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V E T E R I N A R S K I G L A S N I K

nau~ni ~asopisscientific journal

Veterinarski glasnik je ~asopis Fakulteta veterinarske medicine Univerziteta u BeogaduVeterinarski glasnik is published by Faculty of Veterinary Medicine University of Belgrade

VET. GLASNIK Vol. 61 Br. 3-4 Str. 135 - 248 Beograd, 2007.

IZDAVA^:FAKULTET VETERINARSKE MEDICINE UNIVERZITETA U BEOGRADUFACULTY OF VETERINARY MEDICINE UNIVERSITY OF BELGRADEFAKULÃTET VETERINARNOY MEDICINÀ UNIVERSITETA V BELGRADE

SUIZDAVA^: Veterinarska komora SrbijeCOPUBLISHER: Veterinary Chamber of Serbia

GLAVNI I ODGOVORNI UREDNIK – EDITOR IN CHIEF: Vitomir ]upi}

URE\IVA^KI ODBOR – EDITORIAL BOARD:Vojin Iveti}, Milovan Jovi~in, Vera Kati}, Sanja Kova~evi}-Aleksi}, ZoranKuli{i}, Zoran Ra{i}, Horea [amanc, Radoljub Tadi}, Jadranka Tijani},Dragi{a Trailovi}

TEHNI^KI UREDNIK – TECHNICAL EDITOR: Jadranka Tijani}

LEKTORI – LECTORS:Sanja Vrani}, za srpski jezik / for Serbian languageDanijela Gledi}, za engleski jezik / for English languageBo{ko Bo{kovi}, za ruski jezik / for Rusian language

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Publication of this journal is financially supported by:Ministry of Science and Environmental Protection of Republic Serbia

[tampa – Printers: SZR „Simi} Zuhra”, Beograd, Vitanova~ka 15

Adresa ~asopisa:Veterinarska komora Srbije – Veterinarski glasnik, 11000 Beograd,Bulevar oslobo|enja 18, tel/faks 011/2684-597, 2687-475,e-mail: vetksªeunet.yu; www.vetks.org.yu

YU ISSN 0350-2457UDK 619 (05)

VETERINARSKI GLASNIK

VOL. 61

BROJ 3 - 4

STRANA 135 - 248

Beograd 2007.

3-4

V E T E R I N A R S K I G L A S N I KÂSOPIS FAKULTETA VETERINARSKE MEDICINE UNIVERZITETA U BEOGRADU

VET. GLASNIK Vol. 61 Br. 3 - 4 str. 135 - 248 Beograd, 2007.

SADR@AJ – CONTENTS – SODER@ANIE

ORIGINALNI RADOVI – ORIGINAL PAPERS – ORIGINALÃNÀE RABOTÀ

! Caki} P., Paunovi} M, \ikanovi} Vesna, Jakov~ev-Todorovi} Dunja, Simi} V., Kuli{i} Z.:Tipologija i monitoring ekolo{kog statusa teku}ih voda SrbijeTypology and Monitoring of Ecological Status of Moving Waters of SebiaTipologiÔ i monitoring Ìkologi~eskogo statusa teku~ih vod Serbii. . . . . . . 137

STRU^NI RADOVI – PROFESSIONAL PAPERS – SPECIALISTIÊSKIE RABOTÀ

! Büyükünal S. K., Nazli B.: Effect of different electrical stunning methods on meat quality ofmarmara kivircik breed lamb in Turkey RepublicEfekti razli~itih metoda elektri~nog omamljivanja na kvalitet mesa kod MarmaraKivircik jagnjadi u TurskojÕffektì razli~nìh metodov Ìlektri~eskogo obol{eniÔ na ka~estvo mÔsa uMarmara Kiviricik ÔgnÔt v Turcii . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 155

! Prodanov Jasna, Do{en R., Petrovi} T., Lupulovi} Diana, Val~i} M., Pola~ek V.: Zna~aj us-tanovljavanja intrauterinih infekcija virusom klasi~ne kuge svinja u sklopu programasuzbijanja i iskorenjivanja oboljenjaSignificance of Determining Intrauterine Infections with Classical Swine Plague VirusWithin Programme of Curbing and Eradicating this DiseaseZna~enie ustanavlivaniÔ vnutrimato~nìh infekciy virusom klassi~eskoy~umì sviney v sostave programmì preodoleniÔ i iskoreneniÔ zabolevaniÔ . . 163

! Vu~ini} Marijana: Osnovni principi za{tite dobrobiti oglednih ìvotinjaBasic Principles of Experimental Animals Welfare ProtectionOsnovnìe principì ohranì blagosostoÔniÔ opìtnìh ìvotnìh . . . . . . . . . . 173

! Spasojevi} Kosi} Ljubica: Osnovni principi primene i interpretacije Ekg-a kod konjaBasic Principles of ECG Application and Interpretation in HorsesOsnovnìe principì i interpretacii ÕKG u lo{adey . . . . . . . . . . . . . . . . . 183

! Petrov V., Alexandrov M., Lasarova S., Lyutskanov M.: Histopathological Investigations ofRabbit Colibacteriosis, Caused by Enteropathogenic Escherichia coli (O15:H-)Histopatolo{ka istraìvanja kolibakterioze kod kuni}a izazvane enteropatogenombakterijom Escherichia coli (015:H-)Gistopatologi~eskie issledovaniÔ kolibakterioza u krolikov, vìzvannìeÌnteropatogennoy bakteriey Escherichia coli (015:H-) . . . . . . . . . . . . . . . . . . 193

! Tsachev I., Simeonov R., Petrov V.: Infections with Ehrlichia canis and Borrelia burgdorferiin a DogInfekcije bakterijama Ehrlichia canis i Borrelia burgdorferi kod jednog psaInfekcii bakteriÔmi Ehrlichia canis i Borrelia burgdorferi u odnoy sobaki . . . . 201

!Michaylov G., Tsachev I., Petrov V.: Etiological, Clinical and Epidemiological Investigationsof Dermatophytosis among Pets in the Region of Stara Zagora for the Period 2004-2006Etiolo{ka i klini~ko-epidemiolo{ka istra`ivanja prisustva dermatofita kod ku}nihljubimaca u regionu Stara Zagora u periodu 2004-2006. godineÕtiologi~eskie i klini~esko-Ìpidemiologi~eskie issledovaniÔ prisut-stviÔ dermatofitov u doma{nih lÓbimcev v regione StaraÔ Zagora v peri-ode 2004-2006 goda . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 211

!Urumova Valentina, Lyutskanov M.: Ethiological Structure and Epidemiological Monitoringof Bacterial Infections in Industrially Reared Birds in BulgariaEtiolo{ka sruktura i epidemiolo{ki nadzor bakterijskih infekcija kod industrijski ga-jene `ivine u BugarskojÕtiologi~eskaÔ struktura i Ìpidemiologi~eskiy nadzor bakterialÝnìh in-fekciy u promì{lenno razvedënnìh selÝskohozÔystvennìh ptic v Bolgarii 219

! Çetin Ö., Dümen E., Biçer H. H., Koçak Ö.: Ratio of Meat Preparates to Carcass in CattleSlaughtered in IstanbulOdnos preparata mesa u odnosu na trupove goveda klanih u IstanbuluOtno{enie preparata mÔsa v otno{enii tuloviçey krupnogo rogatogo skotaubitìh v Stambule . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 229

DIPLOMIRANI STUDENTI - DOKTORI VETERINARSKE MEDICINE – GRADUATE STUDENTS- DOCTORS OF VETERINARY MEDICINE – DIPLOMIROVANNÀE STUDENTI - DOK-TORÀ VETERINARNOY MEDICINÀ . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 241

UPUTSTVO AUTORIMA - NOTES FOR CONTRIBUTORS. . . . . . . . . . . . . . . . . . . . . . . . . 245

UDK 574.5(497.1)

TIPOLOGIJA I MONITORING EKOLO[KOG STATUSATEKU]IH VODA SRBIJE*

TYPOLOGY AND MONITORING OF ECOLOGICAL STATUS OF

MOVING WATERS OF SEBIA

P. Caki}, M. Paunovi}, Vesna \ikanovi}, Dunja Jakov~ev-Todorovi},V. Simi}, Z. Kuli{i}**

U radu se prikazuju istraìvanja koja se sprovode u cilju primeneOkvirne direktive o vodama Evropske Unije (EU). Podaci o zajedni-cama vodenih organizama, uz abioti~ke parametre, koriste se za defini-sanje tipologije, tip specifi~no referentnih uslova i indeksa ekolo{kogstatusa, {to predstavlja osnovu za ustanovljavanje sistema monitoringaekolo{kog statusa/potencijala voda u Srbiji. Rezultati sprovedenih is-traìvanja koriste se i u nastavku rada na izradi baze podataka o bio-diverzitetu kopnenih voda na podru~ju Srbije. Istraìvanja obuhvataju fi-toplankton, fitobentos, vodene makrobeski~menjake i ihtiofaunu, kaobiolo{ke elemente kori{}ene u procesu primene Okvirne direktive o vo-dama EU. Pored toga, paraziti riba su predmet istraìvanja iz razloga{to nivo parazitiranosti moè zna~ajno da uti~e na strukturu zajednice,te se moè koristiti kao biolo{ki parametar ekolo{kog statusa/potenci-jala. Studija obuhvata sve tipove teku}ih voda. Terenska istraìvanja seobavljaju sa ciljem upotpunjavanja i kompletiranja podataka o pojedi-nim biolo{kim komponentama teku}ih voda.

Klju~ne re~i: tipologija, teku}e vode, referentni uslovi, ekolo{ki indeks,biodiverzitet, biolo{ki elementi kvaliteta, ekolo{ki status

Hidrobiolo{ka istraìvanja koja su se realizovala predstavljaju kom-pleksnu ekolo{ku studiju teku}ih voda Srbije sa ciljem definisanja tipologije i tip

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ORIGINALNI RAD – ORIGINAL PAPER

Uvod / Introduction

* Rad primljen za {tampu 5. 11. 2007. godine** Dr Predrag Caki}, mr Momir Paunovi}, mr Vesna \ikanovi}, mr Dunja Jakov~ev-Todorovi},

Institut za biolo{ka istraìvanja "Sini{a Stankovi}", Beograd; dr Vladica Simi}, Prirodno-mate-mati~ki fakultet, Odeljenje za biologiju, Univerzitet u Kragujevcu; dr Zoran Kuli{i}, Katedra zaparazitologiju, Fakultet veterinarske medicine, Beograd

specifi~no referentnih uslova, kao i formulisanja indeksa ekolo{kog statusa.Tako|e, doprinose procesu formiranja sistema monitoringa ekolo{kog statusa inastavku rada na bazi podataka biodiverziteta vodenih ekosistema Srbije.

Aktivnosti na primeni Direktive o vodama Evropske Unije (DV) u Srbiji,ta~nije, na izradi tipologije i definisanju referentnih uslova za teku}e vode, pokre-nute su po~etkom 2004. godine. U protekle dve godine saradnici Instituta za bi-olo{ka istraìvanja "Sini{a Stankovi}" u Beogradu i Instituta za biologiju i ekologijuPrirodno-matemati~kog fakulteta Univerziteta u Kragujevcu bili su uklju~eni upomenute aktivnosti. Prva faza primene DV obuhvatila je izradu sistema tipologije,primenu sistema na reke ~ije je slivno podru~je ve}e od 4000 km2 i po~etak radana definisanju tip specifi~no referentnih uslova. Zatim se pristupilo primeni siste-ma na vodotoke slivnog podru~ja do 500 km2. Tokom rada prikupljeno je mnogopodataka i biolo{kih uzoraka sa preko 300 lokaliteta sa razli~itih teku}ica {iromSrbije. Definisanje tipova teku}ih voda na podru~ju Srbije jedan je od zadatakakoje treba ispuniti sa ciljem prilago|avanja sistema upravljanja vodama smerni-cama koje su iznete u pomenutoj Direktivi i projektima koji su imali za cilj njenu pri-menu Š1¹.

Tipologija voda prvi je korak u ustanovljavanju sistema u kome sestanje vodenih ekosistema i ekolo{ki status procenjuju u odnosu na referentnostanje koje je specifi~no za svaki tip stani{ta/voda. Iz tog razloga je bitno da seuradi tipologija voda na teritoriji Srbije, kako bi se moglo pristupiti preduzimanjukoraka koji vode ka planiranju, proveri i primeni modernog i efikasnog sistemaprocene statusa vodenih ekosistema.

Tipologija voda, prema konceptu predloènom u Direktivi o vodamaEU Š2¹, predstavlja balans izme|u realne raznovrsnosti voda koja je merljiva krozabioti~ke i bioti~ke karakteristike i nivoa pojednostavljenja odnosa u prirodi. Do-datan izazov u istraìvanju vezanom za tipologiju teku}ih voda predstavlja ~inje-nica da je teritorija Srbije, prema op{tim karakteristikama podru~ja (geografskipoloàj, klima, reljef, geolo{ka struktura, pedolo{ki faktori, istorijski faktori) izra-zito raznovrsna.

Podaci dobijeni ranijim istraìvanjem vodenih makrobeski~menjaka uSrbiji od zna~aja su za ostvarenje ciljeva ove studije. Istraìvanje vodenih mak-robeski~menjaka u Srbiji ima dugu tradiciju, a vodeni ekosistemi opisivani su narazli~ite na~ine, uklju~uju}i abioti~ke faktore i biotu. Detaljan pregled istraìvanjau Srbiji prikazan je u radovima Simi}a Š3¹, Markovi}a Š4, 5¹ i Paunovi}a i sar. Š6¹.

Simi} V. Š3, 7¹, i Simi} S. Š7¹ prikazuju podatke o zajednicama makro-beski~menjaka koji se mogu koristiti u tipologiji reka Srbije. U ovim radovima izne-sene su i prate}e fizi~ke i hemijske karakteristike ispitivanih teku}ica. Kao rezultatsedmogodi{njeg istraìvanja autori defini{u pet grupa teku}ih voda u okviru osamgeografskih regiona.

Klasifikacija/tipologija povr{inskih kopnenih voda preduslov je za de-finisanje sistema procene ekolo{kog statusa okruènja koji je definisan Direkti-vom o vodama Evropske Unije Š2¹. Definisanje tipova teku}ih voda na podru~ju

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Vet. glasnik 61 (3-4) 137 - 153 (2007) P. Caki} i sar.: Tipologija i monitoring ekolo{kogstatusa teku}ih voda Srbije

Srbije jedan je od zadataka koji treba ispuniti sa ciljem prilago|avanja sistemamonitoringa smernicama koje su iznesene u pomenutoj Direktivi Š2¹ i projektimakoji su imali za cilj njenu primenu Š8, 9, 10, 11¹.

Tokom realizacije ovih istraìvanja namera nam je bila da primenimoiskustva ranijih klasifikacija vodenih ekosistema, preporuke iznete u Direktivi o vo-dama Š2¹, ali i da uvaìmo sve posebnosti prirode podru~ja Srbije. Akcenat jestavljen na analizu zajednice vodenih makrobeski~menjaka, kao grupe organi-zama koja je najintenzivnije upotrebljavana u drugim evropskim zemljama prili-kom odre|ivanja tipova i tip specifi~no referentnih uslova za teku}e vode.

Na osnovu plana istraìvanja i raspoloìvih podataka, definisani suslede}i ciljevi istraìvanja kako bi se ostvarila osnovna namera – tipologija/klasi-fikacija teku}ih voda podru~ja Srbije:

– izrada spiska vrsta i komentar o zabeleènoj flori i fauni u pogleduekolo{kih i biogeografskih karakteristika, utvr|ivanje osnovnih osobina zajednica,analiza prostorne dinamike i analiza uticaja hidromorfolo{kih pritisaka (npr. pre-gra|ivanja, naselja) i pritoka na vodene makrobeski~menjake,

– analiza zajednica fitoplanktona, fitobentosa, makrobeski~menjaka iriba sa aspekta sastava i strukture, primena razli~itih biolo{kih i bioti~kih indeksakao biolo{kih elemenata kvaliteta predvi|enih Direktivom o vodama,

– precizno povla~enje granice hidro-faunisti~kih oblasti (ekoregiona)koje odgovaraju op{tim prirodnim i istorijskim karakteristikama podru~ja i lokal-nom rasprostranjenju vodenih organizama,

– podela teritorije Srbije na hidro-faunisti~ke podoblasti, koje su, doodre|enog nivoa, homogene u pogledu karakteristika vodenih organizama (hori-zontalna distribucija hidrobionata) i

– tipologija ispitivanih teku}ica.U nameri da se ostvari pomenuti zadatak, planirane su i slede}e aktiv-

nosti:– priprema i prakti~na provera terenskog upitnika (protokola) za is-

traìvanje vodenih makrobeski~menjaka, koji obuhvata podatke o stani{tu (pra-te}i podaci) i uzorcima,

– pore|enje razli~itih tehnika uzorkovanja vodenih makrobeski~men-jaka,

– pore|enje zajednica vodenih makrobeski~menjaka ispitivanih po-dru~ja i vodotokova,

– izbor referentnih lokaliteta i lokaliteta koji se, prema stepenu antro-pogenog uticaja, mogu okarakterisati kao "bliski prirodnim" ("near-natural" sites) i

– definisanje tip-specifi~no referentnih uslova za teku}e vode.

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Ciljevi istraìvanja / Objectives of investigation

Jedna od aktivnosti je i prikupljanje podataka o hidro-morfolo{kim idrugim prate}im podacima o teku}im vodama podru~ja Srbije. Ovo je definisanotokom realizovanja ovog istraìvanja, jer smo pregledom dostupne literature o pri-rodnim karakteristikama teku}ih voda Srbije ustanovili da odgovaraju}i podacinisu dostupni u okviru jedne publikacije. Najkompletniji pregled teku}ih voda upogledu abioti~kih karakteristika prikazan je u monografiji Gavrilovi}a i Duki}aŠ11¹. Analizom podataka prikazanih u pomenutoj publikaciji uvideli smo da nedo-staju podaci za veliki broj manjih brdsko-planinskih teku}ica koje su za hidro-biolo{ka istraìvanja i najinteresantnije. Zbog toga smo se opredelili za to da radsadrì i prikaz osnovnih abioti~kih karakteristika ispitivanih teku}ica, u nadi da }etaj deo doprineti da se makar jedan deo podataka nalazi sumiran na jednommestu.

Istraìvanja obuhvataju fitoplanktone, fitoperifitone, vodene makro-beski~menjake i ribe, kao biolo{ke elemente {iroko kori{}ene u primeni DV u Ev-ropi. Uz to, istraìvanja parazita riba su tako|e predmet ove studije, jer se nivo pa-razitiranosti moè koristiti za procenu ekolo{kog statusa vodenog sistema.

Terenska istraìvanja imaju za cilj prikupljanje biolo{kog materijala ipodataka o ispitivanom lokalitetu/sektoru. U planiranju terenskih radova akcenatje stavljen na teku}ice koje nisu bile obuhva}ene prethodnim istraìvanjima naadekvatan na~in.

Metodologija koja se primenjuje obuhvata sakupljanje i obradu ma-terijala i prate}ih podataka. Zasnovana je na principima definisanim u DV, rezul-tatima projekata koji su imali za cilj primenu pomenute Direktive Š8¹, kao i na doku-mentima definisanim tokom rada stru~nih grupa Me|unarodne komisije za za{titureke Dunav Š12¹. Pored toga, prilikom definisanja programa istraìvanja u razma-tranje su uzeta iskustva drugih zemalja na polju primene DV Š13¹.

Analiza osobenosti zajednica vodenih organizama podrazumeva sle-de}e momente: broj vrsta, gustina zajednica (kvantitativne i semi-kvantitativnetehnike), diverzitet, kao i polnu i uzrasnu strukturu za pogodne grupe organizama(ribe i re~ni rakovi). Analiza parazita riba obuhvata istraìvanje nivoa i tipa infek-cije, sa ciljem da se odredi relacija izme|u parazitiranosti i ekolo{kog statusavoda.

Detaljna analiza lokaliteta uzorkovanja je zasnovana na terenskomupitniku prilago|enom za potrebe studije i obuhvata prikupljanje podataka o mor-folo{kim osobenostima profila (prose~na, minimalna i maksimalna {irina, pro-se~na i maksimalna dubina, tip podloge, tip korita i na~in grananja) na sektoru odnajmanje 50 m. Analiza zastupljenih tipova podloge vr{i se vizuelnom procenom,u kvadratu 50x50 cm, po sistemu klasifikacije prilago|enom potrebama studije, sciljem da se koristi skala koja opisuje tipove podloge koji realno uti~u na distribu-ciju vodenih makrobeski~menjaka. Kao osnova za modifikaciju sluì skala za kla-

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Metodologija / Methodology

sifikaciju mineralnog supstrata (14, 15): 1) supstrati sitnih ~estica (mulj i vrlo sitanpesak; ~estice nisu vidljive golim okom; <0,125 mm); 2) sitan pesak (~estice vid-ljive golim okom; 0,125-0,5 mm); 3) krupan pesak (0,5-2 mm); 4) {ljunak (2-16mm); 5) oblutak (16-34 mm); 6) kamen (64-256 mm) i 7) stena (>256 mm). Na ter-enu se obavlja i merenje povr{inske brzine vode.

Kriterijumi za izbor lokaliteta uzorkovanja su slede}i: zastupljenost de-lova toka razli~itih karakteristika (vir, preliv, brzak), karakteristi~an lokalitet za sek-tor, raznovrsnost/ujedna~enost stani{ta za makrobeski~menjake, op{te karakteri-stike obodne vegetacije, pristupa~nost terena, uticaj zaga|enja i dr.

Monitoring abiocena obuhvata slede}e pokazatelje: pH, elektri~naprovodljivost, rastvoreni kiseonik i zasi}enost kiseonikom. Istraìvanje obuhvatagotovo celu Srbiju - slivove Velike, Zapadne i Ju`ne Morave, Ni{ave, Ibra, VelikogRzava, Timoka, Save, Kolubare, Tise, Peka, Mlave, Pore~ke reke, kao i materijalprikupljen na glavnom toku Dunava i ekosistemima plavnog podru~ja.

Fitoplankton / Phytoplankton

Fitoplankton, zastupljen brojnim porodicama algi u vodenim eko-sistemima, predstavlja zajednicu sitnih organizama koji ~itav svoj ìvotni ciklusprovode u slobodnoj vodenoj masi ekosistema, izme|u povr{ine i dna, plutaju}i.Ovi organizmi su sposobni za brojne adaptacije koje uslovljava plutaju}i na~in ì-vota. Fitoplankton se razvija u sporoteku}im, ravni~arskim rekama i jezerima, a ni-kada u brzim brdsko-planinskim teku}icama.

Prikupljanje fitoplanktona se vr{i standardnim planktonskim mreà-ma, promera okaca 20-25 ìm. Voda se uzima sa odre|enog broja ta~aka po po-pre~nom profilu teku}ice. Svi uzorci se proce|uju kroz planktonsku mreù iodlaù kao jedan zajedni~ki uzorak Š16¹.

Fitobentos / Phytobenthos

Fitoperifiton ~ine biljni predstavnici koji naseljavaju tvrde delove pod-loge (stene, kamen) i ~vrsto su vezani za njih. Nazivamo ga obra{tajem u kome seuglavnom sre}u predstavnici silikatnih, modrozelenih i zelenih algi. Organizmi koji~ine fitobentos sakupljaju se pomo}u ~etkice ili noà laganim i pa`ljivim skidan-jem, odnosno struganjem sa tvrde povr{ine. Na terenu se prikupljeni materijal od-mah fiksira formaldehidom do finalne koncentracije do 3-4%. Priprema diatomaza determinaciju ura|ena je prema Husted-ovoj metodi Š17¹. Identifikacija jeobavljena kori{}enjem invertnog mikroskopa Nikon, uve}anje 1500x.

Akvati~ni makrobeski~menjaci / Aquatic macroinvertebrate

Akvati~ni makrobeski~menjaci raznovrsna su i biolo{ki heterogenagrupa organizama. Veliki broj vrsta ìvotinja koji obuhvata ova ekolo{ka katego-rija, raznovrsnost akvati~nih stani{ta u Srbiji, kao i relativno mali broj stru~njakakoji se kod nas trenutno bavi vodenim makrobeski~menjacima, doprineli su da suovi hidrobionti slabo ispitani u pore|enju sa stepenom istraènosti u ve}ini sused-

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nih zemalja. S toga, svaki napor u pravcu istraìvanja akvati~nih makrobe-ski~menjaka predstavlja doprinos u procesu upotpunjavanja znanja o vrstamakoje su prisutne u Srbiji i njihovoj ekologiji.

Prikupljanje makrobeski~menjaka obavlja se uz upotrebu ru~nih ben-tosnih mreà promera okca 250 i 500 �m, Sürber-ovim mreàma zahvatnihpovr{ina 0,1 i 0,04 m2 promera okca 250 �m, bentolo{kom dredòm mreèpromera okca 250 �m, bagerima tipa Ekman i Van-Veen, zahvatnih povr{ina 0,04 i0,03 m2, kao i ru~nim sakupljanjem i spiranjem materijala sa razli~itih povr{ina.

Uzorkovanje ru~nim bentolo{kim mreàma vr{i se kombinovanomtehnikom podizanja materijala trzajima nogama sa podloge i njegovim sakuplja-njem u mreù koja je orijentisana u pravcu vodenog toka i ru~nim sakupljanjem sapodloge (kick & sweep teknika), semi-kvantitativnim uzorkovanjem u definisanomvremenskom intervalu, pri ~emu je prikupljano sa svih dostupnih stani{ta, propor-cionalno zastupljenosti stani{ta na sektoru Š16¹.

U slu~aju kori{}enja Sürber-ovih mreà uzorak se sastoji iz tri zahvatapo tipu stani{ta, pri ~emu se vodi ra~una da materijal bude prikupljen sa tipa pod-loge koji je karakteristi~an za sektor Š16¹.

Prilikom kori{}enja bentolo{kih bagera, materijal se ispira kroz sitopromera okca 250 �m, kako bi se odvojile sitne ~estice od uzorka i omogu}ilo od-laganje i kvalitetnije fiksiranje i konzerviranje uzoraka Š16¹.

Determinacija prikupljenih makrobeski~menjaka obavlja se u hidro-biolo{koj laboratoriji kori{}enjem mikroskopa, uve}anja do 90x. Identifikacijaorganizama obavlja se pomo}u adekvatnih klju~eva do najnièg pouzdanog tak-sonomskog nivoa (rod, vrsta) Š19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32,33, 34, 35, 36¹ u laboratoriji Odeljenja za hidrobiologiju Instituta za biolo{ka is-traìvanja "Sini{a Stankovi}", gde se materijal i ~uva.

Ihtiofauna / Ichthyofauna

Primerci riba se izlovljavaju upotrebom mreà staja}ica i povla~nihmreà razli~itog promera okaca (32–50 mm), kao i kori{}enjem agregata Honda(1,5 KW) za elektro-ribolov.

Parazitolo{ka analiza materijala obavlja se na prikupljenim primercimariba. Kod svih jedinki pregledaju se {krge, trbu{na duplja i crevni trakt radi utvr|i-vanja prisustva ekto- i endoparazitskih vrsta. Sve prona|ene vrste Protozoa fik-siraju se metanolom i boje Giemzom nakon su{enja. Ostale prona|ene parazitskevrste prosvetljuju se u laktofenolu i od njih se prave trajni preparati.

Za pregled riba koriste se uobi~ajene parazitolo{ke metode Š37¹. @ivetrematode I akantocefale se opu{taju u destilovanoj vodi na 4oC oko 1 sat i potomfiksiraju u 10% vrelom formalinu. @ive nematode se fiksiraju u vrelom 70% etanolui ~iste u vrelom laktofenolu. Zamrznuti primerci se odmrzavaju u vodi, a zatim fik-siraju u 10% formalinu (trematode i akantocefale), odnosno u 70% etanolu (nema-tode).

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Identifikacija se obavlja na osnovu klju~eva za identifikaciju parazitaŠ38, 39, 40, 41, 42, 43¹.

Na analiziranom uzorku riba radi se ekstenzitet i intenzitet infekcije.

Po obimu istraìvanja potamoplanktona (fitoplankton u rekama), na-jistraèniji vodotokovi u na{oj zemlji su Dunav, Sava i Tisa, dok pojedini manji vo-dotokovi kao {to su Tami{ i Velika Morava skoro da uop{te nisu ra|eni. Naj~e{}epodru~je istraìvanja Dunava i Save je podru~je Beograda (za Savu: [abac-Beograd) re|e drugi delovi reke ili ceo tok kroz Srbiju. Sli~no je i sa Tisom, uz ~in-jenicu da su najobimnija i najkompleksnija istraìvanja vr{ena na delu toka Tisekod Be~eja. Tako|e je neophodno ista}i da se istraìvanja Dunava, uslovno,mogu podeliti na istraìvanje vodotoka pre i nakon izgradnje brana na Dunavukod \erdapa, i istraìvanja podru~ja \erdapa koje nakon izgradnje brana naDunavu ima druga~ije karakteristike fitoplanktona. Istraìvanja fitoplanktona rekeTami{ ra|ena su u vi{e navrata du` celog toka kroz na{u zemlju. Za Veliku Moravuskoro da ne postoje objavljeni podaci. Jedina istraìvanja fitoplanktona Velike Mo-rave ra|ena su 1977.-1979. godine Š6¹.

Prema svojim hidromorfolo{kim i ekolo{kim karakteristikama moè sezaklju~iti da postoje znatne razlike u pogledu kvalitativnog i kvantitativnog sastavafitoplanktona sporoteku}ih, ravni~arskih reka (Dunav, Sava, Tisa, Tami{ i VelikaMorava). Za sve ove reke zajedni~ko je da se kao dominantna grupa u kvalitativ-nom i kvantitativnom pogledu izdvajaju Bacillariophyta, a kao subdominantnaChlorophyta ~iji udeo u sastavu zajednice naro~ito raste u vreme letnjih mesecikada su vodostaji najniì, a temperatura i osvetljenost najve}i Š6¹.

Tokom razmatranja vodenih makrobeski~menjaka upotrebljen jematerijal prikupljan u periodu 1993.-2006. na 356 lokaliteta. Kori{}en materijalobuhvata uzorke sa 192 lokaliteta prikupljen u periodu 2003.-2006. Podaci sa 113lokaliteta dostupni su iz prethodnog perioda (1992.-2003.), a na osnovu istraì-vanja realizovanih u Odeljenju za hidroekologiju i za{titu voda Instituta za biolo{kaistraìvanja "Sini{a Stankovi}" u Beogradu i Odeljenju za hidroekologiju Institutaza biologiju Prirodno-matemati~kog fakulteta Univerziteta u Kragujevcu.

Materijal je prikupljan prete`no u kriti~nim periodima za zajednicu ma-krobeski~menjaka – vreme niskih i visokih voda.

Determinisan je materijal iz 420 uzoraka faune dna i pregledano je uk-upno 40.120 jedinki. Zabeleèno je ukupno 452 taksona vodenih makrobe-ski~menjaka svrstanih u 115 familija, odnosno 239 rodova. Prema broju vrsta,najzna~ajnija grupa na ispitivanim lokalitetima su vodeni insekti (337 taksona),me|u kojima se, prema raznovrsnosti, izdvajaju Trichoptera (95 vrsta), Diptera (78taksona) i Ephemeroptera (73 vrste) Š6¹.

