NALAZ NIVOA SELENA U KRVNOM SERUMU KOD KRAVA U LAKTACIJI

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1 Letopis naučnih radova Godina 35 (2011), broj I, strana UDK: : Originalni naučni rad Original scientific paper NALAZ NIVOA SELENA U KRVNOM SERUMU KOD KRAVA U LAKTACIJI M. Erdeljan, I. Davidov, S. Boboš, M. Radinović, I. Stančić 1 REZIME Mikroelementi su ključne komponente u normalnom funkcionisanju imunološkog sistema i stoga su od ključnog značaja za održavanje imuno-kompetentnosti i optimalnih proizvodnih performansi životinja. Mikroelementi, kao što je selen, direktno utiču na sposobnost odbrane makroorganizma od mikroorganizama, odnosno moduliraju njegovu imunološku odbranu. Nedostatak selena kod zasušenih krava se smatra odgovornim za zadržavanje posteljice. Takođe, manjak selena se vezuje za abortus, visoku učestalost embrio-fetalnog uginuća, slabu fertilnost, povećanu incidencu metritisa, viši nivo opštih infekcija i rođenje mrtve ili slabo vitalne teladi (Smith i Chase, 2008.). Mnogobrojni radovi govore o tome da povećanje koncentracije selena i neki drugih mikroelementa u krvi i drugim tkivima direktno dovodi do smanjenja broja somatskih ćelija u mleku, do smanjenja incidence kliničkih mastitisa i blagog povećanja proizvodnje mleka. Da bi se mogla adekvatno izvršiti suplementacija selena potrebno je utvrditi koliki je raspoloživi nivo selena kod životinje, jer su navedena patološka stanja multikauzalna, a takođe poznati su i toksični efekti selena. Ključne reči: selen, goveda, koncentracija u krvnom serumu UVOD Potpuna uloga selena kao mikroelementra u organizmu nije u potpunosti razjašnjena. Jedna od najvažnijih jasno uspostavljenih funkcija selena, je da je on kofaktor za enzim glutation peroksidazu. Glutation peroksidaze (GPKS) se uglavnom nalaze u citosolu ćelija (70%) i matriksu mitohondrija (30%) (Gropper i sar. 2005). Enzim, GPKS, odgovoran je za katalitičko uklanjanje vodonik peroksida i smanjenje količine drugih peroksida iz tkiva, što je suštinski proces imunološkog sistema. Enzim zahteva selen kao kofaktor inače je njegova aktivnost smanjena. Za selen se može reći da je 1 Dr. vet. med. Mihajlo Erdeljan, asistent; mr Ivana Davidov, asistent; dr Stanko Boboš, red. prof.; mr Miodrag Radinović, asistent; dr Ivan Stančić, docent, Poljoprivredni fakultet, Depatman za veterinarsku medicinu, Novi Sad 92

2 veoma važan kao antioksidans i kao faktor u energetskom metabolizmu fagocitnih ćelija, ali potpun mehanizam njihove modulacije humoralnog imuniteta nije u potpunosti shvaćen. Istraživanja koja su rađena u vezi sa funkcijom selena uglavnom su usmerena na efekte suplementacije selena u hrani, ali je svakako predhodno potrebno utvrditi nivoe selena u krvi kako bi se spoznali pravi efekti suplementacije. Nivoi selena u organizmu se mogu određivati na nekoliko načina, preko aktivnosti enzima u kojima je selen aktivan, koji se mogu odrediti iz tkiva jetre ili direktno iz krvi. Optimalni nivo selena u krvi se kreće u rasponu od 8 do 10 mg/100 ml, Smith i Chase (2008.). U velikom broju istraživanja je pokazano da dopunjavanje selena ispoljava svoj efekat pre svega u patologiji mlečne žlezde i to u vidu smanjenja broja kliničkih mastitisa i u smanjenju broja somatskih ćelija u mleku. Sa druge strane tamo gde su utvrđeni deficiti selena dokazano postoje imunološki problemi u vidu niže koncentracije IgM u krvi sa sniženom funkcijom polimorfonuklearnih leukocita i sa većim brojem opštih infekcija. Takođe javljaju se i reproduktivni problemi u vidu slabijeg fertiliteta, embriofetalnih uginuća, abortusa, rođenja mrtve ili slabo vitalne teladi, zadržavanja posteljice i povećane incidence metritisa. Jedna od studija koje su sprovedena o uticaju selena na zdravlje vimena je izveo Malbe sa sar. (1995.). Ispitivani su efekti ishrane uz dodatak organskog selena u obliku seleniziranog kvasca i neorganskog u obliku natrijum selenita na 100 muznih krava. Obrok je dopunjen sa 0,2 ppm tokom 8 nedelja. Rezultati su pokazali da je selenizirani kvasac dao veći nivo selena u krvi, a da GPKS nivo porastao od 0,22 do 3,0 odnosi 2,3 microkat/g hemoglobina iz kvasca odnosno natrijum selenita. Nivo GPKS u krvi nastavio je da raste i do 10 nedelja nakon što je prestala dopuna selena. Takođe, procenat četvrti kod kojih je utvrđen mastitis je opao zajedno sa brojem somatskih ćelija u mleku. Važno je napomenuti da je testiranje rađeno kod krava sa značajno niskim nivoom selena u krvi. Uticaj dodavanja selena u hrani je ispitivala i Rakić sa sar. (2007.). U ovom istraživanju je osim selena dodavan i vitamin E. Rezultati su pokazali značajnije smanjenje broja somatskih ćelija u mleka tokom laktacije u odnosu na kontrolu. Takođe u kontrolnoj grupi krava je bilo i pojave mastitisa, sa nalazom odnosno uzročnika mastitisa Staph. aureus-a u mleku što nije zabeleženo u drugim eksperimentalnim grupama. Istraživanje koje je sproveo Reffett sa sar. (1988.) ukazuje na to da selen deficitarna telad imaju niže koncentracije IgM u serumu. Slično tome, funkcije polimorfonuklearnih leukocita su smanjene kod krava Gyang i sar. (1994.), odnosno kod životinja koje su hranjene sa selen deficitarnom dijetom u odnosu na kontrolnu grupu životinja koje su primale selen-adekvatnu ishranu. Istraživanja drugih autora Maddox i sar. (1999.) i Spears (2000.) ukazuju na to da endotelne ćelije mlečne žlezde goveda gajene u ćelijskoj kulturi u selen deficitarnom medijumu ispoljavaju poboljšanu adheziju neutrofila kada se stimulišu sa citokinima. Ovi nalazi sugerišu da selen može uticati na neutrofilnu migraciju u tkiva i posledično na inflamaciju. 93

