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Svinje predstavljaju značajnu epizootiološku kariku u održavanju većeg broja infektivnih bolesti od kojih su neke veoma značajne zoonoze. Mali broj vektorima prenosivih zoonoza u našem regionu ne znači da neke od infektivnih bolesti koje su zajedničke životinjama i ljudima, a koje se enzootski i endemski pojavljuju u udaljenim regionima sveta, neće u nekim okolnostima da izazovu epizootije i epidemije i u Evropi. U radu su prikazani mehanizmi održavanja uzročnika infektivnih bolesti, naročito arbovirusa u epizootiološkim ciklusima i vektorima (hematofagnim insektima i artropodama). Prikazane su i odabrane infektivne bolesti koje su za region Evrope „egzotične“ ali kod kojih postoji mogućnost da se prenesu u populacije svinja u našem regionu, a samim tim i da postanu značajan epidemiološki rizik.

West Nile virus (WNV) is a zoonotic mosquito-borne virus classified as family Flaviviridae and genus Flavivirus. The first WNV outbreak in humans in the Republic of Serbia was recorded in 2012. Equids and dogs can show clinical symptoms after WNV infection and are often used as sentinels. This study aimed to (i) give insight into seropositivity for WNV in clinically healthy dog and horse sera in different regions of Serbia and (ii) compare diagnostic value of 'in-house' and commercially available indirect immunofluorescence (IFA) and enzyme-linked immunoassay (ELISA) tests to 'gold standard' virus neutralization test (VNT). Due to cross-reactivity, sera were tested for Usutu virus and tick-borne encephalitis virus in VNT based on the epidemiological data of field presence. Blood sera of dogs (n = 184) and horses (n = 232) were collected from 2011 to 2013. The seropositivity was confirmed by VNT in 36.9 % tested dog sera and 34.9% tested horse sera with highest positivity in regions near two big rivers, while in four dog and seven horse sera, positivity resulted from Usutu virus infection. Comparative results of diagnostic tests in dogs ranged from 18.7 % seropositivity by 'in-house' ELISA to 31.9% by commercially available ELISA. In horses, seropositivity ranged from 36.2% by 'in-house' IFA to 32.5% by commercially available IFA and from 26.3% by 'in-house' IgG ELISA to 20.9% by commercially available ELISA. There were no statistically significant differences according to the McNemar test between 'in-house' and commercially available IFA and ELISA test in horse sera, while the same was not true for two ELISAs used in dog sera (chi(2) = 8.647, p = .003). Established seropositivity in dogs and horses was in accordance with the epidemiological situation and WNV spread in the Republic of Serbia and proven Usutu virus co-circulation. 'In-house' tests remain a valuable tool in early diagnostics of WNV.

Supplementary file 1. Dog and horse sera sampled from 15 locations from 2011 to 2013 for the serological study. Supplementary file 2. Seropositivity in dogs and horses in Serbia in period 2011-2013. Supplementary file 3. The results of „in-house“ and commercial ELISA tests in dog sera. Supplementary file 4. The results of „in-house“ and commercial IFA and ELISA tests in horse sera

Epizootiološke determinante predstavljaju osnov za poznavanje epizootioloških karakteristika pojedinih zaraznih i parazitskih oboljenja, kao i poremećaje zdravlja nezarazne i neparazitske etiologije. U osnovi, radi se o tri determiante, a to su: 1. Prijemčiva vrsta, 2. Uzročni faktor i 3. Spoljašnji faktor. Region Srbije u kome se nalazi Stara planina je izuzetno interesantan sa epizootiološkog aspekta. To se naročito odnosi na površinu koju zahvata opština Dimitrovgad, koja obuhvata delove Stare planine i urbanu sredinu, u čijoj neposrednoj blizini se nalazi jedan od najznačajnijih puteva koji spajaju Evropu sa Istokom kao i na bogatstvo flore i faune kakvo se retko gde sreće u regionu. U odnosu na prijemčive vrste, radi se o regionu u kome se nalazi veći broj vrsta domaćih životinja čija tehnologija uzgoja značajno varira. U odnosu na uzročni faktor, preliminarna ispitivanja su dokazala da na izgled izolovan region, može da predstavlja značajan rezervoar mikroorganizama koji mogu da izazovu epizootije. Istovremeno, a u sadejstvu sa spoljašnjim faktorom, naročito prometom između regiona sveta gde se još uvek pojavljuju egzotične zarazne bolesti životinja, postoji rizik od unosa uzročnika zaraznih bolesti koje se smatraju egzotičnim za našu zemlju i Evropu. Raznovrsnost i variranje elemenata spoljašnje sredine kao epizootiološkog faktora, čine da je procena epizootiološke situacije u regionu Stare planine teška, a analiza rizika od pojave epizootija relativno kompleksna.