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Rezultati rada / Results

Za izradu spiska vrsta kori{}en je celokupni materijal. Tokom analizeprostorne distribucije vodenih makrobeski~menjaka, u slu~aju brdsko-planinskihteku}ica, akcenat je stavljen na uzorke prikupljene u periodu prole}a i ranog leta,kako bi se iz analize eliminisale razlike u zajednici do kojih dolazi usled vremenskedinamike. Ovaj period je izabran kao adekvatan za analizu faune ove grupeteku}ih voda, jer je tada raznovrsnost najve}a Š4, 7, 44, 45, 18, 46, 6¹. Pored toga,identifikacija materijala prikupljenog u periodu prole}a i ranog leta je pouzdanija,jer tada dominiraju krupniji larveni i nimfalni stupnjevi, za razliku od periodakasnog leta i jeseni, kada se beleì dosta larvi u prvom i drugom stadijumurazvi}a. Ova ~injenica naro~ito je uo~ljiva u slu~aju familije Baetidae (Ephemerop-tera) Š18, 46¹. U gotovo svim uzorcima prikupljenim u brdsko-planinskim po-dru~jima tokom septembra i oktobra zabeleèno je dosta predstavnika roda Bae-tis koji su bili sitni i komplikovani za identifikaciju.

Analiza faune ve}ih ravni~arskih reka je, nasuprot brdsko-planinskimteku}icama, vr{ena prete`no u septembaru i oktobru (period niskih voda). Pored~injenice da se uzorci iz velikih reka lak{e prikupljaju u periodu niskih voda,ovakvom izboru je doprinelo i to {to se ve}ina prethodnih istraìvanja (2001. Za-jedni~ko ispitivanje Dunava 1 – JDS1 Joint Danube Survey 1; 2001. Me|unarodnoispitivanje Tise – ITR International Tisza Research; 2004. Istraìvanje Dunava Aq-uaTerra – ADS AquaTerra Danube Survey) obavljala u pomenutom vremenskomintervalu, kao i da je Zajedni~ko ispitivanje Dunava 2 – JDS2 Joint Danube Survey2 planirano za avgust i septembar 2007., ~ime se obezbe|uje poredivost poda-taka.

Zajednice makrobeski~menjaka Dunava i Save u beogradskom re-gionu su analizirane sa 10 lokaliteta i dobijeni rezultati su ukazali na dominantnuulogu grupa Oligochaeta i Gastropoda Š47¹.

Na osnovu dobijenih rezultata o kvalitativnom sastavu zajednica fito-bentosa na lokalitetima istraìvanih teku}ica u razli~itim podru~jima Srbije(isto~ne, centralne, jugozapadnog i zapadnog dela), pra}ena je njihova pros-torna distribucija. Modrozelene alge (Cyanophyta) (33 taksona) su zabeleène na44 lokaliteta na dnu 24 teku}ice. Crvene alge (Rhodophyta) sa svojih osam tak-sona su registrovane na 21 lokalitetu, na dnu devet teku}ica. Alge razdela Xantho-phyta ({est taksona) naseljavaju dno {est teku}ica i konstatovane su na 20 lo-kaliteta. Od zlatnih algi (Chrysophyta) je na dnu istraìvanih teku}ica na|enasamo jedna vrsta (Hydrurus foetidus) i to na dnu ~etiri reke i {est lokaliteta. Zelenealge (Chlorophyta) (22 taksona) su zabeleène na 47 lokaliteta na dnu 18teku}ica. Vrsta Chara vulgaris (Charophyta) je na|ena samo na podru~ju isto~neSrbije u Stanjanskoj reci na dva lokaliteta Š6¹.

Istraìvanja ihtiofaune u Srbiji su sprovedena na 14 profila na rekamaDrini (gornji i donji tok), Dunavu (od 1250 do 1297 r.km., beogradski region, uz-vodno od "\erdapa I", akumulacija "\erdap II", nizvodno od brane "\erdapa II"),Savi ([abac, Obrenovac-Maki{), Velikoj Moravi (Varvarin), Zapadnoj Moravi (selo

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Prijanovi}i – gornji tok, Trstenik – donji tok) i Ju`noj Moravi (Bujanovac i DonjiLjube{) Š6¹.

Analiza parazita riba obuhvata:1) ekologiju parazitoza riba, vodozemaca, ljuskara i meku{aca iz reka,

rezervoara i slatkovodnih jezera Š48, 49¹;2) biologiju, ekologiju, funkcionalnu morfologiju i razvoj parazita u ri-

bama i drugim organizmima u vodi, kao i u pticama vezanim za vodena stani{takoje su zna~ajne u preno{enju parazita riba Š50, 51, 52, 53¹;

3) opis i kvantifikaciju zajednice parazita Š54¹;4) vrste na|enih parazita u zajednici i strukturu zajednice parazita Š54,

55, 56, 57, 58, 59, 60, 61¹;5) biogeografske aspekte koji uti~u na geografsku rasprostranjenost

parazita Š54, 48, 49¹;6) biomonitoring zaga|enosti vode na osnovu zajednice parazita Š48¹;7) faktore koji uti~u na populaciju parazita Š49¹;8) epidemiolo{ke aspekte Š60, 61¹;9) pregled na parazitska oboljenja riba i uticaj parazita na populaciju

doma}ina Š62, 63¹.Zbog velike prisutnosti alohtonih vrsta riba u na{im otvorenim vo-

dama, posebna pa`nja je posve}ena procenjivanju prisustva endoparazita sle-de}ih, a za na{u faunu novih, ribljih vrsta: sivi tolstolobik (Hypophthalmichtys no-bilis), beli tolstolobik (Hypophthalmichtys molitrix), beli amur (Ctenopharyngodonidella), babu{ka (Carassius auratus gibelio), bezribica (Pseudorasbora parva),ameri~ki somi} (Ameiurus nebulosus), sun~anica (Lepomis gibosus) i {ilo (Syng-nathus abaster).

Problem postaje mnogo ve}i uvozom novih ribljih vrsta, naro~ito izazijskih zemalja i SAD, a sa njima skoro potpuno nepoznatih parazitofauna riba.Prilikom istra`ivanja srpskog dela Dunava, na {krgama sivog tolstolobika na|enaje nova parazitska vrsta Copepoda (Sinergasilus polycolpus), koja je ujedno i prvinalaz za parazitofaunu Srbije Š64¹. Na ribljoj vrsti Barbus meridionalis, uhva}enoj ureci Uvac, na|ena je nova parazitska nematoda Philometroides barbi sp. nov.Š65¹.

Ispitivanju parazitske faune riba prirodnih voda (potoci, reke, jezera ibare) mora se posvetiti ve}a pa`nja, ne samo zbog njihovog ekonomskog iprivrednog zna~aja, ve} da bi prou~avanjem ove faune do{li do novih saznanja oribljim parazitima u prirodnim vodenim ekosistemima.

Tipologija teku}ih voda Srbije, pored podataka o fizi~ko-hemijskimkarakteristikama ispitivanih vodotokova, ura|ena je i na osnovu raspolo`ivih ikompletnih podataka o analiziranim zajednicama akvati~nih makrobeski~me-njaka, kao elemenata biolo{kog kvaliteta predvi|enih Direktivom o vodama.

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Diskusija / Discussion

Analiza podataka vr{ena je na izabranim uzorcima akvati~nih mak-robeski~menjaka, u zavisnosti od cilja postupka. U zavisnosti od svrhe analize, iz-abrani su uzorci koji poti~u sa lokaliteta koji se nalaze u okviru reka, ili pojedinihsektora teku}ica, koje su sli~ne prema karakteristikama – sli~na nadmorska visina(ista klasa nadmorske visine) i sli~ne hidro-morfolo{ke karakteristike. Prilikomanalize prostorne distribucije vodenih makrobeski~menjaka kori{}eni su podacikoji poti~u sa lokaliteta koji nisu pod bitnim uticajem zaga|enja. Tako su upotreb-ljeni podaci, uglavnom, sa referentnih lokaliteta ili lokaliteta ~iji se status moèokarakterisati kao "blizak prirodnom" Š6¹. U slu~ajevima kada nisu bili dostupnipodaci sa referentnih lokaliteta, kori{}eni su i podaci sa lokaliteta ~iji se ekolo{kistatus moè preliminarnom procenom okarakterisan kao visok Š6¹.

Za izra~unavanje kori{}enih biolo{kih indeksa kori{}en je AQEM pro-gram Š1¹.

Baza podataka / Database

Baza podataka biodiverziteta vodenih ekosistema Srbije razvijena jena Institutu za biologiju i ekologiju Prirodno-matemati~kog fakulteta Univerziteta uKragujevcu Š66¹.

U periodu od 2000., u okviru redovnih aktivnosti formirana je i redovnoaùrirana Baza podataka o biodiverzitetu vodenih ekosistema Srbije ("ADSer").

Osnovnu strukturu kompjuterske baze podataka (ADSer), ~ine zapisio vrstama hidrobionata i to: makroalgi, makrobeski~menjaka, riba, vodozemaca igmizavaca koje su konstantovane u kopnenim vodama Srbije.

Une{eni podaci o vrstama su dvojakog porekla i to:1. publikovani bibliografski podaci o dosada{njim istraìvanjima gore

pomenutih hidrobionata na podru~ju Srbije i to po~ev{i od 1860. do 2005. go-dine.

2. podaci o istraìvanjima akvati~nih makrobeski~menjaka tokomposlednje 3 godine istraìvanja, a koji nisu do sada publikovani.

Zapisi uneseni u bazu za svaki obra|eni takson su slede}i:1. ta~an naziv taksona,2. identifikacioni broj taksona (ID taksona),3. godina nalaza,4. broj nalaza tokom godine u kojoj je na|en (u~estalost nalaza),5. relativna brojnost taksona izraèna prema slede}oj skali:

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U~e{}e vrste /Specie participation

Oznaka relativne brojnosti /Mark of relative numerousness

Broj individua /Number of specimens

Manje od 3 / Under 3 1 1-10

3-10 2 10-50

10-20 3 50-150

20-40 4 150-500

40-100 5 vise od 500 / more than 500

ne postoji podatak / no data NP ne postoji podatak / no data

6. tip ekosistema gde je vrsta konstantovana,7. podatak o statusu ugroènosti (ako postoji, baza IUCN, zakon o ret-

kim i ugroènim vrstama Republike Srbije),8. podatak o UTM i geografskom poloàju (GPS – podaci) stani{ta

gde je vrste na|ena,9. podaci o autoru i publikaciji.Prema analizi izvr{enoj 01.10.2005. godine baza podataka biodiver-

ziteta vodenih ekosistema Srbije sadrì slede}e:1. ukupan broj zapisa koji je unet u bazu o svim taksonima po eko-

sistemima iznosi 12.443,2. ukupan broj zapisa za makrobeski~menjake po svim ekosistemima

je 8.715,3. ukupan broj zapisa za ribe po svim ekosistemima je 3.587,4. ukupan broj zapisa za vodozemce po svim ekosistemima je 141,5. ukupan broj obuhva}enih vrsta makrobeski~menjaka je 1.657,6. ukupan broj vrsta riba o kojima postoji zapis je 155,7. ukupan broj vrsta vodozemaca o kojima postoji zapis je 32.

Podaci iz aùrirane baze podataka sluè za:1. analizu abioti~ke tipologije teku}ica, koja je izra|ena na osnovu

sistema tipologije predloènog 2004. godine,2. izdvajanje referentnih lokaliteta i lokaliteta ~ije se stanje moè

okarakterisati kao "blisko prirodnom",3. kao i za definisanje granica ekoregiona i4. hidro-ekolo{kih celina na podru~ju Srbije.Kako se ostvarenje aktivnosti na formiranju i popunjavanju ADSer vr{i

od 2000. godine i kako baza sadrì podatke potrebne za nastavak rada na primeniDV, a u oblasti kori{}enja biolo{kih elemenata kvaliteta (BQE), saradnja nausavr{avanju ADSer opravdana je metodolo{ki i ekonomski.

Podela teritorije Srbije na hidrofaunisti~ke oblasti i podoblasti /Division of the territory of Serbia into aquatic fauna areas and subareas

Prema korigovanim granicama, broj ekoregiona na teritoriji Srbijeostaje nepromenjen u odnosu na izvornu verziju Illies-a Š26¹. Teritorija Srbije

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obuhvata 5 ekoregiona (Slika 1). Ekoregion 5 – Dinarski zapadni Balkan; ekore-gion 6 – Helenski zapadni Balkan; ekoregion 7 – Isto~ni Balkan; ekoregion 10 –Karpati; i ekoregion 11 – Panonska nizija.

Izvr{ena je podela Srbije na hidrofaunisti~ke podoblasti na osnovupostoje}ih abioti~kih podataka i informacija o bioti. Od biolo{kih parametarapredvi|enih Direktivom o vodama odabrani su vodeni makrobeski~menjaci, kojise predla`u za multimetrijski sistem procene ekolo{kog statusa.

Za sada je izvr{ena analiza efikasnosti tehnika uzorkovanja; izra|enaje operacionalna lista taksona za makrobeski~menjake; izvr{ena je valorizacija ti-pologije izra|ene na osnovu abioti~kih faktora, a na osnovu rasprostranjenjavodenih makrobeski~menjaka. Tipologija vodotoka na podru~ju Srbije ura|ena jepostepeno kroz analizu rasprostranjenja akvati~nih makrobeski~menjaka u od-nosu na tipolo{ke parametre (nadmorska visina, veli~ina sliva, geologija, pripad-nost ve}em slivu, pripadnost ekoregionu).

Prikupljeni materijal i dobijeni rezultati istra`ivanja su osnov za daljeusavr{avanje tipologije teku}ih voda na podru~je Srbije, kao i za definisanje tipspecifi~no referentnih uslova za izabrane parametre za teku}e vode. Na osnovuanalize zajednica fitoplanktona, perifitona, vodenih makrobeski~menjaka i ihtio-faune radi se na definisanju granica ekoregiona za teritoriju Srbije. U pogledukvaliteta vode, vr{i se definisanje novih indikatorskih grupa i redefinisanje indika-

148


Zaklju~ak / Conclusion

torskih vrednosti za stare grupe bioindikatora. Rezultati se upotpunjuju izradomtestiranja novog ekolo{kog indeksa zasnovanog na Balkanskom bioti~kom in-deksu (BNBI) i primenom ovog indeksa u monitoringu ekolo{kog statusa teku}ihvoda u regionu.

Svakako, zna~ajna karika u sagledavanju ekolo{kog statusa vodaSrbije je i poznavanje parazitofaune riba. Tako|e se nastavlja rad na bazi poda-taka o biodiverzitetu kopnenih voda podru~ja Srbije izra|enoj na Institutu za biolo-giju i ekologiju Prirodno-matemati~kog fakulteta Univerziteta u Kragujevcu.

Namera studije je da se predloì tipologija, tip specifi~no referentniuslovi i indeks ekolo{kog statusa koji bi bili pogodni za dalje pobolj{anje ikori{}enje u usavr{avanju nacionalnog sistema monitoringa i sistema upravljanjavodama.

Rezultati sprovedenih istraìvanja u znatnoj meri doprinose procesuprimene Direktive o vodama EU na podru~ju Srbije.

Prikazane aktivnosti i dobijeni rezultati dosada{njih istraìvanja su de-lom obuhva}eni u okviru Projekta 143023 – "Teku}e vode Srbije - istraìvanje bio-diverziteta i kori{}enje podataka u tipologiji, izradi ekolo{kog indeksa i monitor-ingu ekolo{kog statusa".

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TYPOLOGY AND MONITORING OF ECOLOGICAL STATUS OF MOVING WATERSOF SEBIA

P. Caki}, M. Paunovi}, V. Djukanovi}, D. Jakov~ev-Todorovi}, V. Simi}, Z. Kuli{i}

The paper presents work carried out with the objective of the implementationof the European Union Water Framework Directive. The data on communities of water or-ganisms, together with the abiotic parameters, are used to define the typology, the type ofspecifical reference conditions and the ecological status index, which present a basis forestablishing a system of monitoring the ecological status/potential of the waters in Serbia.The results of the obtained investigations are used also in the continuation of this work inmaking a database on the biodiversity of landlocked bodies of water in the territory of Ser-bia. The investigations include phytoplankton, phytobenthos, aquatic macroinvertebrate,and ichthyofauna, as well as the biological elements used in the process of the implemen-tation of the EU Water Framework Directive. Furthermore, fish parasites are a subject of in-vestigations for the reason that the level of parasite presence could have a significant effecton the structure of the community, and can consequently be used as a biological parame-ter for the ecological status/potential. The study covers all types of moving water. Field re-search is conducted with the aim of compiling complete data on certain biological compo-nents of moving waters.

Key words: typology, moving waters, reference conditions, ecological index, biodiversity,biological elements of quality, ecological status

TIPOLOGIÂ I MONITORING ÕKOLOGIÊSKOGO STATUSA TEKUÎH VODSERBII

P. Caki~, M. Paunovi~, V. Dìkanovi~, DunÔ Âkov~ev-Todorovi~, V. Simi~,Z. Kuli{i~

V rabote pokazìvaÓtsÔ issledovaniÔ, provodimìe s celÝÓ prime-neniÔ PriblizitelÝnoy direktivì o vodah Evropeyskoy Unii (EU). Dannìe osoobçestvah vodÔnìh organizmov, pri abioti~eskih parametrah, polÝzuÓtsÔ dlÔdefinicii tipologii, tip specifi~no referntnìh usloviy i indeksa Ìkologi~e-skogo statusa, ~to predstavlÔet soboy osnovu dlÔ ustanavlivaniÔ sistemì moni-

152


ENGLISH

RUSSKIY

toringa Ìkologi~eskogo statusa / potenciala vod v Serbii. RezulÝtatì prove-dÒnnìh issledovaniy polÝzuÓtsÔ i v prodolènii rabotì na bìrabotke bazìdannìh o bioraznorodnosti suhoputnìh vod territorii Serbii. IssledovaniÔ oh-vatìvaÓt fitoplankton, fitobentos, vodÔnìe makrobezpozvono~nìe, ihtiofau-nu, kak biologi~eskie Ìlementì, polÝzovannìe v processe primeneniÔ Priblizi-telÝnoy direktivì o vodah EU. Pri Ìtom, parazitì rìb predmet issledovaniÔ naosnovanii, ~to urovenÝ parazitirannosti moèt zna~itelÝno vliÔtÝ na strukturusoobçestva, i moèt zna~itelÝno vliÔtÝ na strukturu soobçestva, i moèt polÝ-zovatÝsÔ kak biologi~eskiy parametr Ìkologi~eskogo statusa / protenciala.Issledovanie ohvatìvaet vse tipì teku~ih vod. IssledovaniÔ na meste delaÓtsÔs celÝÓ popolneniÔ i komplektovaniÔ dannìh o nekotorìh biologi~eskih kompo-nentov teku~ih vod.

KlÓ~evìe slova: tipologiÔ, teku~ie vodì, referntnìe usloviÔ, Ìkologi~eskiyindeks, bioraznorodnostÝ, biologi~eskie Ìlementì ka~estva,Ìkologi~eskiy status

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UDK 637.5'3:.513.22

EFFECT OF DIFFERENT ELECTRICAL STUNNINGMETHODS ON MEAT QUALITY OF MARMARA KIVIRCIK

BREED LAMB IN TURKEY REPUBLIC*

EFEKTI RAZLI^ITIH METODA ELEKTRI^NOG OMAMLJIVANJA NA

KVALITET MESA KOD MARMARA KIVIRCIK JAGNJADI U TURSKOJ

S. K. Büyükünal, B. Nazli**

The effects of head-only electrical stunning method were com-pared with the effects of head- to- back electrical stunning method. A to-tal of 90 kivircik breed lambs were randomly allocated immediatelyprior to slaughter to one of three stunning treatments: control group(C), head only group (HO; 1.0 A- AC for 3 s at a frequency of 50 Hz),head to back group (HB; 1,0 A- AC for 3 s at a frequency of 50 Hz) elec-trical stunning. Meat quality was assessed by examining pH, colour asL, a, b values, water holding capacity (WHC) and shear force.

The effect on meat quality was assessed in head-only electricallystunned, head to back electrically stunned and non-stunned lambs.Shear forces were not significantly different between treatments. How-ever colour (L*,a*,b*), water holding capacity (WHC) and muscle ulti-mate pH were found to be significantly higher (P<0.05) between thegroups.

Key words: lamb, electrical stunning, meat, quality

Pre-slaughter stunning is compulsory according to EU Council Direc-tive 93/119 Š4¹. Protecting welfare in the pre-slaughter period, and protectingslaughterhouse workers against animals' reflex movement, lambs are not slaugh-tered without stunning in slaughterhouses Š1, 20¹. At the same time this practicemay affect the meat quality like the type of suckling, lairaging, transport conditionand handling such extrinsic factors are likely to affect meat quality through stress,which decreases muscular glycogen reserves, a process which may lead to high

155

STRU^NI RAD – PROFESSIONAL PAPER

Introduction / Uvod

* Rad primljen za {tampu 30. 01. 2008. godine** S. K. Büyükünal, B. Nazli, Istanbul University Veterinary Faculty, Department of Food Hy-

giene and the Technology, Istanbul, Turkey

ultimate pH of meat Š2¹. Electric stunning is the most common commercially usedmethod in the EU. There are several methods used to apply the electrodes usedwith electrical stunning apparatus like head-only, head-to-brisket and head-to-back Š7¹.

Electrical stunning methods are effective and produce instantaneousinsensibility by the induction of epileptiform activity in the brain Š8¹. The few stud-ies on sheep examine the effect of the type of stunning on bleeding efficiency Š9¹,petechial haemorrhages Š6¹, blood pressure and muscular activity Š16¹, final pHŠ14¹, meat quality parameters such as colour, tenderness, juiciness Š20¹ and meatquality parameters, the presence of haemorrhages and the bone fractures Š18¹but not different electrical stunning methods effects on meat quality parameters.

Animals / @ivotinje

Ninety male lambs of the Kivircik breed from the flock of the Experi-mental Farm of Istanbul University were slaughtered at 5 months of age. Animalswere separated into three group (n = 30 each) for pre-slaughter handling. In thefirst group, the lambs were slaughtered without previous stunning. The secondgroup was electrically stunned by the head only method at 1 A for 3 s. The othergroup was electrically stunned by the head to back method at 1 A for 3 s. Immedi-ately after stunning, the lambs were slaughtered using conventional procedures.The carcasses were chilled at 6oC for 24 h in a conventional chiller.

Measurement / Merenje

Instrumental meat quality was assessed in the M. longissimus dorsibetween 11.12. thoracal vertebrae.

The pH was measured rapidly after carcass dressing (pH 0), 45 min-utes later (pH 45), 4 hours later (pH 4), 24 hours later (pH 24), 48 hours later (pH48) and 72 hours later (pH 72) with a penetrating electrode adapted to a portableHanna HI 8314 electro pH meter.

Twenty-four hours post mortem, the M. longissimmus dorsi was re-moved from the carcass after measuring pH24, and cut into four equal portions.One portion packed in a plastic tray over-wrapped with permeable gas film andstored at 3 oC prior to analysis for initial meat quality. Twenty-four hours post mor-tem water holding capacity (WHC) and colour (as L, a, b values), 72 hours postmortem shear force (SF) were determined.

The rest of the samples were vacuum packed in Cryovac barrier bags.Samples were taken, as required at 24h, 48h and 72h. During this time the bagswere stored at 3oC.

The samples were analysed for pH, colour, WHC as percentage freewater Š20¹. SF was evaluated using an Instron Texture Analyser (model 1140)

156

Vet. glasnik 61 (3-4) 155 - 162 (2007) S. K. Biyikinal and B. Nazli: Effect of differentelectrical stunning methods on meat quality of marmara kivircik breed lamb in Turkey...

Materials and methods / Materijal i metode ispitivanja

equipped with a kremear shear head. Samples were read at 500 kg load cell and50 range.

Data Analysis / Analiza podataka

The data of the meat quality variables were analysed by the SPSSŠ17¹, using the analysis of variance (ANOVA).

Results of the effect of pre slaughter handling (PSH) during ageing onthe meat parameters are presented in Tables 1-4.

Table 1. Effect of pre-slaughter handling and ageing on the pH of meat in MarmaraKivircik lambs /

Tabela 1. Efekat rukovanja pre klanja i zrenja na pH mesa kod Marmara Kivircik jagnjadi

GroupsParameters /

GrupeParametri

C(n=30)x ± s

HO(n=30)x ± s

HB(n=30)x ± s

F-value /F-vrednost

0 minute /0 minuta

7.05 ± 0.050 7.04 ± 0.047 7.03 ± 0.064 2.024ns

45th minute /45 minuta

6.91 ± 0.040a 6.74 ± 0.053c 6.79 ± 0.045b 104.357***

2nd hour /2 sata

6.79 ± 0.050a 6.55 ±0.057c 6.60 ± 0.052b 165.498***

3rd hour /3 sata

6.68 ± 0.060a 6.38 ±0.076c 6.43 ± 0.096b 119.686***

4th hour /4 sata

6.57 ± 0.060a 6.22 ±0.093b 6.26 ± 0.080b 172.557***

8th hour /8 sati

6.33 ± 0.080a 5.91 ± 0.108b 5.83 ± 0.048c 311.081***

12th hour /12 sati

6.05 ± 0.100a 5.70 ± 0.107b 5.64 ± 0.036c 198.139***

24th hour /24 sata

5.83 ± 0.040a 5.79 ± 0.049b 5.79 ± 0.034b 7.582***

48th hour /48 sati

5.92 ± 0.030a 5.89 ± 0.024b 5.86 ± 0.031c 28.508***

72th hour /72 sata

6.00 ± 0.030a 5.98 ±0.027b 6.02 ± 0.044a 11.691***

***: P<0.001 ns: P>0.05

157


Results and discussion / Rezultati i diskusija

Table 2. Meat colour measurement with Minolta /Tabela 2. Merenje boje mesa koriste}i Minoltu

GroupsParameters /

GrupeParametri

K(n=30)x ± s

HS(n=30)x ± s

B(n=30)x ± s

F-value /F-vrednost

L 64.17 ± 2.283a 43.93 ± 2.601c 49.02 ± 4.189b 112.659***

a 16.13 ± 1.169c 18.56 ± 1.077b 21.27 ± 3.041a 16.838***

b 14.53 ± 1.211a 8.92 ± 1.196c 12.72 ± 2.418b 28.142***

***: P<0.001

Table 3. Water holding capacity – WHC measurement /Tabela 3. Kapacitet zadr`avanja vode – merenje WHC

GroupsParameters /

GrupeParametri

K(n=30)x ± s

HS(n=30)x ± s

B(n=30)x ± s

F-value /F-vrednost

24th hour (%) /24. sat (%)

14.10 ± 0.241a 13.83 ± 0.233b 13.80 ± 0.174b 16.685***

5th day (%) /5. dan

17.27 ± 0.264 17.40 ± 0.285 17.41 ± 0.280 2.399ns

8th day (%) /8. dan

20.46 ± 0.355b 23.50 ± 0.292a 23.57 ± 0.301a 939.434***

11th day (%) /11. dan

23.67 ± 0.396a 17.06 ± 0.296b 17.01 ± 0.269b 4154.935***

***: P<0.001 ns: P>0.05

Table 4. Shear force measurement with Instron Texture Analyser (Newton) /Tabela 4. Merenje meko}e mesa koriste}i Instron instrument za analizu teksture (Newton)

GroupsParameters /GrupeParametri

K(n=30)x ± s

HS(n=30)x ± s

B(n=30)x ± s

F-value /F-vrednost

MLD 1801.90 ± 304.942 1705 ± 374.585 1686.4 ± 422.729 0.280ns

ns: P>0.05

158


pH / pH

The values of pH 45 were similar to those found by Paulick et al. Š14¹.During the first 72 hours post mortem except pH 0, pre-slaughter handling affectsthe decrease in pH (Table 1). However other studies in lambs Š19¹ showed no sig-nificant differences in the pH 0, pH 45 and pH 24 with the stunning method. Head-only electrical stunning method had an effect on the final pH of the muscle. From45 minutes onwards significant differences between the different pre-slaughterhandling groups were found in pH values. In the control group the lowest valuewas reached at 24 hours post mortem. On the other hand in the other two groupsthe lowest value was reached at 18 hours post mortem. Our results agree withthose of Vergara and Gallego Š20¹ and Paulick et al. Š14¹, pH decreases more rap-idly, and ageing starts earlier. Petersen and Blackmore Š15¹ found that the in-creased muscle activity in animals stunned electrically is probably responsible fortheir more rapid post-mortem pH decline.

Meat colour / Boja mesa

Meat of the non stunned lambs was brighter (higher L* values) thanthat of electrically stunned groups and the difference between the groups was sig-nificant statistically. Also pre slaughter handling affected a* and b* values (Table2). In similar, studies on domestic birds show that electrically stunned turkeys Š5¹and broilers Š11¹ have redder meat (higher a* values) than those stunned withCO2.

Water holding capacity / Kapacitet zadr`avanja vode

Significant differences were found between the pre slaughter handlinggroups in WHC values except on days 5 (Table 3). In all groups (C, HO and HB),WHC decreased (more water expelled) with storage (14.10 ± 0.241, 13.83 ±0.233 and 13.80 ± 0.174% at 24 h post mortem to 20.46 ± 0.355, 23.50 ± 0.292,23.57 ± 0.301 at 8 days post slaughter, for C, HO and HB, respectively). Thisagrees with the study of Koohmaraie, Whipple and Crouse Š10¹, Moore and YoungŠ12¹ and Vergara and Gallego Š20¹. In the C group the highest percentage of waterexpelled was at 11 days postmortem and 3 days earlier in the HO and HB groups.The tendency to release more water in the C group after 8 days post mortem sug-gests their initial juiciness could be greater.

Shear force / Meko}a mesa

The meat from stunned animals appeared more tender than that of thenon-stunned animals, although the differences were not significant (Table 4). Thedifferences in pH (Table 1) may be the cause of the higher values of shear force inmeat of the non-stunned animals group, due to decreased activity of calpain Š3¹.Another study suggests the activity of calpain depends not only on the amount of

159


enzyme present, but also on the muscle pH Š13¹. Vergara and Gallego Š20¹ did notfind any differences between the stunned and non-stunned lamb meat shear forcevalue.

The effect on meat quality was assessed in head-only electricallystunned, head-to-back electrically stunned and non-stunned lambs. Shear forceswere not significantly different between treatments. However colour (L*,a*,b*),WHC, muscle ultimate pH were found to be significantly higher (P<0.05) betweenthe groups. It is concluded that some of the meat quality parameters are affectedby electrical stunning methods.

ACKNOWLEDGEMENT:This work was supported by Research Fund of the Istanbul University. Project numberT-80/23072002.

