Slide 1

UNIVERZITET U SARAJEVU VETERINARSKI FAKULTET

ZAVOD ZA ZOOTEHNIKU I PERADARSTVO

Šef zavoda: Prof. dr Abdulah Gagić

METODE UZORKOVANJA

– Goletić Teufik, DVM, PhD –

BOLESTI PERADI

SARAJEVO, 2010.

________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
____________________________________________________________________
Slide 2

UNIVERZITET U SARAJEVU VETERINARSKI FAKULTET

METODE UZORKOVANJA

Kolekcija uzoraka – polazna tačka laboratorijskog istraživanja

bolesti životinja.
Svrha

dijagnoza bolesti,
“surveillance” bolesti životinja,
zdravstvena certifikacija,
monitoring odgovora na tretman ili vakcinaciju.

Uzeti uzorci trebaju biti:

5 odgovarajuci,
5 adekvatni u broju i količini,
5 brižljivo upakovani, označeni i
5 dostavljeni u lab na siguran (po okolinu i uzorke) i najbrži
način.

________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
____________________________________________________________________
Slide 3

UNIVERZITET U SARAJEVU VETERINARSKI FAKULTET

POPRATNI AKT

Svi uzorci trebaju biti poprać

popraćeni pismom (POPRATNIM AKTOM) ili
odgovarajuć
odgovarajućim obrascem,
obrascem, koji uključ
uključuje:
7 ime i adresu vlasnika/drž
vlasnika/držaoca kod čije peradi se bolest pojavila,
pojavila,
sa brojem telefona i/ili fax-
fax-a.
7 ime,
ime, poš
poštanski broj i/ili e-mail adre
adres
su, telefon i broj fax-
fax-a poš
pošiljaoca
7 relevantnu anamnezu/historiju bolesti,
bolesti, (treba je staviti u plastič
plastični
omot/foliju izvan transportnog kontejnera)
7 vrsta,
vrsta, provenijenca,
provenijenca, spol,
spol, dob, broj objekta i kod ukrasnih ptica
identitet (oznaka) ptice od koje uzorak potječ
potječe
7 bolest na koju se sumnja i potrebne pretrage
7 lista
ista uzoraka i koriš
korišteni transportni medij (ako je koriš
korišten)

Korištene hemijske olovke ili flomasteri moraju biti otporni na

uvjete pakovanja i transporta, i.e. na vlagu ili smrzavanje.

________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
____________________________________________________________________
Slide 4

UNIVERZITET U SARAJEVU VETERINARSKI FAKULTET

KOLEKCIJA UZORAKA

OD ŽIVIH PTICA
4 Krv, plazmu ili serum

3 treba uzorkovati po moguć

mogućnosti aseptič
aseptično,
no, punkcijom vene
ili srca
3 sa antikoagulansom (EDTA,
EDTA, heparin...) za direktno ispitivanje

kauzalnog agensa (paž

pažljivo promiješ
promiješati što prije),
3 bez antik
antikoagulansa sample),
oagulansa (za serologiju, clotted sample),
3 serumi se mogu čuvati do 7 dana na 4oC, za dužduži period?
4 Feces
3 uzorkovati najmanje 1 g svjež
svježeg fecesa.
fecesa. Alternativni a
ponekad i pož
poželjniji metod je uzimanje briseva kloake,
3 feces je najbolje čuvati i transportovati
transportovati na 4°C.

________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
____________________________________________________________________
Slide 5

UNIVERZITET U SARAJEVU VETERINARSKI FAKULTET

UZORCI OD ŽIVIH PTICA - NASTAVAK

4 Kož
Koža i perje

3 ako je moguć
moguće, uzeti 1 g promijenjenog epitelij
epitelijalnog
alnog tkiva (duboke
strugotine kož
kože),
3 sadrž
sadržaj vezikula (fluid), ukoliko ih ima, treba uzorkov
uzorkovati iz ne
rupturiranih vezikula,
3 folikuli pera (Marekova
(Marekova bolest).
bolest).