Moderan način života, globalna razmena dobara, mnogo češći kontakti i saživot sa kućnim ljubimcima, domaćim i egzotičnim životinjskim vrstama, doveli su do toga da su danas različite vrste životinja češće izložene raznim korona virusima (CoVs). Većina CoVs je specifična za vrstu i ne prenosi se između vrsta. Za mnoge CoVs je utvrđeno da se, nakon uvođenja u prijemčivu populaciju, veoma brzo šire i ostaju endemske uprkos pokušajima da se vakcinacijom i drugim merama spreči njihovo širenje. Većina animalnih CoV je prisutna u prijemčivoj populaciji, uzrokujući bolest koja može da ima blagu do vrlo izraženu kliničku sliku, uključujući i visoku smrtnost. Veoma retko, sojevi animalnih CoVs se prenose na ljude, kada se brzo šire među humanom populacijom u vidu izraženog akutnog respiratornog sindroma (eng. severe acute respiratory syndrome CoV-2; skr. SARS CoV-2). Epizootiološke karakteristike koronavirusnih infekcija su posledica jedinstvenih karakteristika koje mogu da se svrstaju u tri osnovne epizootiološke determinante. Kao prvo, sam virus ima svoje osobine u smislu genetskih karakteristika i održivosti u spoljašnjoj sredini. U odnosu na drugu epizootiološku determinantu odnosno makroorganizam, potrebno je da se naglasi da se korona virusi, češće u odnosu na ostale familije virusa, u prirodi adaptiraju na primarnu vrstu ili čak na novu vrstu životinja. Ovaj tzv "preskok" vrste je pre svega uslovljen biologijom virusa koji često menja antigeni sastav i time ne samo da izbegava imunski odgovor domaćina već u prirodi pronalazi nove vrste za svoje održavanje. Već hiljadama godina spoljašnja sredina doprinosi činjenici da su korona virusi pratioci većeg broja životinjskih vrsta pošto postoje određene odlike spoljašnje sredine (treća epizootiološka determinanta) kao što je deljenje iste ekološke niše od strane različitih vrsta životinja (i čoveka).

Due to the modern way of life, the global exchange of goods, more frequent contacts, and the closer life with pets, domestic and exotic animal species, different species are also more often exposed to various coronaviruses (CoVs). Most CoVs are species-specific and are not transmitted between different species. Many CoVs have been found to spread very rapidly after introduction into the susceptible population and have remained endemic despite vaccination attempts and other measures to prevent their spread. The majority of animal CoV are present in susceptible population and can cause from mild to a severe clinical picture of the disease, including high mortality. Very rarely strains of animal CoVs are transmitted to humans and then spread rapidly among humans like severe acute respiratory syndrome CoV-2 (SARS CoV-2). The epizootiological characteristics of coronavirus infections are a consequence of their unique characteristics that can be classified into three basic epizootiological determinants. First of all, the virus itself has its own characteristics in terms of genetic characteristics and sustainability in the environment. In relation to the second epizootiological determinant, the macro-organism, it is necessary to emphasize that coronaviruses, more often than other families of viruses, adapt to primary species o even to a new species of animal in nature. This, so-called "jumping" the species barriers, is primarily conditioned by the biology of the virus, which often changes the antigenic composition and thus not only avoids the host's immune response but also finds new species in nature for its maintenance. For thousands of years, the environment greatly contributes to the fact that coronaviruses are a companion of populations of many animal species since there are certain characteristics of the environment (third epizootiological determinant) as sharing the same ecological niche by different species of animals (and humans).

Porcine Reproductive and Respiratory Syndrome is a viral disease of swine characterized by reproductive failure of breeding animals and respiratory disorders in all categories. The first PRRS case in Serbia was recorded in 2001 after illegal import of boar semen. PRRS is economically the most important disease due to significant direct and indirect losses. Today, for routine diagnosis of PRRS in infected herds serological methods (ELISA) and molecular methods are used. Although modern diagnostic techniques are very robust, exceptional diversity of the viral strains is often the obstacle for an accurate diagnosis. To estimate the performance of seven different methods for PRRSV genome detection, twenty samples were used. However, none of the methods was able to detect all PRRSV strains. The best sensitivity was obtained by combining two methods. Until today, there is no absolutely accurate test which enables the detection of all circulating strains.

Poslednjih godina je evidentno pogoršanje epizootiološke situacije naročito opasnih zaraznih bolesti, ne samo u Republici Srbiji i regionu Balkanskog poluostrva već i u celom svetu. Afrička kuga svinja postepeno, ali sigurno osvaja evropski kontinent pa su u 2019. godini Slovačka i Srbija po prvi put zabeležile pojavu ove bolesti. Sa druge strane, neprekidno prisustvo slinavke i šapa, boginja ovaca i boginja koza, kao i širenje kuge malih preživara u Turskoj i van njenih granica, pokazatelji su sve većih zahteva za dobrom organizacijom veterinarske struke naše zemlje. Uz bolest plavog jezika, nodularni dermatitis, afričku kugu svinja i bolest Zapadnog Nila, veterinari su suočeni sa pojavom sve većeg broja novih zaraznih bolesti za koje su potrebna znanja kako bi se one na vreme prepoznale i njihovo širenje zaustavilo. Evidentan je porast broja slučajeva tuberkuloze, bruceloze i drugih bolesti za koje se verovalo da su potpuno pod kontrolom. Veterinari su pod stalnim pritiskom pojave ne samo naročito opasnih zaraznih bolesti koje prete regionu, već i onih za koje se verovalo da su na pragu iskorenjivanja. Izbijanje jedne od najopasnijih zoonoza, groznice doline Rift na francuskom ostrvu Majote, važnoj turističkoj destinaciji, ukazuje na to da ne treba zanemariti ni rizik širenja bolesti uveženim slučajevima, ne samo kod ljudi, već i kod životinja., EMERGING DISEASES SIGNIFICANT FOR SERBIA AND THE REGION Over the last few years, there has been an apparent deterioration of epizootiological situation especially in case of emerging diseases, not only in the Republic of Serbia and the Balkan peninsula region, but also worldwide. African swine fever has been gradually but surely spreading across Europe. The disease was recorded for the first time in Serbia 22 XLI INOVACIJE ZNANJA VETERINARA Beograd, 2020 and Slovakia, in 2019. On the other hand, continuous presence of foot and mouth disease, capripox and spreading of peste des petit ruminants in and out of Turkey, clearly indicates the importance of good organisation of veterinary profession in our country. Along with blue tongue disease, nodular dermatitis, African swine fever, West Nile fever, veterinarians are facing increasing number of new contagious diseases which requires knowledge to detect them on time and prevent their spreading. Cases of tuberculosis, brucelosis and other diseases, that have been assumed as kept under control, have significantly increased. Veterinarians are under constant pressure not only by a potential outbreak of emerging diseases that might impose the threat to the region, but also of those considered to have been eradicated. Outbreak of one of the most dangerous zoonosis, Rift Valley fever on the French island of Majote, an important touristic destination, shows that we should not neglect the risk of spreading the disease by imported cases, not only in humans but also in animals.