1. Anil M. H., McKinstry J. L., Wotton S. B.: Electrical stunning and slaughter ofpigs: Guidelines for good welfare assurance. Fleischwirtschaft International, 3, 8-13, 1997.- 2. Devine C. E., Graafhuis A. E., Muir P. D., Chrystall B. B.: The effect of growth rate and ulti-mate pH on meat quality of lambs. Meat Science, 35, 63-77, 1993. - 3. Dransfield E.: Model-ling post-mortem tenderisation- V: Inactivation of calpains. Meat Science, 37, 391-409,1994. - 4. EU Council Directive 93/119. On the protection of animals at the time of slaughteror killing. Official Journal of European Community, 1340/21, 1993. - 5. Fleming B. K., Fron-ing G. W., Beck M. M., Sosnicki A. A.: The effect of carbon dioxide as a preslaughter stun-ning method for turkeys. Poultry Science, 70, 2201-2206, 1991. - 6. Gilbert K. V., Devine C.E.: Effect of electrical stunning on petechial haemorrhages and on the blood pressure oflambs. Meat Science, 7, 197-207, 1982. - 7. Gilbert K. V., Devine C. E., Hand R., Ellery S.:Electrical stunning and stillness of lambs. Meat Science, 11, 45-58, 1984. - 8. Gregory N.G., Wotton S. B.: Sheep slaughtering procedures. I. Survey of abattoir practise. British Vet-erinary Journal, 140, 281-286, 1984. - 9. Kirton A. H., Frazerhurst L. F., Woods E. G., Chrys-tall B. B.: Effect of Electrical Stunning Method and Cardiac Arrest on Bleeding EfficiencyResidual Blood and Blood Splash in Lambs. Meat Science, 5, 347-353, 1981. - 10. Koohma-raie M., Whipple G., Crouse J. D.: Acceleration of postmortem tenderization in lambs andBrahman-cross beef carcasses through infusion of calcium chloride. Journal of Animal Sci-ence, 68, 1278-1283, 1990. - 11. Mohan Raj A. B., Grey T. C., Audsely A. R., Gregory N. G.:Effect of electrical and gaseous stunning on the carcass and meat quality of broilers. Poul-try Science, 31, 725-733, 1990. - 12. Moore V. J., Young O. A.: The Effects of ElectricalStimulation, Thawing, Ageing and Packaging on the Colour and Display Life of LambChops. Meat Science, 30, 131-145, 1991. - 13. Murachi T.: Calpain and calpastatin. TrendsBiochemistry Science, 8, 167-169, 1983. - 14. Paulick C., Stolle F. A., Von Mickwitz G.: Influ-ence od different methods of stunning on meat quality of sheep. Fleischwirtschaft, 68, 3,

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Conclusion / Zaklju~ak

References / Literatura

264-271, 1988. - 15. Petersen G. V., Blackmore D. K.: The effect of different slaughter meth-ods on the post mortem glycolysis of muscle in lambs. New Zealand Veterinary Journal, 30,195-198, 1982. - 16. Petersen G. V., Carr D. H., Davies A. S., Pickett B. T.: The effect of differ-ent methods of electrical stunning of lambs on blood pressure and muscular activity. MeatScience, 16, 1-15, 1986. - 17. SPSS. SPSS for Windows: advanced statistics release 9.0.1.Chicago, USA:SPSS, 1999. - 18. Velarde A., Gispert M., Diestre A., Manteca X.: Effect ofElectrical Stunning on Meat and Carcass Quality in Lambs. Meat Science, 63, 35-38, 2003. -19. Vergara H., Gallego L.: Effect of type suckling and length of lactation period on car-casss and meat quality in intensive lamb production systems. Meat Science, 52, 2, 221-226, 1999. - 20. Vergara H., Gallego L.: Effect of electrical stunning on meat quality of lamb.Meat Science, 56, 345-349, 2000.

EFEKTI RAZLIÎTIH METODA ELEKTRI^NOG OMAMLJIVANJA NA KVALITET MESAKOD MARMARA KIVIRCIK JAGNJADI U TURSKOJ

S. K. Bujukunal, B. Nazli

Efekti metoda elektri~nog omamljivanja samo glave pore|eni su sa efektimaelektri~nog omamljivanja od glave do le|a. Izdvojeno je nasumi~no ukupno 90 jagnjadiKivircik rase neposredno pre klanja radi sprovo|enja jednog od tri vrste tretmana omamlji-vanja: kontrolna grupa (C), grupa gde je tretirana samo glava (HO; 1,0 A-AC za 3 s prifrekvenciji of 50 Hz), grupa tretmana od glave do le|a (HB; 1,0 A-AC za 3 s pri frekvenciji od50 Hz) elektri~nim omamljivanjem. Kvalitet mesa je ocenjen ispitivanjem pH, boje kao vred-nosti L, a, b, kapacitet zadràvanja vode (WHC), i meko}e mesa.

Efekat na kvalitet mesa je procenjen kod jagnjadi koja su elektri~noomamljena u predelu glave, u predelu od glave do le|a, i kod jagnjadi koja nisu omamlji-vana. Meko}a mesa se nije zna~ajno razlikovala me|u tretmanima. Me|utim, boja (L, a, b),kapacitet zadràvanja vode (WHC) i krajnji pH mi{i}a su se razlikovali po grupama, od-nosno bili su zna~ajno vi{i (p<0,05).

Klju~ne re~i: jagnje, elektri~no omamljivanje, kvalitet

161


SRPSKI

ÕFFEKTÀ RAZLI^NÀH METODOV ÕLEKTRI^ESKOGO OBOL[ENIÂ NAKA^ESTVO MÂSA U MARMARA KIVIRICIK ÂGNÂT V TURCII

S. K. Biyikinal, B. Nazli

Õffektì metodov Ìlektri~eskogo obolçeniÔ tolÝko golovì sranenìs Ìffektami Ìlektri~eskogo obolçeniÔ ot golovì do spini. Nami vìdelenonaobum sovokupno 90 ÔgnÔt Kiviricik porodì neposredstvenno do uboÔ radi prove-deniÔ odnogo iz trÒh vidov le~eniÔ obolçeniÔ: kontrolÝnaÔ gruppa (K), gruppa, gdele~ena tolÝko golova (NO; 1,0 A-AS za 3 s pri ~astote ot 50 Hz) Ìlektri~eskimobol{eniem. Ka~estvo mÔsa oceneno ispìtaniem pH, cveta v ka~estve stoimosti L,a, b, moçnostÝ zader`ki vodì (MZV), i mÔgkosti mÔsa.

Õffekt na ka~estvo mÔsa ocenen u ÔgnÔt, Ìlektri~eski obolçÒniìe vpredelah golovì, v predelah ot golovì do spinì, i u ÔgnÔt, ne obolçenì. MÔgkostÝmÔsa ne zna~itelÝno razli~alasÝ me`du le~eniÔmi. Me`du tem, cvet (L, a, b),moçnostÝ zader`ki vodì (MZV) i kraynÔÔ pH mì{c razli~alisÝ po gruppah, to estÝbìli zna~itelÝno bôlÝ{ie (r<0,05).

KlÓ~evìe slova: ÔgnÒnok, Ìlektri~eskoe obolçenie, ka~estvo

162


RUSSKIY

UDK 636.4.09:614.4:616.988.75

ZNAÂJ USTANOVLJAVANJA INTRAUTERINIH INFEKCIJAVIRUSOM KLASI^NE KUGE SVINJA U SKLOPU PROGRAMA

SUZBIJANJA I ISKORENJIVANJA OBOLJENJA*

SIGNIFICANCE OF DETERMINING INTRAUTERINE INFECTIONS

WITH CLASSICAL SWINE PLAGUE VIRUS WITHIN PROGRAMME

OF CURBING AND ERADICATING THIS DISEASE

Jasna Prodanov, R. Do{en, T. Petrovi}, Diana Lupulovi}, M. Val~i},V. Pola~ek**

Ukoliko se suprasna nevakcinisana krma~a inficira virusom kla-si~ne kuge svinja (KKS) dolazi do nastanka intrauterine infekcije fetusa.Posledica infekcije fetusa je pojava viremije. Distribucija virusa u njiho-vim tkivima je sli~na onoj koja se ustanovljava kod postnatalno inficira-nih svinja. Cilj ispitivanja je bio ustanovljavanje intrauterine (transpla-centarne) infekcije u slu~aju pojave KKS u razli~itim periodima supras-nosti kod nevakcinisanih i vakcinisanih krma~a. U okviru dva slu~ajaKKS kod neimunih suprasnih krma~a materijal za ispitivanja je obuhva-tao organe i tkiva fetusa. U tre}em ispitivanom slu~aju KKS, materijal jeobuhvatao krv prasadi pre sisanja kolostruma, koja su bila poreklom odvakcinisanih krma~a, na farmi gde je dijagnostikovana KKS. Uzorcitkiva i krvi prasadi su ispitivani na prisustvo antigena i specifi~nih an-titela protiv virusa KKS imunoenzimskom (ELISA) tehnikom. Iako su ispi-tivanja obavljena na malom broju uzoraka, dobijeni rezultati name}u pi-tanje mogu}nosti nastanka intrauterine infekcije terenskim virusomKKS kod krma~a vakcinisanih K-sojem virusa KKS. Sindrom krma~e-klicono{e i perzistentne infekcije su glavni oteàvaju}i faktori koje jepotrebno dodatno sagledavati u okviru programa suzbijanja i eradika-cije KKS.

Klju~ne re~i: klasi~na kuga svinja, intrauterina infekcija, eradikacija

163


* Rad primljen za {tampu 4. 7. 2007. godine** Mr Jasna Prodanov, istraìva~ saradnik, mr Radoslav Do{en, istraìva~ saradnik, dr Tama{

Petrovi}, istraìva~ saradnik, mr Diana Lupulovi}, istraìva~ saradnik, Nau~ni institut za vet-erinarstvo "Novi Sad"; dr Miroslav Val~i}, vanredni profesor, Fakultet veterinarske medicine,Beograd; mr Vladimir Pola~ek, VSI Kraljevo

Ukoliko se suprasna nevakcinisana krma~a inficira virusom klasi~nekuge svinja (KKS) dolazi do nastanka intrauterine (transplacentarne) infekcije fe-tusa Š9, 11¹. Posledica infekcije fetusa je pojava viremije. Smatra se da je distribu-cija ovog virusa u njihovim tkivima sli~na onoj koja se registruje kod postnatalnoinficiranih svinja Š6¹. Me|utim, iskustva istraìva~a u zemljama koje ve} vi{egodina nastoje da iskorene KKS, ukazuju na sasvim druga~iju situaciju ukoliko sevakcinisane suprasne krma~e izloè infekciji virusom KKS. Istraìvanja obavljenau Meksiku pokazala su da je i pored primene propisanih programa za suzbijanje iiskorenjivanje, neophodna i kontinuirana kontrola farmi svinja vakcinisanih protivKKS Š3, 4¹. Vakcinacija protiv KKS ne obezbe|uje zapat od infekcije, a razlog za tosu razli~itosti imunolo{kog statusa brojnih kategorija svinja koje se mogu na}i nafarmi Š5, 12¹. Kod postnatalno inficiranih prasadi problem su jedinke razli~itogimunolo{kog statusa sa aspekta kolostralne za{tite Š3, 10¹. Sa druge strane,poseban zna~aj imaju transplacentarne infekcije krma~a u razli~itim periodimasuprasnosti. Kod inficiranih vakcinisanih suprasnih krma~a postoji opasnost odnastanka intrauterine infekcije i pra{enja inficirane, imunotolerantne prasadi kodkojih se registruju klini~ki znaci KKS tek u uzrastu od 1-2 meseca Š12¹. Na ovajna~in, mogu} je nastanak i odràvanje endemske infekcije virusom KKS u vakcini-sanom zapatu svinja Š3, 4¹.

U literaturi nema mnogo podataka vezanih za prou~avanje pojave i is-hoda oboljenja nakon infekcije terenskim sojevima virusa KKS kod suprasnihkrma~a razli~itog imunolo{kog statusa. Cilj na{eg ispitivanja je bio ustanovlja-vanje intrauterine (transplacentarne) infekcije u slu~aju pojave KKS u razli~itimperiodima suprasnosti kod nevakcinisanih i vakcinisanih krma~a.

Materijal za ispitivanja je obuhvatao fetuse u okviru dva ispitivanaslu~aja, gde je na osnovu epizootiolo{ke anamneze i klini~kih simptoma kod ne-imunih suprasnih krma~a postavljena sumnja na KKS. Nakon patomorfolo{kogpregleda ukupno 14 plodova, od svakog su uzorkovana tkiva i organi: slezina, bu-breg, tonzile i mandibularni limfni ~vor. U tre}em ispitivanom slu~aju, materijal jeobuhvatao krv uzorkovanu od 10 prasadi neposredno posle pra{enja odnosnopre sisanja kolostruma, na farmi svinja gde je epizootiolo{ki, klini~ki, patomor-folo{ki i laboratorijski dijagnostikovana KKS. Uzorci tkiva i krvi prasadi su ispitivanina prisustvo antigena virusa KKS imunoenzimskom tehnikom (ELISA), dok suuzorci krvnih seruma prasadi ispitivani na prisustvo specifi~nih antitela protiv vi-rusa KKS (ELISA tehnika).

164

Vet. glasnik 61 (3-4) 163 - 171 (2007) Jasna Prodanov i sar.: Zna~aj ustanovljavanjaintrauterinih infekcija virusom klasi~ne kuge svinja u sklopu programa suzbijanja i ...

Uvod / Introduction

Materijal i metode rada / Materials and methods

U okviru prvog ispitivanog slu~aja, klini~kim i patomorfolo{kim pregle-dom nevakcinisane suprasne krma~e koja je uginula pred pra{enje, ustanovljenesu promene karakteristi~ne za KKS (letargija, krvarenja po koì, ta~kasta krvar-enja po bubrezima, hemoragi~ni limfadenitis). Pregledom uterusa ustanovljeno jeprisustvo 4 ploda u levom i 5 plodova u desnom rogu. U predelu perimetrijuma za-beleèna su petehijalna krvarenja, naro~ito izraèna u predelu desnog roga ma-terice. Kod svih plodova ustanovljen je nalaz otoka potko`nog tkiva (pihtijasto-èlatinozni edem u predelu vrata i podvili~ne regije), univerzalni limfadenitis, bu-brezi sa izraènim subkapsularnim, difuznim krvarenjima u kori i meduli, otok peri-renalnog tkiva, opse`na ekhimoti~na krvarenja po epikardu, visceralnom i parie-talnom listu pleure i plu}ima, kao i izraèna hiperemija mo`danog tkiva. Kod poje-dinih plodova ustanovljena su i ta~kasta krvarenja po koì i potko`nom tkivu, dokje jedan plod u levom rogu bio delimi~no mumificiran.

Pregledom organa i tkiva krma~e i plodova, u svim ispitanim uzor-cima, ustanovljeno je prisustvo antigena virusa KKS (ELISA tehnika). Me|utim,pregledom pripadaju}eg dela posteljice istih, prisustvo antigena virusa KKS nijeutvr|eno u dva ispitana uzorka (tabela 1).

Tabela 1. Pregled tkiva i organa i pripadaju}eg dela posteljice plodova na prisustvoantigena virusa KKS (ELISA tehnika) /

Table 1. Fetus tissue and organs and corresponding part of placenta examined for the presence of CSFviral antigen (ELISA)

Materica / Uterus Broj ploda /Number of fetuses

Organi i tkiva /Organs and tissue

Deo posteljice /Part of placenta

Levi rog materice /Left horn of uterus

1 + +

2 + +

3 + –

4 mumifikacija / mummification

Desni rog materice /Right horn of uterus

1 + +

2 + +

3 + +

4 + +

5 + –

(+) pozitivan nalaz; (-) negativan nalaz / (+) positive finding; (-) negative finding

U drugom posmatranom slu~aju, materijal za ispitivanja su bila po-ba~ena prasad poreklom od obolele krma~e. Epizootiolo{kom analizom je us-tanovljeno da krma~a nije vakcinisana protiv KKS i da je u neposrednom okruè-nju dijagnostikovan primarni izvor infekcije. Klini~ki simptomi oboljenja koji su us-tanovljeni kod krma~e nisu bili karakteristi~ni za KKS (blaga apatija, leànje, slab

165


Rezultati rada / Results

apetit). Neposredno nakon zapaànja inicijalnih znakova oboljenja kod iste kr-ma~e, usledio je poba~aj. Na osnovu analize evidencije o ve{ta~kom osemenja-vanju, ustanovljeno je da je poba~aj nastupio 35-40 dana suprasnosti. Patomor-folo{kim pregledom 6 poba~enih plodova ustanovljen je otok potko`ja, petehi-jalna i ekhimoti~na krvarenja po koì u predelu trupa, perinealne regije i repa. Istotako, krvarenja su ustanovljena i na pripadaju}im delovima posteljice. Tri ploda suu momentu pregleda bila delimi~no macerirana. Pregledom organa ustanovljenasu difuzna krvarenja po bubrezima, petehijalna krvarenja po endokardu i he-moragi~ni limfadenitis. U svim ispitanim uzorcima tkiva plodova ustanovljeno jeprisustvo antigena virusa KKS (ELISA tehnika). êtiri dana nakon poba~aja,krma~a je uginula i patomorfolo{kim pregledom ustanovljen je nalaz difuznihkrvarenja na sluzokoì mokra}ne be{ike, univerzalni limfadenitis, a pregledom or-gana i tkiva ustanovljeno je prisustvo antigena virusa KKS.

U okviru tre}eg ispitivanog slu~aja, KKS je registrovana na farmi svinjakapaciteta do 30 krma~a. Epizootiolo{kom anamnezom je ustanovljeno da jeoboljenje prisutno ve} oko 20 dana u objektu zalu~ene prasadi i javlja se kodjedinki uzrasta koji pribli`no odgovara terminu kada se i obavlja zakonom propi-sana vakcinacija protiv KKS. Na osnovu ustanovljenih klini~kih simptoma i pato-morfolo{kih promena postavljena je opravdana sumnja na KKS, {to je i laborato-rijski potvr|eno ustanovljavanjem antigena virusa KKS u organima i tkivima ug-inule zalu~ene prasadi (ELISA tehnika). Kod krma~a koje su bile sme{tene u ob-jektu prasili{ta i kod prasadi na sisi u momentu kada je dijagnostikovana KKS,klini~kim pregledom nisu ustanovljeni znaci bolesti. Pregledom evidencije nafarmi ustanovljeno je da je u proteklom periodu obavljana povremena kontrola efi-kasnosti vakcinacije odnosno laboratorijski je potvr|eno prisustvo specifi~nih an-titela protiv virusa KKS u krvnim serumima krma~a. Me|utim, analizom podatakao vakcinaciji svake krma~e, ustanovljeno je da su pojedine krma~e vakcinisaneKina (K) sojem virusa KKS tokom suprasnosti. S obzirom na to da je odmah posleustanovljavanja infekcije virusom KKS na farmi svinja sproveden program o suzbi-janju oboljenja predvi|en zakonom, samo kod dve krma~e koje su bile nepos-redno pred pra{enje obavljeno je uzorkovanje krvi iz pup~ane vrpce prasadi (ta-bela 2). Na osnovu uvida u evidenciju o vakcinaciji krma~a, ustanovljeno je da jekrma~a br. 14 vakcinisana 26. dana suprasnosti, dok je krma~a br. 22 vakcinisana24. dana suprasnosti. Isto tako, ustanovljeno je da su obe krma~e jo{ u uzrastupraseta dva puta vakcinisane protiv KKS i jo{ jednom kao nazimice. Pregledomuzoraka krvi prasadi pre sisanja kolostruma, prisustvo antigena virusa je us-tanovljeno u oba legla. Me|utim, kod dva praseta krma~e br. 14 isti nije us-tanovljen. Pregledom krvnih seruma prasadi nije ustanovljeno prisustvo spe-cifi~nih antitela protiv virusa KKS. Do trenutka kada je pristupljeno realizacijine{kodljivog uklanjanja zapata svinja (dva dana), kod krma~a i kod prasadi na sisiporeklom iz dva laboratorijski ispitivana legla, nisu ustanovljeni klini~ki znaci KKS.

166


Tabela 2. Rezultati ispitivanja krvi prasadi pre sisanja kolostruma (ELISA tehnika)Table 2. Results of examined blood of piglets before suckling colostrum (ELISA)

Oznaka krma~e /Sow number

Oznaka praseta /Piglet number

Antigen virusa KKS /CSF viral antigen

Antitelaprotiv virusa KKS /Antibodies against CSF virus

br. 22 1 + –

2 + –

3 + –

4 + –

5 + –

br. 14 1 – –

2 + –

3 – –

4 + –

5 + –

(+) pozitivan nalaz; (–) negativan nalaz / (+) positive finding; (-) negative finding

Intrauterine infekcije virusom KKS mogu dovesti do razli~itih posle-dica me|u kojima se naj~e{}e navode: poba~aji, mumifikacija plodova, pra{enjemrtve, avitalne i prasadi sa kongenitalnim tremorom, pra{enje na izgled zdrave, aliperzistentno inficirane prasadi i prasadi kod kojih se mogu ustanoviti i antitela pro-tiv virusa KKS Š9, 11¹. Ishod transplacentarne infekcije zavisi od stadijuma gesta-cije i virulencije virusa koji je izazvao infekciju Š2, 6¹. U okviru prvog ispitivanogslu~aja, ustanovljene patomorfolo{ke promene na uterusu i plodovima su usaglasnosti sa promenama opisanim u literaturi: generalizovan subkutani edem iprisustvo velike koli~ine te~nosti u telesnim {upljinama-ascites, hidrotoraks, pete-hijalna krvarenja na koì i organima Š7, 8¹. Smatra se da ukoliko transplacentarnainfekcija nastupi u zadnjoj tre}ini gestacije, prasad uginjavaju in utero Š2¹. Dobijenirezultati ispitivanja potvr|uju stanovi{te drugih autora Š6, 11¹ da je distribucija i us-tanovljavanje virusa KKS u tkivima in utero inficirane prasadi sli~na distribuciji anti-gena u tkivima i organima svinja postnatalno inficiranih virusom KKS. Me|utim,smatra se da ne nastaje infekcija svih fetusa istovremeno, pri ~emu se procenat fe-tusa koji imunolo{ki mogu da odreaguju na virus uve}ava kako se pove}ava sta-dijumom suprasnosti u kom se odigrala infekcija, {to se moè povezati sa matura-cijom imunog sistema Š2, 8¹. Infekcija virusom KKS se {iri preko epiteliohorijalneplacente, zahvata jedan ili vi{e fetusa, sa posledi~nim {irenjem virusa na susedneplodove Š6¹. U okviru na{ih ispitivanja, iako je antigen virusa KKS ustanovljen usvim plodovima, isti nije ustanovljen u delovima posteljice dva ploda. Smatra seda {to se infekcija odigra kasnije tokom suprasnosti, ve}i je broj neinficirane pra-sadi u okviru inficiranih legala Š8¹, ali u literaturi nema dostupnih podataka koji seodnose na ispitivanje placente inficiranih plodova. înjenica da fetusi koji se na-

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Diskusija / Discussion

laze u isto vreme zajedno u uterusu, mogu biti selektivno inficirani, predstavlja jo{jednu od osobitosti infekcije virusom KKS Š2¹.

U drugom ispitivanom slu~aju, iako klini~ki znaci kod krma~e nisu sasigurno{}u ukazivali na infekciju virusom KKS, na osnovu epizootiolo{kih poda-taka i patomorfolo{kih promena kod poba~enih plodova opravdano je postavlje-na sumnja. Smatra se da prenatalno, u ranoj fazi ontogeneze, virus KKS uti~e nadiferencijaciju organa i dovodi do nastanka malformacija, pa infekcije nastaletokom rane suprasnosti, nose ve}i rizik od nastanka teratogenih efekata Š6, 10¹.Malformacije uklju~uju deformitete razli~itih delova tela (glave, rila, udova i repa) iorgana (bubrega, jetre), kao i centralnog nervnog sistema (mikrocefalus, hidroce-falus, cerebelarna hipoplazija) Š6, 7¹. U okviru na{ih ispitivanja makroskopskimpregledom nisu ustanovljeni deformiteti, ve} samo maceracija pojedinih plodova.S obzirom na to da je krma~a pobacila 35-40. dana suprasnosti i da je u svim ispi-tivanim uzorcima tkiva i organa plodova ustanovljen antigen virusa KKS, moè sepostaviti pitanje duìne vremenskog perioda koji je protekao od infekcije krma~edo infekcije njenih plodova. Frey i sar. Š1¹ smatraju da u slu~aju infekcije slabo ilisrednje virulentnim virusom KKS, infekcija fetusa nastaje u periodu od 13-18.dana nakon infekcije krma~e. Fetusi koji su inficirani tokom prvih 45 dana supras-nosti pokazuju ve}e sklonosti ka prenatalnom uginu}u, {to se klini~ki manifestujeabortusom Š2, 6¹. Ukoliko se prihvati ovakav na~in prora~una, moè se pretposta-viti da je infekcija krma~e nastupila oko 22-25. dana suprasnosti.

Svakako da je sa aspekta suzbijanja i iskorenjivanja KKS, najzna~ajnijitre}i ispitivani slu~aj, gde je ustanovljeno pra{enje kongenitalno inficirane pra-sadi, bez klini~kih znakova oboljenja. Malo je podataka u literaturi koji se odnosena prou~avanje sigurnosti i efikasnosti modifikovanih ìvih vakcina protiv virusaKKS kada se on aplikuje tokom graviditeta ali nisu ustanovljeni neèljeni efektiusled preno{enja vakcinalnog virusa na fetuse Š10, 11¹. Pretpostavlja se da K-sojspre~ava nastanak intrauterinih infekcija jer u organizmu vakcinisane jedinkeskoro u potpunosti prevenira replikaciju virusa KKS Š9, 10¹. Ispitivane krma~e, umomentu kada je KKS dijagnostikovana na farmi, nisu pokazivale klini~ke znak-ove oboljenja. Uprkos ~injenici da su krma~e bile vakcinisane, u ispitivanim uzor-cima krvi prasadi u oba legla ustanovljeno je prisustvo antigena virusa KKS u krvi.Isti~e se i nalaz da kod ispitivanih prasadi nije ustanovljeno prisustvo specifi~nihantitela u serumu pre sisanja kolostruma. To nam moè ukazivati da je od mo-menta infekcije proteklo manje od 20 dana Š8¹.

Kod prasadi inficirane in utero sa terenskim sojevima virusa razli~itevirulentnosti, moè se razviti perzistentna infekcija koja se nakon pra{enja karak-teri{e viremijom, kasnom pojavom bolesti, uginu}em u uzrastu od 2-11 meseci ispecifi~nom tolerancijom imunog sistema za virus KKS Š6, 8¹. [to infekcija nastupiu ranijoj fazi gestacije, to je ve}i broj ustanovljenih perzistentnih infekcija kodprasadi Š2¹. Nakon pra{enja, iz organizma perzistentno inficirane prasadi, virus sekontinuirano izlu~uje {to omogu}ava da se infekcija dalje {iri na prijem~ivejedinke Š2, 8¹. Smatra se da maturacija imunog sistema igra va`nu ulogu u pato-

168


genezi nastanka perzistentnih infekcija, jer virus KKS ima afinitet za }elije imunogsistema, {to moè rezultirati u nastanku alteracija imunolo{ke reaktivnosti Š2¹.Imuni sistem fetusa svinje sti~e sposobnost prepoznavanja antigena virusa KKSizme|u 60. i 75. dana gestacije, ali su perzistentne infekcije uspe{no izazvane ipre i nakon razvoja imunokompetentnosti Š8¹. Me|utim, ukoliko nastupi intrau-terina infekcija, neutralizaciona antitela protiv KKS se sinteti{u samo u terminalnojfazi fetalnog razvoja Š9¹. Kongenitalno inficirana prasad su klini~ki prividno zdravaŠ8¹, imunotolerantna i mogu ìveti duì vremenski period bez razvoja imunogodgovora Š6¹ pri ~emu izlu~uju virus u okolinu sve do trenutka kada nastane tzv.kasna pojava bolesti ("late onset"). Ona se karakteri{e konjunktivitisom, anoreksi-jom, febrom, dermatitisom, dijarejom, lokomotornim poreme}ajima i posterior-nom parezom Š2, 8¹. Faktori koji dovode do "okidanja" i pojave klini~kih znakovanakon dugog inkubacionog perioda nisu poznati Š7, 8¹. Ustanovljeni fenomen senaziva sindrom krma~e-klicono{e ("carrier sow-syndrome") Š9¹ i od velikog jezna~aja u epizootiologiji KKS, jer moè biti uzrok perzistentnog odràvanja àri{tainfekcije. U regionima gde je KKS endemski prisutna postoji opasnost da se infici-rana zalu~ena prasad transportuju na druge farme gde mogu biti uzrok pojaveizbijanja novog àri{ta oboljenja. Rezultati epizootiolo{kih ispitivanja koja suvr{ena u Holandiji, ukazuju na to da su kod vakcinisanih krma~a kongenitalne in-fekcije predstavljale problem i bile su uzrok daljeg {irenja KKS jo{ 4 mesecanakon {to je obavljena vakcinacija priplodnih jedinki K-sojem Š12¹. Postojanjeslabo do srednje virulentnih sojeva virusa, sindrom krma~e-klicono{e i perzis-tentne infekcije su glavni oteàvaju}i faktori koje je potrebno dodatno sagledavatiu okviru programa suzbijanja i eradikacije KKS Š4, 8¹.

Iako su ispitivanja obavljena na malom broju uzoraka, name}e se pi-tanje mogu}nosti nastanka intrauterine infekcije terenskim virusom kod krma~avakcinisanih K-sojem virusa KKS. Dobijene rezultate je svakako neophodno pro-veriti i ispitati u eksperimentalnim uslovima infekcije, jer kada su u pitanju terenskaispitivanja, u tuma~enju rezultata ne mogu biti isklju~eni brojni potencijalni faktorikoji uti~u na pojavu gre{aka pri vakcinaciji. Sa epizootiolo{ke ta~ke gledi{ta,najnepovoljniji ishod intrauterine infekcije je nastanak sindroma krma~e-klicono{e i pra{enje perzistentno inficirane, na izgled zdrave prasadi koja konti-nuirano izlu~uje virus. U ovakvim slu~ajevima klju~ni faktor u eradikaciji je rano ot-krivanje klicono{a.

1. Frey H. R., Liess B., Richter-Reichhelm H. B., Benten K., Trautwein G.: Ex-perimental Transplacental Transmission of Hog Cholera Virus in Pigs. I. Virological and Se-rological Studies. Zbl. Vet. Med. B, 27, 154-164, 1980. - 2. Liess B.: Persistent infections ofhog cholera: a review. Preventive Veterinary Medicine, 2, 109-113,1984. - 3. Morilla A., Car-

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vajal M. A.: Experiences with Classical Swine Fever Vaccination in Mexico. In:Morilla A.,Yoon K. Y., Zimmerman J.J.,eds. Trends in Emerging Viral Infections of Swine,Ames, IowaState Press,162-163, 2002. - 4. Morilla A., Rosales C.: Reemergence of classical swine fe-ver virus in Mexico. In: Morilla A., Yoon K.Y., Zimmerman J.J.,eds. Trends in Emerging ViralInfections of Swine,Ames, Iowa State Press, 149-152, 2002. - 5. Prodanov J., Do{en R.,Val~i} M., Pola~ek V., Petrovi} T., Lazi} S.: Ispitivanje uticaja kolostralnih antitela na razvojpatomorfolo{lih promena posle eksperimentalne infekcije prasadi virusom klasi~ne kugesvinja. Vet.Glasnik 60, 5-6, 323-335, 2006. - 6.Terpstra C.: Hog Cholera: An update of pres-ent knowledge. Br. vet. J. 147, 397-406, 1991. - 7.Trautwein G.: Pathology and pathogene-sis of the disease. In: Classical Swine Fever and Related Viral Infections. Ed: Liess, B.Martinus Nijhoff Publishing, Dordrecht, 27- 49, 1988. - 8. van Oirschot J. T.: Persistent andinapparent infections with swine fever virus of low virulence. Their effects on the immunesystem. Ph.D. Thesis.Universiteit Utrecht, 5-30, 1980. - 9. van Oirschot J. T.: Classical swinefever. In Diseases of Swine, Ed: Straw, B. E., D'Allaire, S., Mengeling, W. L., Taylor, D. J.,Ames, Iowa State University Press, 159-172, 1999. - 10.van Oirschot J. T.: Vaccinology ofswine fever: from lab to field. Vet. Microbiol. 96, 367-384, 2003. - 11.van Oirschot J. T., Terp-stra C.: Hog Cholera Virus. In: Virus infections of porcines. M. B. Pensaert, ed.; New York,Elsever Science Publishers, 113-130, 1989. - 12. Tersptra C., Robijns K. G.: Experience withregional vaccination against swine fever in enzootic areas for limited periods using C-strainvirus. Tijds. Diergeneesk.102, 106-112, 1977.

