IP Popa v 2014 - Variola aviara. Diagnostic si control prin vaccinare

8/12/2019 IP Popa v 2014 - Variola aviara. Diagnostic si control prin vaccinare

1/21

VARIOLA AVIARAVARIOLA AVIARADIAGNOSTIC SI CONTROLDIAGNOSTIC SI CONTROL

PRIN VACCINAREPRIN VACCINARE

Virgilia PopaVirgilia Popa11,, Mirela PopaMirela Popa11, Miliana Petrof, Miliana Petrof11,, Daniela BotusDaniela Botus11, Marian Culcescu, Marian Culcescu11,,

Costin RacoltaCostin Racolta11,, Jenica BucurJenica Bucur22,, Gheorghe StratulatGheorghe Stratulat11

11SN Institutul Pasteur SA,SN Institutul Pasteur SA, 22SC Farmavet SASC Farmavet SA

INTALNIREA TRIMESTRIALA AINTALNIREA TRIMESTRIALA ASOCIETATII MEDICILOR VETERINARI IN PATOLOGIA AVIARASOCIETATII MEDICILOR VETERINARI IN PATOLOGIA AVIARA

SI A ANIMALELOR MICI DIN ROMANIASI A ANIMALELOR MICI DIN ROMANIA

ACADEMIA DE STIINTE AGRICOLE SI SILVICEACADEMIA DE STIINTE AGRICOLE SI SILVICE GHEORGHE IONESCU SISESTIGHEORGHE IONESCU SISESTI

AULA MAGNAAULA MAGNABucuBucurrestiesti 2727 martiemartie 20120144


2/21

V RIOL VI RRIOL VI RManual OIEManual OIE

httphttp

:://www.oie.int/international//www.oie.int/international

--standardstandard

--setting/terrestrialsetting/terrestrial

--manual/accessmanual/access

--

onlineonline// (B307)(B307)

Variola aviara este o boala virala prezenta atat la pasari domesVariola aviara este o boala virala prezenta atat la pasari domestice cat si salbatice (pui, curci,tice cat si salbatice (pui, curci,porumbei, canari, peste 200 de specii aviare)porumbei, canari, peste 200 de specii aviare)

Frecventa preponderent in sezoanele umede si reciFrecventa preponderent in sezoanele umede si reci

19021902 Mark & Sticker; 1908Mark & Sticker; 1908 Cornwarth; 1920Cornwarth; 1920 primele vaccinuriprimele vaccinuri

Boala se transmite lent si este caracterizata printrBoala se transmite lent si este caracterizata printr--oo forma cutanata (forma cutanata (dry poxdry pox / forma variolica/ forma variolica

sau exantematoasa )sau exantematoasa ) si / sau osi / sau o forma difterica (forma difterica (wet pox / forma enantematoasawet pox / forma enantematoasa )) la nivelulla nivelulsuperior al aparatului digestiv si al celui respiratorsuperior al aparatului digestiv si al celui respirator

Pierderi prin mortalitate: 3Pierderi prin mortalitate: 3--5%5% -- 3030--50%; scaderea productiei de oua; intarziere in dezvoltare la50%; scaderea productiei de oua; intarziere in dezvoltare latinerettineret

http://www.hyline.com/aspx/redbook/redbook.aspx?s=5&p=35http://www.hyline.com/aspx/redbook/redbook.aspx?s=5&p=35


3/21

V RIOL VI RRIOL VI R

http://www.cabi.org/isc/?compid=5&dsid=104014&loadmodule=datashehttp://www.cabi.org/isc/?compid=5&dsid=104014&loadmodule=datasheet&page=481&site=144#et&page=481&site=144#


4/21


http://www.oie.int/wahis_2/public/wahid.php/Countryinformation/Ahttp://www.oie.int/wahis_2/public/wahid.php/Countryinformation/Animalsituationnimalsituation


