrough direct percutaneous exposures to blood. 2.m ransfusion or transplant from infected donor.

  
 3.m
2.m High-risk sexual practices.
3.m Health care workers.
4.m |other to child through vertical transmission.
ccupational exposure to blood (needle stick).
4.m atrogenic: unsafe injection.
5.m |ultiple sex partners.
 5.m My commercial D g preparation. 6.m exual/household exposure to anti-HwD +ve contacts.
ym |ost 1 o infections r mild & asymptomatic.
ym erological assays such as A detect Abs toward HwD for diagnosis but cant distinguish e stage of infection.
ym NA-based assays is used for:
 M  
1.m Detection of circulating HwD NA.
2.m |onitoring patients on antiviral therapy.
3.m o genotype HwD isolates.
1.m creening & testing donors of blood, organs, & tissues.
2.m Dirus inactivation of plasma-derived products.
3.m isk-reduction counseling & services.

  4.m nfection control practices: avoiding direct exposure to infected blood:
a.m Anti-HwD +ve persons should not donate any parts of theirs.
b.m Do not share items that might have blood on them (personal care & home therapy).
c.m wover cuts & sores on e body.


ym virus contains HDAg surrounded by HMsAg envelope.
ym t has particles size of 35-37 nm & a buoyant density of 1.24 -1.25 g/m.


 ym genome is circular, -ve ssNA which is 1.7 kb in size.
ym HDD is a defective virus which acquires an HMsAg coat for transmission , however, no homology exists b/w e 2.
ym HDD can be coinfected with HMD (acute infection) or superinfected on top of chronic HMD (severe chronic liver disease).
ym HDD is found t/out e world but with a non-uniform distribution & infects all groups of age.
ym Highest prevalence in taly, e |iddle ast, central Asia, est Africa, & outh America.
ym isk factors:
1.m persons who receive multiple transfusions. persons who r in close contact with them.

  
 2.m D drug abusers.
ym |odes of transmission r similar with HMD but no D & it can also be transmitted perinatally.
ym 2 epidemiological patterns:
1.m n |editerranean countries: HDD is endemic among persons with HMD & most r transmitted by intimate contact.
2.m n non-endemic area: HDD is confined to persons exposed frequently to blood & blood products such as drug addict.
ym 2 patterns:
1.m woinfection pattern: Abs to HDAg develop late in e acute phase of infection & may be of low titre.
 M   2.m uperinfection pattern: usually results in HDD persistence with high level of g|, gG, HDD NA, & HDAg.
ym All markers for HDD replication disappear during convalescene, even HDD Abs may disappear within months to years.
1.m HMD-HDD coinfection: pre or postexposure prophylaxis to prevent HMD infection (HMG &/or Hepatitis M vaccine).

  2.m HMD-HDD superinfection: education to reduce risk behaviors among persons with chronic HMD infection.



ym genus is Hepevirus.
ym virus is transmitted enterically & occurs in epidemic form in developing countries with fecal contamination.



ym viral genome is a +ve sense ssNA 7.6 kb in size.
ym here is possibility of animal to hman transmission.
ym |ost outbreaks occur associated with focally contaminated drinking water.

    ym |inimal person to person contact transmission.
ym wases in non-endemic areas usually have history of travelling to endemic areas.
ym |ostly precautions r for travelers to endemic regions: a void drinking water of unknown purity, uncooked shellfish, &

  uncooked fruits or vegetables not peeled by e travelers themselves.




!
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1.m Dirion: spherical, 80-110 nm in diameter, helical nucleocapsid within icosahedral capsid.
2.m womposition: NA (2%), protein (60%), lipid (35%), & carbohydrate (3%).


 3.m Genome: linear & +ve ssNA, 7-11 kb, diploid, & may be defective & carrying oncogene.
4.m protein: reverse transcriptase contained in side virion.
5.m nvelope: contains glycoprotein & lipid.
1.m ype A:
ym ccur only intracellularly & non-infectious.
 
  
 ym ntracytoplasmic type A particles r 75 nm in diameter & precursors of extracellular type M viruses.
ym ntrecisternal type A r 60 -U0 nm in diameter & unknown entities.

 2.m ype M: 100-130 nm in diameter & contain an eccentric nucleoid.
3.m ype w: e largest group of retroviruses, r U0 -110 in diameter, & contain centrally located nucleoid.
4.m ype D: r 100-120 in diameter, contain an eccentric nucleoid, & exhibit surface spikes shorter than type Ms.
1.m -retrovirus: avian leukosis & sarcoma viruses. 5.m $-retrovirus: fish virus.
2.m -retrovirus: mouse mammary tumor. 6.m pumavirus: foamy degeneration of inoculated

 
  3.m >-retrovirus: mammalian leukemia & sarcoma virus. cells.
4.m '-retrovirus: HD & bovine leukemia virus. 7.m entivirus: HD & neurologic impairment disease.
1.m cotropic viruses: infect & replicate oly in cells from animal of e original host species.
   2.m Amphotropic viruses: exhibit a broad host range (infect natural host & heterologous species as well).
3.m Xenotropic viruses: replicate in some heterologous (foreign cells) but not in cells of e natural hosts.
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ym Are a retrovirus & a member of entivirus genus.
ym Have a cylindrical nucleoid in e mature virion.
ym wontains e 4 genes required for a replicating retrovirus: gag-pro-pol-env.
ym ther additional genes responsible for regulating viral expression & important in disease pathogenesis in vivo:
1.m Tat gene: functions in transactivation & transcriptional activation o f other viral genes.
2.m ev gene: expression of viral structural proteins & facilitation of e export of unsliced viral transcripts from e
nucleus.
3.m Nef gene: increases viral infectivity, facilitate activation of resting T cells, & downregulates e expression o f wD4
& |Hw class 1.
4.m Vpr gene: increases transport of e viral preintegration complex into e nucleus & arrest G2 phase of e cell cycle.
5.m Vpu gene: promotes wD4 degradation.
6.m Vif gene: promotes viral infectivity by suppressing e effects of an inhibitory cellul ar protein present in some
p
p
 human cells.
ym env gene:
m regions of greatest divergence among isolates r localized.
m wodes for e viral envelope proteins.
m product:
1.m wontains binding domain responsible for viral attachment to wD4 molecule & coreceptors.
2.m Determine lymphocytes & macrophages tropism.
3.m warries e major antigenic determinant that elicit neutralizing Abs.
m m product:
1.m wontains both a transmembrane domain that anchors e glycoprotein in e viral envelope.
2.m A fusion domain that facilitates viral penetration into target cells.
ym Host factors important for pathogenesis: age, stress, genetics, & concurrent infection.
ym virus is inactivated:
1.m wompletely by treatment for 10 minutes at room temperature with:
   2.m My extreme pH (pH 1.0-3.0).
3.m My exposure to undiluted bleach for at least 30 seconds (for viruses in clotted or unclotted blood in e needle).
    4.m My paraformaldehyde (for virus in free solution).
5.m n liquids or 10% serum by heating at 56 o w for 10 minutes.
6.m My heating at 68o w for 72 hours (for lyophilized blood products).
1.m Diruses r transmitted by exchange of body fluid.
2.m Dirus persists indefinitely in infected hosts, though it may present in low amount.
   3.m Diruses have high mutation rates.
 
 4.m Dirus infection progresses slowly through specific stages.
5.m Diruses can infect non-dividing terminally differentiated cells.
6.m t may take years for disease to develop.
ym eceptors necessary for HD infection:
1.m eceptors: wD4 molecule on macrophages & cells to gain entry into e cells.
2.m wo-receptors:
m whemokine receptors: ww5 (macrophages-tropic strains) & wXw4 (lymphocytes-tropic strains).


 m co-receptors used by HD for cell entry r found on lymphocytes, macrophages, thymocyte s, on neurons,

p
 & cells on colon & cervix.
ym A dendritic cell-specific lectin, Dw-GN:
m Appears to bind HD-1 but not to mediate cell entry.
m |ay facilitate transport of HD by dendritic cells to lymphoid organs.
m nhance infection of cells.
?  
ym strain can be classified into 3 groups:

   1.m major group |: > U0% of HD-1 infection belongs to | group.
2.m outier group : appears to be restricted to west central Africa.
? 3.m new group N: discovered in 1UU8 in wameroon & is extremely rare.
ym hese 3 groups may represent 3 sep arate introduction of simian immunodeficiency virus into human.
1.m phase 1:
m Acute infection: lasting from infection until seroconversion (onset of detectable virus -specific Abs).
m sually lasts 3-8 weeks.
m Dirus replicate to high titre (usually 5 type).
m woncluded when w develop, seroconversion occurs, & virus load is diminished in e blood.
2.m phase 2:
m Asymptomatic phase which lasts for months to > 15 years.
m wontinuation of viral replication.
m Active immune response & antigenic escape.
p?  ? m woncluded by development of (wD4 + cell < 200/mm 3).
3.m phase 3:
  m Host immune response collapses.
m wlinical progression to AD.
m yncytium inducing (X4 viruses) r detected in most if not all cases.

wD4+ wells
m mmmmmmmmmmmmmmmmm
mmmmmmmmmmmmmmmmmmmmmmmmmmm

Diral load
 1.m 20% of HD infected people become fast progressors:
ym hey lose > 100 wD4 cells /mm 3 / year.
ym wice this percentage in perinatally infected infants, transfusion recipients, & hemophiliacs.
p

  2.m 35% become progressors:
p 
 ym ose 31 wD4 cells /mm 3 / year.
3.m 25% become long term non-progressors:
ym No decrease in wD4.
ym evel of 500 800 wD4 cells /mm 3 / year which is still on e low side of normal.
1.m wD4 lymphocytes & memory cells:
ym cardinal feature: wD4 cell depletion. hich
ym Due to: viral replication in these cells & e death of uninfected cells by indirect mechanism.
ym arly in infection, primary isolate is | tropic, but then later replaced by tropic viruses.
ym At any given time, only small fraction of cell r productively infected & others r killed.
ym survivors r infected & revert to a resting memory state being a long term, stable reservoir for e virus.
ym hen exposed to Ag, they become activated & release infectious viruses.
2.m |onocytes & macrophages:
ym co-receptor of HD on macrophage is ww5 chemokine receptors.
ym n e brain, monocytes & macrophages r highly infected which leads to e development of neuropsychiatric
manifestation in HD infection.
p ?
 ym | tropic strains of HD are important for initial infection.
ym |onocytes & macrophages r major reservoirs for HD in e body because they r relative refractory to wp of
HD so that e viruses survive in these cells & can be transported to e whol e body.
ym nfected macrophages may continue to produce viruses for a long time.
3.m ymphoid organs:
ym n untreated infection & latency state, HD is actively replicating in lymphoid tissues because e
microenvironment in e organ is ideal for e virus.
ym wytokines r released & activate a large pool of cells for e virus.
ym At late stage, e architecture of e lymph node becomes disrupted.
4.m Neural cells:
ym Dirus enter brain via infected monocytes & release cytokines which r toxic to neurons & chemotatctic factors
that lead to infiltration of inflammatory cells into e brain.
ym diseases: HD encephalopathy, peripheral neuropathies, & AD dementia complex.
1.m Activated cells infected by HD:
ym produce virus immediately & die within 1 to 2 days.
ym Half-live: 1.6 days.
ym virus production accounts for e vast majority of virus present in plasma.
?

  2.m Dirus in e plasma:
ym t is estimated that 10 billion HD particles r produced & destroyed each day.
? ym half-live: 6 hours.
3.m HD life-cycle: complete life cycle requires 2.5 days.
4.m esting cells infected by HD:
ym Half-live of at least 5-6 months.
ym present as a major challenge for virus eradication & anti -retroviral therapy.
1.m esistant variants: pre-existing in e host, can be transmitted & evolve.
2.m elective pressure of therapy:
? a.m Drug has limited potency.

  b.m Non-adherence to potent anti-retroviral therapy.
c.m poor absorption or rapid clearance of drugs.
d.m Drugs r not active in cells.
1.m exual contact: male to male, male to female, & female to 3.m perinatal infection:
female. ym ransmission from mother to baby.
2.m Fluid exposure: ym Mreastfeeding.
ym emen, vaginal secretion, breast milk, & blood.

  ym outes of transmission:
a.m njecting drug by using or sharing needles.
b.m ccupational exposure.
c.m ransfusion of blood products.
d.m pen cuts, skin abrasion, & mucous membrane.
1.m General symptoms:
ym wommon symptoms: fatigue, rash, headache, nausea, & night sweat.
ym erious symptoms: prodrome which includes fatigue, malaise, weight loss, fever, dyspnea, chronic diarrhea,
white patches on e tongue, & lymphadenopathy.
ym No treatment: death after 2 years.
2.m plasma viral load:
ym Diral load is of significant prognosis value which reflects e total no. of productively infected cells & their
average burst size.


 ym plasma viral load appears to be e best predictor of long term clinical outcome while wD4 counts r e best
 predictor of short term risk of developing an opportunistic disease.
ym Also important for measurement of antiretroviral drug therapy.
3.m pediatric AD:
ym wlinical findings: lymphoid interstitial pneumonitis, severe oral candidiasis, encelophaty, wasting, generalized
lymphadenopathy, bacterial sepsis, hepatosplenomegaly, diarrhea, & growth retardation.
ym Diral NA load: generally low at birth, then rise rapidly within e 1 st 2 months, & followed by a slow decline
until e age of 2 years (suggesting that has difficulty containing e infection ).
4.m Neurologic disease:
ym diseases: subacute encephalitis, vacuolar myelopathy, aseptic meningitis, & peripheral neuropathy.
 ym AD dementia complex: poor memory, inability to concentrate, apathy, psychomotor retardation, &
behavioral changes.
ym psychology: high risks of psychiatric disorders & e most common is anxiety disorder.
ym pediatric neurology: seizure disorders, progressive loss of behavioral developmental milestones,
encephalopathy, attention deficit disorder, meningitis, & developmental delays.
5.m pportunistic infection: protozoa, fungi, bacteria, & viruses.
6.m wancer:
ym end to be those with a viral cofactor.
ym ncludes non-Hodgkins lymphoma, Kaposis sarcoma, cervical cancer, & anogenital cancer.
ym ffective anti-retroviral drug therapy results in marked decrease in Kaposis sarcoma viruses.
7.m people infected with HD:
ym wan look healthy.
ym wan be unaware of their infection.
ym wan live lifelong productive lives when their HD infection is managed.
ym wan infect people when they engaged in high risk behavior.
1.m Dirus isolation:
ym HD can be cultured from lymphocytes in peripheral blood.
ym Higher titres of virus r found in e patients indicating e HD infection.
ym Diral growth is detected by testing culture supernatant fluids after 7 -14 days for virus reverse transcriptase
activity or for virus-specific Ags.
ym However, they r time consuming, laborious, & r limited to research studi es.
2.m erology or screening HD Ab testing:
ym A/A (reactive) l repeat A/A (reactive) l FA, others e.g. estern Mlot (reactive) l positive for



 HD (sensitivity & specificity exceeding U8%).
   ym Detects e bodys Abs response to HD infection, not detecting e virus.
ym |ost will have detectable Abs within 6-12 weeks after infection, whereas virtually all will be positive within 6
months.
3.m Detection of viral NA or Ags:
ym Amplification assays such as -pw, DNA pw, & bDNA tests r used to detect viral NA.
ym tests can be quantitative, very sensitive, & form e basis for plasma viral load determination.
ym HD NA level r important predictive markers of disease progression & valuable tools with which to monitor
e effectiveness of antiviral therapies.
ym However, e presence of maternal Abs makes serological tests uninformative.
ym reatment:
m Antiviral drug:
1.m wombination therapy:
-m eferred as highly active antiretroviral therapy (HAA).
-m wan suppress viral replication to below limit of detection in plasma & decrease viral loads in
lymphoid tissues which may prolong patient survival.
-m However, it fails to cure HD-1 infection because e virus persists in reservoirs of long -lived, latently
infected cells, including memory wD4 cells.
2.m |onotherapy: rapid emergence of drug resistant mutants.
3.m Zidovudine:
-m educe e transmission of HD from mother to infant.
-m herapy during pregnancy & during e birth process & of e baby after birth to reduce transmission.
m mportant of early diagnosis & testing:
1.m Allows for early treatment to maintain & stabilize e immune system response.
2.m Decreases risk of HD transmission from mother to newborn baby.
3.m Allows for risk reduction education to reduce or eliminate high-risk behavior.
ym prevention:
1.m Daccination: vaccine development is difficult because HD mutates rapidly, is not expressed in all cells that r

   infected, & is not completely cleared by e host after primary infection.
2.m Gene therapy: intracellular immunization genetically alters target cells & makes them resist ant to HD.
p
  3.m HD risk reduction:
a.m Avoid drug & alcohol use to maintain good judgment.
b.m Dont share needles used by others for drugs, tattoos, & body piercing.
c.m Avoid exposure to blood products.
4.m Health education:
a.m Any sexual intercourse should be protected by a condom.
b.m Do not share unsterile needles or syringe.
c.m All women who r potentially exposed to HD should seek HD Abs testing before becoming pregnant.
d.m HD-infected mothers should avoid breastfeeding to reduce transmission of virus to their children.
5.m wontrol measures:
a.m wonsistent & proper use of condom.
b.m oothbrushes, razors, & other implements that could become contaminated with blood should not be
shared with others.
c.m eropositive women or women with seropositive sexual partners because if they pregnant, offspr ing is
high risk of acquiring AD.
d.m Accidents that lead to bleeding should be cleaned with household bleach.
e.m Devices that have punctured e skin should be steam -sterilized by autoclaving before reuse or discarded.
f.m f seropositive, inform medical or dental o fficers.
g.m esting for HD Abs.