SIGNIFICANCE OF DETERMINING INTRAUTERINE INFECTIONS WITH CLASSICALSWINE PLAGUE VIRUS WITHIN PROGRAMME OF CURBING AND ERADICATING

THIS DISEASE

Jasna Prodanov, R. Do{en, T. Petrovi}, Diana Lupulovi}, M. Val~i}, V. Pola~ek

Intrauterine infection of the fetus occurs if a pregnant non-vaccinated sow isinfected with the virus of classical swine plague (CSF). The infection of the fetus results inthe occurrence of viremia and the distribution of the virus in fetal tissue is similar to the dis-tribution which is established in post-natally infected swine. The objective of these investi-gations was to determine intrauterine (transplacental) infection in the event of the appear-ance of CSF in different periods of pregnancy in non-vaccinated and vaccinated sows. Theexamined material were organs and tissue of fetuses within two examined cases of CSF innon-immune pregnant sows. In the third examined case of CSF, the material comprised theblood of piglets before suckling the colostrum, animals originating from vaccinated sows,at a farm in which CSF had been diagnosed. Samples of tissue and blood of the pigletswere examined for the presence of antigens and specific antibodies against the CSF virususing the immunoenzyme technique (ELISA). Even though the investigations were per-formed on a small number of samples, the obtained results raise the question of the possi-bility of the occurrence of intrauterine infection with a CSF field virus in sows vaccinatedwith the C-strain of CSF. The syndrome of a carrier sow and persistent infections are thechief problem factors that need to be considered within the programme of curbing anderadicating classical swine plague.

Key words: classical swine plague, intrauterine infection, eradication

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ENGLISH

ZNAÊNIE USTANAVLIVANIÂ VNUTRIMATO^NÀH INFEKCIY VIRUSOMKLASSIÊSKOY ÛMÀ SVINEY V SOSTAVE PROGRAMMÀ PREODOLENIÂ

I ISKORENENIÂ ZABOLEVANIÂ

Âsna Prodanov, R. Do{en, T. Petrovi~, DiÔna Lupulovi~, M. Val~i~,V. Pola~ek

NaskolÝko suporosnaÔ nevakcinirovannaÔ svinomatka inficiruetsÔvirusom klassi~eskoy ~umì sviney (K^S) prihodit do vozniknoveniÔ vnutri-mato~noy infekcii ploda. Posledstvie infekcii ploda Ôvlenie viremii i dis-tribucii virusa v ih tkanÔh podobna distribucii, ustanavlivaemaÔ u postna-talÝno inficirovannìh sviney. CelÝ ispìtaniÔ bìla ustanavlivanie vnutri-mato~noy (transplacentarnoy) infekcii v slu~ae ÔvleniÔ K^S v razli~nìh perio-dah suporosnosti u nevakcinirovannìh i vakcinirovannìh svinomatok. MaterialdlÔ ispìtaniÔ ohvatìval organì i tkani ploda v ramkah dva ispìtìvanih slu~aÔK^S u neimmunnìh suporosnìh svinomatok. V tretÝem ispìtìvannom slu~ae K^S,material ohvatìval krovÝ porosÔt do sosaniÔ kolostruma, kotorìe bìli prois-ho`deniem ot vakcinirovannìh svinomatok, na ferme, gde diagnosticirovanaK^S. Obraz~iki tkaney i krovi porosÔt ispìtìvanì na prisutstvie antigenov ispecifi~eskih antetel protiv virusa K^S immunoÌnzimnoy (ELISA) tehnikoy. HotÔispìtaniÔ sdelnì na malenÝkom ~isle obraz~ikov, osuçestvlënnìe rezulÝtatìispìtaniÔ nakladivaÓt vopros vozmo`nosti vozniknoveniÔ vnutrimato~noy in-fekcii virusom mestnosti K^S u svinomatok vakcinirovannìh K-{tamom virusaK^S. Sindrom svinomatki-baccilonositelÔ i persistentnoy infekcii glavnìezatrudnÔÓçie faktorì, kotorìe nado dopolnitelÝno zame~atÝ v ramkah pro-grammì preodoleniÔ i iskoreneniÔ K^S.

KlÓ~evìe slova: klassi~eskaÔ ~uma sviney, vnutrimato~naÔ infekciÔ,iskorenenie

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RUSSKIY

UDK 616-092.9:17.023.35

OSNOVNI PRINCIPI ZA[TITE DOBROBITI OGLEDNIH@IVOTINJA*

BASIC PRINCIPLES OF EXPERIMENTAL ANIMALS WELFARE

PROTECTION

Marijana Vu~ini}**

Eti~ka na~ela za{tite dobrobiti oglednih ìvotinja iz podtipa ki~me-njaka nalaù da se njihova upotreba svede na najmanju mogu}u meru,ne samo iz eti~kih, ve} i iz prakti~nih razloga. Ogledi na ki~menjacimamogu da se sprovode samo ako ne postoje odgovaraju}i alternativnimodeli i ako patnja ìvotinja koje se koriste u ogledima moè da seopravda koristima samog in vivo ogleda. U ovom radu, revijalnog karak-tera, izneti su osnovni eti~ki principi kori{}enja oglednih ìvotinja. Tiprincipi su: "3R pravilo", "pravilo 5 sloboda" i "Solna pravilo". Takozvano

"3R pravilo" odnosi se na zamenu oglednih ìvotinja iz podtipa ki~me-njaka alternativnim ìvotinjskim modelima na nièm stupnju evolutiv-nog razvoja, biljnim vrstama, ìvotinjskim i biljnim kulturama }elija itkiva, izolovanim organima, fizi~kim, mehani~kim, hemijskim mode-lima, kompjuterskim simulacijama, smanjenje broja ìvotinja u ogledu iusavr{avanje ogledne procedure do stepena na kojem je mogu}e pot-puno izbe}i ili umanjiti neprijatna telesna i emocionalna iskustva ìvoti-nja u ogledu. "Solna pravilo" se odnosi na procenu biolo{ke pogod-nosti, ponovljivosti i prihvatljivosti in vivo ogleda i alternativnih animal-nih modela.

Klju~ne re~i: ogledne ìvotinje, dobrobit, za{tita, 3R pravilo, petsloboda, Solna principi

Pod za{titom ìvotinja podrazumevaju se sve aktivnosti ~oveka kojimase {titi fizi~ki, psihi~ki i geneti~ki integritet ìvotinja. Pojam dobrobit ukazuje nastepen do kojeg su uslovi ìvota u kojima ~ovek gaji i pod kojima iskori{}ava ìvo-

173

Uvod / Introduction


* Rad primljen za {tampu 29. 11. 2007. godine** Dr Marijana Vu~ini}, vanredni profesor, Katedra za zoohigijenu, Fakultet veterinarske medi-

cine, Beograd

tinje prilago|eni njihovim potrebama Š1¹. Jedna od upotrebnih kategorija, ~ija do-brobit zavisi isklju~ivo od ~oveka, jesu i ogledne ìvotinje.

Izra~unato je da se svake godine u svetu iskoristi od 75 do 100 milionaoglednih ìvotinja u fundamentalnim, primenjenim istraìvanjima, obrazovanju itestiranju razli~itih proizvoda. Samo u evropskim zemljama, broj oglednih ìvoti-nja koji se iskoristi svake godine u iste svrhe dostiè cifru od 10,7 miliona. Najve}ibroj oglednih ìvotinja pripada podtipu ki~menjaka i koristi se u farmaceutskoj in-dustriji za razvoj novih lekova (23%) i ispitivanje vakcina i drugih biolo{kih prepa-rata (21%). Oko 12% oglednih ìvotinja, koje pripadaju podtipu ki~menjaka, isko-risti se svake godine za prou~avanje malignih oboljenja, oko 9% u testovimatoksi~nosti, 2% u ispitivanjima kardiovaskularnog sistema, 1% u obrazovne svrhe i32% u sve druge svrhe. Od svih vrsta koje pripadaju podtipu ki~menjaka najvi{ese koriste razli~iti laboratorijski sojevi mi{eva (44%) i pacova (33%), ptice (10%),ribe (7%), zamorci (2%), kuni}i (1%), dok udeo svih ostalih vrsta ki~menjaka iznosioko 3% Š2, 3¹. Ciljevi svih delatnosti u kojima se iskori{}avaju ogledne ìvotinje suunapre|enje zdravlja i dobrobiti ~oveka, unapre|enje zdravlja, dobrobiti i proiz-vodnih rezultata doma}ih ìvotinja, pronalaènje boljih i efikasnijih na~ina za{titedivljih ìvotinja, posebno ugroènih vrsta, ekolo{ka za{tita, pronalaènje mnogohumanijih i efikasnijih na~ina kontrole {tetnih organizama, provera efikasnosti ikontrole kvaliteta novosisntetisanih i ve} postoje}ih hemijskih jedinjenja, koja sekoriste u razli~ite svrhe, kao i sticanje novih saznanja iz razli~itih oblasti biologije,humane medicine, veterinarske medicine, hemije, poljoprivrede i sl. Biomedicin-ska istraìvanja se obavljaju na razli~itim ìvotinjama: armadilosima (proizvodnjavakcine za lepru), ma~kama (prou~avanje AIDS-a, oboljenja oka i uha i nervnogsistema), ~in~ilama (oboljenja srednjeg uha i gubitak sluha), psima (hirur{ki zah-vati na srcu, hirur{ki zahvati u ortopediji, operacije kuka i drugih zglobova), lasi-cama (virusne bolesti kao {to je influenca), ribama (vid, maligna oboljenja jetre,bakterijske bolesti, termoregulacija, tumori koè), rakovima (poreme}aji motornekoordinacije koje nastaju kod Parkinsonove bolesti ili sifilisa), zamorcima (deficitvitamina C), mi{evima (kancer, procesi starenja, AIDS, imunolo{ka i geneti~ka is-traìvanja, embriotransfer), ne~ovekolikim primatima (kardiolo{ki poreme}aji, ne-urolo{ki poreme}aji, malarija, le~enje imunolo{kih poreme}aja, podudarnost Rhfaktora, meningitis), golubovima (bolesti srca), svinjama (tretman opekotina, za-mena sr~anih valvula), kuni}ima (transplantacija ro`nja~e, efikasnost lekova kojisniàvaju nivo holesterola, spre~avanje razvoja ateroskleroze), pacovima (na~inile~enja paralize nastale usled o{te}enja nerava razli~ite etiologije, testovi efikas-nosti i kontrole kvaliteta lekova, uticaj ishrane na proces starenja, prou~avanjemehanizama odbacivanja tkivnih transplantata), ovcama (prou~avanje efikasnostiopreme koja pomaè uspostavljaju funkcije respiratornih organa novoro|en~adi,prou~avanje razvoja aterovenoznog {anta), puèvima gola}ima (prou~avanjekratkotrajne i dugotrajne memorije) i dr. Danas postoji posebna multidisciplinarnanau~na grana, koja se zove nauka o oglednim ìvotinjama Š3¹. Ova nauka ima dvacilja. Prvi je usavr{avanje kvaliteta ogleda na ìvotinjama. Drugi je za{tita dobrobiti

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Vet. glasnik 61 (3-4) 173 - 181 (2007) Marijana Vu~ini}: Osnovni principi za{tite dobrobitioglednih ìvotinja

oglednih ìvotinja. U ovoj nauci se koriste znanja o biolo{kim osobinama oglednihìvotinja, uslovima sme{taja za ogledne ìvotinje, geneti~koj i mikrobiolo{koj no-menklaturi, podeli i standardizaciji oglednih ìvotinja, preventivi i tretmanu bolestioglednih ìvotinja, eksperimentalnim procedurama, anesteziji, analgeziji, eutana-ziji, alternativnim eksperimentalnim procedurama i eti~kim osnovama rada saoglednim ìvotinjama. Smatra se da svi ki~menjaci poseduju ose}aje sli~ne~oveku, a potvr|eno je da su im neprijatni emocionalni i telesni ose}aji i stanja,kao {to su bol, patnja, dosada, strah i stres zajedni~ke karakteristike Š1¹. Zato pos-toje osnovna na~ela za{tite oglednih ìvotinja, a to su takozvano "3R" pravilo Š4¹ ipravilo "5 sloboda" Š5¹ o kojima je re~ u ovom radu.

Takozvano "3R" pravilo predstavlja osnovu za{tite dobrobiti oglednihìvotinja. Predloìli su ga jo{ 1959. godine Russell i Burch Š4¹ u svojoj knjizi "Prin-cipi humane eksperimentalne tehnike". Skra}enica, 3R, poti~e od engleskih re~ireplacement – zamena, reduction – smanjenje i refinement – usavr{avanje.

Princip zamene podrazumeva zamenu in vivo ogleda na vertebratimain vivo ogledima na invertebratima i in vitro ogledima. Ovaj princip nalaè daogledi na ìvotinjama iz podtipa ki~menjaka mogu da se izvode samo ako ne pos-toje alternativne metode i ako patnja ìvotinja u ogledu moè da se opravda koris-tima in vivo ogleda. Alternative in vivo ogledima su ogledi na ìvotinjskim vrstamana nièm stupnju filogenetskog razvoja (invertebrati), uglavnom na glistama, in-sektima i mikroorganizmima, izolovanim organima, kulturama tkiva i }elija animal-nog i biljnog porekla, ogledi na biljnim vrstama, kompjuterske simulacije i upo-treba razli~itih hemijskih, fizi~kih i mehani~kih modela Š6, 7¹.

Princip smanjenja broja ìvotinja u ogledima, prvenstveno se postièprimenom principa zamene, a potom kori{}enjem najkvalitetnijih oglednih ìvoti-nja, ako je njihova upotreba neminovna, standardizacijom genotipa i mikrobio-lo{kog statusa oglednih ìvotinja, standardizacijom ogledne procedure, prime-nom najpogodnijih statisti~kih metoda za obradu rezultata ogleda i za testiranjeta~nosti nau~ne hipoteze Š8¹ i kori{}enjem istih ìvotinja vi{e puta u razli~itim og-ledima, odnosno izbegavanjem ogleda koji se okon~avaju `rtvovanjem ìvotinja.Ovo zna~i da je nepotrebno izvoditi vi{e puta isti ogled u svrhu potvrde njegoveponovljivosti, kao {to je i nepotrebno obavljati istraìvanja na ìvotinjama za ~ijerezultate ne moè da se uradi ekstrapolacija na ~oveku. Sa druge strane,potrebno je razvijati nove ex vivo i in vitro istraìva~ke metode, pove}avati bio-tehni~ki kvalitet ogleda i usmeriti rad ka dobijanju mnogo relevantnijih rezultata namanjem broju oglednih ìvotinja.

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"3R" pravilo – Zamena i smanjenje broja ìvotinja u ogledima iusavr{avanje ogledne procedure / "Three Rs" - Replacement,reduction, refinement welfare protection

Princip usavr{avanja ogledne procedure podrazumeva izvo|enje og-leda na ìvotinjama na na~in kojim se do najve}e mere smanjuje patnja ìvotinja, apostiè se primenom principa "5 sloboda" Š9¹.

Princip "5 sloboda" Š9, 10, 11, 12, 13¹ nalaè da se svakoj ìvotinji u ek-sperimentu obezbedi:

– sloboda od gladi i è|i, obezbe|ivanjem dovoljne koli~ine kvalitetnehrane i vode, osim u slu~aju kada je oglednom procedurom kontraidikovano, aline u periodu koji moè da ugrozi dobrobit usled gladi i è|i,

– sloboda od neudobnosti, obezbe|ivanjem kvalitetnog sme{tajnogprostora koji pruà ose}aj fizi~ke i termi~ke udobnosti i ose}aj sigurnosti,

– sloboda od bola, bolesti i povreda, primenom odgovaraju}ih pre-ventivnih, profilakti~kih i kurativnih tretmana,

– sloboda od neprijatnih emocionalnih iskusatava i stanja kao {to sustrah, dosada, patnja i stres, primenom odgovaraju}ih medikamenata koji ìvoti-nju osloba|aju neprijatnih telesnih i emocionalnih iskustava i

– sloboda ispoljavanja prirodnih oblika pona{anja i ostvarivanja soci-jalnog kontakta sa ìvotinjama iste vrste, oboga}ivanjem sme{tajnog prostorastrukturnim, socijalnim, auditornim, vizuelnim, taktilnim, olfaktornim i drugim sti-mulusima.

Da bi se ispo{tovala navedena pravila, od nau~nih radnika koji obav-ljaju oglede na ìvotinjama zahteva se odgovornost, koja se obi~no nadovezujena "3R" pravilo (responsibility, eng. = odgovornost), te ono dobija oblik "4R" pravi-la. Pored toga, radi stimulisanja nau~nika na kori{}enje alternativnih modela u is-traìvanjima, testiranjima i u obrazovanju, osmi{ljeno je i drugo "3R pravilo", takoz-vano "Solna pravilo" Š14¹. Ovo pravilo se odnosi na minimum zahteva koje alterna-tivne metode treba da ispune za in vivo oglede i njegova skra}nica tako|e poti~eod engleskih re~i: relevance – biolo{ka prikladnost (odnosi se na biolo{ku rele-vantnost in vitro ogleda u odnosu na in vivo ogled), reliability – pouzdanost (od-nosi se na biolo{ku pouzdanost in vitro ogleda u odnosu na in vivo oglede, od-nosno na ponovljivost ogleda i rezultata u razli~itim uslovima izvo|enja i izme|urazli~itih oglednih institucija) i regulatory acceptability – regularnost prihvatljivosti(odnosi se na realnu pogodnost in vitro testa za odre|enu vrstu istraìvanja). Istimpravilom moè da se tuma~i i biolo{ka prikladnost, pouzdanost i regularnost invivo ogleda na odre|enim animalnim modelima ukoliko ne postoje odgovaraju}ealternative.

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Primena principa "5 sloboda" u za{titi dobrobiti oglednihìvotinja / Five freedoms in experimental animals welfare protection

Solna principi / Solna principles

Za{titi dobrobiti oglednih ìvotinja doprinose i stroge regulative sad-ràne u zakonima o dobrobiti ìvotinja i zakonskim aktima koji se odnose nanau~ne i obrazovne delatnosti. U ve}ini evropskih zemalja, u kojima postoje zako-ni o dobrobiti ìvotinja, posebno poglavlje zakona posve}eno je i za{titi oglednihki~menjaka koji se koriste u nau~ne i obrazovne svrhe i za testiranje razli~itih pro-izvoda. Ovaj deo zakona obavezuje potencijalnog korisnika oglednih ìvotinja daeti~kom komitetu, koji se osniva na institucionalnom, regionalnom i nacionalnomnivou, preda zahtev za izdavanje odobrenja za kori{}enje oglednih ìvotinja izpodtipa ki~menjaka. Preduslov je da osoba koja traì odobrenje za ogled, pose-duje dozvolu za rad sa oglednim ìvotinjama, koja se sti~e poha|anjem stru~nihkurseva a obnavlja se svake godine ili svake tre}e godine kroz kontinuiranuedukaciju o oglednim ìvotinjama Š15¹. Zahtev za izdavanje dozvole za kori{}enjeoglednih ìvotinja treba da sadrì slede}e informacije: cilj i relevantnost ogleda,kompetentnost i iskustvo istraìva~a ili edukatora i ostalog osoblja koje u~estvujeu radu sa oglednim ìvotinjama, obrazloènje za{to se ne koriste alternativne me-tode ve} se ogled izvodi na ki~menjacima, obrazloènje razloga za{to su za ogledizabrane ba{ one vrste ìvotinja za koje se traì dozvola za rad, poreklo oglednihìvotinja, odnosno izvori njihove nabavke, uslovi sme{taja, nege, ishrane i napa-janja ìvotinja u toku ogleda i po zavr{etku ogleda, detaljan opis ogledne proce-dure (vrsta ogleda, u~estalost tretmana ìvotinja u ogledu i trajanje ogledne pro-cedure), pretpostavljeni nivo neprijatnosti ogledne procedure za ìvotinje, upo-treba analgetika i anestetika u ogledu ili drugih metoda i sredstava radi smanjenjastepena neprijatnih emocionalnih i telesnih iskustava (bol, strah, stres, patnja isl.), brzina oporavka ìvotinja posle ogleda i mogu}nost kori{}enja istih ìvotinja uogledima iste ili druge vrste, obrazloènje da li je i za{to je potrebno humano`rtvovanje ìvotinja (kada i kako) i kategorija invazivnosti ogleda na ìvotinjama. Uzavisnosti od stepena naru{avanja fizi~kog, psihi~kog ili geneti~kog integritetaìvotinja, ogledi na ìvotinjama svrstani su u slede}ih 5 kategorija invazivnostiŠ16¹:

– "A" - ogledi na neìvom materijalu, ìvim izolatima (}elijske i tkivnekulture) i na invertebratima.

– "B" - ogledi na ìvotinjama koji ne prouzrokuju ose}aj nelagodnostikod ìvotinja ili je ovaj ose}aj slabog intenziteta. Primeri ovog stepena invazivnostisu eksperimenti na invertebratima sa kompleksnim nervnim sistemom, is-traìvanja na kratkotrajno i ve{to obuzdanim ìvotinjama radi posmatranja i fi-zi~kog ispitivanja (prou~avanje eksterijera, stanja uhranjenosti, morfolo{kog sklo-pa i sl.), intravenska, potko`na, intramuskularna, intraperitonealna, peroralna, in-tratorakalna i intrakardijalna (kategorija "C") aplikacija netoksi~nih materija, akut-na istraìvanja na potpuno anesteziranim ìvotinjama koje se na kraju ogleda`rtvuju eutanazijom, a u toku ogleda ne mogu do}i svesti, tj. prou~avanje trenutne

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Kategorije invazivnosti ogleda na ìvotinjama / Categories ofinvasiveness in animal experiments

smrti, demonstriranje metoda eutanazije posle brzog uvo|enja u besvesno sta-nje, kao {to je predoziranjem anestetika ili dekapitacija posle sedacije ili lake ane-stezije i ogledi koji se zasnivaju na kratkotrajnom gladovanju i èdnovanju ìvoti-nja u vremenu koje odgovara duìni apstinencije u prirodnim uslovima.

– "C" - eksperimenti koji prouzrokuju minimalni stepen stresa ili bola ilisamo kratkotrajni bol. U ovu kategoriju spadaju istraìvanja na ki~menjacima zas-novana na kanulaciji ili kateterizaciji krvnih sudova ili telesnih duplji pod anestezi-jom, male hirur{ke intervencije pod anestezijom, biopsija, laparoskopija, kratko-trajno obuzdavanje radi jednostavne opservacije ili druge vrste ispitivanja, ali kojesu povezane sa minimalnim stresom, kratkotrajni period uskra}ivanja hrane ivode, ali koji prevazilazi period prirodne apstinencije, eksperimenti radi prou~a-vanja pona{anja na svesnim ìvotinjama koje su kratkotrajno obuzdane tako da jekod njih prouzrokovan minimalan stepen stresa ili su obuzdane primenom nepri-jatnih stimulusa. Posle zavr{enih oglednih procedura "C" kategorije kod ìvotinjane sme da se ispolje znaci anoreksije, dehidracije, hiperaktivnost, somnolentnost,pove}ana potreba da leè, intenzivna vokalizacija, poja~ana agresivnost, auto-izolacija, antisocijalno pona{anje ili automutilacija.

– "D" - eksperimenti koji prouzrokuju zna~ajan stepen bola kod ki~me-njaka. Primeri za to su istraìvanja na ki~menjacima koja se zasnivaju na hirur{kimintervencijama pod op{tom anestezijom, posle kojih je mogu} potpun oporavak,indukcija fiziolo{kih ili morfolo{kih anomalija koje za posledicu imaju bol ili stres,izlaganje ìvotinje neprijatnim stimulusima ~ije je izbegavanje nemogu}e, prolon-girano fizi~ko obuzdavanje u trajanju od vi{e ~asova, indukcija promena u po-na{anju, koje dovode do stresa, kao {to su odvajanje mladun~adi od majke, agre-sivnost, izlaganje predatorima, postupci koji prouzrokuju teàk i ireverzibilan bol iprekid senzomotorne organizacije, primena kompletnog Frojdovog adjuvansa,indukcija radijacione bolesti i sl. Eksperimentalne procedure "D" kategorije nesmeju prouzrokovati teàk klini~ki oblik stresa u ~ijoj simptomatologiji preo-vla|uju nepoèljne promene u pona{anju, odsustvo samonege, dehidracija, ab-normalna vokalizacija, prolongirana anoreksija, cirkulatorni kolaps, ekstremna le-targija, nepokretnost, znaci te{ke lokalne i sistemske infekcije i dr.

– "E" - eksperimenti koji se zasnivaju na prouzrokovanju bola jakog in-tenziteta, blizu ili iznad praga tolerancije, neanesteziranih svesnih ìvotinja. Prime-ri su primena mi{i}nih relaksanata ili paralitika, kao {to su sukcinilholin ili drugi ku-rariformni lekovi koji se koriste samo radi obuzdavanja neanesteziranih ìvotinjapri hirur{kim intervecijama, nano{enje te{kih telesnih povreda, opekotina ili trau-ma neanesteziranim ìvotinjama, testovi na toksi~nost i eksperimentalne infekcijeili druge indukovane promene zdravstvenog stanja koje za posledicu imaju ugi-nu}e, poku{aj indukcije psihoti~kog pona{anja ìvotinja, metodi `rtvovanja ìvoti-nje koji nisu propisani od nadle`ne laboratorijske institucije (npr. upotreba strih-nina), indukcija te{kog oblika nepovratnog stresa ili termi~kog {oka. U ve}inizemalja koje priznaju ovu kategorizaciju izvo|enje postupaka "E" kategorije –zabranjeno je.

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U postoje}u regulativu o za{titi dobrobiti oglednih ìvotinja inkorpori-rani su mnogi savremeni eti~ki principi sa ciljem da se pobolj{a kvalitet svih aktiv-nosti u kojima ~ovek koristi ogledne ìvotinje, da se pove}a odgovornost istraì-va~a i edukatora, da se pobolj{a kvalitet proizvoda koji se ispituju na oglednim ì-votinjama, kao i da se obezbedi dobrobit oglednih ìvotinja. Mada su ovi eti~kiprincipi i razli~ito formulisani svi su potekli iz osnovnih principa, odnosno pravilakoja se primenjuju u za{titi dobrobiti oglednih ìvotinja, a to su: "3R pravilo", "prav-ilo 5 sloboda" i "Solna pravilo". Osim po{tovanjem ovih osnovnih pravila, za{titadobrobiti oglednih ìvotinja postiè se i obaveznom kontinuiranom edukacijomosoblja koje u svojim delatnostima koristi ogledne ìvotinje, sticanjem posebnedozvole za rad sa oglednim ìvotinjama i posedovanje odobrenja od eti~kogkomiteta, koji moè biti osnovan na institucionalnom, regionalnom i nacionalnomnivou, za kori{}enje ìvotinja iz podtipa ki~menjaka u istraìvanjima, testiranjima iedukaciji.

1. Vu~ini} Marijana: Pona{anje, dobrobit i za{tita ìvotinja. VKS, Beograd, 388str., 2004. - 2. Van Zutphen L.F.M.: History of animal use. In: Principles of laboratory animalscience (L.E.M. Van Zutphen, V. Baumans and A.C. Beyen, eds.), Elsevier, Amsterdam, 2-5,2001. - 3. Baumans V.: Rev. sci. tech. Off. int. Epiz. 24, 2, 503-514, 2005. - 4. Russell W.M.S.and Burch R.L.: The Principles of Humane Experimental Technique, London, UK: Methuen,238, 1959. - 5. Brambell Committee: Report of the Technical Committee to enquire into thewelfare of animals kept under intensive livestock husbandry systems. Command paper2836. Her Majesty's Stationary Office, London, 1965. - 6. A. Schuppli Catherine and FraserD.: ATLA 33, 487–500, 2005. - 7. Anonimus: The Three Rs declaration of Bologna. In Pro-gress in the Reduction, Refinement and Replacement of Animal Experimentation. Proceed-ings of the 3rd World Congress on Alternatives and Animal Use in the Life Sciences, Bolo-gna, Italy (ed. M. Balls, A-M. van Zeller, & M.E. Halder), Amsterdam, The Netherlands: El-sevier Science B.V., 15, 2000. - 8. Smith A. J., Allen T.: Animal Welfare 14, 347-359, 2005. - 9.Rusche B.: ALTEX 20, Suppl.1, 63-76, 2003. - 10. Mellor D. J., Reid C. S. W.: Concepts ofanimal well-being and predicting the impact of procedures on experimental animals. In: Im-proving the Well-being of Animals in the Research Environment (ed. R.M. Baker, G. Jenkin& D.J. Mellor), Adelaide, Australia: ANZCCART, 3-18, 1993. - 11. Mellor D. J., Stafford K. J.:Australian Veterinary Journal 79, 762-768, 2001. - 12. Rickard D. M.: ATLA 32, Supplement1, 225–227, 2004. - 13. Wells N., Nicholson J.: ATLA 32, Supplement 1, 417–421, 2004. - 14.OECD: Final report of the OECD workshop on harmonization of validation and acceptancecriteria for alternative toxicological test methods. Organisation for Economic Co-operationand Development, Paris, France, 1996a. - 15. FELASA: Principles ans practice in ethical re-view of animal experiments across Europe. A report prepared by the FELASA WorkingGroup on Ethical Evaluation of Animal Experiments, 2005. - 16. CCAC: Categories of Inva-siveness in Animal Experiments. Canadian Council on Animal Care, Constitution Square,Tower II, 315-350 Albert Street, Ottawa, Ontario, CANADA K1R 1B1, 1991.

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BASIC PRINCIPLES OF EXPERIMENTAL ANIMALS WELFARE PROTECTION

Marijana Vu~ini}

Ethical considerations of animal protection and welfare require that the use ofexperimental animals is limited as much as possible. Animal experiments should only beperformed when no alternative is avaliable and when the benefit of the experiment out-weighs the suffering of the animal. This review paper describes the basic principles for theethical use of experimental animals. These are: "Three Rs rule" (replacement, reductionand refinement), "five freedoms" for animals and "Solna principles". "Replacement" meansthe substitution for conscious living higher animals of insentient material. "Reduction"means reduction in the numbers of animals used to obtain information of a given amountand precision. "Refinement" means any decrease in the incidence or severity of inhumaneprocedures applied to those animals which still have to be used. The "five freedoms" are:freedom from hunger and thirst, freedom from adverse environmental impacts, freedomfrom disease and injury, freedom to exhibit normal behaviour and freedom from adversemental states. "Solna principles" state that tests for regulatory purposes need to reflect thefollowing: biological Relevance (meaningfulness and usefulness of a test for a particularpurpose), Reliability (reproducibility of results within and between laboratories), and Regu-latory acceptability (suitability of a test for risk assessment purposes (human health /envi-ronment).

