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Avian Avian ColibacillosisColibacillosis
By:By:
Seyed Ahmad MadaniSeyed Ahmad MadaniResident of Poultry Diseases DivisionResident of Poultry Diseases Division
Dept. of Clinical ScienceDept. of Clinical Science
Faculty of Veterinary MedicineFaculty of Veterinary Medicine
University of TehranUniversity of Tehran
Definition and SynonymsDefinition and Synonyms Localized or Localized or
systemic infection systemic infection Entirely or partlyEntirely or partly Avian Pathogenic Avian Pathogenic
Escherichia coli Escherichia coli (APEC)(APEC)
HistoryHistory
ColisepticemiaColisepticemia Coligranuloma (Hjarre’s dis.)Coligranuloma (Hjarre’s dis.) Air sac dis. (CRD)Air sac dis. (CRD) Coliform cellulitis(inflammatory Coliform cellulitis(inflammatory
process)process) Swollen-head Syndrome (SHS)Swollen-head Syndrome (SHS) Coliform peritonitisColiform peritonitis Coliform SalpingitisColiform Salpingitis Coliform osteomyelitis/synovitisColiform osteomyelitis/synovitis (Turkey osteomyelitis complex)(Turkey osteomyelitis complex) Coliform panophthalmitisColiform panophthalmitis Coliform omphalitis/Yolk sac Coliform omphalitis/Yolk sac
infrctioninfrction
EtiologyEtiologyAvian Pathogenic Avian Pathogenic Escherichia coli Escherichia coli (APEC)(APEC)
Gram negative, non-acid-fast, Gram negative, non-acid-fast, uniform stainng, non-spore-forming uniform stainng, non-spore-forming bacillusbacillus
MacConkey agarMacConkey agar Tergitol-7 agarTergitol-7 agar EMB agarEMB agar 18-4418-44ooC C Biochemical propertiesBiochemical properties
Antigenic StructureAntigenic Structure 167 O, 74 K, 53 H and 17 F167 O, 74 K, 53 H and 17 F O (Somatic) Ag.: LPS (endotoxin)→ Tube O (Somatic) Ag.: LPS (endotoxin)→ Tube
agglutination in 50agglutination in 50ooC for 24 hrC for 24 hr K (Capsular) Ag.: Polysaccharide (1 hr at K (Capsular) Ag.: Polysaccharide (1 hr at
100100ooC)C)Tube agglutinatio→37Tube agglutinatio→37ooC for 2 hrC for 2 hrSlide agglutinationSlide agglutination H (Flagellar) Ag.: motility. Tube H (Flagellar) Ag.: motility. Tube
agglutination → 50agglutination → 50ooC for 2 hrC for 2 hr F (Pilus) Ag.F (Pilus) Ag.
Strain ClassificationStrain Classification
Geographic variationGeographic variation O78, O1, O2 and O35O78, O1, O2 and O35 Untypeable or Rough strains also Untypeable or Rough strains also
occuroccur A study in Spain (1998): only 15% of A study in Spain (1998): only 15% of
strains belonged to serogroups O1, strains belonged to serogroups O1, O2, O35, O36, and O78.O2, O35, O36, and O78.