5/21


http://www.oie.int/en/animalhttp://www.oie.int/en/animal--healthhealth--inin--thethe--world/oieworld/oie--listedlisted--diseasesdiseases--2014/2014/


6/21


Cociu M, Wagner G, Micu N, Tuschak E, Mihaescu G, 1972. Avian poCociu M, Wagner G, Micu N, Tuschak E, Mihaescu G, 1972. Avian pox in a bustardx in a bustard

(Otis tarda(Otis tarda// dropiedropie). (Gefluegelpocken bei einer trappe (Otis tarda).) In: Diseases). (Gefluegelpocken bei einer trappe (Otis tarda).) In: Diseases ofof

Zoo Animals, 14th International Symposium, Wroclaw, Poland. 81Zoo Animals, 14th International Symposium, Wroclaw, Poland. 81--8383


7/21


8/21

VARIOLA AVIARAVARIOLA AVIARA

Cercetari in RomaniaCercetari in Romania

AniiAnii 7070 :: Program national de cercetareProgram national de cercetare

ca urmare a unor episoade grave de boalaca urmare a unor episoade grave de boala

-- sselectia tulpinilor vaccinaleelectia tulpinilor vaccinale-- sselectia schemei de imunizareelectia schemei de imunizare

In 1973In 1973--74 a izbucnit o DV la popula74 a izbucnit o DV la populaie, avicole, fazaneriiie, avicole, fazanerii i la poti la potrnicheria de la Drvarnicheria de la Drvai (TR)i (TR)

unde nu sunde nu s--aa tiut ce tulpinevolueaztiut ce tulpinevolueaz. A fost introdus. A fost introdus n planul prioritar . .de cercetare nan planul prioritar . .de cercetare naional,ional,

care era financare era finanat de M Agriculturii.Sat de M Agriculturii.S--a lucrat cu tulpina COLa lucrat cu tulpina COL i GAL dari GAL dar i cu tulpina Moscovai cu tulpina Moscova

i.... vaccinuri de pe piai.... vaccinuri de pe pia. Temele au durat c. Temele au durat civa aniiva ani.), p.), perioaderioadn care vaccinarea a fostn care vaccinarea a fost

obligatorie.obligatorie.

Dupce apele sDupce apele s--au liniau linitittit n 1980n 1980--82 s82 s--a renuna renunat la folosirea celor doutulpiniat la folosirea celor doutulpini i si s--a adoptata adoptat

programul cu o singurvaccinareprogramul cu o singurvaccinare..

Cea mai spectaculoas

DV a fost la Avicola Bac

uCea mai spectaculoas

DV a fost la Avicola Bac

u --pe fond nevaccinatpe fond nevaccinat n 1988 (noi.n 1988 (noi.-- dec.)dec.)....


9/21

V RIOL VI RRIOL VI Rhttp://www.virology.net/big_virology/BVDNApox.htmlhttp://www.virology.net/big_virology/BVDNApox.html

Virus ADN; genom: 288 kbpVirus ADN; genom: 288 kbp// > 250 gene> 250 gene

PoxviridaePoxviridae

Subfamily ChondropoxvirinaeSubfamily Chondropoxvirinae

OrthopoxvirusOrthopoxvirus AvipoxvirusAvipoxvirus

CapripoxvirusCapripoxvirus

LeporipoxvirusLeporipoxvirus

Parapoxvirus (swinepox, orf )Parapoxvirus (swinepox, orf ) MolluscipoxvirusMolluscipoxvirus