Key words: experimental animal, welfare, protection, 3Rs rule, five freedoms, Solnaprinciples

OSNOVNÀE PRINCIPÀ OHRANÀ BLAGOSOSTOÂNIÂ OPÀTNÀH @IVOTNÀH

MariÔna Vu~ini~

Õti~eskie na~ela ohranì blagostoÔniÔ opìtnìh ìvtnìh iz podtipapozvono~nìh nakladìvaÓt, ~to ih upotreblenie svesti na naimenÝ{uÓ vozmo`-nuÓ meru, ne tolÝko iz Ìti~eskih, uè i iz prakti~eskih pri~in. Opìtì na poz-vono~nìh mogut provoditÝ tolÝko esli ne suçestvuÓt sootvetstuÓ{ie alÝterna-tivnìe modeli i esli stradanie ìvotnìh, polÝzuemìe v opìtah moèt opravdatÝpolÝzami samogo: in vivo opìta. V Ìtoy rabote, obzornogo haraktera, vìnesenì os-novnìe Ìti~eskie principì polÝzovaniÔ opìtnìh ìvotnìh. Õti principì sutÝ:"3R pravilo", "pravilo 5 svobod" i "SolÔna pravilo". Tak nazìvaemoe "3R pravi-lo" otnositsÔ k zamene opìtnìh ìvotnìh iz podtipa pozvono~nìh alÝteranativ-nìmi ìvotnìmi modelÔmi na niz{ey stupeni ÌvolÓcionnogo razvitiÔ, rasti-telÝnìmi vidami, ìvotnìmi i rastitelÝnìmi kulÝturami kletok i tkaney, izo-lirovannìmi organami, fizi~eskimi, mehani~eskimi, himi~eskimi modelÔmi, ko-pÝÓternìmi similÔciÔmi, umenÝ{enie ~isla ìvotnìh v opìte i usover{en-

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RUSSKIY

ENGLISH

stvovaniem opìtnoy procedurì do stepeni na kotoroy vozmo`no vpolne izbeàtÝili umenÝ{itÝ nepriÔtnìe telesnìe i ÌmocionalÝnìe opìtnosti ìvotnìh vopìte. "SolÔna pravilo" otnositsÔ k obosnovìvaniÓ biologi~eskogo blagopriÔt-stviÔ, povtoritelÝnosti i priemlemosti in vivo opìta i alÝternativnìh ani-malÝnìh modeley.

KlÓ~evìe slova: opìtnìe ìvotnìe, blagosostoÔnie, ohrana, 3R pravilo, pÔtÝsvobod, SolÔna principì

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UDK 636.1.09:616.12-073.7

OSNOVNI PRINCIPI PRIMENE I INTERPRETACIJE EKG-aKOD KONJA*

BASIC PRINCIPLES OF ECG APPLICATION AND INTERPRETATION

IN HORSES

Ljubica Spasojevi} Kosi}**

Tokom sprovo|enja fizikalnog pregleda konja, ukoliko se prilikomauskultacije posumnja na aritmiju, elektrokardiografija omogu}avapostavljanje definitivne dijagnoze. Poznavanje osnovnih principa na ko-jima se bazira elektrokardiografija kao dijagnosti~ka metoda, razume-vanje normalne i patolo{ke elektri~ne aktivnosti }elija srca, kao i ge-neze elektrokardiograma, predstavlja neophodne preduslove za pri-menu elektrokardiografije u klini~koj praksi. Klini~ka elektrokardio-grafija kod konja podrazumeva i poznavanje specifi~nosti normalnogsr~anog ritma kod konja, kao i paramentre normalnog Ekg-a kod konja.Interpretacija elektrokardiograma konja obuhvata izra~unavanjesr~ane frekvencije, procenjivanje sr~anog ritma i detekciju prisustvaabnormalnih kompleksa odnosno aritmija. Ovaj rad opisuje karakteris-tike normalnog elektrokardiograma konja i razmatra kako ih treba inter-pretirati.

Klju~ne re~i: Ekg, konj, interpretacija

Elektrokardiografija kao konvencionalna metoda ispitivanja srca, obu-hvata merenje amplitude i vremena razlike potencijala elektri~ne struje, koja jeproizvedena tokom depolarizacije i repolarizacije sr~anih struktura. Elektrokardio-gram (Ekg) je grafi~ki snimak volta`e proizvedene od strane }elija sr~anog mi{i}a,nastale za vreme njihove depolarizacije i repolarizacije.

Elektrokardiografija, prema tome, meri samo elektri~nu aktivnost srca,dok mehani~ke funkcije sr~ane pumpe i valvula nisu obuhva}ene ovom dijag-nosti~kom metodom. Elektrokardiografija je nezamenljiva u pra}enju ritma srca i

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Pojam / Term

* Rad primljen za {tampu 29. 11. 2007. godine** Ljubica Spasojevi} Kosi}, Departman za veterinarsku medicinu, Poljoprivredni fakultet Novi

Sad

dijagnozi aritmija, ali nema zna~aj u proceni funkcija miokarda i dijagnosticiuve}anja srca Š5¹.

Pored konvencionalne elektrokardiografije, koja predstavlja snimanjeelektrokardiografom ìvotinje u stanju mirovanja, kod konja se name}e potrebakori{}enja kontinuiranog dvadeset~etvoro~asovnog (Holter) monitoringa ŠZuccaE. i sar., 2003¹. Holter monitoring je naro~ito indikovan ukoliko postoje podaci opojavi aritmija, sinkope ili ako aritmija ne moè da bude dijagnostikovana tokomodmora i treninga ili Ekg ispitivanjima posle treninga Š12, 14, 17¹.

Elektrokardiografija se zasniva na generisanju elektri~ne struje odstrane srca, provo|enju tako stvorene struje kroz organizam i registrovanju razlikepotencijala elektri~ne struje od strane aparata (galvanometra elektrokardiografa)pomo}u elektroda koje povezuju odre|ene ta~ke na povr{ini organizma sa apara-tom.

Sr~ane mi{i}ne }elije imaju sposobnost generisanja i {irenja elek-tri~ne aktivnosti. Zahvaljuju}i elektri~noj aktivnosti srca, redosled sr~ane aktiv-nosti i kontrakcije je koordiniran.

Srce kao generator / Heart as "generator". Srce funkcioni{e pod de-lovanjem nadraàja (impulsa, depolarizacionog talasa) koji se stvaraju u samomsrcu, ta~nije u sinoatrijalnom ~voru (SA ~vor), sme{tenom u blizini grani~ne linijeprednje {uplje vene sa desnom pretkomorom. Da bi se objasnilo poreklo na-draàja i na~in njegovog sprovo|enja kroz miokard, potrebno je opisati izvesne''elektri~ne'' karakteristike pojedina~ne }elije miokarda i dva osnovna svojstvasr~anog tkiva: automatizam i sprovodljivost.

]elija je polarizovana kada se nalazi u mirovanju, odnosno kada unu-tra{njost sr~ane }elije ima negativan naboj u odnosu na spolja{nju stranu, tako da}elijski potencijal membrane u mirovanju iznosi -90mV. ]elijska membrana u mi-rovanju je skoro nepropustljiva za Na+ i delimi~no propustljiva za K+. ]elija je de-polarizovana ili aktivna kada je unutra{njost }elije pribli`no +15mV u odnosu naspolja{nju sredinu, {to je posledica iznenadnog pove}anja permeabilnosti mem-brane za Na+. Posledica depolarizacije je kontrakcija }elije sr~anog mi{i}a. Repo-larizacija }elije predstavlja brzo isticanje K+ iz }elije (zbog pove}anja propus-tljivosti membrane za K+) i ima za posledicu vra}anje }elije u stanje mirovanja.

Specijalizovani provodni sistem srca ima dve osnovne funkcije: 1.}elije u specifi~nim regionima sistema imaju svojstvo automatizma i mogu da ini-ciraju sr~anu depolarizaciju; 2. ostatak sistema je odgovoran za provo|enje elek-tri~nih impulsa kroz srce na koordiniran na~in.

Sprovodni sistem srca je odgovoran za postojanje odre|enog tokadepolarizacije srca. ]elije sa pove}anim automatizmom ~ine specijalizovanosprovodno tkivo, koje pokazuje nestabilnost membranskog potencijala sa poste-

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Vet. glasnik 61 (3-4) 183 - 192 (2007) Ljubica Spasojevi} Kosi}: Osnovni principi primenei interpretacije Ekg-a kod konja

Osnovni principi elektrokardiografije /Basic principles of electrocardiography

penom, sporom depolarizacijom idu}i prema kraju ciklusa. Sprovodni sistem srcapo~inje SA ~vorom, nastavlja se interatrijalnim snopovoma (internodalnim traktu-sima), AV ~vorom, Hisovim snopom, desnom i levom granom Hisovog snopa izavr{ava se Purkinjeovim vlaknima u miokardu komora. Impuls se {iri po }elijamasprovodnog sistema srca, a zatim i po radnoj muskulaturi (sincicijumu pretko-mora i komora). Brzina provo|enja impulsa je velika sve do AV ~vora, gde seveoma usporava zbog eferentne aktivnosti vagusa, da bi se potom ponovopove}ala preko Hisovog snopa, njegovih grana i u Purkinjeovim vlaknima.Ovakva specijalizovana tkiva brzo provode elektri~ne impulse kroz srce, tako dase sva sr~ana vlakna sincicijuma pretkomora i komora kontrahuju simultano kakobi se proizvela sna`na kontrakcija ŠEdwards N. J., 1987; Bonagura J. D., Reef V.B., 1998¹.

Autonomni nervni sistem uti~e na procese depolarizacije i repolariza-cije. Parasimpatikus smanjuje aktivnost SA ~vora, pove}ava provo|enje impulsapreko pretkomora skra}enjem trajanja akcionog potencijala pretkomora i uspo-rava provo|enje impulsa kreko AV ~vora. Suprotno ovome, simpatikus pove}avasr~anu frekvenciju i skra}uje atrioventrikularno provo|enje impulsa. Trening kodkonja uti~e na prevagu parasimpatikusne nervna aktivnosti Š13¹. Parasimpatikus-na aktivnost dominira tokom odmora i kod konja ~esto varira sa promenamakrvnog pritiska ŠGedde L.A. i sar, 1965, cit. Bonagura J.D., Reef V.B., 1998¹.Izraèna sinusna aritmija i AV blok drugog stepena koji se ~esto susre}e kod nor-malnih konja prouzrokovani su menjanjem tonusa vagusa i sluè da reguli{u ar-terijski krvni pritisak u mirovanju (odmoru).

Organizam kao provodnik / Organism as conductor. Organizam sepona{a kao homogeni zapreminski provodnik i mesta postavljanja elektrodabitno uti~u na dobijeni snimak.

Geneza elektrokardiograma (odvodi i elektrokardiograf) / Genesis of

electrocardiogram (leads and electrocardiograph). Odvodi predstavljaju elektrode kojepovezuju odre|ene ta~ke povr{ine organizma sa elektrokardiografom. Elektro-kardiograf ne predstavlja ni{ta drugo do galvanometar koji otkriva, poja~ava i reg-istruje promene u voltaì.

Svaki odvod, prema tome, predstavlja pogled na depolarizaciju i repo-larizaciju sr~anih struktura iz razli~itog ugla. Sistem odvoda koji se koristi kodkonja je sli~an onome koji se koristi kod ljudi, pasa i ma~aka, mada se koriste ineki modifikovani odvodi kao {to je odvod baza-apeks srca (vidi tabelu 1).

Kada se talas depolarizacije kre}e prema pozitivnoj elektrodi, koja senalazi na povr{ini tela, pomo}u elektrokardiografa se registruje pozitivan otklon.Negativan otklon se registruje, ako se talas depolarizacije udaljava od ove ta~ke.Otklon se ne pojavljuje ako talas depolarizacije ''putuje'' pod pravim uglom u od-nosu na pomenutu ta~ku.

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Tabela 1. Sistem odvoda kod konja /Table 1. Leads system for horses

Odvodi u frontalnoj ravni / Leads in frontal plane

Bipolarni odvodi / Bipolar leadsOdvod I = levi prednji ekstremitet (+) i desni prednji ekstremitet (–) /Lead I = left front extremity (+) and right front extremity (-)Odvod II = levi zadnji ekstremitet (+) i desni prednji ekstremitet (–) /Lead II = left back extremity (+) and right front extremity (-)Odvod III = levi zadnji ekstremitet (+) i levi prednji ekstremitet (–) /Lead III = left back extremity (+) and left front extremity (-)Unipolarni augmentirani odvodi / Unipolar augmented leadsOdvod aVR = desni prednji ekstremitet (+) u odnosu na levi prednji i zadnji ekstremitet (–) /Lead aVR = right front extremity (+) against left front and back extremity (-)Odvod aVL = levi prednji ekstremitet (+) u odnosu na desni prednji i levi zadnji ekstremitet (–) /Lead aVL = left front extremity (+) against right front and left back extremity (-)Odvod aVF = levi zadnji ekstremitet (+) u odnosu na desni prednji i levi prednji ekstremitet (–) /Lead aVF = left back extremity (+) against right front and left front extremity (-)

Prekordijalni odvodi / Precordial leads

Baza – apeks odvod = levi apeks srca (+) i desni jugularni `ljeb (–) /Base-apex lead = left heart apex (+) and right jugular channel (-)V3 – u blizini apeksa srca (+) /V3 – near heart apex (+)V10 – iznad me|uskapularnog prostora /V10 – above interscapular space

Depolarizacija pretkomora predstavljena je na Ekg-u kao P-talas. Vre-me koje protekne od po~etka depolarizacije pretkomora do po~etka depolariza-cije komora ozna~eno je kao PQ ili PR interval. Ovaj interval se sastoji od P-talasa(period provo|enja impulsa kroz pretkomore) i PR (PQ) segmenta (provo|enjeimpulsa kroz AV ~vor). Depolarizaciju komora elektrokardiograf beleì kao QRSkompleks. QRS kompleks moè normalno da ispolji nekoliko varijacija oblika, {toje posledica ~injenice da razli~iti odvodi snimaju razli~it pogled na tok depolariza-cije komora.

Tokom repolarizacije komora registruje se T-talas sa otklonom prili~nomale visine i obima promene voltaè. Otklon koji predstavlja repolarizaciju pretko-mora (Ta) se ~esto zabeleì kod konja, naro~ito pri brìm frekvencijama ŠBona-gura J.D., Reef V.B., 1998; Bonagura JD, 1985, Bonagura JD, Miller MS, 1985¹.

Tokom sprovo|enja fizikalnog pregleda konja, ukoliko se prilikomauskultacije posumnja na aritmiju, elektrokardiografija omogu}ava postavljanjedefinitivne dijagnoze. Sa druge strane, zna~ajnost elektrokardiografskog nalazamora da se interpretira u svetlu klini~kih simptoma koje ìvotinja ispoljava Š11, 3¹.

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Klini~ka elektrokardiografija / Clinical electrocardiography

Tehnika snimanja Ekg-a / ECG filming technique

Najbolje mesto na kojem se moè napraviti Ekg snimak dobrog kval-iteta je boks u kojem konj boravi, jer je u svom okruènju ìvotinja najmanjeuzbu|ena. U toku snimanja Ekg-a, potrebno je da pacijent ima suv dla~ni pokri-va~ i da se nalazi na izolovanoj suvoj povr{ini. Snimanje se vr{i na ìvotinji kojastoji. Preporu~uje se kori{}enje gumenih prostirki na kojima ìvotinja treba dastoji, da bi se smanjile elektri~ne smetnje. Elektrode se spajaju za koù pacijentapomo}u aligator {tipaljki ili se koriste metalne elektrode sa gumenim kai{evimakoji se zategnu na mesta postavljanja elektroda, prema navedenom sistemu od-voda, koji se koristi kod konja (u tabeli 1 je prikazan sistem odvoda koji se koristikod konja). Da bi se pove}ala sprovodljivost od koè na elektrode, na koù sestavlja elektrodna pasta ili alkohol.

Uobi~ajeno ba`darenje instrumenta se vr{i pri 1 cm = 1 mV, {to zna~ida svaki mali kvadrat od 1 mm na milimetarskom papiru predstavlja 0,1 mV na ver-tikalnoj osovini papira. Pored ovoga, moè da se koristi i druga~ije ba`darenje in-strumenta (1 mV = 2 cm ili 1 mV = 0,5 cm). Preporu~uje se za konje upotrebabrzine od 25 mm/s pri snimanju Ekg-a. Zna~enje upotrebe ove brzine snimanja jeu tome da svaki mali kvadrat od 1 mm predstavlja na horizontalnoj osovini vremeod 0,04 s. Trajanje pojedinih oblika elektri~ne aktivnosti srca moè da se odredibrojanjem kvadrata i mnoènjem njihovog broja sa 0,04s Š5¹.

Interpretacija Ekg-a / Interpreting ECG

Snimak elektrokardiograma treba sistemati~no pregledati, {to podra-zumeva odre|ivanje frekvencije, ritma, odre|ivanje srednje elektri~ne osovinedepolarizacije, merenje parametara Ekg-a i njihovo upore|ivanje sa referentnimvrednostima (tabela 2).

Tabela 2. Vrednosti parametara normalnog Ekg-a kod konja /Table 2. Normal ECG parameter values in horses

Parametar /Parameter

Maksimalna vrednost /Maximal value

Frekvencija / Frequency 28 – 44/min

P talas / P wave 0,16 s

PR interval / PR interval 0,38 – 0,48 s

QRS kompleks / QRS complex 0,14 s

R-talas (I odvod) / R wave (lead I) 0,8 – 1,1 mV

R-talas (II odvod) / R wave (lead II) do 2,2 mV / up to 2.2 mV

QT interval / QT interval 0,575 s

Osovina / Axis 0- 100o

Postoji nekoliko metoda za odre|ivanje sr~ane frekvencije. Najbrìna~in je da se broj 1500 podeli brojem kvadrati}a (na milimetarskoj hartiji Ekg

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trake) izme|u dva susedna R-talasa (ako se za snimanje koristi brzina od25 mm/sec.).

Normalni ritam srca je sinusni ritam, ali kod pojedinih ìvotinjskih vrstapored normalnog sinusnog ritma i drugi ritmovi se susre}u kod potpuno zdravihjedinki. Normalni sinusni ritam je regularni ritam poreklom iz sinusnog ~vora. Nje-gove karakteristike na Ekg-u su postojanje P-talasa, QRS kompelksa i T-talasa zasvaki otkucaj srca. Sinusna aritmija je regularno iregularni ritam i sinhrona je sarespiracijama (ubrzanje frekvencije srca pri inspirijumu i usporavanje frekvencijesrca pri ekspirijumu). Kod konja se pored normalnog sinusnog ritma i sinusne arit-mije, AV blok prvog i drugog stepena, vagusnog porekla smatraju normalnim rit-mom. Sinusna aritmija i AV blok su uslovljeni delovanjem vagusa, pa prema tome,trening ili uzbu|enje vode prelasku u regularni sinusni ritam Š13, 15, 10¹.

Sistem odvoda u frontalnoj ravni treba pregledati da bi se proverilasrednja elektri~na osovina srca (srednja elektri~na osovina depolarizacije iliprose~ni talas depolarizacije). Srednja elektri~na osovina srca se izraàva ustepenima, zato {to svaki odvod nosi odre|enu vrednost stepeni u frontalnoj ravni(odvod I ima vrednost 0o, odvod II – 60o, odvod III – 120o, odvod aVR – (-150o),aVL – (-30o), aVF - 90o). Izra~unati srednju elektri~nu osovinu srca u stvari zna~iprona}i odvod u kome QRS kompleks pokazuje najve}u neto pozitivnu vrednost.Gruba procena pravca osovine depolarizacije komora tj. QRS kopleksa moè seizvr{iti najbrè tako {to se na|e odvod gde QRS kompeks ima najve}u netopozitivnu vrednost. Osovina se pruà ka pozitivnom polu tog odvoda. Osovinasrca je kod `drebadi i jednogodi{naka varijabilna i ~esto je orjentisana kranijalno,dok je kod ve}ine odraslih konja orjentisana levo i kaudalno. Abnormalne devija-cije osovine se susre}u kod kardiomegalije, cor pulmonale, poreme}aja pro-vo|enja impulsa i poreme}aja elektrolita Š19¹.

P-talas Ekg-a je posledica depolarizacije pretkomora. Normalno seproces depolarizacije pretkomora kre}e sa desna na levo i kranio-kaudalno, {to bizna~ilo da bi P-talas trebalo da bude pozitivan u I, II i aVF odvodu. P talas moè dabude nazubljen (oblika slova M), jedinstven, bifazi~an (negativno/pozitivan) ili ~akpolifazi~an. U okviru istog odvoda Ekg-a mogu da postoje razli~ite forme P-talasa ida njihov izgled varira od talasa do talasa. Ovo je normalna, fiziolo{ka pojava kodkonja i ukazuje na lutaju}i pejsmejker. Tokom tahikardije skra}uje se trajanje P ta-lasa, on postaje visok i {iljat i pra}en je izraènom repolarizaciojom pretkomora,{to se na Ekg-u vidi kao vidljivi pretkomorni T-talas (Ta) u okviru PR intervala. Ova~injenica elektrokardiografsku dijagnozu uve}anja pretkomora ~ini veomate{kom. U slu~aju negativno-pozitivnog P-talasa inicijalni vrh je prouzrokovandepolarizacijom srednje i kaudalne tre}ine desne pretkomore, dok je drugi vrhposledica aktivacije pretkomornog septuma i medijalne povr{ine leve pretkomoreŠ2, 4¹.

PR interval se meri od po~etka P-talasa do po~etka QRS kompleksa, aima zna~enje depolarizacije pretkomora i provo|enje impulsa do AV ~vora. Inter-val se menja sa promenom sr~ane frekvencije, i promene PR intervala su u korela-

188


ciji sa promenama krvnog pritiska. Kod konja se javlja fiziolo{ki AV blok prvog ste-pena, odnosno usporeno provo|enje impulsa kroz AV ~vor, pa je odre|ivanjemaksimalnih normalnih vrednosti PR intervala veoma te{ko. Ka{njenje ili ne-mogu}nost provo|enja impulsa sa pretkomora na komore predstavlja atrioven-trikularni blok. Ovakav poreme}aj provo|enja impulsa je klasifikovan kao AV blokprvog, drugog ili tre}eg stepena. AV blok prvog stepena postoji kada PR intervalprevazilazi normalne vrednosti, odnosno kada impuls kasni, ali se jo{ uvek pre-nosi preko AV sprovodnog sistema i aktivira komore, prouzrokuju}i QRS na elek-trokardiogramu. Drugi stepen AV bloka zna~i da se impuls povremeno ne provodiod pretkomora na komore. Ovakav poreme}aj provo|enja impulsa se na elektro-kardiogramu dijagnostikuje pri postojanju P-talasa koga ne slede QRS-T kom-pleksi Š15, 18¹.

Morfologija QRS kompleksa je varijabilna kod konja. QRS komplekspredstavlja depolarizaciju komora i, zavisno od odre|enog odvoda, neke kompo-nente mogu da ne budu prisutne. Ne treba zaboraviti da razli~iti odvodi depolari-zacioni talas ''vide'' iz razli~itog ugla. Kompletna penetracija sprovodnog sistema uzidove obe komore srca kod konja omogu}ava simultanu aktivaciju obe komore.Ovo je i razlog za{to se iz {irine QRS kompleksa i amplitude R zupca kod konja nemo`e izvesti zaklju~ak o dimenzijama leve i desne komore ŠHamlin R.L. i sar.,

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Slika 1. Ekg konja snimljen pri brzini od 25 mm/sec., pri ba`darenju instrumenta od10 mm = 1 mV, u perifernim odvodima (I, II, III, aVR, aVL, aVF) i baza-apeks prekordijal-nom odvodu. Interpretacija Ekg: frekvencija 39/min., ritam sinusni, osovina ~60o, P =0,12 s, PR = 0,28 s (II odvod) i 0,36 s (baza-apeks), QRS<2,2 mV (II odvod), QT =0,48 s. Ekg dg: normalni elektrokardiogram konja /

Figure 1. ECG in a horse recorded at a rate of 25 mm/sec., at instrument calibration of 10 mm = 1 mV, in pe-ripheral leads (I, II, III, aVR, aVL, aVF) and apex base in precordial lead. ECG interpretation: frequency39/min., synus rhythm, axis ~60o, P = 0.12 s, PR = 0.28 s (lead II) and 0.36 s (base-apex), QRS.<2,2 mV(lead II), QT=0.48 s. ECG dg: normal horse electrocradiogram

baza-apeksodvod /base-apexlead

periferniodvodi /Peripheral leads(I, II, III, aVR,aVL, aVF)

RP

Q T

R

PR T

S

1964; Hamlin R.L, Smith C.R., 1965; cit. Bonagura J.D., Reef V.B.,1998¹. Prose~navrednost amplitude R-talasa u odvodu II kod zdravih trka~kih konja je 0,8 – 1,1 mV.Vrednosti preko 2,2 mV amplitude R-talasa u odvodu II i preko 1,7 mV u odvodu I,obi~no predstavljaju abnormalne vrednosti. Povremeno se, me|utim, kod zdravihkonja mo`e nai}i na vrednosti koje prevazilaze navedene grani~ne vrednosti Š4, 7,8¹.

T-talas predstavlja repolarizaciju komora. Vektor-T talasa se ~estoprostire prema pozitivnom polu odvoda III, {to zna~i da se pozitivan T-talas javlja uodvodu III. Potrebno je ista}i da nema jedinstvenog mi{ljenja i stava kada je re~ oelektrokardiografskom zna~enju i vrednosti T-talasa kod konja. Devijacija ST seg-menata kod konja sa hipovolemijom ili {okom ukazuje na ishemiju miokarda, dokse uve}anje T-talasa mo`e razviti kod hipoksije miokarda ili hiperkalijemije. Ek-stremna labilnost T-talasa ~ini interpretaciju promena T-talasa prili~no te{kom.Postoje dokazi da T-talasi menjaju polarnost u zavisnosti od faze treninga, pri~emu talasi u prekordijalnim odvodima postaju pozitivni i za{iljeni (amplituda T- -talasa >1 mV) Š2, 1, 9¹.

QT interval po~inje od po~etka QRS kompleksa i traje do kraja T-ta-lasa, a od malog je klini~kog zna~aja. Interval se skra}uje sa pove}anjem sr~anefrekvencije Š16¹.

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D. W., Gabel A. A.: Effects of training on resting and postexercise ECG in standardbredhorses, using a standardized exercise test. Am J Vet Res, 37, 12, 1485-1488, 1976. - 17.Scheffer C. W. J., Robben J. H., Sloet van Oldruitenborgh-Oosterbaan M. M.: Continuousmonitoring of ECG in horsses at rest and during exercise. Veterinary Record, 7, 371-374,1995. - 18. Vibe-Petersen G., Nielsen K.: Electrocardiography in the horse (A report of find-ings in 138 horses). Nord Vet Med, 32, 3-4, 105-121, 1980. - 19. White N. A., Rhode E. A.:Correlation of electrocardiographic findings to clinical disease in the horse. J Am Vet MedAssoc 164, 1, 46-56, 1974. - 20. Zucca E., Ferrucci V., DiFabio V., Croci C., Ferro E.: The useof electrocardiographic recording with Holter monitoring during treadmill exercise to evalu-ate cardiac arrhythmias in racehorses. Veterinary research communications 27 Suppl. 1,811-814, 2002.

BASIC PRINCIPLES OF ECG APPLICATION AND INTERPRETATION IN HORSES

Ljubica Spasojevi} Kosi}

If an arrhythmia is suspected on cardiac auscultation, electrocardiography willmake posible a definitive diagnosis. Knowledge of the basic principles of the electrocardi-ography as diagnostic method, understanding of the normal and pathologic cardiac cellelectrical activity, as well as genesis of the electrocardiogram, represent the prior conditionfor applied electrocardiography in clinical practice. Clinical electrocardiography in horsesimplies knowledge of the normal equine heart rrhythm and ECG parameters values. Inter-pretation of the electrocardiogram include determination of heart rate and rrhythm and de-tection of the presence of abnormal complexes. This article described caracteristics of thenormal equine ECG and discused their interpretation.

Key words: ECG, horse, interpretation

OSNOVNÀE PRINCIPÀ PRIMENENIÂ I INTERPRETACII ÕKG U LO[ADEY

LÓbica Spasoevi~ Kosi~

V te~enie provedeniÔ fizi~eskogo osmotra lo{adey, naskolÝko priauskulÝtacii usomnitsÔ na aritmiÓ, ÌlektrokardiografiÔ davaet vozmo`nostÝopredelenie okon~atelÝnogo diagnoza. Poznanie osnovnìh principov na kotorìhbaziruetsÔ ÌlektrokardiografiÔ kak diagnosti~eskiy metod, ponimanie normalÝ-noy i patologi~eskoy elektri~eskoy aktivnosti kletok serdca, kak i geneza elek-trokariogrammì, predstavlÔet soboy neobhodimìe predvaritelÝnìe usloviÔ dlÔprimeneniÔ Ìlektrokardiografii v klini~eskoy praktike. Kline~eskaÔ Ìlektro-kardiografiÔ u lo{adey podrazumevaet i poznanie specifi~nosti normalÝnogoserde~nogo ritma u lo{adey, slovno i parametrì normalÝnogo ÕKG-a u lo{adey.

191


RUSSKIY

ENGLISH

InterpretaciÔ Ìlektrokardiogrammì lo{adey ohvatìvaet is~islenie serde~noy~astotì, ocenka serde~nogo ritma i detekcii prisutsviÔ nenormalÝnìh komplek-sov to estÝ aritmiÔ. Õta rabota opisìvaet harakteristiki normalÝnoy Ìlektro-kardiogrammì lo{adey i rassmatrivaet kak ih nado interpretirovatÝ.

KlÓ~evìe slova: ÕKG, lo{adey, interpretaciÔ

192


UDK 639.112.2.09:616.981.48

HISTOPATHOLOGICAL INVESTIGATIONS OF RABBITCOLIBACTERIOSIS, CAUSED BY ENTEROPATHOGENIC

ESCHERICHIA COLI (O15:H-)*

HISTOPATOLO[KA ISTRA@IVANJA KOLIBAKTERIOZE KOD KUNI]A

IZAZVANE ENTEROPATOGENOM BAKTERIJOM ESCHERICHIA COLI

(015:H-)

V. Petrov, M. Alexandrov, S. Lasarova, M. Lyutskanov**

The histopathological alterations in the intestinal tract of recentlyweaned rabbits with experimental and spontaneous Eschercihia coli(O15:H–) infection were followed. A considerable shortening and thick-ening of well epithelized intestinal villi were observed, whose tips, aftera Warthin-Starry staining, were profusely colonized with coliform bacte-ria. The observed pathological pattern was a permanent finding in suchinfections and could be used as a pathognomic feature in the differen-tial diagnosis of spontaneous diarrhoeic syndromes. The adhesion ofcolibacteria to enterocytes, together with the data from the bacteriologi-cal studies (isolation, identification, determination of the O-serogroupaffiliation and the biochemical behaviour) allowed the assignment ofisolates to the group of enteropathogenic E. coli (EPEC). Only when re-quired, more detailed diagnostical procedures as PCR, could be per-formed.