epizootiological and epidemiological role of the golden jackals (Canis aureus) as potential reservoirs for zoonotic vector-borne pathogens, for the first time in Serbia. Jackal spleens and collected ticks were tested by molecular methods (qPCR, conventional PCR, RFLP, sequencing) for the presence of vector-borne pathogens from the genera: Babesia spp., Rickettsia spp., Anaplasma spp., Ehrlichia spp., Francisella spp., Bartonella spp., Leishmania spp., and species from Borrelia burgdorferi sensu lato complex and Coxiella burneti. A total of 216 jackals originating from 10 localities throughout the territory of the Republic of Serbia were analyzed. Beside that, 118 ticks collected from jackals were also tested. The collected ticks belonged to one of three species: Ixodes ricinus (54,2%), Dermacentor reticulatus (39,8%) i Haemaphysalis concinna (5,9%). The presence of DNA of Babesia canis with the prevalence of 4.2% (9/216), Anaplasma phagocytophilum (0.9%, 2/216) and Leishmania spp. (6.9%, 15/216) was confirmed by molecular methods in the spleen of the tested jackals. In ticks collected from jackals, the presence of DNA from two Babesia spp. (Babesia canis, Babesia microti), Anaplasma phagocytophilum and three species from the Borrelia burgdorferi s.l. complex (Borrelia garinii, Borrelia lusitaniae, Borrelia valaisiana) was confirmed..., Istraživanjem sprovedenim u okviru ove doktorske disertacije, po prvi put u Srbiji, analiziran je epizootiološko-epidemiološki značaj šakala (Canis aureus) kao potencijalnih rezervoara za uzročnike zoonoza koje se prenose vektorima. Molekularnim metodama (qPCR, konvencionalni PCR, RFLP, sekvenciranje) u slezinama šakala i krpeljima sakupljenim sa šakala, dokazivano je prisustvo vektorima prenosivih patogena iz rodova: Babesia spp., Rickettsia spp., Anaplasma spp., Ehrlichia spp., Francisella spp., Bartonella spp., Leishmania spp., i vrsta u okviru Borrelia burgdorferi sensu lato kompleksa i Coxiella burneti. Ukupno je analizirano 216 šakala poreklom sa 10 lokaliteta širom teritorije Republike Srbije. Na prisustvo patogena ispitano je i 118 sakupljenih krpelja koji su pripadali jednoj od tri vrste: Ixodes ricinus (54,2%), Dermacentor reticulatus (39,8%) i Haemaphysalis concinna (5,9%). Molekularnim metodama u slezinama šakala potvrđeno je prisustvo DNK Babesia canis sa prevalencijom od 4,2% (9/216), Anaplasma phagocytophilum (0,9%, 2/216) i Leishmania spp. (6,9%, 15/216). U krpeljima sakupljenim sa šakala potvrđeno je prisustvo DNK dve vrste Babesia (Babesia canis, Babesia microti), Anaplasma phagocytophilum i tri vrste Borrelia burgdorferi s.l. kompleksa (Borrelia garinii, Borrelia lusitaniae, Borrelia valaisiana)...