 

"
7!
"


   
ym From e family orthomyxoviridae & causes respiratory tract infection.
ym 3 immunologic types divided by differences exhibited in e nucleocapsid (Np) & matrix (|) proteins:
1.m A (man & animal): has subtypes determined by antigenic variation
in e surface glycoprotein, HA & NA.
2.m M (man & animal). No cross reaction b/w e 3.
3.m w (man only).
ym Dirion: spherical, pleomorphic, 80-120 nm in diameter with helical, U nm nucleocapsid.
ym womposition: NA (1%), proteins (73%), lipid (20%), & carbohydrate (6%).
ym Genome: -ve sense segmented (A & M: 8, w: 7) ssNA of 13.6 kb for e overall size.
ym proteins: contains U structural proteins & 1 non -structural protein:
m nucleoprotein associated with viral DNA to form ribonucleoprotein (Np) that assumes e helical
configuration & forms e viral nucleocapsid.


 m 3 large proteins, pM1, pM2, & pA r bound to viral Np for NA transcription & replication.
m matrix (| 1) protein forms e shell underneath e viral lipid envelope & important for particle morphogenesis
& is e major component of e virion.
ym nvelope:
m |ade of lipid derived from e cell surround e virus particle.
m 2 virus encoded glycoprotein, hemagglutinin (HA) & neuraminidase (NA) r inserted into e envelope &
exposed as spikes about 10 nm long on e surface of e particle.
m D/t HA & NA, envelope becomes e important Ag.
m | 2 ion channel & N2 proteins r also present in e envelope.
ym D/t segmented nature of e NA, e virus u ndergoes genetic reassortment which results in sudden changes in viral
surface Ags which contribute to e epidemiological features of influenza & difficulty in developing vaccine.
ym nfectivity is destroyed by lipid solvents, protein denaturants, formaldehyde , acidic environment, & irradiation.
ym nfluenza virus infections occur worldwide & cause annual outbreaks which occur in waves & of variable intensity.
ym pidemiology according to immunogenic types:
1.m ype A:
m ndergoes both antigenic shift & drifts involving e genomic reassortment of HA & NA proteins. As a
result, e population has no immunity against e new strain.
m wauses pandemics (massive epidemics) which can sweep across e continents & around e world. his
occurred in 1U18 (H1N1), 1U57 (H2N2), & 1U68 (H3N2).
m pidemics arise through minor antigenic drifts as a result of mutation which tend to be 2 -3 years.
2.m ype M:
m ometimes causes epidemics which have a longer interepidemic period of 3 -6 years d/t antigenic drift.
m Does not undergo antigenic shift.
3.m ype w: e least significant because it causes mild sporadic respiratory disease but not epidemic influenza.

    ym urveillance for influenza outbreaks is necessary to identify e early appearance of e new strain s. solation of a virus
with an altered HA in e late spring during a mini epidemic signals a possible epidemic e following winter.
ym Avian influenza:
m influenza viruses do not appear to undergo antigenic change in e birds d/t birds life span.
m his means that genes that caused previous influenza p andemics in human still exist unchanged in e aquatic
bird reservoir.
m nfluenza virus of e ducks multiply in e living cells lining e intestinal tract & r shed highly in fecal matters.
m vidence supports that pigs serve as mixing vessels for reassortants as t heir cells contain receptors recognized
by both human & avian viruses. school-age children r e predominant vectors of transmission.
m f given enough opportunities, e highly pathogenic H5N avian strain could spread and sustain among
humans either by reassortment or by adaptive mutation. his can cause a devastating influenza pandemic.
ym eroarcheology shows that e highest Ab levels in a particular age group reflect dominant Ags of e virus responsible
for childhood infections of e group.



ym HA protein binds virus particles to susceptible cells & e major Ag against neutralizing Abs.
ym esponsible for e continual evolution of new strains & subsequent influenza epidemics.
ym Able to agglutinate Mws under certain conditions.
ym mechanism: a short signal sequence at e aa terminal inserts e polypeptide into l e signal is removed l e
HA protein is cleaved into 2 subunits, HA1 & HA2 l HA2 anchors e HA molecule in e membrane with a short

"

 hydrophilic tail extending into e cytoplasm l oligosaccharide residues r added at several sites l e HA molecule
is folded into a complex structure l 5 antigenic sites (exposed on e surface of e structure) on HA molecule exhibit
extensive mutation.
ym HA spike one virus particle is a trimer composed of 3 intertwined HA1 & HA2 dimers.
ym cleavage of HA protein is necessary for e virus particle to be infactious & is mediated by cellular protease.
ym nfluenza virus remain confined to e respiratory tract bec ause e enzymes r common there.
ym spike of NA is a tetramer composed of 4 identical monomers & contains 4 active (catalytic) sites.
ym NA functions at e end of e viral replication:

"

 1.m Acts as a sialidase enzyme which removes sialic acid from glycoconjugates.
2.m t facilitates release of virus particles from infected cell surface during e budding process.
3.m t helps prevent self aggregation of virions by removing sialic acid residues from viral glycoproteins.
4.m t helps e virus negotiate through e muc in layer in e to reach e target epithelial cells.
1.m Antigenic drift:
ym .e. e minor antigenic changes.



 ym D/t e accumulation of point mutations in e gene resulting in aa changes in e protein.

 
 ym equences changes can alter antigenic sites on e molecule such that a virion can escape recognition by e host
immune system.
ym immune system also acts as selection force that allows new antigenic variants to expand.
 ym A variant must sustains 2 or > mutations before a new strain emerges.
2.m Antigenic shift:
ym .e. major antigenic changes in HA & NA results in a new subtype.
ym eflects e drastic changes in e sequence of a viral surface protein which causes epidemics.
ym segmented genomes of influenza viruses reassert readily in doubly infected cells.
ym mechanism involves genetic reassortment b/w human & avian influenza which is possible only for influanze
virus type A.
ym Dirus attaches to cell surface sialic acid via e receptor site located on HA l virus particles r internalized within
endosomes (receptor-mediated endocytosis) l fusion b/w viral envelope & cell membrane (uncoating) l viral
Nps r released d/t low pH in endosome l acid pH causes conformational change in HA & brings HA2 fusion
peptide in contact with e membrane l |2 ion channel protein permits e entry of ions in e endosome into e virus


  particle l viral nucleocapsids r released into cytoplasm & nucleus l transcription & replication by virus encoded
polymerase l nucleocapsids r assembled in e nucleus & moved out to cell surface l HA & NA r synthesized in
, modified, & assembled into trimers & tetramers & r inserted into plasma membrane l maturation by
budding l NA removes terminal sialic acids from cellular & virus surface glycoproteins l release of free virus
particles.
ym Dirus spreads from person to person through air borne droplets or by contact with contaminated hands or surface.
ym nfection is successful if e cough reflex, mucous secretion, & neutralizing gA fail to prevent to remove e virus from
respiratory epithelial cells.
ym Diral NA lowers e viscosity of e mucous film in , laying bare e cellular surface receptors & promoting e spread of
virus containing fluid to e lower portion of e tract.

    ym incubation period is from 1-4 days depending upon e size of viral dose & e immune status of e host.
ym Diral shedding starts e day preceding onset of symptoms, peaks within 24 hours, remains elevated for 1 -2 days, &
declines over e next 5 days.
ym nfection causes cellular destruction & desquamation of superficial mucosa of e which results in edema &
mononuclear infiltration, but does not infect e basal layer of e epithelium.
ym Diral damage to lowers its resistant to 2 o bacterial invaders such as staphylococci & many more.
1.m ncomplicated influenza:
ym Appear abruptly & include chills, headache, & dry cough followed by high fever (3 -5 days), generalized
muscular aches, malaise, & anorexia.
ym espiratory symptoms last for another 3 -4 days while e cough, coryza, & weakness may persist for 2 -4 weeks
after major symptoms subside.
ym hese symptoms r characterized of type A & M, while type w only causes common cold illness.
ym n pediatrics, G manifestations such as vomiting r common, while febrile convulsion & otitis media can also
occur. ype A is an important cause of croup in children under 1 year old of age.