Key words: colibacteriosis, enteropathogenic E. coli (EPEC)

During the last three decades, a number of investigators have empha-sized the role of enteropathogenic Escherichia coli (EPEC) as a leading bacterialagent in diarrhoeic diseases in rabbits during the first post weaning weeks. Thisgroup of colibacteria, categorized as a pathovar, is distinguished with the absence

193


Introduction / Uvod

* Rad primljen za {tampu 12. 7. 2007. godine** Vladimir Petrov, Faculty of Veterinary Medicine, Trakia University, Stara Zagora, Bulgaria; M.

Alexandrov, S. Lasarova, Institute of Experimental Pathology and Parasitology, BulgarianAcademy of Sciences, Sofia, Bulgaria; M. Lyutskanov, Faculty of Veterinary Medicine, TrakiaUniversity, Stara Zagora, Bulgaria

of toxin production and invasiveness. The pathogenic effect of EPEC is realizedthrough alterations in the cytoskeleton of enterocytes Š30, 16, 17¹.

The epidemiology of the disease, the methods of diagnostics, the al-ternatives for control and eradication have been investigated Š18, 4, 23, 24¹.

Via both scanning and transmission electron microscopy, it was evi-denced that the most important micromorphological changes consisted in the ad-hesion of smaller or larger groups of colibacteria on the enterocytes and the M-cells on Payer's patches. As a result of this adherence and the consequentchanges in the cytoskeleton of the intestinal epithelium, the intestinal villi becomeconsiderably shortened and the intestinal mucosa – thinner Š14, 22, 31, 9¹.

This kind of studies are still limited, and in Bulgaria, have not been per-formed up to the present. That is why our objective was to follow the light micro-scopic features of the intestinal tract in rabbits with experimental and spontane-ous E. coli (O15:H-) infection.

Animals and experimental design / @ivotinje i eksperimentalni plan

In this study, 30 rabbits were included, divided into two groups as fol-lows:

Group I included 15 animals with experimental colibacteriosis pro-voked by the highly pathogenic E. coli U83/39 (serotype O15: H-) strain.

Group II consisted of 15 rabbits, offspring of does, experimentally in-fected during a previous experiment with the same E. coli strain. Part of these ani-mals exhibited signs of spontaneous intestinal diarrhoea.

All animals from both groups, participating in the experiment, wereweaned at the age of 30 days and did not shed Eimeria spp. oocysts at the time ofweaning.

E. coli strain / Soj E.coliThe experimental colibacteriosis was induced by an enteropatho-

genic E. coli U83/39 (O15: H-) strain, kindly provided by Dr. J. E. Peeters, from theNational Institute of Veterinary Research, Brussels, Belgium. It served to prepare abacterial suspesion with a density of 3 x 107 c.f.u./ml.

Bacteriological studies / Bakteriolo{ka istra`ivanja

Rectal swab samples were analyzed for detection of the causing agentin the rabbits from group I as well as for the determination of the presence of en-teropathogenic E. coli in the rabbits with spontaneous disease (group II) thatmanifested an intestinal indigestion. The routine bacteriological techniques forisolation and classification of enterobacteria as well as of identification of colibac-teria were utilized Š29¹. The O-serogroup affiliation of E. coli isolates was deter-mined Š29¹.

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Vet. glasnik 61 (3-4) 193 - 200 (2007) V. Petrov et al.: Histopathological investigations ofrabbit colibacteriosis, caused by enteropathogenic Escherichia coli (O15:H-)


Histological studies / Histolo{ka istra`ivanja

After euthanasia of rabbits by the 3rd-4th day of the clinical manifesta-tion of the disease, ileal loops were obtained for a histological study after ligationfrom both ends, filling and placement in 10% buffered formalin for 72-96 h. Fromthem, rings with a height of 5 mm were prepared, embedded in paraffin, 6-8 �msections were cut and stained with haematoxylin/eosin and according to Warthin--Starry Š11¹, using routine histological techniques.

The observations were performed on a transmitted light NU-2 micro-scope.

In animals with experimental E. coli U83/39 (O15: H-) infection, a con-siderable shortening and thickening of well epithelized intestinal villi was ob-served (Fig. 1), whose tips, after Warthin-Starry staining, appeared massivelycolonized with coliform bacteria (Fig. 2) and at numerous sites, their propria wasinfiltrated with pseudoeosinophils (Fig. 3).

In rabbits, offspring of EPEC-infected does, apart from the histopa-thological changes specific for colibacteriosis, a weaker or more severe infectionwith Eimeria spp. was also present (Fig. 4).

195


Results and discussion / Rezultati i diskusija

Figure 1. Ileum of a rabbit with experimental E. coli U83/39 (O15: H-) infection. Stronglyshortened, thickened and well epithelized intestinal villi with hyperaemic propria anddisintegrated epithelium on the tips. Hematoxylin/eosin staining

Slika 1. Ileum kuni}a sa eksperimentalnom infekcijom E.coli U83/39 (O15: H-). Veoma skra}ene, zadebljane i do-bro epitelizovane intestinalne resice sa hiperemi~nom propriom i disintegrisanim epitelom na vrhovima.Bojenje hematoksilin eozinom

According to the present studies in rabbits with experimental andspontaneous E. coli (O15:H-) infection, the histological alterations due to EPECare related to the atrophy of intestinal villi, vacuolization, increased amount of cy-toplasm, impaired structure of the superficial epithelial layer of the intestinal mu-cous coat, presence of desquamated cells in the lumen. These injuries accordingto a number of authors take place gradually and consecutively, occurring initiallyat a cell level Š30, 16, 17¹.

196


Figure 2. Ileum of a rabbit with experimental E. coli U83/39 (O15: H-) infection. An intestinalvillus, colonized by coliform bacteria. Warthin-Starry staining

Slika 2. Ileum kuni}a sa eksperimentalnom infekcijom E. coli U83/39 (O15: H-). Intestinalna resica koloniziranakoliformnim bakterijama. Bojenje po Wartin-Stari metodi

Figure 3. Ileum of a rabbit with experimental E. coli U83/39 (O15: H-) infection. The propria isinfiltrated with pseudoeosinophils. Hematoxylin/eosin staining

Slika 3. Ileum kuni}a sa eksperimentalnom infekcijom E. coli U83/39 (O15: H-). Propria je infiltriranapseudoeozinofilima. Bojenje hematoksilin eozinom

According to Milon (1996) this process occurs in three stages:– Attachment / Vezivanje. Initially, the bacteria are loosely attached to en-

terocytes via plasmid-determined fimbrial adhesins, called bundle forming pili(BFP).

– Effacement / Brisanje. The alterations in the cytoskeleton are resultingfrom the action of several functional proteins, synthetized after the attachment. Anumber of chromosome genes, located adjacently and forming the so-called“pathogenicity island” or "locus of enterocyte effacement" (LEE), are responsiblefor their synthesis.

– Cytoskeleton reorganization / Citoskeletna reorganizacija. The actin under-goes a change from filamentous into globular and this results in an altered entero-cyte cytoskeleton. The change consists in the formation of pedestal-like struc-tures and effacement of the microvilli.

On the other side, it was shown that the changes in membrane pro-teins blocked the transport systems of the colonized enterocyte, impairing the ab-sorption of water, the excretion and absorption of ions (Law 1994). The histopa-thological picture of this process is manifested through the rounding of epithelialcells and subsequent disorganization of the entire superficial layer of enterocytesin affected areas.

197


Figure 4. Ileum of a rabbit with spontaneous E. coli (O15: H-) infection. Distinctly shortened,thickened intestinal villi with single enterocytes, infected with Eimeria spp. (arrows).The tips of the villi are covered with coliform bacteria. Warthin--Starry staining

Slika 4. Ileum kuni}a sa spontanom infekcijom E. coli (015:H-). Jasno skra}ene, zadebljane intestinalne resice sapo jednim enterocitom, inficirani Eimeria spp. (strelice). Vrhovi resica su prekriveni koliformnim bakteri-jama. Bojenje po Vartin-Stari metodi

In some cases, ovoid bacteria on the epithelial surface, but not in thecell's cytoplasm or the deeper layers of the mucous coat, could be observed.

In some parts of the lamina proprià, weak to moderate oedema and in-filtration with pseudoeosinophils was observed. Also, hyperaemia and mucosalhaemorrhages were noticed Š5, 20, 21¹.

Similarly to our studies, after the special Warthin--Starry staining andin other reports in EPEC involvement in rabbits using scanning electron micros-copy, small or bigger clusters of colibacteria, tightly attached to the surface of en-terocytes and particularly on enterocytes, covering the Payer’s patches withoutpresence of diffuse adherence, have been observed Š31, 9, 22, 14¹. It seemed thatthis pathological pattern was a permanent finding in such infections and could beused as a pathognomic feature in field cases, indicative of the need for further di-agnostic studies.

The detection of Eimeria spp. oocysys in some animals is most proba-bly a co-infection, due to the lack of application of coccidiostatics, but it could besupposed that the simultaneous Eimeria spp. infection aggravated the pathologi-cal features of colibacteriosis.

1. Blanco J. E., Blanco M., Blanco J., Mora A., Balaguer L., Cuervo L., Balsalo-bre C., Muñoa F.: Prevalence and characteristics of enteropathogenic Escherichia coli witheae gene in diarrhoeic rabbits. Microbiology and Immunology, 41, 77-82, 1997. - 2. BodonL. and Prohaszka: Isolation of an adenovirus from rabbits with diarrhoea. Acta Vet. Acad.Scient. Hung., 28, 247-255, 1980. - 3. Borriello S. P. and Carman R. J.: Association ofiota–like toxin and Clostridium spiroforme with both spontaneous and antibiotc – associ-ated diarrhea and colitis in rabbits. J. Clin. Microbiol., 17, 414-418, 1983. - 4. CamguilhemR.: Isolement d’une souche d’Echerichia coli (sérogrupe O103) responsible d’entérite coli-bacillaire du lapin en engraissement. Rev. Med. Vet. (Toulouse) 136, 61-68, 1985. - 5. Can-tey J. R. and R.K. Blake: Diarrhea due to Escherichia coli in the rabbit: a novel mechanism.J. Infect. Dis., 135, 454-462, 1977. - 6. Eckert J., Taylor M., Licois D., Coudert P., CatchpoleJ., Bucklar H.: Identification of Eimeria and Isospora species and strains. Morphologicaland biological Characteristics. In: Biotechnology. Guedelines of Techniques in coccidiosisResearch. (Eckert J., Braun R., Shirley M.W., Coudert P., Ed) pp 103-119. Office for officialpuplications of the European communities. Luxemburg, 103-119, 1995. - 7. Finazzi G., Car-deti G., Pacciarini M. L., Losio M., Tagliabul S.: Characterization of strains Escherichia coliisolated from rabbits with enteritis in Lombardia and Emilia-Romagna during 1997-1999. 7th

World Rabbit Congress, 2000. - 8. Fries A. S.: Tyzzer’s diseaseand importance of inappar-ent infecion in biomedical research. Doctoral tesis at the Royal Veterinary and AgriculturalUniversity, Copenhagen, Denmark, 1981. - 9. Inman L. R. and J. R. Cantey: Specific Adher-ence of Escherichia coli (Strain RDEC-1) to Membranous (M) Cells of the Peyer's Patch inEscherichia coli Diarrhea in the Rabbit, The Journal of Clinical Investigation, 71, 1-8, 1983. -10. Jerse A. E., Yu J., Tall B. D., Kaper J. B.: A genetic locus of enteropathogenic Escheri-chia coli necessary for the production of attaching and effacing lesions on tissue culturecells. Proc. Natl. Acad. Sci. USA 87, 7839-7843, 1990. - 11. Kerr D. A.: Improved Warthin-Starry method for tissue sections; Am J Clin Pathol, 8, 63-67, 1938. - 12. Laplance J. P.: Letransit digestif chez les monogastiques. III. Comportement (prise de nourriture – caecotro-phie), motricité et transit digestifs, et pathogénie des diarrhées chez li lapin. Ann. Zootech.

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27, 25-265, 1978. - 13. Law D.: Adhesion and its role in the virulence of enteropathogenicEscherichia coli. Clin. Microbiol. Rev. 7, 152-173, 1994. - 14. Licois D., Reynaud A.,Federighi M., Gaillard-Martinie B., Guillot J. F., Joly B.: Scannig and transmission electronmicroscopic study of adherence of Echerichia coli O103 enteropatogenic and/or entero-haemorrhagic strain GV in enteric infection in rabbits. Infect. Immun., 59, 10, 3796-3800,1991. - 15. Löliger H. C.: Erkrankungen des Darmesbeim Kaninchen. Ursahen und Bekäm-fungsmöglichkeiten. In: Memoria del II congreso mundial de cunicultura, Barcelona. II. Nu-tricion pathologia, 265-282, 1980. - 16. Milon A. Weaned rabbit colibacillosis: a model forstudy of enteropathogenic Escherichia coli (EPEC). 6th World Rabbit Congress, Toulouse,1996. - 17. Nataro J. P. and Kaper J. B.: Diarrheagenic Escherichia coli Clin. Microbiol. Rev.,11, 142-201, 1998. - 18. Okerman L., Lintermans P., Coussement W., Devrise L. A.: Escheri-chia coli ne produisant pas d’enterotoxine comme agents d’entérite chez le lapin avant leseverage. Recl. Méd. Vét. 158, 467-472, 1982. - 19. Peeters J. E.: Coccidioseen zijn bestijd-ing in de industriële konijnenteelt. Proefschrift licentie zoötechinie, RUG, faculteit dier-deneeskunde, Gent., 1-24, 1981. - 20. Peeters J. E., Geeroms R., Glorieux B.: ExperimentalEscherichia coli enteropathy in weanling rabbits; clinical manifestations and pathologicalfindings. J. Clin. Path., 94, 521-528, 1984a. - 21. Peeters J. E., Phol P., Charlier G.: Infectiousagents associated with diarrhoea in comercial rabbits; a field study. Ann. Rech. Vet. 15, 24-29, 1984b. - 22. Peeters J. E., Charlier G. J., Raeymaekers R.: Scannig and transmissionelectron microscopy of attaching effeacing Escherichia coli in weanling rabbits. Vet. Pathol.22, 54-59, 1985. - 23. Peeters J. E.: Etiology and pathology of diarrhoea in weanling rabbitsSeminar in the community programme for the coordination of agriculture research, 6-7 no-vember, Edited by Teresa Auxilia, 127-137, 1986. - 24. Peeters J. E. and Geeroms R.: Traite-ment et eradication de la colibacillose (serotype O15:K-:H-) sur le terrain. Juurnees de larecherenche cunicole 12-13 decembre, Paris. Communication, 1, 1-8, 1990. - 25. PenteadoA. S, Ugrinovich L. A., Corrêa S. da S., Oliveira M. N., de Castro A. F. P., de Ávila F. A.: De-teccão do gene eae em amostras de Escherichia coli (AEEC) isoladas de coelhos de cri-acões do estado se São Paulo, Brasil., Ars Veterinaria, 17, 3, 207-212, 2001. - 26. PenteadoA. S., Urginovich L. A., Blanko J., Blanco M., Blanco J. E., Mora A., Andrade J. R. C., CorrêaS. S., Pestana de Castro A. F.: Serobiotipes and viruilence genes of Escherichia coli stainsisolated from diarrheic and healthy rabbits in Brazil. Vet. Microbiol. 89, 41-51, 2002. - 27.Petric M., Middleton P. J., Grant C., Tan J. T., Hewitt C. M.: Lapine rotavirus: preliminry stud-ies of epizootology and transmission. Can. J. Comp. Med., 42, 143-147, 1978. - 28. PrescotJ. F.: Intestinsl disorders and diarrhoea in the rabbit. Veterinary Bulletine, 48, 457-480, 1978.- 29. Quinn P. J., Carter M. E., Markey B. K., Carter G. R.: Clinical Veterinary microbiology.Harcourt Publishers Limited. 209-237, 1999. - 30. Rosenshine I., Donenberg M. S., Kaper J.B., Finalay B. B.: Signal transduction between enteropathogenic Escherichia coli and epi-telial cells; EPEC induces tyrosine phosphorylation of host cell proteins to initiate cyto-skeletal rearrangements and bacterial uptake. EMBO J. 11, 3551-3560, 1992. - 31.Takeuchi A.: Scannig and transmission electron microscopic study of Escherichia coli O15(RDEC - 1) enteric infection in rabbits. Infect. Immun., 19, 686-694, 1978. - 32. Wescott R.B.: Helmint parasites. In: Biology of the laboratory rabbit. Eds. S. H. Weisbroth, R. E. Flattand A. L. Krauss. Academic Press, New York, pp. 317-326, 1974. - 33. Whitney J. C.: A re-view of non-specific enteritis in the rabbit. Laboratory Animal Science, 10, 209-221, 1976.

199


HISTOPATOLO[KA ISTRA@IVANJA KOLIBAKTERIOZE KOD KUNI]A IZAZVANEENTEROPATOGENOM BAKTERIJOM ESCHERICHIA COLI (015:H-)

V. Petrov, M. Aleksandrov, S. Lasarova, M. Ljutskanov

Pra}ene su histopatolo{ke promene na intestinalnom traktu tek zalu~enih ku-ni}a sa eksperimentalnom ili spontanom infekcijom bakterijom Escherichia coli (O15:H-).Zapaèna su znatna skra}enja i zadebljanja dobro epitelizovanih intestinalnih resica, na~ijim vrhovima su uo~ljive bogato kolonizirane koliformne bakterije, posle bojenja poVartin-Stari metodi.Uo~ena patolo{ka {ema predstavlja konstantni nalaz kod takvih infek-cija i mogla bi se upotrebiti kao patognomska karakteristika kod diferencijalne dijagnozespontanih sindroma dijareje. Adhezija kolibakterija na enterocite, zajedno sa podacima izbakteriolo{kih studija (izolacija, identifikacija, odre|ivanje pripadnosti O-serogrupi i bio-hemijsko pona{anje) omogu}ilo je da se odredi da izolati pripadaju grupi enteropatogeneE.coli (EPEC). Detaljnije dijagnosti~ke procedure, kao {to je PCR, mogu se vr{iti samo uko-liko je neophodno.

Klju~ne re~i: kolibakterioza, enteropatogena E. coli (EPEC)

GISTOPATOLOGIÊSKIE ISSLEDOVANIÂ KOLIBAKTERIOZA UKROLIKOV, VÀZVANNÀE ÕNTEROPATOGENNOY BAKTERIEY

ESCHERICHIA COLI (015:H-)

V. Petrov, M. Aleksandrov, S. Lazarova, M. LÓtskanov

Sleènì gastopatologi~eskie izmeneniÔ na intestinalÝnom traktetolÝko otlu~ennìh ot materi krolikov s ÌksperimentalÝnoy ili spontannoy in-fekciey bakteriey Escherichia coli (015:H-). Zame~enì znatnìe sokraçeniÔ iutolçeniÔ horo{oe Ìpitelizovannìh intestinalÝnìh Ôzì~kov, na ~Ýih verhahzame~enì bogato kolonizirovannìe koliformnìe bakterii, posle kra{eniÔ poVartin-Starìy metodu. Zeme~ena patologi~eskaÔ shema predstavlÔet soboy kon-stantnìe rezulÝtatì u takih infekciy i mogla bì upotrebitÝsÔ kak patologi~es-kaÔ harakteristika u differencialÝnogo diagnoza spontannìh sindromov diarei.AdgeziÔ kolibakteriy na Ìnterocitì, vmeste s dannìmi iz bakteriologi~eskihtrudov (izolÔciÔ, identifikaciÔ, opredelenie prinadle`nosti 0-serogruppe ibiohimi~eskoe povedenie) dalo vozmo`nostÝ opredelitÝ, ~to izolÔtìprinadleàt gruppe Ìnteropatogennoy E. coli (EPEC). Bolee detalÝnìe diag-nosti~eskie procedurì, kak PCR, mogut sover{itsÔ tolÝko poskolÝku neobhodimo.

KlÓ~evìe slova: kolibakterioz, ÌnteropatogennaÔ E. coli (EPEC)

200


SRPSKI

RUSSKIY

UDK 636.7.09:616.98(497.2)

INFECTIONS WITH EHRLICHIA CANIS AND BORRELIABURGDORFERI IN A DOG*

INFEKCIJE BAKTERIJAMA EHRLICHIA CANIS I BORRELIA

BURGDORFERI KOD JEDNOG PSA

I. Tsachev, R. Simeonov, V. Petrov**

A clinical case of Ehrlichia canis and Borrelia burgdorferi infec-tions in a 5year-old male German Shepherd is described. Clinical, sero-logical, necropsy and histopathological examinations supporting thediagnosis have been performed.

Key words: Ehrlichia canis, Borrelia burgdorferi, dog, necropsyfindings, histopathological findings

Canine monocytic ehrlichiosis (CME) is a tick-borne disease, causedby Ehrlichia canis. Ehrlichiae are Gram negative, obligate intracellular bacteriawith a size of 0.5-1.5 µm. A principal vector of their distribution is the brown dogtick Rhipicephalus sanguineus Š1¹. Recently published data from 2006 are un-questionably proving the zoonotic character of the disease, observed amongpeople in Venezuela Š2¹.

CME is nosogeographically related to Africa, Asia, the Middle East,Europe, North and South America. The data for its prevalence in the Mediterra-nean basin are as follow: Italy 13.5%, Greece 21.8%-41.2%, France 46.4%, Israel63%, Spain 2.3-66.7%, Tunisia 72%, Turkey 74% Š3, 4, 5, 6, 7, 8, 9, 10, 11, 12¹.

In Bulgaria, Ehrlichia canis was both clinically and serologically de-tected for the first time by Tsachev in a kennel near Plovdiv Š13¹, and later retro-spectively, in many regions in South and North Bulgaria – Stara Zagora, Yambol,Plovdiv, Burgas, Blagoevgrad, Silistra, Varna. Veliko Tarnovo, Pleven, Montanaand Rousse Š14,15¹.

Lyme borreliosis (LB) is a zoonotic tick-borne, naturally focal, multi-systemic disease with dermatological, neurological, cardiovascular, arthropathic

201


Introduction / Uvod

* Rad primljen za {tampu 12. 7. 2007. godine** Ilia Tsachev, R. Simeonov, Vladimir Petrov, Faculty of Veterinary Medicine, Trakia University,

Stara Zagora, Bulgaria

and other manifestations. Ticks are vectors of the infection, and the principaltransmissor is Ixodes ricinus. This tick species is widely distributed in Bulgaria andaccording to the data of Hristova et al., 2003 Š16¹ it is up to 20% infected with LBagents.

The etiological agent of borreliosis is Borrelia burgdorferi sensu latocomplex. On the basis of DNA/DNA hybridization, ribotyping and 16S ribosomalsequencing of isolates, the strains Borrelia burgdorferi senso stricto, B. afzelii andB. garinii have been specified.

At present, borreliosis in humans and dogs is highly prevalent all overthe world, including in Bulgaria. For 1990-2002, 3261 cases of diseased people in26 regions have been reported Š17¹. For the region of Stara Zagora, the clinicallyrecorded cases for 1998-2002 are 75 Š18¹.

The scientific reports about the incidence of LB among dogs in Bul-garia are very few – only three. The first one describes a clinical case of erythema-tous form in an 8-year-old Boxer Š19¹. The authors isolated the causative agent,performed an antibiogram and treated the dog with amoxicillin and consequently– with doxycycline and sulphonamides. The second report was a seroepidemiol-ogical one and comprised a population of 106 dogs, 499 cows and 512 sheepfrom 13 regions, endemic for human LB. The results showed the highest sero-prevalence in sheep (56.25%) followed by cows (54.31%) and the lowest in dogs –22.6% Š20¹. In a study of 16 dogs using ELISA, Martinov et al., 2006 Š21¹, observeda significant result only in one dog whereas by indirect immunofluorescence –only 2 seropositive out of 46.

In March 2005, a 5-year old, 30-kg German Shepherd named Buck,was referred to a clinic in Stara Zagora. The state of the dog was poor – enhancedheart rate, body temperature of 39.9 oC, adynamia, weakness, depression, apa-thy, tachypnea, dehydratation, anorexia, myalgia, slightly enlarged popliteallymph nodes, pale mucous coats and swelled hindlimbs. A 2-day therapy withglucose, vitamin C, novocaine, degan, dexamethasone and lincospectin was ap-plied. No blood laboratory analysis was done.

Due to the lack of remission by the 3rd day, euthanasia was performedaccording to the owner's will. From the history it became clear that the dog lived inthe settlement of Dve Mogili, a region of Stara Zagora, together with 2 other dogs.One year ago it was vaccinated against canine distemper, hepatitis, parvovirosis,leptospirosis and rabies. Several times, ticks were observed on the dog's skin.

Prior to the euthanasia of Buck, blood for analysis was obtained in anEDTA-containing vacutainer and separately, in another vacutainer for blood se-rum separation. The other two dogs living in the same yard were also sampled.The following analyses in the blood serum were performed:

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Vet. glasnik 61 (3-4) 201 - 210 (2007) I. Tsachev et al.: Infections with Ehrlichia canis andBorrelia burgdorferi in a dog


Monocytic ehrlichiosis, Lyme disease and dirofilariosis / Monocitna

erlihiosa, lajmska bolest i dirofilarioza. ELISA /IDEXX Snap® 3DXTM Test, USA wasused. The result is qualitatively visualized (+/-), without detecting the antibodyload. The diagnostic value of the used test was very high – specificity for all threediseases – 100%; sensitivity: for E. canis – 99%, for dirofilariosis – 98%; for B. burg-dorferi – 92% Š22, 23, 24¹.

Ehrlichiosis / Erlihioza. An indirect immunofluorescence technique/IFA/ was used Š25¹, that is still a gold standard for ehrlichiosis Š26¹. A formal inacti-vated suspension (2.106/ml) of cells infected with E. canis (Synbiotics Europe,France) and monospecific rabbit anti-canine FITC- labeled IgG (Sigma, Ger-many) was used as the antigen. Serum working dilutions of 1:100 to 1:1600 wereused, and the result was detected on an immunofluorescent microscope LOMO(x 200) (Fig. 1). Titres higher than 1/100 were accepted as significant Š27¹.

Leptospirosis / Leptospiroza. An analysis for the presence of leptospi-rosis was done using the reaction of microagglutination-lysis (RMAL). Taking intoconsideration the current etiological image of the disease Š28¹, apart from the con-ventional L. canicola and L. icterohaemorhagiae, L. pomona and L. tarassovi werealso included (Regional Diagnostic and Research Veterinary Institute, Stara Za-gora). Titres of 1/100 were accepted as significant Š29¹.

Blood smears were prepared from the EDTA blood and stained ac-cording to Romanovski-Giemsa.

After the euthanasia, a gross pathology examination was done andsamples for histological study were obtained. The materials included mesenteriallymph nodes, spleen, stomach, ileum, urinary bladder and brain. Imprint prepara-tions were prepared from lymph nodes and stained with Hemacolor®. The histo-logical specimens were fixed in 10 % neutral formalin and embedded in paraffin,

203


Figure 1. An indirect positive immunofluorescenceSlika 1. Indirektna pozitivna imunofluorescencija

cut on a paraffin microtome with a thickness of cross sections of 4 µm, and stainedwith haematoxylin/eosin (H/E).

Serological results / Serolo{ki rezultati. The ELISA testing of Buckshowed the following results: (+) for ehrlichiosis, (+) for Lyme disease and (–) fordirofilariosis. The possibility to determine quantitatively the antibody titre in ehrli-chiosis induced us to perform an IFA test, that yielded a titre of 1:800.

The testing for leptospirosis to L. canicola, L. icterohaemorhagiae, L.pomona and L. tarasovi was negative (–) at 1:50. It was performed because lep-tospirosis was suspected on the basis of the observed weak icterus and the ne-croses of the tongue.

Of the other two dogs that lived together with Buck, one reacted posi-tively for LB, whereas the other was LB-negative. Both dogs were negative for ehr-lichiosis, dirofilariosis and leptospirosis.

The blood smears obtained from Buck and the other two dogs werenegative for babesiosis and haemobartonellosis.

Gross pathology findings / Ukupni patolo{ki nalazi. The inspection ofthe body revealed icteric visible mucous coats and hindlimb swellings. The pe-ripheral lymph nodes were slightly enlarged. After dissection of body cavities, amarked haemorrhagic diathesis was observed. The haemorrhages (petechiaeand ecchymoses) were primarily located on gastrointestinal, genitourinary andcardiac mucosae (Fig. 2). On the tongue’s borders, symmetrical necroses wereobserved (Fig. 3). The lungs were oedematous, and the chest cavity – filled withabout 0.5 L yellow-reddish fluid. The bronchial and mediastinal lymph nodes were

204


Results / Rezultati

Figure 2. The haemorrhages (petechiae and ecchymoses) on epicardiac mucosaeSlika 2. Hemoragije (petehije i ehimoze) na epikardijalnoj sluzoko`i

slightly enlarged and reddened. The same alterations, but more severe, were ob-served in the mesenterial lymph nodes (Fig. 4). The spleen was enlarged andshowed macroscopic signs of chronic venous hyperaemia (Fig. 5). The kidneyswere significantly enlarged and spattered with spot-like haemorrhages (Fig. 6).The urinary bladder was with a distended wall and its mucous coat was with multi-ple petechial haemorrhages. After dissection of the skull, a marked venous hyper-aemia of cerebral blood vessels was revealed.

205


Figure 3. Tongue’s borders - symmetrical necrosesSlika 3. Rubovi jezika – simetri~ne nekroze

Figure 4. Enlargement and reddening of mesenterial lymph nodes (Lymphadenitis simplex)Slika 4. Uve}anje i crvenilo mezenterijalnih limfnih `lezda (limfadenitis simpleks)

Histopathological findings / Histopatolo{ki nalazi. The histologicalexamination of the kidneys, liver and lymph nodes, showed a proliferation of lym-phoreticular and plasmatic cells. In the spleen, multiple necrotic foci and sidero-cytes were observed. In the gastric and intestinal propria, epithelial celldesquamation, haemorrhages and infiltration with neutrophils were present. Inthe liver, centrolobular necroses and Kupffer cell activation were revealed. In thekidneys, haemorrhages, degeneration of epithelial cells of renal tubules and pro-liferation into renal glomerules, mainly from plasmatic cells were shown. Haemor-

206


Figure 5. SplenomegaliaSlika 5. Splenomegalija

Figure 6. Petechial haemorrhages on renal cortexSlika 6. Petehijalne hemoragije na renalnom korteksu

rhages and mononuclear proliferation were also observed in the propria of the uri-nary bladder. Some areas of the lymph nodes and the spleen were necrotic. Thebrain hemispheres revealed vascular hyperaemia, oedema and lymphocytic-plasmatic proliferations.

Cases of mixed infection with zoonotic agents in the dog, especiallywith vector-borne ones, have been present for a long time, but are very few Š30¹.Data from Israel showed 73% co-infections with more than three transmissiveagents: Babesia canis, Babesia gibsoni, Ehrlichia canis, Ricketsia conorii, Boreliaburgdorferi, Bartonella visonii Š7¹. It was established that 85% of dogs with hepato-zoonosis were seropositive for E. canis as well; there is also evidence for cases ofco-infections with Ehrlichia platys, Borrelia burgdorferi and Leishmania infantumŠ31, 32, 34¹. Studies on leptospirosis and leishmaniasis performed in Greece, alsohave shown mixed infections – occult and clinical Š29¹. The latest information re-ports a certain increase in the prevalence of vector-borne infections Š34¹.