4 Jaja

3 najmanje 10 jaja po jatu ptica (preporuč

(preporučljivo 30 jaja po jatu),
3 jaja je najbolje čuvati na sobnoj temperaturi (18 – 22 oC).
C).

________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
____________________________________________________________________
Slide 6

UNIVERZITET U SARAJEVU VETERINARSKI FAKULTET

OD UGINULIH PTICA

Tkivo (općenito)

pluća mozak slezena

trahea bubrezi jetra

zračne vreće pankreas

proventriculus, intestinum, cekalne tonzile ..

________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
____________________________________________________________________
Slide 7

UNIVERZITET U SARAJEVU VETERINARSKI FAKULTET

"AMBIJENTALNI" I UZORCI HRANE

4 Razlozi uzorkovanja
3 za monitoring
monitoring higij
gijene,
ene, ili
3 kao dio dijagnoze i nadzora bolesti .

4 "Ambijentalni" uzorci se obič

obično uzimaju sa
5 poda ili prostirke,
5 površ
površine ventilacionih otvora, i
5 sistema za hranjenje i napajanje .
Navedene vrste uzoraka su posebno važne za kontrolu valionica i
klaonica
5 hrana (iz hranilica)
5 voda (iz pojilica,
pojilica rezervoara, te prirodnih i vješ
vještač
tačkih izvora
snabdijevanja

________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
____________________________________________________________________
Slide 8

UNIVERZITET U SARAJEVU VETERINARSKI FAKULTET

VELIČINA UZORKA

4 Za klinič
klinički manifestnu bolest uzeti uzorci moraju biti
representativni

4 Za razvijanje "surveillance"
surveillance" programa,
programa, te za monitoringa
monitoringa zdravlja
ptica koriste se opć
opći statistič
statistički metodi
metodi uzorkovanja. Za navedene
svrhe mož
može se koristiti i program FreeCalc raspolož
raspoloživ na:
na:

http://www.ausvet.com.au/content.php?page
http://www.ausvet.com.au/content.php?page=
=res_software#freecalc
Calc

________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
____________________________________________________________________
Slide 9

UNIVERZITET U SARAJEVU VETERINARSKI FAKULTET

SALMONELOZA

OIE - Salmonellosis (salmoneloza) je infektivna bolest ljudi i životinja

uzrokovana mikroorganizmima dva specijesa roda Salmonella -
Salmonella enterica and S. bongori.
bongori.

________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
____________________________________________________________________
Slide 10

UNIVERZITET U SARAJEVU VETERINARSKI FAKULTET

7 Zakonska osnova
Direktiva vijeća EU 92/117/EEC (razmatra protektivne mjere protiv određenih zoonoza
i zoonotskih agenasa)

7 uzorke za bakteriološke pretrage prikupljati po mogućnosti aseptično prije bilo kakvog

tretmana (antibiotskog),
7 po mogućnosti uzorkovanje izvršiti tokom akutne faze bolesti ili odmah nakon uginuća
ptice,
7 kod intenzivno držane peradi “ambijentalni“ uzorci (prostirka, prašina, brisevi površine
poda, feces), sa ekonomskog aspekta mogu biti najjeftiniji i adekvatni za identifikaciju
inficiranih jata,
7 u lab dostaviti ako je moguće reprezentativan broj bolesnih ili “svježe“uginulih ptica

7 izbjeći unakrsnu kontaminaciju uzoraka u transportu do laboratorija

________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
____________________________________________________________________
Slide 11

UNIVERZITET U SARAJEVU VETERINARSKI FAKULTET

7 Kompletan uzorak mora sadrzavati:

5 1/d pilići: podlošci transportnih kutija, pilići uginuli u transportu,

5 pilići dobi 4 sedmice (brojleri) ili 2 sedmice prije prebacivanja u

eksploatacione objekte (neslice): skupni uzorak fecesa "pripremljen"
od pojedinačnih uzoraka "svježeg " mase ne manje od 1 g,
5 Roditelska jata: uzorke uzimati najmanje svake druge sedmice tokom
perioda nesenja:

3 skupni uzorak fecesa "pripremljen" od pojedinačnih uzraka "svježeg"

fecesa (mase ne manje od 1 g),
3 iz valionice – skupni uzorak mekonijuma (≈ od 250 pilića), po roditeljskom
jatu, valionički ostatak od 50 jaja (unutar čije ljuske se nalazi uginuli embrij)
ili 50 leševa 1/d pilića po roditeljskom jatu.