 2.m pneumonia:
 ym lethal impact of influenza epidemic is death d/t pneumonia & cardiopulmonary diseases.
ym pneumonia can be result of viral, 2 o bacterial, or combination of both.
ym ncreased mucous secr etion helps carry agent to  & is attributed to loss of ciliary clearance, dysfunction of
phagocytic cells, & provision of a rich bacterial growth medium by e alveolar exudates which leads to
bacteria superinfection.
3.m eyes syndrome:
ym s an acute encephalopathy of children & adolescents b/w 2-16 years old.
ym cause is unknown but it is a complication of type A & M & varicella -zoster virus.
ym mmunity, i.e. Abs against HA & NA is long-lived & subtype-specific.
ym esistance to initiation of infection is related to Abs against HA, whereas decreased severity of disease &
decreased ability to transmit virus to contacts r related to Abs against NA.
ym erum Abs persist for months or years, but gA r for shorter duration.
  ym riginal antigenic sin: subsequent infections or immunization reinforce e Abs to response to e 1 st subtype of
influenza experienced earlier.
ym w| is responsible for e clearance of an established infection. wytotoxic cells response is cross -reactive & can be
directed against both internal proteins & surface glycoprotein.
1.m Detection of Ags: a rapid diagnosis can be made by e detection of influenza Ag from nasopharyngeal aspirates &
throat washings by F & A.
2.m Dirus isolation: virus may be readily isolated from nasopharyngeal aspirates & throat swabs & cultured in
 M   embryonated eggs or |DwK cells.
3.m erology: a retrospective diagnosis may be made by serology via wF type -specific, HA & N subtype & strain
specific, & A for type-specific diagnosis.
4.m |olecular amplification test such as pw.
1.m reatment:
a.m Amantidine: effective against influenza A if given early in e illness. However, resistance emerges rapidly.
b.m imantidine: similar to amantidine but fewer neurological side effects.
c.m ibavirin: effective against both influenza A & M.
d.m Neuraminidase inhibitors: highly effective with fewer side effects. esistance towards it emerges slowly.
  2.m prevention:

  a.m Daccination:
ym nactivated split/subunit vaccines r available against influenza A & M.
ym vaccine is normally trivalent, consisting of one A H3N2 strain, one A H1N1 strain, & one M strain.
b.m prophylaxis: Amantidine can be used for those who r allergic to e vaccine (embryonated egg) r during period
before e vaccine takes effect.

 
 $!
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ym nder e paramyxoviridae family whose members r all initiate infection via respiratory tract.
ym Dirion: spherical, pleomorphic, > 150 nm in diameter with helical nucleocapsid of 13-18 nm.
ym womposition: NA (1%), protein (73%), lipid (20%), & carbohydrate (6%).
ym Genome:
m inear, non-segmented, -ve sense, ssNA which is non-infectious & of 15 kb in size.
m No-segmented genome negates any opportunity for frequent genetic reassortment which causes e virus to be
antigenically stable.
m Do not undergo antigenic drift as a result of mutation introduced during replication because nearly all aa
involved in structural & functional roles.
ym 6 structural proteins:
m 3 proteins r complexed with e viral NA: e nucleoprotein (N or Np) that forms e helical nucleocapsid of 13 -18
nm in diameter.


 m his represents e major internal protein & 2 other larger proteins which involved in e viral polymerase activity
functioning in NA transcription & replication.
m 3 proteins participate in viral envelope: matrix (|) protein underlies e viral envelo pe which has an affinity for
both N & e viral surface glycoprotein & essential for virion assembly.
ym nvelope:
m |ade up of lipid which surrounds e nucleocapsid.
m tudded with 8-12 nm spikes of 2 different transmembrane glycoproteins.
ym Glycoproteins:
m Help differentiate e various genera in e family.
m larger tetramer glycoprotein (HN or H or G) is responsible for attachment to e host cells.
m (F) glycoprotein mediates membrane fusion & hemolysisn activities.
m pneumoviruses appear to contain 2 additional small envelope proteins (|2-1 & H).
ym Divided into 2 subfamilies & 7 genera whose most of e members r monotypic & antigenically stable.
ym genera:
1.m espirovirus: 2 serotypes of human parainfluenza viruses.
2.m ubulavirus: 2 other parainfluenza virus & mump virus.

 
  3.m Avulavirus: avian parainfluenza virus.
4.m |orbillivirus: measles virus.
5.m Heipavirus: zoonotic paramyxoviruses.
6.m pneumovirus: respiratory syncytial viruses & pneumonia virus.
7.m |etapneumovirus: human respiratory pathogen.
ym Dirus attaches to host cell via hemagglutinin glycoprotein (wD46 or wD150 molecules) l virion envelope fuses
with e cell membrane by e action of e fusion of glycoprotein F1 cleavage product l release of viral nucleocapsid


  directly into cell l mNA transcripts r made in e cytoplasm by viral NA polymerase l viral proteins r
synthesized in e cytoplasm l viral glycoproteins r synthesized & glycosylated in e secretory pathway l
progeny nucleocapsids form in e cytoplasm & migrate to cell surface l virus matures by budding.

  
ym major cause of  disease in young children & r widely distributed geographically.
ym ype 3 is most prevalent during e 1 st year of life while infection with type 1 & 2 occur at a lower rate.
ym ype 3 is endemic with some increase during spring while type 1 & 2 tend to cause epidemics during fall or winter.




 ym einfections r common t/out childhood & in adults & results in mild .
ym Diruses r ransmitted by direct person -to-person contact, large droplet aerosols & thr ough both nose & eyes which
spread readily in a population.
ym incubation period: 5-6 days.
ym wan also cause nosocomial infection in pediatric wards in hospitals.
ym Diral replication is limited to respiratory epithelia & viremia is rare.
ym infection involves only nose & throat resulting in harmless common cold syndrome.
ym f involves e larynx & upper trachea, croup characterized by respiratory obstruction d/t swelling of e larynx &
related structures presents.
ym f spread deeper to lower trachea & bronchi , pneumonia or bronchiolitis occur.
  ym Dirus shedding is 1 week after onset of illness & persistent shedding especially in children may facilitate e viral
spread.
ym everity is determined by: 3.m mmune status of e patients.
1.m usceptibility of e protein to cleavage by proteases. 4.m Airway hyperreactivity.
2.m production of inappropriate protease by host cells. 5.m production of virus-specific g during
primary infection.
1.m mmunocompetant patients:


 ym ype 1 & 2: rhinitis, pharyngitis, laryngotracheitis, croup, bronchiolitis, & pneumonia.
ym ype 3: severe illness of croup & febrile illness.



ym most common complication: otitis media.
2.m mmunocompromised patients: susceptible to severe infection with mortality rates in M| transplantation of <20%.
1.m Detection of Ags: a rapid diagnosis can be made by e detection of parainfluenza Ag from nasopharyngeal
 aspirates & throat washing.


 2.m Dirus isolation: virus may be readily isolated from nasopharyngeal aspirates & throat swabs.
3.m erology: a retrospective diagnosis made by serology, wF is most widely used.
1.m wontact isolation precautions to manage nosocomial outbreaks: restriction of visitors, isolation of infected patients,


  & growning & hand washing by medical personnels.

!

 2.m antiviral ribavirin for treatment of immunocompromised patients with .
3.m Moth subunit vaccine & live attenuated type 3 virus vaccine for prevention.
 