Newly emerged serogroups: O18, O115, Newly emerged serogroups: O18, O115, O116, O132O116, O132
Strain ClassificationStrain Classification
0
5
10
15
20
25
30
35
40
O78K80 O2K1 O111K58
1344
1357
1380
1344: 1344: O78K80O78K80, , O88H10, O2K1 O88H10, O2K1 and O111K58and O111K58
1357: 1357: O78K80O78K80, , O111B4 and O111B4 and O119B14O119B14
1380: 1380: O78K80O78K80, , O128K67, O2K1 O128K67, O2K1 and O124K72and O124K72
Characterization of Characterization of APECAPEC
BiotypingBiotyping Phage TypingPhage Typing Antibiotic ResistanceAntibiotic Resistance Multilocus Enzyme Electrophoresis (MLEE)Multilocus Enzyme Electrophoresis (MLEE) Whole Cell Protein ProfilingWhole Cell Protein Profiling RibotypingRibotyping RFLP/Southern blotRFLP/Southern blot Pulse Field Gel Electrophoresis (PFGE)Pulse Field Gel Electrophoresis (PFGE) RFLP-PCRRFLP-PCR REP-PCR(repetitive extragenic palindromic)REP-PCR(repetitive extragenic palindromic) ERIC-PCR (enterobacterial repetitive ERIC-PCR (enterobacterial repetitive
intergenic consensus)intergenic consensus) Randomly Amplified Polymorphic DNA Randomly Amplified Polymorphic DNA
Analysis Analysis (RAPD-PCR)(RAPD-PCR)
Characterization of APECCharacterization of APEC
Virulence and PathogenesisVirulence and PathogenesisEmbryo lethalityEleven 12-day-old chicken embryo
Inoculation via allantoic cavity100 cfu
2-day mortality
<10%→nonvirulent10-29%→intermediate >29%→virulent
Virulence and Virulence and PathogenesisPathogenesis
Generalizing from Mammalian pathogensGeneralizing from Mammalian pathogens Correlational Studies (Occurrence of Correlational Studies (Occurrence of
factors)factors)1.1. PhenotypingPhenotyping
2.2. Genotyping → PCR → Multiplex PCRGenotyping → PCR → Multiplex PCR In-vitro Studies In-vitro Studies (cell or organ culture/expression of virulence (cell or organ culture/expression of virulence
factors)factors) Recombinant or Knock-out mutant Recombinant or Knock-out mutant
pathogenicitypathogenicity
Virulence Factors & Virulence Factors & PathogenesisPathogenesis
Adhesion: F1fimbriae, F11(P) fimbraieAdhesion: F1fimbriae, F11(P) fimbraie Colonization: F1, P, Curli fimbriae, TshColonization: F1, P, Curli fimbriae, Tsh Penetration and Invasion: Motility Penetration and Invasion: Motility
(flagella)(flagella) Phagocytosis resistance: P, K1 Ag., O78 Phagocytosis resistance: P, K1 Ag., O78
Ag.Ag. Serum/Complement resistance: K1 Serum/Complement resistance: K1
capsule, LPS (O78), OMP (Iss, Trat, capsule, LPS (O78), OMP (Iss, Trat, OmpA)OmpA)
Iron acquisition: Aerobactin (Iron acquisition: Aerobactin (iuciuc))
Type 1 fimbriaeType 1 fimbriae
Gene: fim F1 Serotype, Proteinaceous surface
appendages Thread-like appearance D-mannose sensitive ► bind to many types of
eukaryotic cells
Adhesion:Express in Trachea&lungs&airsacs but not in other internal organs
Involving in Phagocytosis
P fimbriaeP fimbriae
Gene: Gene: pappap Pyelonephritis-associated pili or Pili
Blood Ag. F11 serotypeF11 serotype Receptors are in internal organs (not Receptors are in internal organs (not
in trachea)in trachea)
→ → Maybe involved in SepticemiaMaybe involved in Septicemia D-mannose resistantD-mannose resistant
Curli fimbriaeCurli fimbriae
Binding to MHC-I, extracellular Binding to MHC-I, extracellular matrix&Serum Prmatrix&Serum Pr
Survival outside the host & colonizationSurvival outside the host & colonization Most isolates are curliatedMost isolates are curliated Thin, Coiled, hair-like surface appendagesThin, Coiled, hair-like surface appendages Gene: Gene: csgcsg Colonization, Internalization to eukaryotic Colonization, Internalization to eukaryotic
cells & Coagulopathy due to fibronectin cells & Coagulopathy due to fibronectin interactioninteraction
Serum ResistanceSerum Resistance
K1 capsule or other capsular Ag.K1 capsule or other capsular Ag. Smooth lipopolysaccharides (O78)Smooth lipopolysaccharides (O78) Most on ColV plasmid and Conjugated R Most on ColV plasmid and Conjugated R