YatapoxvirusYatapoxvirus Subfamily EntomopoxvirinaeSubfamily Entomopoxvirinae

Unclassified virusesUnclassified viruses

FowlpoxFowlpox

CanarypoxCanarypox

PigeonpoxPigeonpox

etcetc

GenotipuriGenotipuri

NANA

-- europeaneuropean


10/21

V RIOL VI RRIOL VI RV CCINURI VII MONOV LENTE CCINURI VII MONOV LENTE

http://www.poultrymed.comhttp://www.poultrymed.com


11/21

V RIOL VI RRIOL VI RV CCINURI VII MONOV LENTE CCINURI VII MONOV LENTE

Nr. crt. Tara Firma Tulpina Nr. crt. Tara Firma Tulpina

1 Africa de Sud Onderstepoort 31 Japonia Sinsui

2 Argentina Boehringer Ingelheim Beaudette 32 Japonia Nisseken Beaudette

3 Argentina Rosenbusch 33 Japonia Nisseken Beaudette

4 Australia Intervet FPV-085a 34 Korea Dae Sung

5 Australia LAH 35 Korea de Sud KBNP 2775

6 Brazilia Merial 36 Korea de Sud Cavac

7 Brazilia Merial 37 Mexic Boehringer Ingelheim Beaudette

8 Brazilia Biovet 38 Mexic Bio-Zoo9 Brazilia Biovet 39 Olanda Intervet WP

10 Brazilia Hertape Calier 40 Romania Pasteur Col-OF

11 Brazilia Fort Dodge 41 Romania Pasteur GAL-MP1

12 China Qingdao Yebio Bioengineering Quail - adapted 42 Romania Romvac Col-81 pigeon

13 Croatia Veterina 43 Romania Romvac MP1

14 Franta Merial DCEP 25 44 Rusia Avivac

15 Franta Laprovet 45 Serbia Vet. Medicine Inst.16 Germania LAH HP-B 46 Spania Hipra

17 India

Inst. of Anim. Health and Vet.

Biologicals 47 SUA Intervet

18 India Hester 48 SUA Merial Select

19 India Indovax 49 SUA Merial Select

20 India Ventri 50 SUA SPAH (Shering Plough)

21 India Bio Med BM 51 SUA SPAH (Shering Plough)

22 Indonesia Medion M-92 52 SUA LAHI Pigeon Pox Virus

23 Indonesia Vaksindo Satwa Nusantara 53 SUA LAHI

24 Indonesia Vaksindo Beaudette 54 SUA Zoetis

25 Iordania Jovac HP-2 55 SUA Zoetis Chicken

26 Iran

Razi Vaccin and Serum

Research Inst. 56 SUA Ceva Cutter

27 Israel Biovac 57 Venezuela Cala

28 Israel Abic 58 Vietnam Navetco Weybridge

29 Italia Fatro

30 Italia Iso Brescia

http://www.poultrymed.comhttp://www.poultrymed.com


12/21

V RIOL VI RRIOL VI RV CCINURI VII MONOV LENTECCINURI VII MONOV LENTE

IN ROM NIN ROM NI

Nr. crt. Firma Tulpina1 ABIC Biological Laboratories TEVA Lts Komarov

2 Ceva Phylaxia Veterinary Biologicals Co. Ltd Cutter

3 Intervet International BV Gibbs

4 Intervet International BV WP

5 Laboratorios Hipra SA FPV 92

6 Lohmann Anmal Health GmbH HP-B

7 Merial DCEP 25

8 Pasteur Col-OF

9 Pasteur Gal-MP1

10 Pasteur IP72 pigeon

11 Romvac Col-81 pigeon

12 Romvac Gal-MP1

http://www.icbmv.ro/ro/nomenclatorhttp://www.icbmv.ro/ro/nomenclator--produseproduse


13/21


http://partnersah.vet.cornell.edu/avianhttp://partnersah.vet.cornell.edu/avian--atlas/search/lesion/417atlas/search/lesion/417


14/21

V RIOL VI RRIOL VI RIncluzii intraIncluzii intra -- citoplasmaticecitoplasmatice

(corpusculi)(corpusculi) BollingerBollinger

http://www.museum.osakafuhttp://www.museum.osakafu--u.ac.jp/html/en/material/detail.php?id=1362u.ac.jp/html/en/material/detail.php?id=1362