3 Dalje studije su pokazale da će dopunjavanje ishrane kod mlečnih krava, sa odgovarajućim nivoima selena može poboljšati aktivnost glutation peroksidaze i optimizovati odbranu organizma od bolesti (Sordillo 2005.). Drugi autori, Weiss i Hogan (2005.), pak navode da dopunjavanje ishrane mlečnih krava sa odgovarajućim nivoima selena (0,3 ppm suve materije) smanjuju prevalencu, ozbiljnosti i trajanje mastitisa, kao i broj somatskih ćelija. Pored toga, oni navode da organske forme selena imaju bolje efekte od neorganskih izvora. Nedostatak selena kod zasušenih krava se smatra odgovornim za zadržavanje posteljice. Broj zadržanih posteljica je smanjen, u istraživaju Kornel univerziteta u Ohaju, kada su goveda koja su imala nedostatak selena dobila dopunski selena (50 mg) i vitamin E (680 IU) injekciono 20 dana pre teljenja ili su hranjene sa dodatkom od 1 mg selena po danu. Zaključeno je da bi se zadržavanje posteljice moglo smanjiti dodatkom selena kada, i samo kada je stanje izraz nedostataka selena i nije vezano za infekcije, blizanački graviditet, mlečnu groznicu ili neke druge faktore koji povećavaju učestalost ovog stanja (Smith i sar ). Manjak selena takođe je vezan za abortus, visoku učestalost embrio-fetalnog uginuća, slabu fertilnost, povećanu incidencu metritisa, viši nivo opštih infekcija i rođenje mrtve ili slabo vitalne teladi. Nivoi selena u krvi ovih goveda uglavnom su bili ekstremno niski - manje od 5 mg/100 ml (Smith i sar ). Smatra se da je nedostatak selena vezan za zemljište koje je u pojedinim područjima deficitarno u selenu, tako da su posledično i žitarice uzgojene na ovim zemljištima siromašne u selenu. MATERIJAL I METODE RADA U ogledu su učestvovale krave u laktaciji, klinički zdrave, od kojih je jedna sveže oteljena. Ukupno je uzeto 24 uzoraka od isto toliko krava na jednoj farmi u Vojvodini. Krv je uzimana iz repne vene uz primenu principa asepse i antisepse. Prilikom vađenja krvi korišćeni su vakutajneri uz odgovarajuću iglu. Serum je izdvajan tokom 24h držanjem uzoraka na sobnoj temperaturi. Nivo selena je u krvnom serumu određivan metodom gasne hromatografije. REZULTATI ISTRAŽIVANJA I DISKUSIJA Analizom uzoraka krvnog seruma od 24 ispitivane krave dobijen je sledeći sadržaj selena u krvi koji je prikazan u tabeli 1, odnosno grafički (Graf 1). Nivo selena u krvi je izražen u. Utvrđeno je da su sve krave koje su učestvovale u u istraživanju imale manjak selena u krvi. Kod četiri krave je to drastično nisko (ispod 3 ), u odnosu na preporučenu vrednost koja iznosi 8-10, a tek kod tri krave je neznatno ispod donje granice, odnosno iznad 7. Prosečna vrednost nivoa selena kod ispitivanih krava iznosi 5,