plasmidplasmid Outer Membrane Proteins (OMPs)Outer Membrane Proteins (OMPs)
1.1.Plasmid coded Plasmid coded traT → traT → TraT proteinTraT protein
2.2.Chromosaomal coded Chromosaomal coded ompA ompA → OmpA → OmpA
3.3.Plasmid codedPlasmid coded iss iss → Iss proteine → Iss proteine (increased serum survival)(increased serum survival)Correlation/Causing/Confounding
Coliform CellulitisColiform Cellulitis (Inflammatory Process)(Inflammatory Process)
Synonyms: Avian cellulitis, inflammatory Synonyms: Avian cellulitis, inflammatory process, infectious process and IP process, infectious process and IP
Sheets of serosanguinous to caseated, Sheets of serosanguinous to caseated, fibrinoheterophilic exudate in subcutaneous fibrinoheterophilic exudate in subcutaneous tissuestissues
History: Randall et al. (1984)History: Randall et al. (1984) Economic significance: Economic significance:
↑condemnations, ↑downgrading, ↑labor.↑condemnations, ↑downgrading, ↑labor. In 1996: 2.6 million broilers were condemned = 30% of total In 1996: 2.6 million broilers were condemned = 30% of total
condemnationcondemnation
In 1991: $20 million In 1998: $80 millionIn 1991: $20 million In 1998: $80 million
EtiologyEtiology
1. Pseudomonas aeruginosa
2. Proteus vulgaris3. Enterobacter
agglomerans4. Pasteurella
multocida5. Strep. dysgalactiae6. Aeromonas sp.
E. coliE. coli
Same serogroups as
APECO1, O2, O78
•Virulence properties are the same
•Usually produce colicin & aerobactin•Cellulitis isolates have greater ability to produce cellulitis lesion
Presence of endemic populations of cellulitis-
associated E .coli in the broiler house
environment
EpidemiologyEpidemiology
Environment Birds
E. coli
Coliform Ccellulitis
Breed•Fast growing•Heavy broiler strains•Genetic → Strength of skin•Aggresiveness•Poor feathering
Feathering↓
Scratching↑Sex
Males↑Feather growth weight aggressiveness
Stocking Density
•↑Skin scratches•↑Stress•↑Contact
Litter
FeedingAA↓(Cys&Met), ↑Energy(E/P ratio, fat deposite), Vit. E, Vit. A, Zinc
PathobiologyPathobiology
Host: Older chicken are more likely Host: Older chicken are more likely to develop cellulitis.to develop cellulitis.
No Antemortem Clin. Signs → No Antemortem Clin. Signs → Lesions are discovered at processing Lesions are discovered at processing by inspectorsby inspectors
Gross Pathology: Gross Pathology: Unilateral
Skin colorNormal YelowRed-brown
Size 1-10 cm
Scratch orScabsOn theSkin
Subcutaneous Edma
ExudateMus. Hemo.
Fibrinous orCaseous plaque
In Mus. Or Subcutis(waffle skin)
PathogenesisPathogenesis
Skin trauma
ScratchesCorrelation with
Systemic Colibacillosis
Association withAscites Valgus-varus
Leg deformity
Confounding ?Sex
Breed
Large abdomenCommon risk factors (rapid
growth)
6-12 hr PI → exudate18-24 hr PI → Caseous
plaqueLesions prsist 3 W PI
Age> 3 weeks
Dx/Tx/Control/PreventionDx/Tx/Control/Prevention
DxTime of OccurrenceTransportation/Rearing house
No TxManagement
andEnvironment
(risk factors)
MontoringAt Processing:CondemnationAseptic Culture
Cost benefit
Swollen Head SyndromeSwollen Head Syndrome SHS is an acute to subacute cellulitis involving SHS is an acute to subacute cellulitis involving
the periorbital & adjacent sc tissues of the the periorbital & adjacent sc tissues of the headhead
Host: Chicken (broilers, broiler breeders & Host: Chicken (broilers, broiler breeders & commercial layers), Turkey and Guinea fowlcommercial layers), Turkey and Guinea fowl
Pathogenesis: Pathogenesis:
Clin. Signs & Lesions: Edematous swelling over Clin. Signs & Lesions: Edematous swelling over the eye, gelatinous edema of the facial skin & the eye, gelatinous edema of the facial skin & periorbital tissues & caseous exudate in the periorbital tissues & caseous exudate in the conjuctival sac, facial sc tissues & lacrimal conjuctival sac, facial sc tissues & lacrimal gland.gland.