    
 
ym Distribution is worldwide & is e major pediatric respiratory tract pathogen.
ym ubgroup A infections appear to cause > severe illness than subgroup M infections.
ym Diruses r spread by large droplets & direct contact with e eyes & e nose as e main portal of entry.




 ym Although virus is very labile, it can survive on environmental surfaces for up to 6 hours.
ym n tropical areas, e virus epidemics may coincide with rainy seasons.
ym virus can also cause nosocomial infections in nurseries & on pediatric hospital wards. ransmission occurs
primarily via hospital staff members.
ym virus can also cause symptomatic disease in healthy young adults in crowded conditions like in military camps.
ym eplication occurs initially in epithelial cells of e nasopharynx.
ym Dirus may be spread into e lower respiratory tract & causes bronchiolitis & pneumonia.
ym here is lymphocyte migration, resulting in peribronchiolar infiltration, submucosal tissues become edematous &
plug consisting of mucus, cellular debris, & fibrin occlude e smaller bronchioles.
ym incubation period b/w exposure & onset of illness is 3 -5 days.
  ym Diral shedding may persist for 1-3 weeks from infants & young children with e presence of high viral titre, whereas
adults shed for only 1-2 days.
ym patients with impaired w| may become persistently infected with e virus & shed virus for months.
ym nfants at risk of severe infection:
1.m nfants with congenital heart disease.
2.m nfants with underlying pulmonary disease especially bronchopulmonary dysplasia.
3.m mmuocompromised infants d/t congenital immunodeficiency disease.
ym wlinical features range from inapparent infection or common cold through pneumonia in infants to bronchiolitis in


 very young babies with wheezing.
ym progression of symptoms may be very rapid, culmi nating in death.



ym einfection is common in both children & adults, but usually is limited the upper respiratory tract.
ym wan also cause otitis media.
1.m Detection of Ags: a rapid diagnosis can be made by e detection of D Ags from nasopharyngeal aspirates &
 throat washing.


 2.m Dirus isolation: virus may be readily isolated from nasopharyngeal aspirates & throat swabs.
3.m erology: a retrospective diagnosis made by serology, wF is most widely used.


  1.m Aerosolized ribavirin can be used for infants with severe infection & for those at risk of severe disease.
2.m No vaccine yet available.

!

 3.m D gs can be used to protect infants at risk of severe D disease.



ym |umps occur worldwide where e cases appear t/out e year in hot climates & peak in winter & spring.
ym utbreaks occur where crowding favors dissemination of e virus.
ym pidemics may occur in army camp.




 ym |umps is primarily an infection of children where it reaches e highest incidence in children ag ed 5-U years.
ym |umps is contagious & most susceptible persons can acquire infection from e close member.
ym |odes of transmission: direct contact, air borne droplets, & fomites contaminated with saliva or urine.
ym overall mortality rate for mumps is low, mostly d/t encephalitis.
ym Humans r e only natural hosts for mumps virus.
ym 1o replication occurs in nasal or upper respiratory tract epithelial cells.
ym Diremia then disseminates e virus to e salivary glands & other major organ systems.
 
ym incubation period ranges from 2-4 weeks but is typically 14-18 days.
ym Dirus is shed in saliva from about 3 days before & U days after e onset of salivary gland swelling.
ym Dirus frequently infects e kidneys & viruria may persist for up to 14 days after e onset of clinical symptoms.
ym > 1/3 of all mumps infections r subclinical including e majority of infections in children under 2 years old.
ym welling of e gland: 50% of patients.
ym A prodromal period of malaise & anorexia is followed by rapid enlargement of parotid glands with pain.
ym wN involvement: aseptic meningitis & meningoencephalitis (bith resolve w/o sequelae).


 ym pacreatitis is reported in 4% of cases.


 ym Genital system:
m estes & ovaries may be involved.
m rchitis: 20-50% of men with mumps (lack of elasticity of tunica albuginea does not allow inflamed testis to
swell, e complication is extremely painful).
m Atrophy of testis may occur as s result of pressure necrosis.
m |umps oophoritis occurs in 5% of wo men.
1.m solation & identification of virus:
ym amples r from saliva, wF, & urine.
ym |onkey kidney cells r preferred for viral isolation.
 ym For rapid diagnosis, F using mumps-specific antiserum can be used to detect mumps virus Ags.
ym wp of mumps: cell rounding, giant cell formation, & hemadsorption.



2.m erology:
ym positive g| Abs strongly suggests recent infection.
ym ignificant increase in gG Abs b/w acute & convalescent specimens.
ym |ethods used: A or H.
1.m mmunization with attenuated live mumps virus vaccine to reduce mumps associated morbidity & mortality.


  ym |umps vaccine is in combination with measles & rubella: ||.

!

 ym 2 doses of || r recommended for school entry.
2.m nfected students & health care worker should be excluded from work until U days after e onset of parotitis.
  M   
ym |easles is higly contagious, a single serotype, no animal reservoir, & infection confers lifelong immunity.
ym prevalence & age incidence r related to population density, economic, & environmental factors, & e use of ||.
ym |odes of transmission: respiratory routes & hematogenous transplacental transmission.





ym A continuous supply of susceptible individuals is required for e virus to p ersist in e community.
ym |easles is endemic t/out e world.
ym pidemics recur regularly every 2-3 years & tend to occur in late winter & early spring.
ym Humans r e only natural hosts for measles virus.
ym pathogenesis: entry via l multiplies locally l infection spreads to e regional lymphoid tissue l further
multiplication occurs l 1o viremia disseminates e virus l virus replicates in D system l 2o v viremia seeds e
 
epithelial surfaces of e body (skin, , & conjunctiva) l focal replication.
ym wp: multinucleated giant cells with intranuclear inclusion body in lymphoid tissue t/out e body.
ym incubation period: 8-12 days but may lasts up to 3 weeks in adults.
ym 2 phases of measles infection:
1.m prodromal period of catarrhal stage:
m asts for 2-4 days.
m wharacterized by fever, sneezing, coughing, running nose, eye redness, Kopliks spots, & lymphopenia.
m cough & coryza reflects an intense inflammatory reaction involving e mucosa of e .
m Kopliks spots: pathognomonic for measles, small, bluish-white ulcerations on e buccal mucosa opposite e
lower molars. he spots contain giant cells & viral Ags & appear about 2 days before rash.
m fever & cough persist until e rash appears & then subside within 1 -2 days.
2.m xanthematous stage:
m asts for 5-8 days.
m moculo-popular skin rash:
-m tarts on e head & then spreads progressively to e chest, e trunk, & down e limbs.
-m Appears as light pink, discrete macula-papules that coalesce to form blotches, becoming brownish in
5-10 days.
-m fading rash resolves with desq uamation.


 m |odified measles:
-m ccurs in partially immune persons such as infants with residual maternal Abs.


 -m incubation period is prolonged.
-m prodromal symptoms r diminished.
-m Kopliks spot r usually absent & rash is limited.
ym complications of measles:
1.m most common complication is bacterial superinfcetion which causes otitis media (5 -U% of cases), sinusitis,
pneumonia, & sepsis.
2.m Giant cells pneumonia in patients with deficiencies in w| which d/t unrestrained viral replication & has a high
fatality rates.
3.m Acute postinfectious encephalitis: 1: 1000 which is very serious autoimmune demyelinating disease (no virus
production in e wN).
4.m ubacute sclerosing panencephalitis:
m 1: 300000.
m disease begins insidiously 5-15 years after a case of measles.
m wharacterized by progressive mental deterioration, involuntary movements, muscular rigidity, & coma.
m sually fatal within 1-3 years after onset.
m xhibits high titers of measles Ab in wF & serum & defective measles virus in brain cells.
1.m Dirus detection: can be detected directly in epithelial cells in respiratory secretions & urine.
2.m solation & identification:
ym amples: nasopharyngeal & conjunctival swabs, blood samples, respiratory secretion, & urine.
 ym wp: multinucleated giant cells containing both intranuclear & intracytoplasmic inclusion bodies.


 ym hell using fluorescent Ab staining.
3.m erology:
ym Depends on a fourfold rise in Ab titer b/w acute -phase & convalescent-phase sera.
ym A, H, & Nt tests.
1.m Ditamin A treatment has decreased mortality & morbidity.
2.m |easles virus is susceptible in vitro to inhibition by ribavirin.
3.m Daccination for measles:
ym |onovalent form.
ym n combination with live attenuated rubella vaccines (|).


  ym ive attenuated rubella & mumps vaccine (||).

!

 ym Daccine induced Abs persist for up to 33 years.
ym wontraindications:
m pregnancy.
m Allergy to eggs or neomycin.
m mmune compromise (except for AD patients).
m ecent administration of gs.

 
 "
!
"

   
ym An acute febrile illness characterized by a rash & lymphadenopathy that affect children & young adults.
 ym ubella virus is a member of ogaviridae family & e sole member of genus ubivirus.
ym ubella is worldwide in distribution.
ym nfection occurs t/out e year with a peak incidence in e springs.

    ym pidemics occur every 6-10 years with explosive pandemics every 20 -25 years.
ym nfection is transmitted by e respiratory route, but rubella is not contagious as measles.