________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
____________________________________________________________________
Slide 12

UNIVERZITET U SARAJEVU VETERINARSKI FAKULTET

AVIJARNA INFLUENCA

Zakonska osnova
Direktiva Vijeća EU 92/40/EC i 2005/94/EC

Avijarna influenca predstavlja infekciju peradi uzrokovanu bilo

kojim influenca A virusom čiji je IVPI kod pilića dobi šest sedmica
veći od 1.2 ili infekciju sa influenca A virusima subtipa H5 ili H7
koji posjeduju specifičan aminokiselinski slijed na mjestu
cijepanja hemaglutinina [HA]

________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
____________________________________________________________________
Slide 13

UNIVERZITET U SARAJEVU VETERINARSKI FAKULTET

Uzorci

⌧ od živih ptica – trahealni i kloakalni brisevi ili ,kao alternativa,

odgovarajući uzorak svježeg fecesa.

⌧ od uginulih ptica - intestinalni sadržaj (feces) ili kloakalne i

oro-faringealne briseve, uzorke tkiva pluća, traheje, zračnih vreća,
crijeva, slezene, bubrega, mozga, jetre i srca (pojedinačno ili kao
"pool“).
⌧ uzorci krvi:
od svih ptica (jata sa < od 20 ptica), i
od 20 ptica (veća jata),

⌧ uzorci krvi trebaju "odstajati“ radi koagulacije i izdvajanja

SERUMA KOJEGA TESTIRAMO

________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
____________________________________________________________________
Slide 14

UNIVERZITET U SARAJEVU VETERINARSKI FAKULTET

NJUKASLSKA BOLEST - NEWCASTLE DISEASE

Zakonska osnova
Direktiva vijeća EU 92/66/EEC:
' Newcastle disease‘
disease‘ predstavlja infekciju peradi uzrokovanu bilo kojim
avijarnim sojem Paramyxovirusa tip 1 sa intracerebralnim index-
index-om
patogeniteta (ICPI) kod dan starih pilić
pilića već
većim od 0,7.

Uzorci
Od bolesnih ptica:

Kloakalni brisevi (ili feces), i

trahealni brisevi ,

Od "svjež
svježe" uginulih ptica:

feces ili intestinalni sadrž

sadržaj,
tkivo mozga, trahea, pluć
pluća, jetra, slezena, i
drugi primjetno afektirani organi i tkiva,

________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
____________________________________________________________________
Slide 15

UNIVERZITET U SARAJEVU VETERINARSKI FAKULTET

Korisni linkovi:

http://www.oie.int/eng/normes/mcode/en_chapitre_2.1.1.htm
(CHAPTER 2.1.1. CRITERIA FOR LISTING DISEASES)
http://www.oie.int/eng/normes/mcode/en_chapitre_2.7.5.htm
(CHAPTER 2.7.5. FOWL TYPHOID AND PULLORUM
DISEASE)
http://www.oie.int/eng/normes/mmanual/A_00011.htm
(CHAPTER I.1.1. SAMPLING METHODS)
http://www.oie.int/eng/normes/mcode/en_chapitre_3.4.1.
htm#chapitre_3.4.1. (HYGIENE AND DISEASE SECURITY
PROCEDURES IN POULTRY BREEDING FLOCKS AND
HATCHERIES)
http://www.who.int/csr/disease/avian_influenza/en/
http://www.who.int/csr/disease/influenza/influenzanetwork/en/
http://www.wpro.who.int/wr/chn/
http://www.oie.int/eng/en_index.htm

________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
____________________________________________________________________
Slide 16

UNIVERZITET U SARAJEVU VETERINARSKI FAKULTET

THANK YOU

________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
____________________________________________________________________