Nosema ceranae je микроспоридијa којa паразитира код Европске медоносне пчеле, Apis mellifera, и доминантна је врста рода Nosema у југоисточној Европи. У зависности од степена инфекције и периода године, овај ендопаразит може имати далекосежне последице по пчеле, њихово здравље и продуктивност. Последњих година посебно је истраживан утицај N. ceranae на имунитет пчела а закључци појединих истраживања издвајају ефекат супресије гена значајних за имунитет пчела, односно доказују имуносупресивне последице инфекције врстом N. ceranae. Терапија ноземозе подразумева употребу антибиотика фумагилина, који има доказану ефикасност у сузбијању инфекције али његови споредни ефекти могу представљати проблем за саме пчеле и квалитет пчелињих производа, те је стога је његова примена у многим земљама ограничена. Имајући ово на уму константно се трага за природним и синтетским компонентама које би биле замена за фумагилин. Тестиране су многе супстанце, а међу онима које су показале најбољи ефекат издвајају се екстракти биљака и алги и синтетске витаминско-минералне мешавине. У овом раду је у кавезном експерименту тестирано пет препарата: (1) антибиотик фумагилин, (2) биљни екстракт тимол; и три адитива у исхрани пчела: (3) комерцијални препарат Beewell AminoPlus који представља витаминско-аминокиселинску мешавину, (4) комерцијални препарат Medenko forte који садржи екстракте храстове коре, пелена и жалфије и (5) полисахаридни екстракт гљиве Agaricus blazei. Пчеле су у свим третираним групама трећег дана живота инфициране спорама N. ceranae, а различите групе су третиране препаратима од првог, трећег и шестог дана. Код третираних пчела је праћен ефекат на преживљавање пчела, степен Nosema инфекције, нивое експресије гена значајних за имунитет пчела (абецин, дефензин, хименоптецин, апидецин и вителогенин) и ниво оксидативног стреса праћен кроз активност антиоксидативних ензима (каталазе, супероксид дизмутазе и глутатион С-трансферазе) и концентрације малондиалдехида..., Apis mellifera, and the dominant Nosema species in Southeast Europe. Depending on the infection level it could have significant consequences on bees, their health and productivity. The impact of N. ceranae on honey bees’ immunity has been particularly investigated in recent years. Some conclusions of this research underline N. ceranae-induced suppression of immune-related genes, proving its immunosuppressive impact. Treatment for nosemosis includes the use of the antibiotic fumagillin. Fumagillin has proven effect in Nosema control, but its side effects may pose a problem for the bees and the quality of their products, which is why its application in many countries have been limited. Hence, researchers are constantly looking for natural and synthetic components which could be suitable substitutes for fumagillin. Many products have been tested and plant/algae extracts and synthetic vitamin-mineral complexes showed the best effects. This laboratory (cage) experiment included testing of five products: (1) antibiotic fumagillin (2) a plant extract - thymol, and three dietary supplements: (3) a commercial amino acid and vitamin complex named Beewell AminoPlus, (4) a commercial mixture of oak bark, sage and absinth extracts named Medenko forte, and (5) a polysaccharide extract of Agaricus blazei mushroom. Bees in all treatment groups were infected with N. ceranae spores on day 3, and treated with the listed products from day 1, 3 or 6 after emergence. Bee survival, Nosema loads, levels of immune-related genes (abaecin, apidaecin, hymenoptaecin, defensin and vitellogenin) expression and the level of oxidative stress, assessed through the activity of antioxidative enzymes (catalase, glutathione S-transferase and superoxide dismutase) and the concentration of malondialdehyde, were measured in all experimental groups. The obtained values were compared with those of control (noninfected and infected) groups...

Istraživanjem sprovedenim u okviru ove doktorske disertacije, po prvi put u Srbiji, analiziran je epizootiološko-epidemiološki značaj šakala (Canis aureus) kao potencijalnih rezervoara za uzročnike zoonoza koje se prenose vektorima. Molekularnim metodama (qPCR, konvencionalni PCR, RFLP, sekvenciranje) u slezinama šakala i krpeljima sakupljenim sa šakala, dokazivano je prisustvo vektorima prenosivih patogena iz rodova: Babesia spp., Rickettsia spp., Anaplasma spp., Ehrlichia spp., Francisella spp., Bartonella spp., Leishmania spp., i vrsta u okviru Borrelia burgdorferi sensu lato kompleksa i Coxiella burneti. Ukupno je analizirano 216 šakala poreklom sa 10 lokaliteta širom teritorije Republike Srbije. Na prisustvo patogena ispitano je i 118 sakupljenih krpelja koji su pripadali jednoj od tri vrste: Ixodes ricinus (54,2%), Dermacentor reticulatus (39,8%) i Haemaphysalis concinna (5,9%). Molekularnim metodama u slezinama šakala potvrđeno je prisustvo DNK Babesia canis sa prevalencijom od 4,2% (9/216), Anaplasma phagocytophilum (0,9%, 2/216) i Leishmania spp. (6,9%, 15/216). U krpeljima sakupljenim sa šakala potvrđeno je prisustvo DNK dve vrste Babesia (Babesia canis, Babesia microti), Anaplasma phagocytophilum i tri vrste Borrelia burgdorferi s.l. kompleksa (Borrelia garinii, Borrelia lusitaniae, Borrelia valaisiana)..., epizootiological and epidemiological role of the golden jackals (Canis aureus) as potential reservoirs for zoonotic vector-borne pathogens, for the first time in Serbia. Jackal spleens and collected ticks were tested by molecular methods (qPCR, conventional PCR, RFLP, sequencing) for the presence of vector-borne pathogens from the genera: Babesia spp., Rickettsia spp., Anaplasma spp., Ehrlichia spp., Francisella spp., Bartonella spp., Leishmania spp., and species from Borrelia burgdorferi sensu lato complex and Coxiella burneti. A total of 216 jackals originating from 10 localities throughout the territory of the Republic of Serbia were analyzed. Beside that, 118 ticks collected from jackals were also tested. The collected ticks belonged to one of three species: Ixodes ricinus (54,2%), Dermacentor reticulatus (39,8%) i Haemaphysalis concinna (5,9%). The presence of DNA of Babesia canis with the prevalence of 4.2% (9/216), Anaplasma phagocytophilum (0.9%, 2/216) and Leishmania spp. (6.9%, 15/216) was confirmed by molecular methods in the spleen of the tested jackals. In ticks collected from jackals, the presence of DNA from two Babesia spp. (Babesia canis, Babesia microti), Anaplasma phagocytophilum and three species from the Borrelia burgdorferi s.l. complex (Borrelia garinii, Borrelia lusitaniae, Borrelia valaisiana) was confirmed...

