Immunization Of Broiler Chickens Against Necrotic Enteritis . - USDA
Immunization of broilerchickens against necroticenteritis: Progress andpossibilitiesJohn Prescott, Ravi Kulkarni
Necrotic enteritis Small intestinal infection of broiler chickens Caused by Clostridium perfringens Serious infection, through mortality and morbidity 6 billion dollar disease High cost of subclinical disease and of prevention byantibiotics Need to stop preventive use of medically-importantantibiotics
Immunization of broiler chickensagainst necrotic enteritis The challenges Can we immunize? What is the basis of immunity? What are the important antigens? What is an ideal vaccine? How can we deliver vaccine antigens? How should we test vaccines? Future possibilities
The challenges Complex disease Understanding the basis of protective immunity Immunizing in the face of maternally-derivedimmunity Identifying key antigen(s) Defining the best systems to test vaccines Safety, efficacy, robustness, cost Field versus lab testing
Major advance in NE research2008 Keyburn et al. identifieda novel toxin, NetB, thatplays a key role indevelopment of NE.5
FishmealProteinleakageCoccidiosisWholewheatHigh Cp –StarchNetB vestrainsGutstasisWet litterTpeLenhancesvirulenceWholewheatNetBMajor factors predisposing to NENE
NE strains are unique NE genetic loci highly conserved, stronglycorrelated with disease netB critical, but more complex than this Several chromosomal loci associated with netBpositive isolates
Lepp et al.,2010NELoc-2(11.2 kb)Extracellular solutebinding protein7- -hydroxysteroiddehydrogenaseResolvaseGlycosyl hydrolaseCotH proteinVTC domain superfamilytubulin/FtZ, GTPaseresolvaseVTC domain superfamilytubulin/FtZ, GTPaseSigma factor SgiINELoc-3(5.6 kb)(42 kb)Β lac domainMarR regulatorMGA ining proteintransposaseChitinase BChitinase ALeukocidin ILeukocidin IICAAX proteaserecombinasemembrane proteinmembrane proteinF5/8 type C domainphosphodiesterasesortasesurface proteinpeptidoglycan boundcell-wall proteindiguanylate cyclaseswim zinc fingerABC transporterSignal peptidase IIdentification of NE-specific lociNELoc-1Putative rPlasmidHypothetical protein8
Lepp et al.,2010NELoc-2(11.2 kb)Extracellular solutebinding protein7- olvaseGlycosyl hydrolaseCotH proteinVTC domain superfamilytubulin/FtZ, GTPaseresolvaseVTC domain superfamilytubulin/FtZ, GTPaseSigma factor SgiINELoc-3(5.6 kb)(42 kb)Β lac domainMarR regulatorMGA ining proteintransposaseChitinase BChitinase ALeukocidin ILeukocidin IICAAX proteaserecombinasemembrane proteinmembrane proteinF5/8 type C domainphosphodiesterasesortasesurface proteinpeptidoglycan boundcell-wall proteindiguanylate cyclaseswim zinc fingerABC transporterSignal peptidase INE locus 1: Mucin colonization,degradationNELoc-1Putative rPlasmidHypothetical protein9
Lepp et al.,2010NELoc-2(11.2 kb)Extracellular solutebinding protein7- -hydroxysteroiddehydrogenaseResolvaseGlycosyl hydrolaseCotH proteinVTC domain superfamilytubulin/FtZ, GTPaseresolvaseVTC domain superfamilytubulin/FtZ, GTPaseSigma factor SgiINELoc-3(5.6 kb)(42 kb)Β lac domainMarR regulatorMGA ining proteintransposaseChitinase BChitinase ALeukocidin ILeukocidin IICAAX proteaserecombinasemembrane proteinmembrane proteinF5/8 type C domainphosphodiesterasesortasesurface proteinpeptidoglycan boundcell-wall proteindiguanylate cyclaseswim zinc fingerABC transporterSignal peptidase INE locus 1: Tissue adhesionNELoc-1Putative rPlasmidHypothetical protein10
Lepp et al.,2010NELoc-2(11.2 kb)Extracellular solutebinding protein7- -hydroxysteroiddehydrogenaseResolvaseGlycosyl hydrolaseCotH proteinVTC domain superfamilytubulin/FtZ, GTPaseresolvaseVTC domain superfamilytubulin/FtZ, GTPaseSigma factor SgiINELoc-3(5.6 kb)(42 kb)Β lac domainMarR regulatorMGA ining proteintransposaseChitinase BChitinase ALeukocidin ILeukocidin IICAAX proteaserecombinasemembrane proteinmembrane proteinF5/8 type C domainphosphodiesterasesortasesurface proteinpeptidoglycan boundcell-wall proteindiguanylate cyclaseswim zinc fingerABC transporterSignal peptidase INetB toxin damageNELoc-1Putative rPlasmidHypothetical protein11
Can we immunize?