Upper Resp.Viral Infect.
Avian PneumovirusIBV
Ammonia
E. ColiVia
SinusesNasal cavity
Eustachian tube
Inflammatoryexudate
Salpingitis/Peritonitis Salpingitis/Peritonitis (Adults)(Adults)
↓↓Egg production & Sporadic Mortality in Egg production & Sporadic Mortality in Commercial Layers and BreedersCommercial Layers and Breeders
The most common causes of mortality in The most common causes of mortality in lay in laying henslay in laying hens
Other host: esp. female duck & geeseOther host: esp. female duck & geeseConfused with egg binding Confused with egg binding Route of Infection:Route of Infection:Predisposing factors:Predisposing factors:Pathology: Pathology:
1.Ascending from Cloaca2.Spreading from air sacculitis( but more frequently in young birds)
3.?! Hematogenic due to septicemia
1. Heavy egg production→relaxing the sphincter between vagina & cloaca
2. Mucosal Viral Infection (IB)
3. Mycoplasmosis
1.Distention of thin-walled oviduct with single or multiple masses of caseous exudate in the general form of the oviduct
2.Exudate ismalodorous and laminated & often contain central egg, shells and/or membranes
▼Peritonitis
(with or without salpingitis)
Coliform Omphalitis/Yolk Coliform Omphalitis/Yolk Sac InfectionSac Infection
Definition/Synonyms:Definition/Synonyms: The Commonest Cause of Mortality The Commonest Cause of Mortality
during 1during 1stst Week Week Etiology:Etiology: Route of Infection: Route of Infection:
Inflammation of the NavelInflammation of the umblicusNavel illMushy Chick Disease
1.E. coli2.Proteus spp.3.Bacillus spp (esp. B.
cereus)4.Staphylococci5.Pseudomonas spp.6.Clostridia
•Contamination of unhealed Navel◄ Fecal Contamination of eggs•in ovu contamination ◄Oophoritis/Salpingitis/Artificial Insemination•Intestinal contamination•Hematogenic
Coliform Omphalitis/Yolk Coliform Omphalitis/Yolk Sac Infection (cont.)Sac Infection (cont.)
Dead in Shell (late in incubation)Dead in Shell (late in incubation) Death Shortly after hatchingDeath Shortly after hatching Incidence↑ after hatching & ↓ about Incidence↑ after hatching & ↓ about
6 days with occasional losses up to 3 6 days with occasional losses up to 3 w.w.
Infection with low virulent: no Infection with low virulent: no mortalitymortality
1.Retained caseated Yolk2.↓ Weight gain
RednessSwellingEdemaSmall abscessesAbdominal distensionHyperemiaSkin& Body wall lysisWet and dirty chicksMushy chicksDehydrationVisceral goatEmaciatioPericarditis > 4 days
Deprivation of Nutrient & AbsAbsorption of Toxins
Spread of E. coli Colisepticemia
Adhesion to Intestines or Other Visceral Orgas
StrangulationDeath
Treatment is not recommended
ColisepticemiaColisepticemia
Pathologic Stages:Pathologic Stages:
Origin of Infection:Origin of Infection: 2 & 12 weeks (4-9)2 & 12 weeks (4-9)
1.Acute Septicemia2.Subacute
Polyserositis3.Chronic
Granulomatosis1.Respiratory –origin2.Enteric – origin3.Neonatal
Colisepticemia4.Acute Sep. of
Layers
ColisepticemiaColisepticemia Green discoloration following exposure to Green discoloration following exposure to
airair Odor ← Indole productionOdor ← Indole production Atrophy of FB or Inflammation ≠ IBDAtrophy of FB or Inflammation ≠ IBD Pericarditis/ Myocarditis: Cloudiness of Pericarditis/ Myocarditis: Cloudiness of
Pericardial sac+ edema of epicardium+ Pericardial sac+ edema of epicardium+ Fibrinous exudate→ exudate Fibrinous exudate→ exudate organization(7-10 days)→ Constrictive organization(7-10 days)→ Constrictive PericarditisPericarditis
PeritonitisPeritonitis PerihepatitisPerihepatitis
Prevention/Control/Prevention/Control/TreatmentTreatment
Egg & Laying Egg & Laying ManagementManagement: : Contamination/Collecting/Nest/Floor Contamination/Collecting/Nest/Floor eggs/Broken/Hatchery(esp. after 18)eggs/Broken/Hatchery(esp. after 18)
Nutritional Nutritional Management Management : ↑Pr, ↑Se, : ↑Pr, ↑Se, ↑Vit. E, ↓Deprivation, ↓Water ↑Vit. E, ↓Deprivation, ↓Water contamination, ↓food contamination contamination, ↓food contamination (mash>pellet)(mash>pellet) ,rodent feces ,rodent feces
Health Health Management Management : IB, ND, Mg, IBD, : IB, ND, Mg, IBD, Aflatoxicosis, Ammonia, Dust, Aflatoxicosis, Ammonia, Dust, TRT, TRT, THE.THE.