4 Tabela 1. Sadržaj selena u krvnom serumu Table1. Selenium level in blood serum Red. broj Sadržaj selena Red. broj Sadržaj selena Red. broj Sadržaj selena 1 1, , , , , , , , , , , , , , , , , , , , , , , , Selen Graf 1. Sadržaj selena u krvnom serumu - distribucija Graph 1. Selenium level in blood serum - distribution ZAKLJUČAK Na osnovu nama dostupne literature, dodavanje selena je preporučljivo u proizvodnji mleka. Može se preporučiti adekvatna dijeta koja treba da sadrži odgovarajući nivo selena u suvoj materiji. Kod nekih grla, potrebno je da se izvori selena dopune sa injekcijama selena kako bi se održavao adekvatni nivo selena u krvi iznad preporuče- 95

5 nog od U ovim stadima gde su nivoi selena izuzetno niski, a gde se koriste injekcioni preparati za ubrzani povratak nivoa selena u normalu dodaci u hrani mogu pružiti dovoljno selena za održavanje odgovarajućeg nivoa u krvi krava. Testovi krvi se svakako preporučuju da bi se utvrdio status selena kada god u zapatu postoje problemi sa reprodukcijom, imunološkim odgovorom ili patologijom mlečne žlezdem. Unošenje obrokom selena mora biti pod kontrolom kako zbog mogućnosti nedovljne suplementacije tako i zbog mogućih toksični efekata na životinju. LITERATURA 1. Gropper, S.S., J. Smith, and J. Groff. (2005.) Advanced Nutrition and Human Metabolism: Glutathione peroxidase. 4th ed. Wadsworth. Belmont, CA Gyang, E.O., J.B. Stevens, W.G. Olsen, S.D. Tsitsamis and E.A. Usenik. (1994.) Effects of selenium-vitamin E injection on bovine polymorphonucleated leukocyte phagocytosis and killing of Staphylococcus aureus. Am. J. Vet. Res. 45: Maddox, J. F., K. M. Aheme, C. C. Reddy and L. M. Sordillo. (1999.) Increased neutrophil adherence and adhesion molecule mrna expression in endothelial cells during selenium deficiency. J. Leukocyte Biol. 65: Malbe, M., M. Klaassen, W. Fang, V. Myllys, M. Vikerpuur, K. Nyholm, S. Sankari, K. Suoranta and M. Sandholm. (1995.) Comparisons of selenite and selenium yeast feed supplements on Se-incorporation, mastitis and leukocyte function in Se-defecient dairy cows. J. Vet. Med. A. 42, Rakić, M., Kirovski D., Vujanac I., Dimitrijević B., Džmura G., Pudlo P. (2007.) Uticaj glukagona, vitamina E i selena na zdravlje mlečne žlezde i količinu proizvedenog mleka. Ishrana, reprodukcija i zaštita zdravlja goveda. Banja Kanjiža. 6. Reffett, J. K., J. W. Spears and T. T. Brown. (1988.) Effect of dietary selenium on the primary and secondary immune response in calves challenged with infectious bovine rhinotracheitis virus. J. Nutr. 118: Smith R.D., Chase L.E., : Nutrition and reproduction, Dairy Integrated Reproductive Management, (2008.) Cornell University livepoul/dirm14.pdf 8. Sordillo, L. M. (2005.) Bovine Mammary Immunity and Mastitis Susceptibility: Impact of Oxidant Stress in Transition Cows. Proc. CA ARPAS Conf, Coalinga, CA. October. 9. Spears, J. W. (2000.) Micronutrients and immune function in cattle. Proc. Nutr. Soc. 59: Weiss, W. P. and J. S. Hogan Effect of selenium source on selenium status, neutrophil function, and response to intramammary endotoxin challenge of dairy cows. J. Dairy Sci. 88:

6 FINDING OF SELENIUM LEVELS IN BLOOD SERUM OF LACTATING COWS by M. Erdeljan, I. Davidov, S. Boboš, M. Radinović, I. Stančić SUMMARY Trace elements are key components in the normal functioning of the immune system and therefore are crucial for maintaining immune competence and optimal performance of production animals. Trace elements such as selenium, directly affect the ability of macroorganisms to defense against microorganisms or modulate its immune response. Selenium deficiency in dry cattle is liable for the retention of the placenta. Also, selenium deficiency is associated with abortion, high frequency of embryo-fetal mortality, low fertility, increased incidence of metritis, a higher level of general infection and birth of dead or weak vital calves (Smith and Chase, 2008.). Numerous papers show that increasing concentrations of selenium and some other micronutrients in the blood and other tissues directly leads to a reduction in the number of somatic cells in milk, to reduce the incidence of clinical mastitis and a slight increase in milk production. In order to make adequate selenium supplementation it is necessary to determine what is the available level of selenium in animals, because the mentioned pathological conditions have multiple causes, also there are known the toxic effects of selenium. Key words: selenium, cattle, concentrations in blood serum Primljeno: Prihvaćeno:

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