The percentage of ixodid ticks, infected with Borrelia and Ehrlichia isrelatively low, varying from <1% to 6% Š35¹. Experimental studies in mice showedthat these agents were transmitted independently one from the other Š36¹. TheLyme borreliosis and the human granulocytic ehrlichiosis is a rarely seen combi-nation – they accounted for 3-15% of tick-borne infections in Connecticut and Wis-consin Š37¹. The serological data from the ELISA and IFA tests showed an infec-tion with E Ehrlichia canis and Borrelia burgdorferi, that could be prior or current.

It is known that 50-90% of animals, seropositive for LB, are asympto-matic. Less than 5% manifest clinical signs of the disease. During the acute phase,fever, lethargy, lymphadenopathy, lameness, are most commonly exhibited. In ourpatient, no lameness was observed.

The clinical signs in ehrlichiosis is more various, but the fever, depres-sion, lethargy, anorexia, weight loss, pale mucosae, hindlimb swellings, are all ac-companying signs Š38¹. Ecchymoses on mucous coats could also be present andwere observed by us as well. The region of origin of Buck and the other two dogs(Stara Zagora region) was with a proved 15% seroprevalence of E. canis Š14¹.

According to most investigators, the most important gross pathologyfinding in ehrlichiosis was the enlargement of the spleen and the liver, the gastro-intestinal and genitourinary haemorrhages and pulmonary oedema Š33, 39, 40,41, 42¹. The same changes could be observed in LB, but very frequently, arthritis,uveitis, myocarditis could also be present; and in cases of generalized infection –dermatitis as well Š23, 43, 44, 45¹. The petechial bleedings in kidneys, hepatic ne-croses and lymph nodes’ and splenic enlargements were more characteristic forthe acute form of leptospirosis, whereas the uraemic syndrome was more typicalfor the chronic course of the disease due to the developing interstitial nephritisŠ46, 47¹. The similar pathoanatomical and histopathological findings in these dis-

207


Discussion / Diskusija

eases makes their exact diagnostics difficult, thus necessitating the use of otherlaboratory diagnostic methods.

Our clinical, epizootological, serological, pathoanatomical and histo-pathological data allowed us to diagnose monocytic ehrlichiosis. The serologicalresults showed also a Borrelia burgdorferi infection. It could not be said for surewhether it was current or prior, because no attempts at isolation or PCR were per-formed. We assume that in this case, a carrier was more likely present, becauseBorrelia burgdorferi antibodies were also present in one of the two dogs that livedtogether with the patient, without clinical signs of the disease.

Tick-borne infections in both men and animals are endemic for manyregions of the world, including Bulgaria. The continuously appearing new datashowed that these diseases are widely distributed all over the world and becauseof this, are systematically and thoroughly studied. The correct identification of thedetailed etiological study of their reservoirs and vectors, would result in a moresuccessful control of their prevalence.

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INFEKCIJE BAKTERIJAMA Ehrlichia canis I Borrelia burgdorferi KOD JEDNOG PSA

I. Tsachev, R. Simeonov, V. Petrov

Opisan je klini~ki slu~aj infekcije bakterijama Ehrlichia canis i Borrelia burg-dorferi kod 5-godi{njeg Nema~kog ov~ara. Izvr{ena su klini~ka, serolo{ka i histopatolo{kaispitivanja, kao i nekropsija koji potvr|uju dijagnozu.

Klju~ne re~i: Ehrlichia canis, Borrelia burgdorferi, pas, nalazi nekropsije, histopatolo{kinalazi

INFEKCII BAKTERIÂMI Ehrlichia canis i Borrelia burgdorferi U ODNOYSOBAKI

I. Tsahev, R. Simeonov, V. Petrov

Opisan klini~eskiy slu~ay infekcii bakteriÔmi Ehrlichia canis i Bor-relia burgdorferi u 5-letney Nemeckoy ov~arki. Sover{enì klin~eskie, serologi~es-kie i gistologi~eskie ispìtaniÔ, slovno i nekropsiÔ, kotorìe podtver`daÓt di-agnoz.

KlÓ~evìe slova: Ehrlichia canis, Borrelia burgdorferi, sobaka, rezulÝtatì nekropsii,gistopatologi~eskie rezulÝtatì

210


SRPSKI

RUSSKIY

UDK 636.7/.8.09:616-002.828(497.2)

ETIOLOGICAL, CLINICAL AND EPIDEMIOLOGICALINVESTIGATIONS OF DERMATOPHYTOSIS AMONG PETSIN THE REGION OF STARA ZAGORA FOR THE PERIOD

2004-2006*

ETIOLO[KA I KLINI^KO-EPIDEMIOLO[KA ISTRA@IVANJA

PRISUSTVA DERMATOFITA KOD KU]NIH LJUBIMACA U REGIONU

STARA ZAGORA U PERIODU 2004-2006. GODINE

G. Michaylov, I. Tsachev, V. Petrov**

The etiological structure of dermatophytoses in pets (dogs andcats) was analyzed using routine mycological techniques. For this pur-pose, 268 speciments obtained from both dogs and cats with sus-pected dermatophytoses were investigated. The specimens includedcrusts, hairs and injured tissues of skin lesions.

The mycological study comprised a native microscopy and inocu-lation on specific nutrient media followed by isolation and species iden-tification.

A retrospective analysis was performed on the basis of history,clinical symptoms and the mycological studies. It was found that thepresence of positive samples in dogs and cats was 18.6% and 13.2%,respectively.

Most commonly, Microsporum canis was isolated in 92.3% of ca-nine and 100% of feline specimens. It was also found that animals agedunder 1 year were more susceptible to the examined mycological infec-tions. No significant gender-dependent differences have been ob-served.

Key words: Dermatophytoses, M. canis, susceptibility, dogs, cats

Dermatophytoses are superficial fungal infections, affecting the kera-tin tissues (nails, hair) and the horny layer (Stratum corneum) of the skin. The

211


Introduction / Uvod

* Rad primljen za {tampu 12. 7. 2007. godine** Gosho Michaylov, Ilia Tsachev, Vladimir Petrov, Faculty of Veterinary Medicine, Trakia Univer-

sity, Stara Zagora, Bulgaria

fungi, causing these infections in animals, are known as dermatophytes and be-long to the Microsporum and Trichophyton genera. Depending on the main hostsand the natural areal, the various species of both genera are classified as zoo-philic, anthropophilic and geophilic. Zoophilic species are always pathogenic foranimals, but a large part of them affect people as well. Anthropophilic species in-fect humans and, more rarely, animals. Geophilic species inhabit the soil andserve as a reservoir of infection for both humans and animals. More than 20 der-matophyte species are described as causing dermatophytosis in cats and dogsŠ13¹. The principal agents are M. canis, M. gypseum and T. mentagrophytes Š5,14¹. In over 95% of cases in cats, M.canis was found to cause dermatophytosisŠ10¹. Although this organism is not a natural resident of the feline hair coat Š8¹, it isbelieved that cats are a natural reservoir for it Š10, 14¹. In dogs, the percentage ofdermatophytoses varies from 4 tî 10% Š14¹, but some studies report a higherprevalence Š2, 11¹. In cats, dermatophytosis is encountered almost twice as manytimes Š14¹. The prevalence of dermatophytosis varies according to the climate,the temperature, the humidity, the rearing, the presence of natural reservoirs of in-fection Š6¹.

Young animals and especially these under 1 year of age are more fre-quently affected Š6, 3¹.

The aim of the present study was to determine the etiological structureand some epidemiological parameters of dermatophytosis among dogs and catsin the region of Stara Zagora.

Animals / @ivotinjeFor a 2-year period (March 2004 – April 2006) clinical samples were

obtained from 268 animals (224 from dogs and 44 from cats) with tentative diag-nosis of dermatophytosis and analyzed in our laboratory. The samples were ob-tained from animals referred to the Clinic of Infectious Diseases or sent by privateveterinarians. For each case, the history, clinical signs, age, gender, breed andway of rearing were recorded.

Specimen collection / Sakupljanje uzoraka

The specimens were collected by obtaining epidermal skin scrapesand hairs at the boundary of skin lesions. The sampling areas were previouslytreated with 70% ethanol. The specimens sent by private veterinary clinics wereplaced in individual sterile plastic containers.

Laboratory methods / Laboratorijske metode

1. microscopic examinationAll samples were studied for the presence of fungi by direct micros-

copy in 20% NaOH solution. Taking into consideration that the arthrospores of

212

Vet. glasnik 61 (3-4) 211 - 218 (2007) G. Michaylov et al.: Etiological, clinical andepidemiological investigations of dermatophytosis among pets in the region of ...


zoophilic dermatophytes were from the ectothrix type, a part of specimens werestudied in paraffin.

2. cultural investigationsFor cultivation, 3 types of nutrient media were used: Sabouraud’s dex-

trose agar (Difco), Mycosel agar (Difco) and DTM (Difco). All three media con-tained cycloheximide (0.5 mg/ml) and chloramphenicol (0.05 mg/ml). The speci-mens were inoculated in standard Petri’s dishes at 27oC for 3 weeks.

The species identification of isolates was done on the basis of macro-and micromorphological traits according to Tilton and McGinnis (1987); Rippon(1988).

Statistical analysis / Statisti~ke analize

The data were statistically processed using the �2 test. Values of

P<0.05 were considered as statistically significant.

The investigation included 224 dogs and 44 cats with clinical signs,specific for dermatophytosis. Forty eight animals (17.9%) were positive for der-matophytosis. Dermatophytes were isolated in 42 out of the 224 canine (18.6%)and 6 out of the 44 (13.6%) feline samples (Table 1).

Table 1. Results from microscopy and microbial culturing of samples /Tabela 1. Rezultati mikroskopske analizei mikrobialne kultivacije uzoraka

Male /Mu`jaci

Female /@enke

Total /Ukupno

Microscopy /Mikroskopija

Microbial culturing /Mikrobialna kultivacija

Number ofsamples /Broj uzoraka

%Number ofsamples /Broj uzoraka

%

Dogs /Psi

108 116 224 48 21.4 42 18.6

<1 year /<1 godine

47 37 84 31 26.0 31 26.0

>1 year />1 godine

65 77 142 9 12.7 11 15.6

Cats /Ma~ke

18 26 44 8 18.2 6 13.6

<1 year /<1 godine

8 10 18 3 16.6 4 22.2

>1 year />1 godine

11 15 26 2 7.6 2 7.6

The table shows that the prevalence of dermatophytes in animalsyounger than 1 year of age was higher (P<0.001). Table 2 presents the seasonaldistribution of studied specimens.

213


Results / Rezultati

Table 2. Seasonal distribution of studied specimensTabela 2. Raspored ispitivanih uzoraka po godi{njim dobima

Spring / Prole}e Summer / Leto Autumn / Jesen Winter / Zima

Number ofsamples % Number of

samples % Number ofsamples % Number of

samples %

Dogs / Psi

Total number ofsamples / Ukupanbroj uzoraka

56 25 45 20 62 27,6 61 27,4

Positive afterculturing / Pozitivniposle kultivacije

9 16 8 17,7 15 24,2 10 16,3

Cats / Ma~ke

Total number ofsamples / Ukupanbroj uzoraka

9 20.4 8 18,1 13 29,5 14 32

Positive afterculturing / Pozitivniposle kultivacije

1 11,1 1 12,5 2 15,3 2 14,2

No statistically significant differences were observed with regard topositive results, but the highest percentage of positive samples was established inOctober and November. M.canis was present in 92.3% of canine and 100% of fe-line samples. In the other canine samples, T. mentagrophytes was recovered.

Investigations on dermatophytosis have been performed in manycountries all over the world: Ì.canis, Ò.mentagrophytes and M.gypseum are re-ported as principal etiological agents in dogs and cats Š8, 7, 14¹. In our studies, weobserved M.canis and Tr.mentagrophytes in dogs and only M.canis in cats. Thesedata are in accordance with results obtained in the United Kingdom, whereM.canis and Tr.mentagrophytes together account for about 95% îf isolates Š14¹.

In the present study, the percentage of positive results for all studiedpatients with tentative diagnosis of dermatophytosis, was 17.9%. similar resultsare reported in the UK Š14¹, Brazil Š2¹. The percentage of positive samples amongdogs was 18.6%. This is in disagreement with available literature data, where thispercentage ranged between 4 and 10%. In dogs, compared to cats, the preva-lence of dermatophytosis is lower Š4, 12¹. This could be explained by the fact thatcats are kept individually as pets. Only in one case, where 3 cats shared the samehouse, the disease was observed in all three after contact with stray cats. Moreo-ver, the infection was also discovered in the owner (Fig.1).

All these observations point out that stray cats are a natural reservoirof the infection Š9¹ and that M.canis is a highly contagious and infectious agent.

214


Discussion / Diskusija

There are no statistically significant data about the gender- and breed-related sus-ceptibility, but the incipience of the diseases in animals younger than 1 year is in-dicative. The high sensitivity of adolescent animals to the infection could beprobably due to immune-related factors.

There were no statistically significant data about the seasonal inci-dence of these infections, but the percentage of positive samples was higher inOctober and November. This was probably related to recent climatic changes inour country.

The clinical signs are various and include the typical circular skin le-sions with erythema, desquamation, hair loss with a marked demarcation line(Fig. 2), as well as the involvement of diffuse areas with signs of squamous derma-titis and partial hair loss. That is why, laboratory methods are essential for the

215


Figure 1a. M. canis infection in dog /Slika 1a. Infekcija M. canis kod psa

Figure 1b. M. canis infection in cat /Slika 1b. Infekcija M. canis kod ma~ke

Figure 2a. M. canis infection in twenty fiveold woman /

Slika 2a. Infekcija M. canis kod dvadesetpetogodi{nje`ene

Figure 2b. M. canis infection in twenty fiveold woman /

Slika 2b. Infekcija M. canis kod dvadesetpetogodi{nje`ene

proper diagnosis. The analysis of data from the microscopy and the microbial cul-turing showed more positive results in the first method– 48 vs 42 in dogs and 8 vs6 in cats, respectively.

Various artifacts could yield false positive results throughout the mi-croscopic investigation, and that is why, by now, the microbial culturing is a "goldstandard". On the other hand, some saprophytic fungi such as Alternaria, Asper-gillus, Penicillium etc. (Fig. 3) grow faster and could be responsible for false nega-tive results. Therefore, an exact diagnosis necessitates their combined use.

1. Biberstein E.L.: Dermatophytes. In: Review of Veterinary Microbiology. EdsE.L. Biberstein and Y.C.Zee.. Blackwell Scientific Publications, Boston, 272-285, 1990. - 2.Brilhante R. S. N. et al.: High rate of Microsporum canis feline and canine dermatophytosisin Northeast Brazil; Epidemiological and diagnostic features, Mycopathologia, 156, 303-308, 2003. - 3. Cafarchia C., Romito D., Sasanelli M., Lia R., Capelli G., Otranto D.: The epi-demiology of canine and feline dermatophytoses in southern Italy. Mycoses, 47, 11-12,508-513, 2004. - 4. Caretta G., Mancianti F., Ajello L.: Dermatophytes and keratinophilicfungi in cats and dogs. Mycoses, 32, 620-626, 1989. - 5. Levis D. C. et al.: Epidemiologyand clinical fetures of dermatophytosis in dogs and cats at Lousiana State Univeristy, 1981-1990. Vet Dermatol 2, 51, 1991. - 6. Mancianti F., Nardoni S., Cecci S., Corazza M., TacciniF.: Dermatophytes isolated from symptomatic dogs and cats in Tuscany, Italy during a 15-year-period. Mycopathologia, 156, 16, 13-18, 2002. - 7. Marchisio V. F., Gallo M. G. Tullio V.:Dermatophytes from cases of skin disease in cats and dogs in Turin, Itay, Mycoses, 38, 239-244, 1995. - 8. Moriello K. A., De Boer D.J.: Fungal flora of the coat pet cats. American Jour-nal of Veterinary Research, 52, 602-606, 1991a. - 9. Moriello K. A.: Treatment of dermato-phytosis in dogs and cats: review of published studies. Vet. Dermatol, 15, 2, 99-1007, 2004.

216


Figure 3a. Fungial colinies of saprophiticfung-six day Aspergilus on DTM agar

Slika 3a. Kolonije saprofitnih gljivica - {esti dan kul-ture Aspergilus na DTM agaru

Figure 3b. Fungial colinies of saprophiticfung - seven da culture of Penecilliumon DTM agar

Slika 3b. Kolonije saprofitnih gljivica - sedmi dankulture Penicillium na DTM agaru


- 10. Muller G. H., Kirk R. A., Scott D. W.: Fungal deseases. In: Small Animal Dermatology,4th edn. W.B. Saunders, Philadelphia, 295-346, 1989. - 11. Pinter L.: Epidemiological andclinical features of dermatophytoses in dogs and cats in Croatia between 1990 and 1998.Veterinarski Archiv 69, 5, 261-270, 1999. - 12. Romano C., Valenti L., Barbara R.: Dermato-phytes isolated from asymptomatic stray cats. Mycoses, 40, 471-472, 1997. - 13. Scott D.W., Miller W. H., Griffin C. E.: Muller and Kirk’s Small Dermatology, 6th edn. Philadelphia;W.B.Saunders, 340, 2001. - 14. Sparkers A. H.: Epidemiological and diagnostic features ofcanine and feline dermatophytosis in the United Kingdom. Vet Rec. 133, 57,1994.

ETIOLO[KA I KLINI^KO-EPIDEMIOLO[KA ISTRA@IVANJA PRISUSTVADERMATOFITA KOD KU]NIH LJUBIMACA U REGIONU STARA ZAGORA

U PERIODU 2004-2006 GODINE

G. Michaylov G, I. Tsachev, V. Petrov

Istraìvanje je obuhvatilo analizu prisustva dermatofita kod ku}nih ljubimaca(pasa i ma~aka) kori{}enjem standardnih mikolo{kih tehnika. Ispitano je ukupno 268 uzo-raka pasa i ma~aka sa sumnjom na prisustvo dermatofita. Kori{}eni su uzorci dlake ìvoti-nja, kraste, kao i promenjena mesta na koì. Ura|en je nativan mikroskopski pregled, akori{}ene su i hranljive podloge u cilju identifikacije i izolacije uzro~nika. Uzev{i u obzirklini~ke simptome, istoriju bolesti kao i pomenuta mikolo{ka ispitivanja, utvr|eno je da su18,6% ispitivanih pasa i ma~aka bili pozitivni i 13,2% sumnjivi. Microsporum canis je izolo-van iz 92,3% pse}ih i 100% uzoraka ma~aka. Ustanovljeno je da su ìvotinje mla|e odgodinu dana bile prijemljivije na infekciju, dok razlike u polovima nisu imale zna~aja za is-traìvanje.

Klju~ne re~i: dermatofitoza, M.canis, psi, ma~ke

ÕTIOLOGIÊSKIE I KLINIÊSKO-ÕPIDEMIOLOGIÊSKIEISSLEDOVANIÂ PRISUTSTVIÂ DERMATOFITOV U DOMA[NIH

LÁBIMCEV V REGIONE STARAÂ ZAGORA V PERIODE 2004-2006 GODA

G. Michaylov G, I. Tsachev, V. Petrov

Issledovanie ohvatilo analiz prisutstviÔ dermatofitov u doma{nihlÓbimcev (sobak i ko{ek) polÝzovaniem standardtnìh mikologi~eskih tehnik.Nami ispìtano sovokupno 268 obraz~ikov sobak i ko{ek s somneniem na pri-sutstvie dermatofitov. PolÝzovanì obraz~iki {ersti ìvotnìh, korostì, slovnoi izmenÒnnìe mesta na koè. Sdelan nativnìy mikroskopi~eskiy osmotr, a polÝ-zovanì i pitatlÝnìe sredì s celÝÓ identifikacii i izolÔcii vozbuditelÔ.PrinÔv vo vnimanie klini~eskie simptomì, istoriÓ bolezni slovno i upomÔnutìemikologi~eskie ispìtaniÔ, utverÌdeno nami, ~to 18,6% ispìtannìh sobak i ko{ekbìli poloìtelÝnìe i 13,2% somnitelÝnìe. Microspoum canis izolirovan iz

217


SRPSKI

RUSSKIY

92,3% soba~Ýih i 100% obraz~ikov ko{ek. Nami ustanovleno, ~to `ivotnìe mo-lo`e odnoga goda bìli bolee priemlemìe na infekciÓ, poka raznicì v polah neimeli zna~eniÔ dlÔ issledovaniÔ.

KlÓ~evìe slova: dermatofitoz, M. canis, sobaki, ko{ki

218


UDK 619:616.98:636.5.033(497.2)

ETHIOLOGICAL STRUCTURE AND EPIDEMIOLOGICALMONITORING OF BACTERIAL INFECTIONS ININDUSTRIALLY REARED BIRDS IN BULGARIA*

ETIOLO[KA SRUKTURA I EPIDEMIOLO[KI NADZOR BAKTERIJSKIH

INFEKCIJA KOD INDUSTRIJSKI GAJENE @IVINE U BUGARSKOJ

Valentina Urumova, M. Lyutskanov**

The study was performed on the 9 largest poultry farms and hatch-eries in Bulgaria during the period from 2001 to 2006.

Eight hundred thirty three samples from birds have been investi-gated – seven hundred ten from gallinaceous and one hundred twentythree from waterfowl. The samples have been taken from corpse mate-rial (liver, spleen, joint, air sack, small intestine) as well from dead em-bryo, cloacal and nasal swabs. They were analyzed using routine labo-ratory or the semi-automatic system of identification CRYSTAL for en-terobacteriae and staphylococci, as well as using the API-20 NE sys-tem for nonfermentative bacteria.

A postitive microbial finding was current in 67.8% of samples fromdead chick embryos. The isolates belong to 13 different microbial spe-cies. A total of 14 species were isolated from the corpse material, nasaland cloacal swabs. Their specific presence is age dependent. Similarresults have been found for waterfowl.

Key words: ethiology, bacterial diseases, reared birds, routinemethods, diagnostic

Bacterial diseases account for a considerable share of infectious pa-thology in intensively reared birds. Their successful control requires a profoundknowledge of their etiological structure, especially the dynamics, as well as their

219


Introduction / Uvod

* Rad primljen za {tampu 12. 7. 2007. godine** Valentina Urumova, Mihni Lyutskanov, Department of Veterinary Microbiology, Infectious

and Parasytic Dyseases, Faculty of Veterinary Medicine, Trakia University, Stara Zagora, Bul-garia

epidemiological features. For some microbial species there is a real problem ofestablishing routine methods of diagnostics.

The aim of the present study was to elucidate the etiological structureand the dynamics of manifestation of different bacterial infections in industriallyreared birds – poultry and waterfowl, in some of the big farms in Bulgaria with re-gard to the optimization of prevention and control procedures.

The epidemiological studies were performed on 9 farms – 6 poultryfarms and 3 waterfowl farms using various rearing technologies.

A total of 833 samples were analyzed, including 710 from gallina-ceous birds and 123 from waterfowl. The samples were obtained from dead em-bryos, from newly hatched chickens and ducklings, as well as from growing birdsat a different age in order to monitor the dynamics of the etiological structure.

In order to determine the etiological factor, bacteriological investiga-tions were carried out in samples as follows:

– Dead embryo samples – every sample covered 12 embryos; lung tis-sues and air sack.

– Corpse material samples – every sample covered 6 corpses.From each corpse ventricle blood, liver, spleen, marrow, and in case of

arthritis a joint sample were investigated. Lung tissue samples and air sack sam-ples were taken in case of indication of mycoplasma. Lung tissue samples weretaken in case of indication of ornitobacteria. Small intestine samples were investi-gated in case of indication of clostridia.

Bacteriological investigations also included:– Cloacal samples received from chickens and fowls originating from

the same farms.The samples were bacteriologically analyzed using routine laboratory

techniques for isolation, identification and typization. A part of the microbial iso-lates were identified by the semi-automatic system of identification CRYSTAL forenterobacteriae and staphylococci, as well as via using API-20 NE system.

The bacteriological analysis of 355 samples from dead chick embryosrevealed a positive microbial finding in 241 (67.8%) samples whereas no bacteriawere present in 32.3 % of samples.

The obtained isolates included representatives of 13 microbial spe-cies (Fig. 1). Most commonly, representatives of the Enterobacteriaceae family,headed by Escherichia coli (23.2%) were isolated from dead chick embryos. This

220

Vet. glasnik 61 (3-4) 219 - 227 (2007) Valentina Urumova and M. Lyutskanov: Ethiologicalstructure and epidemiological monitoring of bacterial infections in industrially reared ...


Results / Rezultati

was valid for all surveyed flocks. The second prevalence was that of the Proteusgenus (16.1% ). Members of Enterobacter spp. wåre found in 5.7% of samples.The other enterobacteriae included Klebsiella spp. and Citrobacter spp.

Gram positive cocci (staphylococci, streptocicci and enterococci)were recovered relatively rarely – only in single samples.

Ps. aeruginosa isolates were detected considerably more frequently –this organism was evidenced in 4.5% of samples.

Bacteriological findings in newly hatched broiler chickens (0-10 days of age) /Bakteriolo{ki nalaz kod tek izleglih brojler pili}a (starosti 0-10 dana)

The bacteriological investigations of 171 carcasses of broilers chick-ens at the age of 1 - 10 days, originating from 5 different farms, yielded findings in-cluding members of 9 various microbial species, shown on the figure 2.

Again, in newly hatched chickens, Esherichia coli was detected in36.8% of studied samples. Similarly to the picture with chick embryos, the secondplace was held by representatives of the Proteus genus (16.8%).

The third prevalence was that of Salmonella spp. representatives, ob-served in 7.0% of samples. As the cause of death in newly hatched broiler chick-ens, members of Enterobacter spp. and Ps. aeruginosa, were next detected in4.7% and 4.1% respectively îf studied carcasses, but not in all studied batches.

221


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Esherichia coli

Proteus spp.

Providentia spp.

M organella morganii

Salmonella spp.

Klebsiella spp.

Citrobacter spp.

Enterobacter spp.

Staphylococcus spp

Streptococcus spp.

Enterococccus spp.

Pseudomonas spp.

Ps. aeruginosa

No findings

Figure 1. Percentages of bacterial isolates from dead chick embryos originating from5 flocks (farms) /

Slika 1. Procenat bakterijskih izolata kod uginulih embriona pili}a koji poti~u od 5 zapata (farmi)

Bez nalaza /

Bacteriological findings in broiler chickens at the age of 10 - 30 days /Bakteriolo{ki nalaz kod brojler pili}a starih 10-30 dana

With the increase of age, the relative proportion of samples with apositive bacterial finding decreased. Sterile samples accounted for 72.9%. Themicrobial isolates in the other 27.1% consisted of representatives of 6 species.The most commonly isolated species in this age group was again E. coli, presentin 20.6% of studied samples.

222


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Figure 2. / Slika 2.

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Figure 3. Percentages of bacterial isolates from dead broiler chickens at the ageof 10-30 days /

Slika 3. Procenat bakterijskih izolata kod uginulih brojler pili}a starih 10-30 dana

An interesting finding in this age group was the detection of anaerobicbacteria from the Clostridium genus (4.7%). In this case, there was an outbreak ofnecrotic enteritis in 3 different batches in one farm and in a single batch from another farm. The other microbial species, including the salmonellae observed in 3carcasses from one farm, could be considered as accidental.

Bacteriological findings in broiler chickens older than 30 days /Bakteriolo{ki nalaz kod brojler pili}a starijih od 30 dana

In broiler chickens older than 30 days, the species diversity of bacteriacontinued to decrease. Four microbial species were evidenced, among themMycoplasma spp. This was the typical age for infection of birds with mycoplasmaeand as expected, they were associated with colibacteria.

Bacteriological findings in newly hatched ducklings (0-7 days of age) /Bakteriolo{ki nalaz kod tek izleglih pa~i}a (starosti 0-7 dana)

The bacteriological analysis of samples obtained from dead ducklingsduring the first 7 post hatching days showed isolates belonging to 7 microbialspecies (Fig. 5).

It could be seen that 55.6% îf carcasses contained bacteria that couldbe determined as etiological agents.

223


65.3

4.94.95.8

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30

40

50

60

70

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Figure 4. Percentages of bacterial isolates from dead broiler chickens older than 30 days /Slika 4. Procenat bakterijskih izolata kod uginulih brojler pili}a starijih od 30 dana

Bez

nalaza

Similarly to broiler chickens, Escherichia coli was the most prevalentspecies in newly hatched ducklings – in 21.6% of samples. Then followed speciesfrom the Proteus-Providentia-Morganella group.The greatest part of isolates werefrom carcasses of just hatched ducklings and birds up to the age of 3 days, sug-gesting their vertical transmission.

The more frequent detection of Ps. aeruginosa (7.5%) should be em-phasized – a twofold higher percentage compared to the findings in newlyhatched broiler chickens at the same age. Furthermore, the data were almostcomparable for all farms surveyed.

Bacteriological findings in growing ducklings (7-21 days of age) /Bakteriolo{ki nalaz kod pa~i}a u rastu (starosti 7-21 dan)

For this age group, the microbial findings included members of 5 mi-crobial species, presented in Fig. 6.

In growing ducklings, the species diversity of bacterial etiologicalagents decreased compared to that in newly hatched ones. Again, Escherichiacoli (18.8%) was dominant, although compared to the first age group, its relativeproportion was reduced. The share of salmonella-induced infections, althoughnot high (12.6%), deserved attention because of the high lethality and the risk of apermanent carriership. Special attention should be paid to the isolation of 4 sam-ples of Riemerella anatipestifer, the agent of the bacterial polyserositis. The infec-tion was found in 5 batches of different origin in 3 different farms.

224


21.6

13.7

6.8

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2

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40.4

0 5 10 15 20 25 30 35 40 45 50

Esherichia coli

Proteus spp.

Providentia spp.

Salmonella spp.

Citrobacter spp.

Enterococccus spp.

Ps. aeruginosa

No findings

Figure 5. Percentages of bacterial isolates from dead ducklings at the age of 0-7 days /Slika 5. Procenat bakterijskih izolata kod uginulih pa~i}a starih 0-7 dana

Bez nalaza

Bacteriological findings in growing ducklings – 3-6 weeks of age /Bakeriolo{ki nalaz kod pa~i}a u rastu – starosti 3-6 nedelja

The analyses proved the existence of 3 microbial species: Pasteurellamultocida (18.2%), Salmonella Typhimurium. (9.1%) and Riemerella anatipestifer(9.1%).

The detection of Pasteurella multocida causing avian cholera was notunusual in waterfowl rearing whereas the presence of Riemerella anatipestiferconfirmed the increasing importance of this type of infection. This was also accen-tuated by the fact that the agent was isolated also from 4 rats from the affectedfarm. Up to now, similar data were not available in our country.

1. Bacterial infections occupy an important place in the pathology ofindustrially reared birds – both chicken and waterfowl species. A significant spe-cies diversity of etiological agents, most obvious in dead chick embryos andnewly hatched chickens and ducklings, was established.

2. With the advancing of age, the number of encountered microbialspecies in chickens and waterfowl decreased and this was valid for all studiedfarms and flocks.