Nodularni dermatitis (ND) je akutno, subakutno ili hronično virusno oboljenje goveda i bivola. Prvi slučaj nodularnog dermatitisa u Srbiji je laboratorijski potvrđen 07.06.2016. godine. Najviše žarišta je bilo registrovano u Pčinjskom okrugu. Za sprečavanje širenja bolesti, celokupna populacija goveda je vakcinisana živom atenuiranom vakcinom od Neethling soja (OBP, Južnoafrička Republika). Cilj ove doktorske disertacije je ispitivanje aktivno i pasivno stečenog imunskog odgovora protiv virusa nodularnog dermatitisa kod goveda i novorođene teladi metodom virus neutralizacionog testa (VNT), tehnikom indirektne imunofluorescencije (IFAT) i komercijalnim ID Screen® Capripox double antigen Multi-species-ELISA testom proizvođača IDvet (Monpelje, Francuska). Pored seroloških ispitivanja, kod vakcinisanih životinja ispitivano je i prisustvo vakcinalnog i terenskog soja virusa metodom Real-Time PCR. Materijal za ispitivanje je sakupljen na teritoriji opštine Kraljevo u selu Vrdila, gde nije bilo prijavljenih slučajeva nodularnog dermatitisa. Uzorkovanje je sprovedeno tri puta u toku 2017. godine i to pre revakcinacije, mesec dana nakon revakcinacije i pet meseci nakon revakcinacije. Za ispitivanja aktivnog imunskog odgovora krvni serumi su sakupljeni od 79 životinja pre i mesec dana nakon revakcinacije i od 74 životinje pet meseci nakon revakcinacije. Za utvrđivanje specifičnosti ELISA testa ispitani su krvni serumi od 20 svinja seropozitivnih na Suipoxvirus. Pasivni imunski odgovor je ispitan pregledom krvnog seruma i kolostruma od 17 krava (na dan teljenja) i krvnog seruma od 20 teladi (0. i 14. dana starosti). Za ispitivanje karakteristika ELISA testa za ispitivanje mleka sakupljeno je pojedinačno i zbirno mleko od krava u laktaciji i to od 52 krave sa 12 gazdinstava pre revakcinacije, od 49 krava sa 11 gazdinstava mesec dana nakon revakcinacije i od 53 krave sa 15 gazdinstava pet meseci nakon revakcinacije. Kao negativna kontrola u cilju validacije ELISA testa ispitano, Nodular dermatitis (ND) is acute, subacute or chronic viral disease of cattle and buffalo. First laboratory confirmed ND outbreak in Serbia occurred on 07 June 2016. The most of the outbreaks registered was in Pčinjski district. In response to prevent spreading of the disease, vaccination of all cattle population was applied using of live attenuated Neethling vaccine (OBP, South Africa). Goal of this doctoral dissertation was investigation of active and passive acquired immunity against ND in cattle and new-born calves using of virus neutralization test (VNT), indirect fluorescence antibody test (IFAT) and commercially available ID Screen® Capripox double antigen Multi-species-ELISA test from IDvet (Montpelier, France). Next to serological investigation, presence of vaccine and field nodular dermatitis virus (NDV) strain was investigated using of Real- Time PCR method. Material for this study was collected in Vrdila village, Kraljevo municipality area with no previous reported ND outbreaks. Sampling was performed three times in 2017, before revaccination, one month post re-vaccination and five months post re-vaccination. For serological investigation of active acquired immunity blood serum samples were collected out of 79 cattle before and one month post re-vaccination and out of 74 cattle five months post re-vaccination. Blood serum samples from 20 swine seropositive to Suipoxvirus were used for investigation of ELISA specificity. Passive acquired immunity was investigated from blood serum and colostrums samples out of 17 cows (on day of calving) and from blood serum samples out of 20 new-born calves (0 and 14 days old). For investigation of characteristics of ELISA test for milk samples, individual and bulk milk samples were collected from cows which were in lactation, out of 52 cows and 12 farms before re-vaccination, 49 cows and 11 farms one month after re-vaccination and out of 53 cows and 15 farms five months post re-vaccination. As for negative control with t