Passive immunity: NetVax vaccine for layersprovides maternal antibody for broilers“Toxoid A”:supernatantantigensMortality in 2228-day-oldbroilers
Passive immunization rNetBversus “toxoid” (Keyburn et al., 2013)Day 21 chicksDay 14 chicks
Key findings from passiveimmunization Yes, you can immunize Immunization with “PlcC [alpha-toxoid]” (netBnegative) gives good but incomplete protection rNetB plus “toxoid” rNetB alone older chicks Useful strategy but not in later broiler production Need active immunization
Can we activelyimmunize?
Active immunization against C.perfringens in NE: Key lab findings Secreted proteins crucial Supernatants vary in protective ability Several different antigens provide reasonableprotection Intestinal mucosal IgY IgA important Protection depends on challenge severity; system? Mixed antigens or chimeric proteins often betterprotection than individual antigens
What is the basis ofimmunity?
What is the basis of immunity? Not understood in detail, Th2 and Th17 cytokinemediated Antibodies to secreted virulence factors (PlcC,NetB, zinc metalloprotease) important Antibodies to secreted housekeeping(“moonlighting?”) proteins important Many different antigens provide some protectionexperimentally Effect may be by impairing bacterial growth ratherthan neutralizing toxin (IgY)
Defined antigens with value inimmunization against NEAntigenRoleValue*Reproducibility**Alpha toxin (PlcC)Virulence? Phospholipase . NetB toxoidVirulence . Zinc metalloproteaseVirulence? Mucin degradation . Fructose biphosphate Housekeeping-Moonlighting?aldolase, FBAAdhesion? . PilusVirulence (collagen adhesion) Pyruvate ferrodoxin oxidoreducatase,PFORHousekeeping Glyceraldehyde-3-phosphateoxidoreductase, GPDHousekeeping . Phosphoglyceromutase, PGMHousekeeping . Elongation factor-TuHousekeeping Endo-β-N-acetylgluosaminidaseHousekeeping0 . *Overallprotection, 25-49%, , 50-74%, , 75%; **, , 4 studies; , 3 studies,
NE lesion scores in chickens immunized IM with CPproteins and different challenge severityPlcCSevere challengeKulkarni et al.,2007
Vaccination with recombinantNetB toxoid (Keyburn et al., 2013)
Protection of broiler chickens against NEafter SC immunization with PlcC-NetBchimeric toxoidHunter et al., 2019a
What is an ideal vaccine? Safe, effective, cost-effective, profitable Easily administered: in ovo or in drinking water Robust under field conditions Local intestinal immunity important, so best iforally administered 100% protection
How can we deliverimmunization?
Live attenuated oralSalmonella vectoredvaccines
Salmonella vaccine vectors“Regulated expression” of foreign antigens; “delayedattenuation”, so virulent at time of infection;programmed lysis so disappear{BglII/BamHI} 0EcoRV 2656NdeI 2345PPtrctrc {NcoI/BspHI} 56MCSMCbla SSS5ST1T25ST1T2asd pYA3493asdpYA33422890 bp3.0kbEcoRV 1926XbaI 1789pBR oripBR oriBglII 659Improved plasmid vectorsexpressing CP antigens
Different Salmonella vectors expressingPlcC have different efficacy****Intestinal lesion scores of birds immunized three times, aged 1, 7 and 14 days, withSalmonella vaccine vectors (χ3987, χ9241, χ9853, χ9945, χ11442, χ11445, vectoronly control group (χ3987), expressing the C. perfringens PlcC, and challenged atday 28 with C. perfringens CP4.Prescott et al., unpublished
Impact of immunization with Salmonellavectored PlcC and/or NetB toxoids onpercentage of broiler chickens with severelesionsJiang et al., 2015
Mean intestinal lesion scores in broilersimmunized with single Salmonella vaccineexpressing different antigensGroupExperiment 1Experiment cC-fba-netB2.53.13.04.01.5*0.8**1.9*0.9**fba-0.7** Hunter et al., 2019b
Live attenuated C. perfringensmutants for oral vaccination? No.MutantMean SD (% control)Number studiesPhosphodiesterase190 603netB95 313Diguanylate cyclase901CP1-3475 ABC transporter 931Zinc metalloprotease1Prescott et al., unpublished108
Future possibilities
Potential avirulent live oral vaccinevectors for necrotic enteritisVectorAntigenMouse protectionAuthorLactobacillus caseiPLC (alpha toxoid) Gao, 2019Lactobacillus caseiPLC Alimolaei, 2017Lactobacillus caseiPLC Song, 2018Bacillus subtilissporesPLC Hoang, 2008
in ovo immunization? One study of efficacy Eimeria profilin and rNetB Partial protection against experimental NE Adjuvant important Far more work needed
How should we testvaccines experimentally?