ImmunizationImmunization
Inactivated Vaccines:Inactivated Vaccines:
Live Vaccines:Live Vaccines:
Subunit Vaccines: Subunit Vaccines:
• Protection against homologous Serogroups
• Passive Protection in progeny
• Poly/Monovalent• Adjuvant• O78, O2, O1• Ultrasonic/Heat/
Formalin
•Naturally non-pathogenic piliated strain(BT-7) in chicken>14 days on even heterologous strains•J5: incomplete endotoxin•carAB mutant of O2 strain (↓Arg&pyrimidine)•Recombinant: O78/F1 on attenuated S. typhimurium•Iss protein(2006)
•Type 1 fimbriae(FimH)
Chlortetracycline 20%Chlortetracycline 20%
Water Soluble PowderWater Soluble PowderMechanism: Inhibition of Aminoacil Mechanism: Inhibition of Aminoacil transferase attaching to 30S ribosometransferase attaching to 30S ribosomeDrinking water: 80-100 gr/100 lit. 5-7 dDrinking water: 80-100 gr/100 lit. 5-7 dFood: 1500-2000gr/ton. 5-7dFood: 1500-2000gr/ton. 5-7dContra.: Layers (↓shell thickness), renal & Contra.: Layers (↓shell thickness), renal & hepaic insufficiencyhepaic insufficiencyDrug interaction: Erythromycin, Drug interaction: Erythromycin, Enrofloxacin, Neomycin, NaCo3, Ca, Mg, Enrofloxacin, Neomycin, NaCo3, Ca, Mg, Fe.(12h in hard water)Fe.(12h in hard water)Withdrawal: 5-7dWithdrawal: 5-7dPack: 500gr/ 1 kgPack: 500gr/ 1 kg
Chlortetracycline 50%Chlortetracycline 50%
Premix powderPremix powder Prevention dose: 200-400 gr/ tonPrevention dose: 200-400 gr/ ton Treat.: 600-800gr/ ton, 1000 gr/tonTreat.: 600-800gr/ ton, 1000 gr/ton Pack.: 5 kgPack.: 5 kg
Chlortetracycline 20%Chlortetracycline 20%
Water soluble powderWater soluble powder 500 gr/200 lit, 3-5 d500 gr/200 lit, 3-5 d With.: 7dWith.: 7d Pack.: 500 grPack.: 500 gr
Oxytetracycline 20%Oxytetracycline 20%
Water soluble powderWater soluble powder Prev.: 30 gr/ 100 lit, 3 dPrev.: 30 gr/ 100 lit, 3 d Treat.: 60-120 gr/ 100 lit, 5 dTreat.: 60-120 gr/ 100 lit, 5 d Pack.: 500 grPack.: 500 gr
Oxytetracycline 10%Oxytetracycline 10%
Premix powderPremix powder Prevent.: 500-1000 gr/ tonPrevent.: 500-1000 gr/ ton Treat.: 1000-2500 gr/ tonTreat.: 1000-2500 gr/ ton Pack.: 5 kg Pack.: 5 kg With.: 7 dWith.: 7 d
Oxytetracycline 50%Oxytetracycline 50%
Premix powderPremix powder Prevent.: 100-200 gr/ tonPrevent.: 100-200 gr/ ton Treat: 200-300 gr/ tonTreat: 200-300 gr/ ton
Tetracycline HCl 20%Tetracycline HCl 20%
Water soluble powderWater soluble powder 500 gr/ 200 lit, 3-5d500 gr/ 200 lit, 3-5d Pack.: 500 grPack.: 500 gr