  M 
ym Neonatal, childhood, & adult infections occur t/out e mucosa of e upper respiratory tract.
ym nitial viral replication in e respiratory tract l multiplication in cervical lymph nodes l viremia develops after 7-
U days & lasts until e appearance of Abs on about day 13 -15 days.
 
ym After e rash appears, e virus remains detectable only in e nasopharynx where it may persist for several weeks.
ym ne attack of e disease confers a lifelong immunity because rubella gG persists for life.
ym mmune mother transfer Abs to their offspring who r then protect ed for 4-6 months.
ym Megins with malaise, low-grade fever, & a morbilliform rash appearing on e same day.


 ym rash starts on e face, extends over e trunk & extremities, & rarely lasts > than 3 days.


 ym ransient arthralgia & arthritis r commonly seen in adults, especially women.
ym are complications include thrombocytopenia & penic purpura & encephalitis.
1.m solation & identification of virus:
ym pecimen r obtained from nasopharyngeal or throat swabs.
 ym Darious lines of monkey or rabbit origin may be used for cell culture.
ym Diral Ags can be detected by F 3-4 days postinoculation.



2.m erology:
ym Detection of gG is evidence of immunity.
ym H & A tests are a standard serological test for rubella.


  1.m ubella is a mild, self-limited illness, & no specific treatment is indicated.
2.m GD is injected into e mother cannot protect e fetus against rubella.

!

 3.m || vaccine: to prevent congenital infection with lifelong immunity & less side effects.

 M   

ym |aternal rubella associated with rubella infection during pregnancy may result in infection of e placenta & fetus.
ym infection may lead to deranged & hypoplastic organ development resulting in structural anomalies in e
newborn.
ym etratogenic effects depend on e age of pregnancy:
  1.m 1 st trimester: abnormalities in e infant in 85% of cases.
2.m 2nd trimester: detectable defects r found in 16% of infants.
3.m After 20 weeks of gestation: birth defects r uncommon.
m earlier in pregnancy infection occurs, e greater e damage to e fetus.
ym At birth, virus is usually detected in pharyngeal secretion, multiple organs, wF, urine, & rectal swabs.
ym Diral excretion may last for 12-18 months after birth, but e level of shedding gradually decreases wi th age.
ym wan be divided into 3 broad categories:
1.m ransient effects in infants.
2.m permanent manifestations that may be apparent at birth or become recognized during e 1 st year.
3.m Developmental abnormalities that appear & progress during childhood & adolescence.



ym classic triad of rubella: cataract, cardiac abnormalities, & deafness.



ym nfants may develop transient symptoms of growth retardation, rash, hepatosplenomegaly, jaundice, &
meningoencephalitis.
ym wN involvement: > global & may lead to mental retardation with problems in balance & motor skills which may
require institutionalization.


  ym No specific treatment but can be prevented by childhood immunization with ||.

!




 
 
!
"


   
ym hey are two major groups in this virus class. hey are:
$&"!#/ &!$ 1.m nterovirus inhabit inside the human alimentary canal.
2.m hinovirus mainly at the nose and throat.m
ym he picornaviruses are nonenveloped (naked), small (22 to 30 nm) icosahedral virions resistant to lipid solvents
ym he virus capsid is composed of 60 copies each of four viral proteins Dp1 4, which form a quasi = 3 icosahedral shell.
ym he NA strand consists of approximately 7,500 nucleotides and is covalently bonded to a noncapsid viral protein (Dpg)
at its 5i end and to a polyadenylated tail at its 3i end.
ym his genome NA serves as an mNA and initiates the synthesis of virus macromolecules. hus, this virus is a positive
sense virus.
&"/ &/"' ym nterovirus and some rhinoviruses are stabilized by magnesium chloride against thermal inactivation

!+!&!$ ym nteroviruses have a buoyant density in wswl of about 1.33 to 1.34 g/ml
ym Human rhinoviruses are about 1.38 to 1.42 g/ml.
ym he viral capsid gives the picornaviruses their characteristic shap e and size and protects the infectious viral NA from
hostile environments and host ribonucleases.
ym nteroviruses can survive for long periods in organic matter and are resistant to the low pH in the stomach (pH 3.0 to
5.0). My contrast, rhinoviruses are la bile at this pH range.
ym picornaviruses are inactivated by pasteurization, boiling, formalin, and chlorine.
ym he picornaviridae family contains U genera:
1.m nterovirus:
a.m polioviruses.
b.m woxsackieviruses ( Group A & M).
c.m choviruses.

0(* (&!$ d.m nteroviruses.
2.m hinovirus: includes more than 100 antigenic types.
3.m Hepatovirus: Hepatitis A.
4.m parechovirus: parechovirus.
5.m Aphthovirus: Foot-and-mouth disease virus of cattle.
6.m wardiovirus: ncephalomyocarditis virus of rodents.
ym ite: replication cycle occurs in the cytoplasm of cells :
he virion attaches to specific receptor on the plasma membrane

eceptor binding triggers a conformational change in the virion which results in release of the viral NA into the cell cytos ol

Genome-linked protein (Dpg) is removed from the viral NA and associates with the ribosomes

ranslation occurs by a cap-independent mechanism, using the internal ribosome entry site ( ) downstream from the 5
end of the viral genome

he infecting viral NA is translated into a polyprotein that contains both proteins and essential replication proteins

NA-dependent NA polymerase are synthesized
'0 (&!$
Diral NA strand is copied

he complementary strand serves as a template for the synthesis of a new plus strand

he coat precursor protein p1 is cleaved to form aggregates of Dp0, Dp3 and Dp1

hen adequate concentration is reached, these promoters assemble into pentamers that package plus -stranded Dpg-
NA to form provirions

he provirions are not infectious until a f inal cleavage changes Dp0 to Dp4 and Dp2

he virus are released when the cell disintegrates m

$&'"!1"/"!/

!0!1"/
m t is acute infectious disease
$&"!#/ &!$ m ts serious form affects the wN
m he destruction of motor neurons in the spinal cord results in flaccid paralysis
General properties m ypical nterovirus
m hey are inactivated when heated at 55 w for 30 min
m |g2+, 1 mol/ prevents its inactivation

"!'"&'!* Animal usceptibility m polioviruses have a very restricted host range

& Growth of Dirus
&2'"/ m |ost strains will infect monkeys when inoculated directly into the brain or the spinal cord
m poliovirus requires a primate-specific membrane receptor for infection, thus, polio virus only
infect those of the primates and higher animals
Antigenic properties m here are three antigenic types of poliovirus
 portal of entry mouth

primary multiplication of the virus takes place in the oropharynx or intestine

he virus is regularly present in the throat and in the stools before the onset of illness

ne week after infection there is little virus in the throat

Dirus continues to be excreted in the stools for several weeks even though high antibody levels are present in the blood

Antibodies to the virus appear early in the disease, usually before paralysis occur

(&2!%'$'

(&2!0!%) he virus first multiplies in the tonsils, the lymph nodes of the neck, peyers patches and the small intestines

he wN are invaded by the dissemination of the virus via hematogeneous spread

poliovirus can spread along the axons of peripheral nerves to the wN, where it continues to progress along the fibres of the
lower motor neurons, thus involve the spinal cord or the brain.

he virus invades certain types of nerve cells, and in the process of its intracellular multiplication it may damage or
completely destroy these cells

pAAY

ym ncubation period: 7 14 days.

ym |ost common form of the disease

|ild Disease ym |inor illness
ym wharacterized by fever, malaise, drowsiness, headache, vomiting, constipation, sore throat

0$ (0 Nonparalytis
ym he disease lasts for 2 10 days
*$#$% poliomyelitis
(Aseptic |eningitis) ym tiff neck and severe headache
ym Flaccid paralysis
paralytic poliomyelitis
ym ncoordination secondary to painful spasms of nonparalyzed muscles may also occur
progressive ym |uscle wasting
postpoliomyelitis ym t is not a consequence of persistent viral infection, but rather a result of physiologic and
|uscle Atrophy aging changes in paralytic patients already burdened by loss of neuromuscular functions
ym hroat swabs.
ym pecimen should be kept frozen.
(3(%$! ym wp appear in 3 6 days.
ym pw.
ym equire molecular techniques to differentiate between the wild and the vaccine type.
ym mmunity is permanent to the virus type causing infection.
ym Antibody mediated.
++/$&) ym passive immunity is transferred from mother to child : maternal antibodies gradually disappear within 6 months.
ym Dirus-neutralizing antibodiesm forms soon after exposure to the virus, often before the onset of illness, and persists for life:
the Dp1 surface protein of poliovirus is responsible for the formation of the neutralizing antibodies .
ym poliomyelitis has three epidemiologic phases:
1.m ndemic.
2.m pidemic.
3.m Daccine era
#'+!0!%) ym he prevaccine pattern include the improves systems of hygiene and sanitation in cooler climates .
ym he diseases occur in all age groups, but more prevalent in children .
ym Adult is less susceptible to the infection because of the acquired immunity.
ym Humans are the only known reservoir hosts .m
ym Moth live-virus and killed-virus vaccine are available:
1.m alk vaccine ( formalinized vaccine) is prepared from virus grown in monkey kidney cultures .
2.m Killed-virus vaccine: induces humoral antibodies but does not induce local intestinal immunity. hus, the virus can
still replicate inside the gut.
3.m ral live attenuated vaccine:
m he live polio vaccine infects, multiplies, and immunizes.
m he vaccine induce the production of g |, g G & g A antibodies in the intestines (systemic and gut immunity) .