15/21

TESTE PCR FPVESTE PCR FPVMetoda rPCR cu SybrGreenMetoda rPCR cu SybrGreen

GelGel--electroforeza de controlelectroforeza de control

postpost--amplificare a testului deamplificare a testului de

amplificare in timp real pentruamplificare in timp real pentru

virusul difterovirusul diftero--varioleivarioleiaviare.Linia 1: martor negativaviare.Linia 1: martor negativ

(NFW); linia 2: 465(NFW); linia 2: 465--nodul; linianodul; linia

3: 4663: 466 nodul; linia 4: 465nodul; linia 4: 465

proba comasata tampon creierproba comasata tampon creier

+ tampon nazo+ tampon nazo--faringian; liniafaringian; linia

5: 4665: 466 proba comasataproba comasata

tampon creier + tampon nazotampon creier + tampon nazo--

faringian; linia 6: proba vaccin,faringian; linia 6: proba vaccin,

dilutie 1/10; linia 7: probadilutie 1/10; linia 7: proba

vaccin, dilutie integrala; linia 8:vaccin, dilutie integrala; linia 8:

standard ADNstandard ADN PCR MarkerPCR Marker

(Sigma); linia 9: Martor pozitiv(Sigma); linia 9: Martor pozitiv

Col; linia 10: Martor pozitivCol; linia 10: Martor pozitivGal.Gal.

ExtracExtracie ADN: cu trusa QIAamp DNA Mini Kit (Qiagenie ADN: cu trusa QIAamp DNA Mini Kit (Qiagen -- 51304).51304).

PCR VDV pentru genaPCR VDV pentru gena dTKdTK : amorse PopaV 2004; amestec de: amorse PopaV 2004; amestec de

amplificare 2x Brilliant II Sybr Green qPCR Master Mix, Agilent;amplificare 2x Brilliant II Sybr Green qPCR Master Mix, Agilent;

amplificator spectrofotometric Mx3005P, Stratagene.amplificator spectrofotometric Mx3005P, Stratagene.

Martor pozitiv: tulpina Gal (Instittutul Pasteur); tulpina ColMartor pozitiv: tulpina Gal (Instittutul Pasteur); tulpina Col(Institutul Pasteur).(Institutul Pasteur).

Martor negativ: apa ultrapura PCR sterila (Promega)Martor negativ: apa ultrapura PCR sterila (Promega)


16/21

TESTE PCRESTE PCR

http://www.ncbi.nlm.nih.gov/BLAST/Blast.cgihttp://www.ncbi.nlm.nih.gov/BLAST/Blast.cgi

http://www.ncbi.nlm.nih.gov/tools/primerhttp://www.ncbi.nlm.nih.gov/tools/primer--blast/primertool.cgi?ctg_time=1395403164&job_key=emGRjQaHblast/primertool.cgi?ctg_time=1395403164&job_key=emGRjQaH

r6OQjpSC9aym_O6BlO39nonor6OQjpSC9aym_O6BlO39nono


17/21

TESTE PCR FPVESTE PCR FPV

PCR core P4b FPVPCR core P4b FPV--like. Kitul folosit: High Fidelitylike. Kitul folosit: High FidelityPCR Master/Roche. Produsul PCR: 578bpPCR Master/Roche. Produsul PCR: 578bp

L,marker de greutate molecularaL,marker de greutate moleculara--50bp DNA50bp DNA

ladder/Fermentas; 1ladder/Fermentas; 1-- sample 7425;2sample 7425;2-- sample 7425;sample 7425;

33-- Vaccin comercial; 4Vaccin comercial; 4-- TL Col; 5TL Col; 5-- TL GAL; 6TL GAL; 6--

Pigeon;7Pigeon;7-- marker negativmarker negativ

core4b FPVlike : 578 bpfpv 241 ext:fpv 241 ext:

-- 1000 bp1000 bp-- origine nordorigine nord--americana;americana;

-- 2200 bp2200 bp-- origine europeanaorigine europeana

PCR gena fpv241ext. Kitul folosit: High Fidelity PCR Master/RochPCR gena fpv241ext. Kitul folosit: High Fidelity PCR Master/Roche.e.