Colistin 3000000Colistin 3000000
3,000,000 Colistin sulphate/ gr. Water 3,000,000 Colistin sulphate/ gr. Water Soluble powderSoluble powder
Mech.: Polymixins bacteriocidals. Mech.: Polymixins bacteriocidals. Gram negative. Intestinal absorption↓Gram negative. Intestinal absorption↓
Water: 100 gr / 500 lit, 5-7dWater: 100 gr / 500 lit, 5-7d With.: 2 dWith.: 2 d Drug synergism: Potentiated Sulfa, Drug synergism: Potentiated Sulfa,
Tetracyclines, Chloramphenicle, Tetracyclines, Chloramphenicle, Erythromycin, Bacitracin, FlorfenicolErythromycin, Bacitracin, Florfenicol
Enrofloxacin 10%Enrofloxacin 10%
Oral SolutionOral Solution Mech.: Inhibition of DNA-gyrase Mech.: Inhibition of DNA-gyrase
activityactivity 1-1.5 lit / 1000 lit, 5 d1-1.5 lit / 1000 lit, 5 d Drug inter.: Chloram., Tetracyclines, Drug inter.: Chloram., Tetracyclines,
Macrolids, Al, Ca, Fe, Mg, Mn, Mineral Macrolids, Al, Ca, Fe, Mg, Mn, Mineral supplementsupplement
With.: 2 dWith.: 2 d Pack: 1 litPack: 1 lit
Flumequine 20%Flumequine 20%
Oral solutionOral solution 2w>: 25 ml/ 100 lit, 3-5 d2w>: 25 ml/ 100 lit, 3-5 d 2w<: 50 ml/100 lit, 3-5d2w<: 50 ml/100 lit, 3-5d Conta.: Layers at productionConta.: Layers at production Drug interaction: Sulfonamides, Drug interaction: Sulfonamides,
Trimet., Nitrofourans, CuSO4, Tetra.Trimet., Nitrofourans, CuSO4, Tetra. With.: 2-3dWith.: 2-3d Pack.: 1 litPack.: 1 lit
Florfenicol 10%Florfenicol 10%
Oral solutionOral solution 1 lit/ 1000 lit, 5 d1 lit/ 1000 lit, 5 d Contra.: Layers at productionContra.: Layers at production With.: 5 dWith.: 5 d Pack.: 1 litPack.: 1 lit
Lincomycin+SpectinomyLincomycin+Spectinomycincin
A Lincosamide + an AminoglycosideA Lincosamide + an Aminoglycoside 50 S + 30 S50 S + 30 S 150 gr/ 200 lit, 3 d150 gr/ 200 lit, 3 d Drug inter.: macrolides, Chloramph., Drug inter.: macrolides, Chloramph.,
Flor.Flor. With.: 2 dWith.: 2 d Pack.: 150 grPack.: 150 gr
Potentiated SulfaPotentiated Sulfa Methasol 48% (sultrim)Methasol 48% (sultrim) Sulfadiazin + TrimetoprimSulfadiazin + Trimetoprim Oral suspensionOral suspension Prevent.: 200 ml/ 800 lit, 3dPrevent.: 200 ml/ 800 lit, 3d Treat.: 200 ml/ 600 lit, 3-5 dTreat.: 200 ml/ 600 lit, 3-5 d Drug inter.: Vits B, Penicilins, Drug inter.: Vits B, Penicilins, Contra.: layers and breeders at Contra.: layers and breeders at
productionproduction With.: 5 dWith.: 5 d Pack.: 200 ml bottlePack.: 200 ml bottle
The endThe end