"'1'$&!$ ym Moth killed-virus and live-virus vaccine induce antibodies and protect the wN from subsequent invasion by wild virus
ym imiting factors r e nterference:m

!$&"!0 m f the alimentary tract is infected by other enterovirus at the time the vaccine is given, the establishment of polio
infection and immunity may be blocked.
m his phenomenon usually occurs at places where enterovirus infection is common.
m pregnancy is neither an indication for nor a contraindication to required immunization.
m ive-virus vaccine should not be administered to immunodeficient or immunosuppressed individuals or their
household contacts.
m nly killed-virus (alk) vaccine is to be used.
m here are no antiviral drugs are pr epared for the treatment of poliovirus .

  K

!
"


   
ym A large subgroup of the enterovirus
ym hey are divided into two groups
A & M
ym Having different pathogenicity potential for mice
ym hey produce variety illnesses to human, such as aseptic meningitis, respiratory and undifferentiated febrile illnesses
$&"!#/ &!$ ym Group A: Herpangina (vesicular pharyngitis), hand-foot-and-mouth disease & acute hemorrhagic conjunctivitis
ym Group M: pleurodynia, myocarditis, pericarditis, severe generalized disease of infants
ym A number of A and M serotypes can give rise to meningoencephalitis and paralysis
ym woxsackie virus group M are the most commonly identified causative agents of viral heart disease in humans.
ym he coxsackie virus are more pathogenic than ech ovirus

"!'"&'!* ym wertain strain can grow in monkey kidney cell culture

ym ome Group A strains grow in human amnion and human embryonic lung fibroblast cells
&2'"/
ym Dirus has been recovered form the blood at initial stages of infection in humans

(&2!%'$'
ym Dirus is also found in the stool for 5 6 weeks

(&2!0!%) ym Dirus distribution is similar to that of other enterovirus
ym ncubation period = 2 U days
ym wlinical manifestation = diverse
Disease wausative Dirus wlinical |anifestations
m Fever, malaise, headache, nausea, abdominal pain are
m all types of Group M coxsackie common early symptoms
Aseptic meningitis
virus m |ild muscle weakness suggestive for paralytic
m some strains of Group A poliomyelitis
coxsackie virus m patients almost recover completely from nonpolio virus
paresis
m Abrupt onset of fever and sore throat with discrete
vesicles on the posterior half of the palate, pharynx,
Herpangina m some Group A woxsackie virus tonsils or tongue
m elf limited
m Frequent in small children
m ral and pharyngeal ulcerations
m Desicular rash of the palms and soles that may spread
to the arms and legs
Hand-foot-and-
mouth disease m woxsackie virus A16 m Desicles heal without crusting (clinically differentiated
from the vesicles of herpesvirus and poxvirus)
m he virus can be isolated from stool, pharyngeal
secretions and vesicular fluid.

0$ (0 m Fever, stabbing chest pain that lasts for 2 days to 2
weeks
$#$%
pleurodynia m Group M viruses m preceded by malaise, headache and anorexia
m Abdominal pain occurs in half of the cases
m ecover is complete bur relapses are common
m t is a serious disease
m |ay cause inflammation of the heart or the
pericardium
|yocarditis m Group M viruses m |ay cause permanent heart damage to any age
group
m persistent viral infections of the heart may also occur,
sustaining chronic inflammation
espiratory ract
m A number of woxsackie virus m wause common colds & undifferentiated febrile
nfections illnesses
m xtremely fatal disease
m imultaneous viral infections of various organs
Generalized Disease m n severe cases myocarditis or pericarditis can occur
of nfants m Group M viruses first 8 days of life
m wan be preceded by a brief episode of diarrhea and
anorexia
m wan be acquired transplacentally
ym Although the G is the primary site of replication for enterovirus, they do not cause marked disease there.
ym wertain Group A virus are responsible for diarrhea in children

(3!"(&!") m Diruses can be isolated from throat washings during the first few days of illnes s and the from
ecovery of Dirus the stools the first few weeks
(%$! m woxsackievirus A21 can be found in the nasal secretions
 m n aseptic meningitis, strains can be recovered from the wF as well as the G
Nucleic Acid m everse transcription pw are reactive (detect many serotypes) and specific
Detection m eal time pw
m Neutralizing antibodies appear early during the course of infection, tend to be specific for
erology the infecting virus and persist for years
m erum antibodies can be detected by F
ym Neutralizing antibodies are transferred from mother to child
++/$&) ym Adults tend to have more types of antibodies against woxsackievirus than do children, indicating that multiple
experiences with these viruses are common and increasingly so with age
ym woxsackievirus are more frequently encountered during the summer and early fall
#'+!0!%) ym |any children develop antibodies against these viruses which indicates infection

!$&"!0 ym here are no antiviral drugs or vaccines available for the treatment of woxsackievirus





!
"


   
ym chovirus (nteric wytopathic Human rphan viruses) .
$&"!#/ &!$ ym hey primarily infect the human enteric tract .
ym o establish etiologic association of an enterovirus with disease, the following criteria are used:
1.m here is much higher rate of recovery of virus from patients with the disease than from healthy individuals of the
same age and socioeconomic level in the same area at the same time
2.m Antibodies against the virus develop during the course of infection
3.m he virus can be isolated from body fluids or tissues manifesting the lesions
ym |any echovirus infections are associated with aseptic meningitis
ym ashes are more common in young children

0$ (0 nterovirus wlinical manifestation
$#$% ym whief cause of acute hemorrhagic conjunctivitis
ym Has a sudden onset of subconjuctival hemorrhage
nterovirus 70 ym |ost common in adults
ym womplete recovery is a rule
ym he virus is highly communicable and spreads rapidly under crowded or unhygienic conditions
ym presents with meningitis, encephalitis, and paralysis resembling poliomyelitis
nterovirus 71
ym |ain causes of wN disease
ym t is impossible in an individual case to diagnose an echovirus infection on clinical grounds.
ym However, in epidemic situations echovirus must be considered
1.m ummer outbreaks of aseptic meningitis
2.m ummer epidemics, especially in young children, of a febrile illness with rash
(3(%$! ym Diagnosis depend on:
1.m ab test
2.m Nucleic acid detection assays, pw
3.m Diral isolation from throat swabs, rectal swabs, wF
ym nfection with 2 or more viruses may occur .
ym ame like other enterovirus
#'+!0!%) ym sually occur during summer and early autumn
ym High prevalence in young children
ym Avoid contact with patients exhibiting acute febrile illness esp, those with a rash

!$&"!0 ym here are no antiviral drugs or vaccine available for the treatment of any enterovirus diseases


!
"


   
ym wommon cold viruses which can be isolated from nasal, throat and oral secretions .
$&"!#/ &!$ ym wause upper respiratory tract infections with common cold syndrome .
ym here are more than 100 species
ym Human rhinoviruses can be divided into two groups : major & minor receptor groups

0(* (&!$ m |ajor receptor groups they use the intercellular adhesion molecule-1 (wA|-1) as a receptor
m |inor receptor groups utilize the low-density lipoprotein receptor (D) family
ym hinoviruses are picornavirus similar to enterovirus
ym Differ from enterovirus by having buoyant density in wswl = 1.40 g/ml and acid labile
General properties ym Dirions are unstable under the pH 5 6
ym womplete inactivation at pH 3.0

"!'"&'!* ym hinoviruses are more thermostable than enteroviruses
&2'"/ Animal ym hese viruses are only infectious for humans, gibbons and chimp anzees
usceptibility & ym |ost grow better at 33 w which is similar to the temperature of the nasopharynx in human,
Growth of Dirus than at 37 w
Antigenic property ym |ore than 100 serotypes are known
 he virus enters the upper respiratory tract high titers of virus in nasal secretions as early as 2 4 days after
infection

eplication is limited to the surface epithelium of the nasal mucosa

(&2!%'$'
dema & infiltration of the nasal mucosa

(&2!0!%)
Nasal secretion increases in quantity
ym hinoviruses rarely causes lower respiratory tract infection
ym ometime after infection, virus titers fall even though the infection persists!
ym ncubation period = 2 days 4 days