Nosema ceranae is a microsporidian endoparasite of the European honey bee, Apis mellifera, and the dominant Nosema species in Southeast Europe. Depending on the infection level it could have significant consequences on bees, their health and productivity. The impact of N. ceranae on honey bees’ immunity has been particularly investigated in recent years. Some conclusions of this research underline N. ceranae-induced suppression of immune-related genes, proving its immunosuppressive impact. Treatment for nosemosis includes the use of the antibiotic fumagillin. Fumagillin has proven effect in Nosema control, but its side effects may pose a problem for the bees and the quality of their products, which is why its application in many countries have been limited. Hence, researchers are constantly looking for natural and synthetic components which could be suitable substitutes for fumagillin. Many products have been tested and plant/algae extracts and synthetic vitamin-mineral complexes showed the best effects. This laboratory (cage) experiment included testing of five products: (1) antibiotic fumagillin (2) a plant extract - thymol, and three dietary supplements: (3) a commercial amino acid and vitamin complex named Beewell AminoPlus, (4) a commercial mixture of oak bark, sage and absinth extracts named Medenko forte, and (5) a polysaccharide extract of Agaricus blazei mushroom. Bees in all treatment groups were infected with N. ceranae spores on day 3, and treated with the listed products from day 1, 3 or 6 after emergence. Bee survival, Nosema loads, levels of immune-related genes (abaecin, apidaecin, hymenoptaecin, defensin and vitellogenin) expression and the level of oxidative stress, assessed through the activity of antioxidative enzymes (catalase, glutathione S-transferase and superoxide dismutase) and the concentration of malondialdehyde, were measured in all experimental groups. The obtained values were compared with those of control (noninfected and infected) gr, Nosema ceranae je mikrosporidija koja parazitira kod Evropske medonosne pčele, Apis mellifera, i dominantna je vrsta roda Nosema u jugoistočnoj Evropi. U zavisnosti od stepena infekcije i perioda godine, ovaj endoparazit može imati dalekosežne posledice po pčele, njihovo zdravlje i produktivnost. Poslednjih godina posebno je istraživan uticaj N. ceranae na imunitet pčela a zaključci pojedinih istraživanja izdvajaju efekat supresije gena značajnih za imunitet pčela, odnosno dokazuju imunosupresivne posledice infekcije vrstom N. ceranae. Terapija nozemoze podrazumeva upotrebu antibiotika fumagilina, koji ima dokazanu efikasnost u suzbijanju infekcije ali njegovi sporedni efekti mogu predstavljati problem za same pčele i kvalitet pčelinjih proizvoda, te je stoga je njegova primena u mnogim zemljama ograničena. Imajući ovo na umu konstantno se traga za prirodnim i sintetskim komponentama koje bi bile zamena za fumagilin. Testirane su mnoge supstance, a među onima koje su pokazale najbolji efekat izdvajaju se ekstrakti biljaka i algi i sintetske vitaminsko-mineralne mešavine. U ovom radu je u kaveznom eksperimentu testirano pet preparata: (1) antibiotik fumagilin, (2) biljni ekstrakt timol; i tri aditiva u ishrani pčela: (3) komercijalni preparat Beewell AminoPlus koji predstavlja vitaminsko-aminokiselinsku mešavinu, (4) komercijalni preparat Medenko forte koji sadrži ekstrakte hrastove kore, pelena i žalfije i (5) polisaharidni ekstrakt gljive Agaricus blazei. Pčele su u svim tretiranim grupama trećeg dana života inficirane sporama N. ceranae, a različite grupe su tretirane preparatima od prvog, trećeg i šestog dana. Kod tretiranih pčela je praćen efekat na preživljavanje pčela, stepen Nosema infekcije, nivoe ekspresije gena značajnih za imunitet pčela (abecin, defenzin, himenoptecin, apidecin i vitelogenin) i nivo oksidativnog stresa praćen kroz aktivnost antioksidativnih enzima (katalaze, superoksid dizmutaze i glutation S-transferaze) i koncentracije malondialdehida.

Cilj ove doktorske disertacije je identifikacija i molekularna karakterizacija sojeva herpesvirusa konja poreklom iz uzoraka od konja sa teritorije Republike Srbije i Republike Srpske (BiH) kao i arhiviranih sojeva EHV-1 Katedre za mikrobiologiju Fakulteta veterinarske medicine Univerziteta u Beogradu. Arhivirani sojevi EHV-1 su liofilizati pomenutog virusa izolovani osamdesetih godina prošlog veka iz uzoraka pobačenih fetusa konja, kao i iz uginule novorođene ždrebadi sa ergele „Ljubičevo“. Ispitivani su uzorci poreklom od ukupno 137 nevakcinisanih konja sa ergela i iz privatnog sektora koji su obuhvatali 112 uzoraka nosnih briseva i 100 uzoraka organa - submandibularnih limfnih čvorova, slezine, produžene moždine i kičmene moždine prikupljenih od 25 konja. Prikupljeni uzorci nosnih briseva i organa konja su primenom klasičnih i molekularnih metoda virusološke dijagnostike ispitivani na prisustvo konjskih herpesvirusa 1, 4, 2 i 5 (EHV-1, EHV-4, EHV-2 i EHV-5), dok su arhivirani sojevi ispitivani primenom molekularnih metoda u cilju molekularne karakterizacije. Posle izolacije i identifikacije virusa na kulturi tkiva primenom testa virus neutralizacije, direktne imunofluorescencije i metode Nested multiplex PCR od ukupno 212 ispitivanih uzoraka organa i nosnih briseva konja prisustvo konjskog herpesvirusa 1 je utvrđeno u 72 uzorka, dok je metodom izolacije virusa na kulturi ćelija sa identifikacijom primenom metode Nested multiplex PCR iz prethodno navedenih uzoraka konjski herpesvirus 5 detektovan u 7 uzoraka, a konjski herpesvirusi 4 i 2 nisu izolovani ni iz jednog uzorka nosnih briseva i organa konja. Primenom metode Nested multiplex PCR za ispitivanje prisustva nukleinskih kiselina konjskih herpesvirusa 1, 4, 2 i 5 direktno u uzorcima ispitivanog materijala (organa i nosnih briseva konja) detektovano je ukupno 162 uzorka pri čemu je EHV-1 detektovan u 153 uzorka, EHV-4 u 11, EHV-2 u 4, a EHV-5 u 23 uzorka ispitivanog materijala. Od ukupnog broja pozitivnih uzora, The aim of this PhD thesis was the identification and molecular characterization of equine herpesviruses from horses originating from the Republic of Serbia and The Republic of Srpska (Bosnia and Herzegovina), as well as archivated EHV-1 strains in the possession of The Department of Microbiology, Faculty of Veterinary Medicine, University of Belgrade. Archivated EHV-1 strains were isolated during abortion storms that occured on „Ljubičevo“ stud farm during the 1980s. Samples originating from 137 unvaccinated horses from stud farms and private breeders, i.e. 112 nasal swab samples and 100 organ samples from 25 horses (submandibular lymph nodes, spleen, spinal cord and medulla) were examined for the presence of equine herpesviruses 1, 4, 2 and 5 (EHV-1, EHV-4, EHV-2 and EHV-5) using standard and molecular virological methods. Archivated EHV-1 samples were examined by molecular methods in order to perform their genetic characterization. Equine herpesvirus 1 was isolated and identified by virus neutralization test, direct immunofluorescence and Nested multiplex PCR in 72 samples. Equine herpesvirus 5 was isolated from 7 examined samples and identified by Nested multiplex PCR, whilst the isolation of EHV-4 and EHV-2 was not successful. The direct examination of the presence of equine herpesviruses 1, 4, 2 and 5 was performed by Nested multiplex PCR directly from the samples of organs and nasal swabs and confirmed 162 positive samples of which 16.67% accounted for mixed infections with multiple equine herpesviruses. Equine herpesvirus 1 was confirmed in 153 samples, EHV-4 in 11, EHV-2 in 4 and EHV-5 in 23 samples, whilst EHV-2 and EHV-4 were only detected in mixed infections with EHV-1 and/or EHV-5. Partial gB gene nucleotide sequences of identified EHV-1 strains were 98 to 100% homologous amongst each other and with sequences from GenBank whilst their phylogenetic analysis showed grouping with strains from Turkey, UK, USA and Japan...