Produsii PCR: 2200bp/FPV si CNPV European; 1000bp/FPV si CNPVProdusii PCR: 2200bp/FPV si CNPV European; 1000bp/FPV si CNPV

American L,marker de greutate molecularaAmerican L,marker de greutate moleculara--200bp DNA200bp DNA

ladder/Fermentas; 1ladder/Fermentas; 1-- sample 7425;2sample 7425;2-- sample 7425;sample 7425; 33-- VaccinVaccin

comercialcomercial; 4; 4-- TL Col; 5TL Col; 5-- TL GAL; 6TL GAL; 6-- Pigeon;7Pigeon;7-- marker negativmarker negativ


18/21

TESTE PCR FPVESTE PCR FPVfpv 241 ext:fpv 241 ext:

-- 1000 bp1000 bp-- origine nordorigine nord -- americana;americana;

-- 2200 bp2200 bp-- origine europeanaorigine europeana

Marker de greutate molecularaMarker de greutate moleculara--200bp DNA200bp DNA

ladder/Fermentas; 1ladder/Fermentas; 1-- 465 nodul; 2465 nodul; 2-- 466 nodul; 3466 nodul; 3-- 465 tp465 tp

(cr+far.); 4(cr+far.); 4-- 466 tp (cr+far.) 5466 tp (cr+far.) 5-- Vaccin comercial; 6Vaccin comercial; 6-- TLTL

COL; 7COL; 7-- TL GAL; 8TL GAL; 8-- marker negativmarker negativ


19/21

ConcluziConcluzi ii

Testele PCR, insotite de analize prin restrictie enzimatica (RE)Testele PCR, insotite de analize prin restrictie enzimatica (RE),, ss--auau

dovedit fiabile si adecvate in diagnosticul variolei aviare, semdovedit fiabile si adecvate in diagnosticul variolei aviare, semnalandnaland

prezenta tulpinilor salbatice virale de gaina (Fowlpox),prezenta tulpinilor salbatice virale de gaina (Fowlpox), dede genotipgenotipeuropeaneuropean la pasari crescute in Romaniala pasari crescute in Romania

Selectia vaccinurilor utilizate pentru profilaxia specifica trebSelectia vaccinurilor utilizate pentru profilaxia specifica treb

uie sa tinauie sa tina

cont de acest faptcont de acest fapt

Prezenta tulpinilor salbatice impune adoptarea schemei de vaccinPrezenta tulpinilor salbatice impune adoptarea schemei de vaccinareare

cu doua administrari de vaccin, cu tulpini virale atenuate, adeccu doua administrari de vaccin, cu tulpini virale atenuate, adecvatevate

varstei pasarilor, si structuri genetice si antigenice care sa avarstei pasarilor, si structuri genetice si antigenice care sa acoperecopere

profilurile tulpinilor din teren si deci sa induca protectie fatprofilurile tulpinilor din teren si deci sa induca protectie fata de acesteaa de acestea

((schemaschema clasicaclasica pentru Romania,pentru Romania, Col si GalCol si Gal).).


20/21

Multumiri medicilor veterinari si fermierilor colaboratori

Multumiri Dnei Conf. Dr. Eugenia Avram, FMV Spiru Haret / Cercetator stiintific I, I.Pasteur

Multumiri Dlui Conf. Dr. Mihai Danes, USAMV Bucuresti / Cercetator stiintific I, I.Pasteur

Amorsele testului rPCR au fost designate in cadrul

Ctr. MEC Biotech 03-PP-456 / 2003-2006

Testele PCR FPV au fost dezvoltate in cadrul programului CD al I.Pasteur


21/21

Documents

IP Popa v 2014 - Variola aviara. Diagnostic si control prin vaccinare