3. In chicken fowl at all ages, the Escherichia coli sp., causing embryo-nal death and severe septicaemiae in newly hatched and growing chickens andlocal infections in older birds, was absolutely dominant.

4. The frequent isolation of Proteus-Providentia-Morganella (PPM),whose role in chick embryo pathology is known, implied a revision of the ap-

225


0 5 10 15 20 25 30 35 40 45 50

Esherichia coli

Proteus spp.

Salmonella spp.

Riemerella anatipestifer

Enterococccus spp.

No findings

Figure 6. Percentages of bacterial isolates from dead ducklings at the age of 7-21 days /Slika 6. Procenat bakterijskih izolata kod uginulih pa~i}a starih 7-21 dan

Bez nalaza

Reviewing and conclusions / Pregled i zaklju~ci

proach to them throughout their isolation from newly hatched and growing chick-ens. This was generally applicable also for Ps. aeruginosa related infections.

5. In growing chickens, clostridial and mycoplasmatic infections wererecognized as problematical.

6. The investigations did not confirm the anticipated more frequentisolation of microbial species as O. rhinotracheale, Bordetella avium, Kingellakingii, Pasteurella gallinarum, but this could be due to the relatively low number oftested farms and the rather short period of the survey.

7. In ducks, the repeated detection of pasteurellae as well as of theRiemerella anatipestifer species deserved special attention. The isolation of thelatter from rats was also an important epidemiological finding with regard to thepossibility for the formation of carriership in rodents, combined with accidental oreven permanent shedding.

1. Asplin F. D.: A septicaemic disease of ducklings. Vet. Rec. 67, 854-858,1955. - 2. Bangun A., Tripathy D. N., Hanson L. E.: Studies of Pasteurella anatipestifer: Anaproach to its classification. Avian Dis. 25, 326-337, 1981. - 3. Brogden K. A., Rhoades K.R., Rimler R. B.: Serologic types and physiologic characteristics of 46 avian Pasteurellaanatipestifer cultures. Avian Dis. 26, 891-896, 1982. - 4. Charlton B. R.: Bordetella aviumand Ornithobacterium rhinotracheale from Calofornia poultry submission. Proc 48th West-ern Poultry Disease Conference, 80, 1999. - 5. Cooper C. F., Hung P. E., Chang Y. F.: Mo-lecular characterization of a plasmid isolated from Riemerella anatipestifer. Avian Pathol.27, 339-345, 1998. - 6. Frommer A., Bock R., Inbar A., Zemer S: Muscovy ducks as a sourceof Pasteurella anatipestifer infection in turkey flocks. Avian Pathol 19, 161-163, 1990. - 7.Hafez H. M.: Current status on the role of Ornithobacterium rhinotracheale (ORT) in respira-tory disease complexes in poultry. Archiv fuer Gefuergelkunde, 60, 208-211, 1996. - 8. Hel-fer D. H., Helmboldt C. F.: Pasteurella anatipestifer infection in turkey. Avian Dis. 21, 712-715, 1977. - 9. Loh H., Teo T. P., Tan H.: Serotypes of Pasteurella anatipestifer isolates fromducks in Singapore: A proposal of new serotypes. Avian Pathol. 21, 453-459, 1992. - 10. Pa-thanasophon P., Swada T., Tanticharoenyos T.: New serotypes of Riemerella anatipestiferisolated from ducks in Thailand. Avian Pathol. 24, 195-199, 1995. - 11. Piechula K., Pohl S.,Mannheim W.: Phenotypic and genetic relationships of so-called Moraxella (Pasteurella)anatipestifer to the Flavobacterium/Cytophaga group. Vet. Microbiol. 11, 261-270, 1986.

ETIOLO[KA STRUKTURA I EPIDEMIOLO[KI NADZOR BAKTERIJSKIH INFEKCIJAKOD INDUSTRIJSKI GAJENE @IVINE U BUGARSKOJ

Valentina Urumova, M. Lyutskanov

Istra`ivanja su vr{ena na 9 najve}ih `ivinarskih farmi u Bugarkoj u periodu od2001. do 2006. godine.

226



SRPSKI

Ispitano je ukupno 833 uzoraka ìvine – 710 uzoraka ìvine i 123 uzorakavodenih ptica. Uzorci su uzeti sa le{eva (jetra, slezina, zglob, vazdu{ne kese, tankogcreva), kao i sa uginulih embriona, brisevi sa kloake i iz nosa. Analize su vr{ene koriste}irutinske laboratorijske metode ili polu-automatski sistem identifikacije CRYSTAL za enero-bakterije i stafilokoke, kao i API-20 NE sistem za nefermentne bakterije.

Pozitivan nalaz mikroba utvr|en je kod 67,8% uzoraka sa uginulih embrionapili}a. Izolati poti~u od 13 razli~itih vrsta mikroba. Izolovano je ukupno 14 vrsta sa uzorakale{eva, iz briseva nosa i kloake. Njihovo specifi~no prisustvo je u zavisnosti od starostiìvine. Ustanovljeni su sli~ni rezultati kod vodenih ptica.

Klju~ne re~i: etiologija, bakterijske bolesti, gajene ptice, rutinske metode, dijagnostika

ÕTIOLOGIÊSKAÂ STRUKTURA I ÕPIDEMIOLOGIÊSKIY NADZORBAKTERIALÃNÀH INFEKCIY U PROMÀ[LENNO RAZVEDËNNÀH

SELÃSKOHOZÂYSTVENNÀH PTIC V BOLGARII

Valentina Urumova, M. Liutskanov

IssledovaniÔ sover{enì na 9 naibolÝ{ih pticevod~eskih ferm vBolgarii v periode ot 2001 do 2006.

Nami ispìtano sovokupno 833 obraz~ika selÝskohozÔystvennìh ptic -710 obrz~ikov selÝskohozÔystvennìh ptic i 123 obraz~ika vodnìh ptic. Obraz~i-ki vzÔtì s trupov (pe~enÝ, selezënka, sustav, vozdu{nìe sumki tonkoy ki{ki),slovno i s okoletìh Ìmbrionov, mazki s kloaki i iz nosa. Analizì sover{enì,polÝzuÔ rutinnìe laboratornìe metodì ili polu-avtomati~eskaÔ sistema identi-fikacii KRISTAL dlÔ Ìnterobakteriy i stafilokokkov, i API-20 NE sistema dlÔnefermentnìh bakteriy.

PoloìtelÝnìe rezulÝtatì mikrobov utver`denì u 67,8% obraz~ikovs okoletìh Ìmbrionov cìplÔt. IzolÔtì proishodÔt iz 13 razli~nìh vidov mikro-bov. Izolirovano nami sovokupno 14 vidov s obraz~ikov trupov, iz mazkov nosa ikloaki. Ih specifi~eskoe prisutstvie v zavisimosti ot starosti selÝskohozÔyst-vennìh ptic. Nami ustanovlenì podobnìe rezulÝtatì u vodnìh ptic.

KlÓ~evìe slova: ÌtiologiÔ, bakterialÝnìe bolezni, razvedënnìe pticì,ruti~nìe metodì, diagnostika

227


RUSSKIY

UDK 637.5.07'62(560.118)

RATIO OF MEAT PREPARATES TO CARCASS IN CATTLESLAUGHTERED IN ISTANBUL*

ODNOS PREPARATA MESA U ODNOSU NA TRUPOVE GOVEDA

KLANIH U ISTANBULU

Ö. Çetin, E. Dümen, H. H. Biçer, Ö. Koçak**

This study was performed to determine the relationship betweenthe ratio of the bones and valuable meat preparates to the carcass, theage and the sex parameters in Holstein and Swiss Braun race cattlewhich are widely breeded in our country.

The half and quarter carcasses of the cattle that are salughtered inIstanbul were used as working material. The carcasses were separatedinto 4 groups as above 3 years of age group (n=24), under 3 years ofage group (n=46), male group (n=53) and female group (n=17). To-tally 140 carcasses were evaluated.

According to the obtained results, hind quarter weight, fillet, loin,rump, tranche, sirloin, round, flank, shank, brisket, fore loin, sticking,chuck and total bones parameters were significantly different at(p<0.001) level between above the 3 years of age and under the 3years of age group. Between the same groups the sirloin tip parameterwas significantly different at p<0.01 level. At the parameters of legweight, shank and half carcasses there was no significant differencebetween the groups.

We could not determine any significant differences in the percent-age ratio of all meat parameters to the carcass between the groups ofabove 3 years of age and under 3 years of age.

In the male and female groups, all the parameters except loin, legweight and shank were significantly different between the 2 groups.Hind quarter and shank parameters were significantly different atp<0.05 level, round parameter was significantly different at p<0.01level, and the other valuable meat preparates were significantly differ-ent at p<0.001 level.

229


* Rad primljen za {tampu 7. 9. 2007. godine** Ömer Çetin, Emek Dümen, Istanbul University School of Veterinary Medicine Dept. of Food

Hygiene & Technology, Avcilar,Istanbul; H. Hakan Biçer, Real Hipermarketler Zinciri A. S.;Ömür Koçak,Istabul University School of Veterinary Medicine Dept. of Animal Brreding &Husbandry, Avcilar, Istanbul

Rump parameter was evaluated as significantly different atp<0.05 level between male and female groups. The other parameterswere not significantly different between the mentioned groups.

Key words: ratio, bones, meat, carcass, age, sex, Holstein race cattle,Swiss Braun race cattle

Meat is one of the most essential foodstuff for human nutrition be-cause of its composition and organic contents. The essential aminoacids that areincluded in meat proteins are needed to form numerous body functions of humanbeings, and especially of children Š12, 14, 18, 24¹.

The production of red meat and its products are generally provided bylarge farms in our country. The meat and the Fish Cooperation, slaughterhousesthat belong to municipalities and integrated meat companies are the main provid-ers of meat and its products Š9, 16¹. Unfortunately, grading processes cannot beapplied perfectly except in The meat and the Fish Cooperation and some privateslaughterhouses in our country Š5, 16, 17¹. The meat quality of the large farm ani-mals differs according to the different individual properties such as race, age, sex,fat ratio and etc. Furthermore, during the time interval from slaughtering to con-suming the meat and / or its products some quality criterions may be changedŠ13, 26¹.

The average weight of the carcasses of the local and half-bred racelarge farm animals that are slaughtered is 140 kg. and 200 kg. respectively Š9, 19¹.In the developed countries these values differ from 477 kg. to 500 kg. Š10, 11¹. Be-cause of the low weight ratio of the carcasses of the local races, breeders gener-ally prefer half – bred races.

The prices of the meat preparates differ depending on their qualities.Fore loin, fillet, rump and tranche are known as valuable meat preparates and theprices of these products are naturally higher than the lower quality meat prepa-rates such as flank and other friers Š6, 20¹. Besides, the valuable meat preparatescan be processed technologically and sold in the form of meat products such aspastirami and smoked meat, which are highly preferred by the Turkish consumersŠ23¹.

Generally, the meat preparates that are obtained from the carcasses,are utilized according to the processing abilities of the meat and the meat's value.However, some integrated private meat companies can offer the valuable meatpreparates as "fresh meat" to the market after applying suitable technological pro-cesses to the products and the "fresh meat" market is becoming a new sector withthe help of technological processes Š1, 7, 22, 26¹.

230

Vet. glasnik 61 (3-4) 229 - 240 (2007) Ö. Çetin et al.: Ratio of meat preparates to carcassin cattle slaughtered in Istanbul

Introduction / Uvod

In this study, the objective was to determine the ratio of the valuablemeat preparates and the bones that are obtained from carcasses of half-bred racelarge farm animals to the carcasses according to the animals' age and sex.

The carcasses of two half - bred races (Holstein and Swiss Braun)were used as working material. The carcasses were obtained from differentslaughterhouses located in Istanbul city.

The carcasses were separated into 4 groups as the above 3 years old(n=24), under 3 years old (n=46), male (n=53) and female (n=17). Totally 140carcasses were evaluated.

Assigment of the races / Odre|ivanje rasa

Slaughterhouse documents were evaluated as base documents in theprocess of the assignment of the races.

Assigment of the ages / Odre|ivanje uzrasta

In the process of the age assignment slaughterhouse documentswere evaluated as base documents. Besides, the projecting parts of thoracal,lumbal and sacral vertebras were examined if they were in the form of cartilage orbone for verification of the slaughterhouse documents Š8, 25¹.

Assigment of sex / Odre|ivanje pola

In the process of the sex assignment, slaughterhouse documentswere evaluated as base documents. Besides, development of the neck musclesand chest parts of the carcasses were examined for verification. In addition, Mus-culus gracilis, Canalis inguinalis testical fat tissues, the angle of Symphisis pelvisand Tuberculum pubicum were examined, too Š25¹.

Prime Cutting / Krupno se~enje

The carcasses were cut along the median line and half carcasses wereobtained. Half carcasses were cut again, from the level of 12th and 13th costa andthe front and the back quarter carcasses were obtained. Quarter carcasses werekept in suitable warehouse conditions at +4 oC for 3 days for aging processes. Af-ter aging was completed, all the quarter carcasses were weighed one by one Š3, 4,13¹.

Detail Cutting / Detaljno se~enje

From the back quarter of the carcasses, rump, round, tranche, sirloin,sirloin tip and shank, from the lumbal part of the carcasses fillet, loin and from the

231


Material / Materijal

Method / Metode

front part of the carcasses preparates were obtained. Then all the preparates wereweighed by using sensitive balance and percentage calculations of the prepa-rates to the carcass that the evaluates preparate belonged to, were performed foreach preparate one by one Š24, 25¹.

Statistical Analysis / Statisti~ka analiza

The samples obtained from the groups were evaluated statistically.Statistical analysis was perfomed at SPSS version 10.0 Š21¹.

The meat preparates and the bones that were obtained from quartercarcasses were rated to the half and whole carcass weight that the meat prepa-rates and the bones belonged. The results are shown in Table 1, Table 2, Table 3and Table 4.

Table 1. The amount of meat preparates and bones in carcasses of the under 3 years ofage and above 3 years of age groups of the front and back quarters (kg.)

Tabela 1. Koli~ina preparata mesa i kostiju trupova `ivotinja starih manje od 3 godine i vi{e od 3 godine saprednjih i zadnjih ~etvrti (kg)

Back quarter carcassmeat preparates /

Preparati mesa sa zadnje ~etvrti

Group /Grupa

Number (n) /Broj

Mean /Srednjavrednost

St. Mean Error /Standardnadevijacija

Hind Quarter /Zadnja ~etvrt ***

12

4624

84,1592,69

± 0,86± 0,89

Fillet /File ***

12

4624

2,452,71

± 0,03± 0,04

Loin /Slabina ***

12

4624

4,575,02

± 0,05± 0,06

Rump /Krsta ***

12

4624

8,709,81

± 0,12± 0,15

Tranche /[nicla ***

12

4624

9,6310,59

± 0,10± 0,13

Sirloin /Bubre`njak ***

12

4624

4,144,50

± 0,05± 0,05

Round /But ***

12

4624

6,577,30

± 0,11± 0,08

Sirloin Tip /Vrh bubre`njaka **

12

4624

7,578,10

± 0,10± 0,15

Flank /Potrbu{ina ***

12

4624

9,2010,10

± 0,09± 0,11

Shank /Kolenica ***

12

4624

5,505,93

± 0,08± 0,08

232


Results / Rezultati

nastavak tabele 1.

Fore quarter carcass meatpreparates /

Preparati mesa sa prednje ~etvrti

Group /Grupa

Number (n) /Broj



Leg /Noga

12

4624

112,0198,63

± 21,94± 1,02

Brisket /Grudi ***

12

4624

16,4117,87

± 0,16± 0,20

Fore Loin /Ledja ***

12

4624

6,286,94

± 0,08± 0,08

Sticking /Donji deo vrata ***

12

4624

14,6715,79

± 0,13± 0,29

Chuck /Greben ***

12

4624

17,4719,21

± 0,15± 0,19

Shank /Kolenica

12

4624

4,404,66

± 0,12± 0,06

Amount of total bone and halfcarcasses /

Ukupna koli~ina kostiju i polutke

Group /Grupa

Number (n) /Broj



Total Bones /Ukupno kostiju ***

12

4624

31,6134,52

± 0,16± 0,18

Half carcass /Polutka

12

4624

196,17191,32

± 22,15± 1,85

***p<0.001, **p<0.01, *p<0.05Group 1: Below 3 years of age / Grupa 1: Ispod 3 godine starostiGroup 2: Above 3 years of age / Grupa 2: Iznad 3 godine starosti

Table 2. The percentage ratio of meat preparates and bones in carcasses of the under 3years of age and above 3 years of age groups of the front and back quarters

Tabela 2. Procentualni odnos preparata mesa i kostiju prednjih i zadnjih ~etvrti trupova `ivotinja starihispod 3 godine i preko 3 godine



Group /Grupa

Number (n) /Broj

Mean(% value) /

Srednjavrednost


Hind Quarter /Zadnja ~etvrt %

12

4624

47,37348,45

± 0,89± 0,13

Fillet /File %

12

4624

1,371,41

± 0,26± 0,10

Loin /Slabina %

12

4624

2,572,62

± 0,04± 0,01

Rump /Krsta %

12

4624

4,895,13

± 0,10± 0,05

Tranche /[nicla %

12

4624

5,425,53

± 0,10± 0,04

233


nastavak tabele 2.

Sirloin /Bubre`njak %

12

4624

2,352,35

± 0,04± 0,01

Round /But %

12

4624

3,693,81

± 0,08± 0,02

Sirloin Tip /Vrh bubre`njaka %

12

4624

4,264,23

± 0,09± 0,06

Flank /Potrbu{ina %

12

4624

5,185,27

± 0,09± 0,02

Shank /Kolenica %

12

4624

3,093,09

± 0,06± 0,01



Group /Grupa

Number (n) /Broj

Mean(% value) /

Srednjavrednost


Leg /Noga %

12

4624

52,6351,55

± 0,89± 0,13

Brisket /Grudi %

12

4624

9,239,34

± 0,17± 0,03

Fore Loin /Ledja %

12

4624

3,533,62

± 0,07± 0,01

Sticking /Donji deo vrata %

12

4624

8,268,24

± 0,15± 0,11

Chuck /Greben %

12

4624

9,8410,04

± 0,18± 0,03

Shank /Kolenica %

12

4624

2,252,24

± 0,07± 0,01



Group /Grupa

Number (n) /Broj

Mean(% value) /

Srednjavrednost


Total Bones /Ukupno kostiju %

12

4624

17,7918,04

± 0,17± 0,03

Group 1: Below 3 years of age / Grupa 1: Ispod 3 godine starostiGroup 2: Above 3 years of age / Grupa 2: Iznad 3 godine starosti

234


Table 3. The amount of meat preparates and bones in carcasses of male and femalegroups of the front and back quarters (kg.)

Tabela 3. Koli~ina preparata mesa i kostiju sa prednjih i zadnjih ~etvrti trupova mu`jaka i `enki (kg)



Group /Grupa

Number (n) /Broj



Hind Quarter /Zadnja ~etvrt ***

12

5317

84,4692,48

± 0,90± 1,01

Fillet /File ***

12

5317

2,452,71

± 0,03± 0,04

Loin /Slabina ***

12

5317

4,635,03

± 0,05± 0,05

Rump /Krsta ***

12

5317

8,849,86

± 0,12± 0,18

Tranche /[nicla ***

12

5317

9,7810,57

± 0,11± 0,16

Sirloin /Bubre`njak ***

12

5317

4,194,51

± 0,05± 0,06

Round /But **

12

5317

6,697,29

± 0,10± 0,08

Sirloin Tip /Vrh bubre`njaka

12

5317

7,688,02

± 0,09± 0,20

Flank /Potrbu{ina ***

12

5317

9,3210,14

± 0,10± 0,12

Shank /Kolenica *

12

5317

5,565,93

± 0,08± 0,08



Group /Grupa

Number (n) /Broj



Leg /Noga

12

5317

91,8298,33

± 0,88± 1,06

Brisket /Grudi ***

12

5317

16,6517,82

± 0,17± 0,22

Fore Loin /Ledja ***

12

5317

6,376,96

± 0,07± 0,10

Sticking /Donji deo vrata *

12

5317

14,9015,66

± 0,14± 0,36

Chuck /Greben ***

12

5317

17,7419,20

± 0,17± 0,21

Shank /Kolenica

12

5317

4,434,65

± 0,11± 0,06



Group /Grupa

Number (n) /Broj



Total Bones /Ukupno kostiju ***

12

5317

32,0634,45

± 0,17± 0,17

Half carcass /Polutka ***

12

5317

171,29190,81

± 1,75± 1,98

Group 1: Male / Grupa 1: Mu`jaci ; Group 2: Female / Grupa 2: @enke

235


Table 4. The percentage ratio of meat preparates and bones in carcasses of the male andfemale groups of the front and back quarters

Tabela 4. Procentualni odnos preparata mesa i kostiju sa prednjih i zadnjih ~etvrti trupova mu`jaka i `enki



Group /Grupa

Number (n) /Broj

Mean(% value) /

Srednjavrednost


Hind Quarter /Zadnja ~etvrt %

12

5317

48,1948,47

± 0,10± 0,15

Fillet /File %

12

5317

1,401,42

± 0,01± 0,01

Loin /Slabina %

12

5317

2,612,64

± 0,01± 0,01

Rump /Krsta %

12

5317

5,005,16

± 0,04± 0,06

Tranche /[nicla %

12

5317

5,515,53

± 0,02± 0,05

Sirloin /Bubre`njak %

12

5317

2,362,36

± 0,01± 0,01

Round /But %

12

5317

3,773,81

± 0,04± 0,02

Sirloin Tip /Vrh bubre`njaka %

12

5317

4,334,20

± 0,03± 0,08

Flank /Potrbu{ina %

12

5317

5,255,31

± 0,01± 0,02

Shank /Kolenica %

12

5317

3,133,10

± 0,02± 0,02



Group /Grupa

Number (n) /Broj

Mean(% value) /

Srednjavrednost


Leg /Noga %

12

5317

51,8751,33

± 0,10± 0,15

Brisket /Grudi %

12

5317

9,399,34

± 0,02± 0,04

Fore Loin /Ledja %

12

5317

3,593,64

± 0,02± 0,02

Sticking /Donji deo vrata %

12

5317

8,408,20

± 0,02± 0,16

Chuck /Greben %

12

5317

10,0110,06

± 0,03± 0,03

Shank /Kolenica %

12

5317

2,502,43

± 0,05± 0,01



Group /Grupa

Number (n) /Broj

Mean(% value) /

Srednjavrednost


Total Bones /Ukupno kostiju %

12

5317

18,0818,05

± 0,03± 0,03

Group 1: Male / Grupa 1: Mu`jaci ; Group 2: Female / Grupa 2: @enke

236


According to the obtained results, significant differences were deter-mined between carcasses of the above 3 years of age and under 3 years of ageand male and female groups. Cold carcass weights, valuable meat preparatesand bone ratios parameters were analysed.

According to our study, the average of the carcass weights of the be-low 3 years of age, above 3 years of age, male and female groups were deter-mined as 392.34 kg, 382.64 kg, 354.58 kg ve 381.62 kg respectively. In a studyperformed by Fiems et al. Š11¹, it was indicated that the cold carcass weight valuesof double-muscled Belgian Blue race cattle as from 469.7 kg. to 500.8 kg. Accord-ing to another study of Fiems et al. Š10¹, the average weight values of non - preg-nant and non - lactating female cattle was about 447.0 kg.

Between the groups of above 3 years of age and under 3 years of age,we determined significant differences for all the parameters except shank. Signifi-cant difference at (p<0.001) level were found in the hind quarter weight, fillet, loin,rump, tranche, sirloin, round, flank, brisket, fore loin, sticking, leg, chuck andweight of the among leg muscles parameters. Sirloin tip parameter was also sig-nificantly different at (p<0.01) level. In the parameters of leg weight and shank, theresults were higher in the above 3 years of age group than the under 3 years ofage group, however the differences were not significant between the two groups.

The percentage ratio of all valuable meat preparates to its carcass wasnot significantly different between the groups of above 3 years of age and under 3years of age. However, in general, the percentage ratio of the valuable meat pre-parates to its carcass was higher in the group of above 3 years of age than the per-centage ratio of the valuable meat preparates to its carcass in the group of under 3years of age.

Between the male and female groups, significant differences were de-termined for all parameters except sirloin tip, leg weight and shank. Hind quarter,shank and sticking parameters were significantly different between the groups at(p<0.05) level, round significantly different between the groups at (p<0.01) leveland the other valuable meat preparates were significantly different between thegroups at (p<0.001) level.

According to the results of the parameters in question, rump was sig-nificantly different at (p<0.05) level between the male and the female groups. Theother parameters were not evaluated as significantly different between thegroups.

At the mean values of percentage bone ratio to the carcass parame-ters, our findings were 18.04% in the group of above 3 years of age, 17.79 % in thegroup of under 3 years of age, 18.08% in the group of male and 18.05% in thegroup of female. Altuntas Š2¹ points out that the mean values of percentage boneratio to the carcass parameters are between 14.90% and 15.46% at Simental racecattle. According to Yücesan Š26¹, the percentage ratio of bones to the carcasses

237


Discussion and Conclusion / Diskusija i zaklju~ak

at local and half-bred races are 19.5% and 18.8% respectively. Özbeyaz et al. Š19¹indicate the percentage ratio of bones to the carcasses at Brangus hybrid malecalves, Simental melezi erkek danalarda ve limozin melezlerinde are 16.0%,16.2% and15.9 % respectively. According to Koçak et al. Š15¹, the bone ratio val-ues of 11 -15 month old Holstein male cattle to the carcass are between 16.4% -18.48%.

In this study the results show that the ratios of the bones and the valu-able meat preparates to the carcass were higher in the above 3 years of age groupthan in the under 3 years of age group, but the same ratio values in the male andfemale groups differed similarly from each other.

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ODNOS PREPARATA MESA U ODNOSU NA TRUPOVE GOVEDA KLANIH UISTANBULU

O. Cetin, E. Dumen, H. H. Bicer, O. Kocak

Istraìvanja su izvr{ena da bi se ustanovio odnos izme|u kostiju i vrednih de-lova mesa s obzirom na trup, uzrast i pol kod hol{tajn i {vajcarskih sme|ih rasa goveda ~ijeje uzgajanje {iroko rasprostranjeno u na{oj zemlji.

Polovine i ~etvrtine trupova goveda koje se kolju u Istanbulu su kori{}ene kaoradni materijal. Trupovi su podeljeni na 4 grupe, i to: starosna grupa iznad 3 godine (n=24),ispod 3 godine (n=46), grupa mu`jaka (n=53), i grupa ènki (n=17). Ispitano je ukupno140 trupova.

Na osnovu dobijenih rezultata, parametri za teìnu zadnje ~etvrti, file, slabinu,krsta, {niclu, bubre`njak, but, potrbu{inu, kolenicu, grudi, pe~enicu, donji deo vrata, gre-ben, i ukupne kosti zna~ajno (p<0,001) su se razlikovali izme|u starosnih grupa, odnosnoìvotinja iznad i ispod 3 godine starosti. Izme|u istih grupa, parametri za vrh slabine bili suzna~ajno razli~iti (p<0,01). Kod parametara za teìnu noge, kolenice i polutke nije bilozna~ajnih razlika izme|u grupa.

Nisu ustanovljene zna~ajne razlike izme|u procentualnog odnosa svih para-metara mesa u odnosu na trupove izme|u grupa iznad i ispod 3 godine starosti.

Izme|u grupa mu`jaka i ènki utvr|ene su zna~ajne razlike kod svih parame-tara osim za slabinu, teìnu noge i kolenice. Zna~ajne razlike (p<0,05) su registrovaneizme|u parametara za zadnju ~etvrt i kolenicu, tako|e zna~ajna razlika (p<0,01) kod para-metara za but, a i ostali preparati mesa su se tako|e zna~ajno razlikovali (p<0,001).

Ustanovljena je zna~ajna razlika (p<0,05) kod parametara za krsta izme|ugrupa mu`jaka i ènki. Ostali parametri se nisu zna~ajno razlikovali izme|u pomenutihgrupa.

Klju~ne re~i: odnos, kosti, meso, trup, starost, pol, Hol{tajn rasa goveda, {vajcarskasme|a rasa goveda

239


SRPSKI

OTNO[ENIE PREPARATA MÂSA V OTNO[ENII TULOVIÇEY KRUPNOGOROGATOGO SKOTA UBITÀH V STAMBULE

O. Cetin, E. Diten, H. H. Biker, O. Kocak

IssledovaniÔ sover{enì, ~tobì ustanovilosÝ otno{enie sredi ko-stey i stoÔçih ~astey mÔsa v otno{enii tuloviça, vozrasta i pola u hol{tayn i{veycarskih burìh porod krupnogo rogatogo skota ~ÝÒ razvedenie {iroko raspros-traneno v na{ey strane.

Polovinki i ~etvertinki tuloviçey krupnogo rogatogo skota, ubi-vaemìe v Stambule, polÝzovannìe v ka~estve rabo~ego materiala. Tuloviçarazdelenì na 4 gruppì, a imenno: vozrastnaÔ gruppa sverh 3 goda (n=24), niè 3 goda(n=46), gruppa samcov (n=53), i gruppa samok (n=17). Nami ispìtano sovokupno 140tuloviçey.

Na osnove polu~ennìh rezulÝtatov, parametrì dlÔ vesa zadney ~et-verti, file, bok, krestec, çnicelÝ, po~elnaÔ ~astÝ, okorok, brÓ{inu, koleninu,grudÝ, hrebtinu, ni`nÔÔ ~astÝ {ei, grebenÝ, i sovokupnìe kosti zna~itelÝno(r<0,001) razli~alisÝ sredi vozrastnìh grupp, otnositelÝno ìvotnìh sverh iniè 3 goda vozrasta. Sredi takih è grupp, parametrì dlÔ verha boka bìlizna~itelÝno razli~nìe (r<0,01). U parametrov dlÔ vesa nogi, koleninì i polov-inki ne bìlo zna~itelÝnìh raznic sredi grupp.

Ne ustnovlenì zna~itelÝnìe raznicì sredi procentnogo otno{eniÔvseh parametrov mÔsa v otno{enii tuloviçey sredi grupp sverh i niè 3 godavozrasta.

Utver`denì zna~itelÝnìe raznicì sredi grupp samcov i samok u vsehparametrov krome dlÔ boka, vesa nogi i koleninì. Suçestvovala zna~itelÝnaÔraznica (r<0,05) sredi parametrov dlÔ zadney ~etverti i koleninì, takèzna~itelÝnaÔ raznica (r<0,01) u parametrov dlÔ lÔ`ki, a i ostalÝnìe parametrìmÔsa takè zna~itelÝno razli~alisÝ (r<0,001).

Ustanovlena zna~itelÝnaÔ raznica (r<0,05) u parametrov dlÔ krestecsredi grupp samcov i samok. OstalÝnìe parametrì ne zna~itelÝno razli~alisÝsredi upomÔnutìh grupp.

KlÓ~evìe slova: otno{enie, kosti, mÔso, tuloviçe, vozrast, pol, Hol{taynporoda krupnogo rogatogo skota, Swis Braun poroda krupnogorogatogo skota

240


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243

Vet. glasnik 61 (3 - 4) 241 - 243 (2007) Diplomirani veterinari

yu issn 0350-2457 veterinarski glasnik udk 619 (05) · nica da je teritorija srbije, prema op{tim...

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