najznačajnijih oboljenja živine izazvano virusom iz familije Birnaviridae, rod Avibirnavirus. Bolest izaziva visoku stopu morbiditeta i mortaliteta, smanjenje produktivnosti i dovodi do imunosupresije. Kontrola bolesti se sprovodi primenom biosigurnosnih mera, poštovanjem tehnoloških normativa i imunizacijom živine. Vakcine protiv IBD su kategorizovane kao klasične (blage, intermedijalne i „vruće“), imunski kompleks vakcine i vektor vakcine. U Srbiji se još uvek u rutinskoj praksi ne primenjuju vakcine na bazi „imunskog kompleksa“ i rekombinantne vektorske vakcine. U ovom radu su ispitivani i poređeni efekti primene klasične vakcine i rekombinantne vektorske vakcine protiv IBD, kroz analizu humoralnog i celularnog imunskog odgovora, praćenjem nivoa, ujednačenosti i brzine stvaranja imunskog odgovora, pregleda burzi Fabrici, merenjem B/B indeksa i zaštite koju vakcine pružaju nakon veštačke (challenge) infekcije virusom infektivne burzalne bolesti. Rezultati ispitivanja pokazuju da u grupi pilića vakcinisanih rekombinantnom vektorskom vHVT13 vakcinom, nakon veštačke infekcije vvIBD nije bilo obolelih ni uginulih pilića. Kod grupa pilića vakcinisanih živim atenuiranim vakcinama („intermedijarna“; „intermedijarna plus“) konstantovani su klinički simptomi bolesti i uginuća. Takođe, praćenjem indeksa burze (B:BR i B:BI) utvrđena je statistički značajna razlika u korist grupe vakcinisane vHVT13 vakcinom. Serološki monitoring sproveden je simultano sa dva ELISA kit kompleta: unapređenim ELISA IBD plus test i klasičnim ELISA IBD kitom. Klasičnim ELISA testovima nije moguće sprovoditi monitoring posle vakcinacije vektor vakcinom jer oni ne detektuju u dovoljnoj meri anti VP2 antitela. Utvrđen je znatno viši titar antitela meren unapređenim IBD plus testom u grupi vakcinisanoj vektor vakcinom (ne manje od 5000) u odnosu na grupe pilića vakcinisane živim atenuiranim vakcinama protiv IBD. Rezultati pregleda burzi, monitoring krvi i veštačka infekcija pokazuju da su vektor vakci, Infectious bursal disease (IBD) is one of the most significant viral diseases in poultry production caused by Avibirnavirus, Birnaviridae. Disease is characterized by high morbidity and mortality rate, as well as low productivity and marked immunosupression. Disease control is carried out by application of biosecurity measures, following of technical procedures and active immunization of chicken. IBD vaccines are categorized as classical (mild, intermediate and intermediateplus), immunocomplex vaccines and recombinant vector vaccines. Routine practice in Serbia includes application of classic vaccines, while immunocomplex and recombinant vaccines are not in significant use. Objective of this experiment was to compare effects of classic and recombinant vaccines against IBD by analyzing level, homogeneity and development agility of humoral and cellular immune response, inspection of bursa, calculation of body/bursa index (B/B index) and protection after challenge test. Obtained results showed no diseased or mortality in the group of chickens vaccinated with vHVT13 after challenge with vvIBD virus. Disease symptoms and death occurred in control groups vaccinated with live attenuated vaccines (intermediary, intermediary plus). Statistically significant difference in B.BR and B.BI index was noticed in group vaccinated with vHVT13 when compared to control group. Serological testing was conducted with two distinct ELISA kits: classic ELISA IBD kit and ELISA IBD plus kit. Classic ELISA kit can not be used for monitoring of flocks vaccinated with recombinant vector vaccine because it does not detect antiVP2 antibodies in significant measure. ELISA IBD plus kit detected significantly higher titer in experimental group (not less than 5000) compared to groups vaccinated with classic vaccines. Results of flow citometry of peripheral blood show no decrease in IgY level neither after vaccination nor after artificial infection. There is no statistical difference in IgY level among