Reproducing necrotic enteritis experimentallyNetB(critical)TpeL(for enhanced disease)Age of CP brothcultureCP inoculationconditions: numbers,method of deliveryCoccidial infection:virulent, vaccineInductionof NEChange to high animalprotein dietat challengeShojadoost et al., 2011Immunosuppression:IBD vaccineWheat - barley- based diet
Assessment of vaccine efficacy:Important issues No standard model to reproduce NE: Does Eimeriabias results? No challenge strain(s) criteria No standard assessment system No standard challenge severity No “gold standard” vaccine comparison6 point NE scoring system:Keyburn et al., 2008
The future Immunization has promise in reducing NE Very useful adjunct in control Oral attenuated Salmonella with “mixed” antigenspromising Need for field testing attenuated Salmonellavaccines Explore other avirulent vaccine vectors in ovo immunization plus oral vaccine boost?
Acknowledgements Canadian Poultry Research Council-Agriculture and Ari-FoodCanada, Poultry Industry Council, OMAFRA Roy Curtiss, Ken Roland, Arizona State University
Passive immunity: NetVax vaccine for layers provides maternal antibody for broilers Mortality in 22-28-day-old broilers "Toxoid A": supernatant antigens . Passive immunization rNetB . Active immunization against C. perfringens in NE: Key lab findings Secreted proteins crucial
Feedmaster Classic Broiler Range Ideal for the smaller producer who sells live broilers to the market. It is a cost-effective range with a moderate growth rate. 35 Day Model Number Of Chicks Classic Broiler Starter Classic Broiler Grower Classic Broiler Finisher Total Feed Day-old- 13.5 Days 500 g 13.5 Days - 3 Days before slaughter 3 Days .
and Influenza Immunization Summit (NAIIS) is pleased to announce the recipients of the 2016 Immunization Excellence Awards. There are six categories of recognition: overall influenza season activities, influencer, "immunization neighborhood" champion, corpo-rate campaign, adult immunization champion, and adult immunization publication award.
The average annual producer price for broiler was R15.46/kg in 2013, which was R0.48c higher than 2012. The decline in 2010 might be due oversupply of broiler meat responding to the FIFA world cup held in South Africa and the low economic growth. Generally the price of broiler meat increased significantly mainly due to the increased input costs.
immunization against α-syn can occur in one of two forms, active or passive immunity [18]. Active immunization involves stimulating the immune system to produce anti-bodies against toxic α-syn conformations, while passive immunization involves administering anti- α-syn antibodies to the patient, which confers temporary protection against the .
Production segments involved in the production and processing of broilers 1. Broiler breeders 2. Hatchery 3. Grow-out farms a) Grow-out farms are where broiler chickens are raised. b) Grow-out houses are specialized buildings that allow age-appropriate control of temperature, humidity, food
Broiler diets are formulated to provide the energy and nutrients essential for health and efficient broiler production. The basic nutritional components required by the birds are water, amino acids, energy, vitamins and minerals. Feedmaster provides the following: All our products are specifically formulated to achieve the desired
In Zambia there are both small-scale broiler farmers, who keep a few birds only for their own consumption, and large-scale industrialized broiler farmers, who keep several thousands of chickens at a time, and all sizes of farms in between. Light and its availability are very important in poultry production, especially for broilers. The
Pendidikan Akuntansi FKIP Universitas Sebelas Maret. Penetapan profil dan learning outcome ini dimaksudkan untuk membantu pemerintah dalam menyiapkan guru akuntansi yang bermutu menurut persepsi mahasiswa, alumni, dosen, pengguna lulusan, Asosiasi Profesi, dan pengambil keputusan. Sumber data penelitian ini adalah 96 orang mahasiswa, 248 orang alumni, 15 orang dosen, 15 orang pengguna lulusan .