0$ (0
ym Acute illness usually lasts for 7 days
$#$% ym Nonproductive cough may persists for 2 3 weeks
ym neezing, nasal obstruction, nasal discharge, sore throat
ym Headache, mild cough, malaise, chilly sensation
ym here is little of no fever
)+&!+ ym he nasal and nasopharyngeal mucosa become red and swollen
ym he sense of smell become less keen
ym econdary bacterial infection may produce acute otitis medis, sinusitis, bronchitis, pneumonitis, esp in children
ym Neutralizing antibodies to the infecting virus develops in serum and secretions of most person
++/$&) ym Antibodies develop 7 21 days after infection
ym he appearance of neutralizing antibodies in nasal secretions is parallel to the serum antibodies
ym this disease occurs throughout the world
ym n temperate climates the highest attack rates are in early fall and late spring
ym prevalence rates are lower in summer
#'+!0!%) ym |ode of transmission close contact, by means of virus-contaminated respiratory secretions
ym hinoviruses can survive in the environment for several hours
ym n a single outbreak, multiple serotypes of rhinoviruses can cause infections
ym No specific prevention methods are available
ym he development of vaccine against rhinoviruses are very unlikely because it is difficult to grow the virus in a culture
"'(&+'$& media, the fleeting immunity and the different serotypes that cause infection

!$&"!0 ym Antiviral drugs are thought to be the most likely control measure for rhinoviruses because of problem in developing the
vaccine
ym A 5- day course of high doses of intranasal interferon -





7
 





!"&!$ '&(0
he total inactivation (killing) of all forms of all microbial life in terms of the organisms ability
&'"04(&!$ to reproduce

$*' &!$ he killing or removal of organisms capable of producing an infection

2'+ (0 A substance that is used to kill organisms in inanimate surfaces but it is too toxic to be applied
directly to tissues.
#$*' &($&
'*$&!$ he prevention of sepsis (putrefaction), either by removal or killing of the organisms ( -cidal) or
$&' by opposing their growth (-static)

2'+ (0
A substance that can be applied topically to living tissues
($&'& 
'%'"+$% emoval of microbes from a limited area

($&4(&!$ ower microbial counts on eating utensils

t is estimated by plotting the bacterial count on a semi -log scale against the time of exposure
'&2!# to the antimicrobial action
1.m t is either logarithmic or exponential curve
m ne log decrease indicates U0% of the population is killed

"!'"&'!* 2.m he initial lag or shoulder may be due to either multihit inactivation dynamics or to
spore germination
'(/"'+'$&!* '(&2"(&'
/"1' 3.m he final lag is due to either to the presence of mixed population of sensitive or
'(&2 resistant organism or to protection from the antimicrobial action e.g. the presence of
organic material.

)'!* ingle-hit inactivation- damaging one target molecule is enough to kill the organism
$( &1(&!$
|ulti-hit inactivation- damaging more than one target is necessary to inactivate an organism.

m Number of microbes
( &!"(**' &$% m nvironment e.g. organic matter, temperature, biofilms
 ($&+ "!3(0 m ime of exposure
m |icrobial characteristics
&"'(&+'$&
&!$!* m Alternation of membrane permeability
$&+ "!3(0 m Damage to proteins
m Damage to nucleic acid
%'$&
&'"04(&!$

2) (0+'&2!#
2'+ (0+'&2!#
m Dry heat m thylene oxide
)'!* 1.m Heat
m |oist heat m Meta propiolactone
&'"04(&!$ m Aldehydes
2.m onizing radiation Gamma rays m Formaldehyde
m Glutaraldehyde
m Filtration removes microbe
m ow temperature inhibits microbial growth
m efrigeration
m Deep freezing
' 2($+ m yophilization m All are alkylating agents
m High pressures denatures protein
m Desiccation prevents metabolism
m smotic pressure cause plasmolysis
m adiation damages DNA
1.m thylene oxide
m Advantages:
m Highly microbicidal
m High penetrating power
m t is used to sterilize heat
m Kills organisms by oxidation
")2'(& sensitive equipments
m .g. by direct flaming,incinerators, hot air oven
m Disadvantages:
m An explosive gas
m Highly toxic gas
m Need a special chamber to
perform sterilization.
2.m Meta propiolactone
m Autoclave m An alkylating agent that hydrolyze
very quickly to acrylic acid
m A device that produces steam at high pressures and
temperature in a closed chamber e.g.121 w m sed to sterilize serum and other
'&2!# m he type of the autoclave depends on how the air is biological agents
removed. m sed to kill microorganisms in the
a)m Downward displacement autoclave preparation of killed antigen
!&2'(& b)m High prevacuum autoclave vaccines.
m Advantages : 3.m Aldehydes
a)m ocal release of latent heat of evaporation m Good cidal activity even against
at points of steam condensation on the spores
articles being sterilized m Moth aldehydes are toxic and may
b)m Good penetration of steam cause hypersensitivity
m Disinfect fibre-optic endoscopes
onizing adiation ( gamma radiation)
m Are lethal to DNA
m Great penetrating power, even of sealed packs
m ources: radioactive isotopes, such as woblt -60
m Disadvantages: expensive and need extra precaution

&'"04(&!$
"! '
nvolves the steps:
1.m Designing the sterilization process
2.m esting the process
3.m esting the process
m nitial contamination level
M!3/"#'$ m ncreased bioburden means increased time required to achieve a given level of sterility assurance
m hus, precleaning before sterilization is important.
'%$$%&2' m t is the probability that the organism may survive through a sterilizing process
m he required level is dependent on the intended use of the articles
&'"04(&!$ &'"0&) m 10-6 is required for aseptic procedures
"! ' (/"($ ' m 10-12 is required for canned food
m 10-3 is required for the articles that are going to come in contact with the skin or the mucus
 membrane

(&'!* m o calculate the time required to achieve a certain le vel of sterility assurance we must know the
decimal reduction time (D value)
3! #(0( &!$ m he time required to reduce the microbial population by U0% at a certain temperature

'&$%&2'
2) (0 m emperature and pressure measurement on record charts
 "! ' &'&$% m Air leak test for pre-vacuum autoclaves

 m Mrownes tubes
m he tubes that are put in the pack to be sterilized
m he chemical liquid indicator will change color from red to green when exposed to
the required temperature for the required time

2'+ (0 m Mowie-Dicks est
&'&$% m placing autoclave sheets with markings that change color in the pack and look for a uniform
color change in the test sheet indicating the appropriate penetration of steam to the packs
'&$%&2' center.
"! ' m his test measures the sterilizers ability to remove all the air from the chamber in a pre -
determined period of time.

M( &'"!0!% (0 m pore strips of the heat resistant Bacillus staerothermophilus

m hey are included in different parts of the autoclave in a pack
!"'&'& m After the completion of the sterilization process, they are cultured in suitable broth.

$*' &!$ $&'

35' &1' m o reduce the count of pathogenic organisms in a potential source of infection to below that required to cause infection

0'($$% m educes the number of organism

#'%'"+$% m emoves organic matter, dirt, grease that might protect the organism from inactivation

m pasteurization
m Holder type process - 63w for 30 minutes
m Flash process - 72w for 20 sec
m Moiling at 100w
m wan kill most vegetative organism within one minute
m Mut spores might need more than 20 minutes
'(&$% m ow temperature steam disinfection
m t is more effective if the steam is combined with formaldehyde.
m hydillization
m sed for heat sensitive culture media
m t is a successive procedure
m pls read notes,he3

$*' &!$3) m |ost efficient wavelength at 260nm

0%2& m o decontaminate open spaces and surfaces
)' 2) (0+'&2!# m Disadvantages poor penetration.

1.m Filtration heat-labile liquids

a)m Depth filter
m wonsists of granular material bonded into thick layer containing
twisted channels of small diameter
b)m |ilipore membrane filters
0&"(&!$ m porous membranes made of cellulose acetate or nitrate,
polycarbonate etc
m hey replaced the depth filters

2.m Filtration of air using HpA filter

m sed in operating theatres or in microbial safety hoods
$*' &!$($# m wleaning is a prerequisite before chemical disinfection because organic matter and dirt can
($&'6&2 interfere with their action and help protect the organism against inactivation
m Few chemicals are sporicidal.
2'+ (0