Bruceloza pasa je bolest poznata preko četiri decenije, ali i pored toga ne postoje standardizovani dijagnostički protokoli, kao ni generalni dogovor o najprikladnijem dijagnostičkom testu. Svaka laboratorija definiše sopstvene kriterijume. Ovakva raznovrsnost testova i nedostatak jasno definisanih protokola dovodi do teškoća u interpretaciji rezultata seroloških testova u različitim laboratorijama. Iz tog razloga, cilj ove doktorske disertacije obuhvatio je unapređenje dijagnostike primenom preporučenih i novih, sopstveno pripremljenih testova. Jedan od zadataka ove disertacije je bilo i ispitivanje upotrebljivosti Bruce-ladder multiplex PCR metode za ispitivanje kliničkih uzoraka odnosno tkiva uterusa i testisa pasa. Sakupljen je materijal (krv, testisi i materice) od 225 nevlasničkih pasa i to 145 ženki i 80 mužjaka. Dobijeni rezultati su pokazali da je od ukupno 225 ispitanih uzoraka, 33 ili 14,67% krvnih seruma imalo merljiv titar antitela u 2-ME TAT. Najniži ispitivani titar od 1/50 imalo je 13 krvnih seruma ili 5,78%, kod 8 krvnih seruma ustanovljen je titar od 1/100 ili 3,55%, dok je titar od 1/200 imalo 12 uzoraka seruma ili 5,33%. Primenom klasičnih bakterioloških metoda, B. canis je izolovana iz tri uzorka homogenizata tkiva reproduktivnih organa (1,33%) i to iz 2 uzorka poreklom od mužjaka i jednog uzorka poreklom od ženke. Jedan izolat je poticao od serološki negativnog psa. Od 225 uzoraka ispitanih Bruce-ladder multiplex PCR metodom, pozitivna reakcija je ustanovljena kod dva (0,88%). Pre formulacije indirektnog ELISA testa, određivane su koncentracije proteina. Elektroforetska analiza antigena dobijenih toplotom i ultrazvukom kao i denzitometrijska kvantifikacija su pokazale da je u antigenu dobijenom toplotom, najzastupljenija frakcija molekulske mase 10,95 kDa sa učešćem od čak 43,12% koja odgovara R-LPS-u brucela. Ista frakcija je u antigenu dobijenom ultrazvukom bila zastupljena sa 11,56%, odnosno u količini koja je bila 3,7x manja..., The scientific community has known about canine brucellosis for over four decades, even so, there are no standardized diagnostic protocols, nor a general agreement on the most appropriate diagnostic test. Each laboratory defines its own criteria. This variety of tests and the lack of clearly defined protocols lead to difficulties in interpreting the results of serological tests in different laboratories. For this reason, the goal of this doctoral dissertation has been to improve diagnostics using recommended and new, self-prepared tests. One of the tasks of this dissertation was to examine the usability of the Bruce-ladder multiplex PCR method for testing clinical samples, i.e. dog uterine and testicular tissues. The material (blood, testicles and uteruses) was collected from 225 dogs without owners, 145 female and 80 male dogs. The results obtained showed that from a total of 225 tested samples, 33 or 14,67% of blood sera had measurable antibody titer in 2-ME TAT. 13 or 5,78% blood sera had the lowest tested titer of 1/50, in 8 or 3,55% blood sera a titer of 1/100 was determined, while 12 serum samples or 5,33% had the titer of 1/200. By applying classic bacteriological methods, B. canis was isolated from three samples of homogenized reproductive organ tissue (1,33%), 2 from samples originating from males and one specimen originating from a female. One isolate originated from a serologically negative dog. Of the 225 samples assayed using the Bruce-ladder multiplex PCR method, a positive reaction was established in two (0,88%). Protein concentrations were determined prior to the formulation of the indirect ELISA test. The electrophoretic analysis of antigens retrieved using heat and ultrasound, as well as the densitometric quantification, showed that the antigen retrieved using heat had the most prevalent molecular weight fraction of 10,95 kDa with a participation of as much as 43,12% that corresponds to Brucella R LPS. The same fraction was represented in the antig

The infectious bursal disease virus (IBDv) is widespread in poultry flocks all around the world. Various biotypes have emerged and because of that, adequate management practices and vaccination of chicks are of paramount importance for the protection against field strains. One day old Lohmann Brown chicks were vaccinated with intermediate vaccines and the recombinant VAXXITEK HVT-IBDv vaccine formulation, and challenged at 48 days of life with the very virulent IBDv (vvIBDv) strain CH/99. The best protection (100%) was achieved with the recombinant vaccine administered by the subcutaneous or intramuscular route at a day old, while intermediate and intermediate plus vaccines protected 80% of birds from clinical symptoms. The highest bursa body ratio (5.33, 3.50 and 4.12) was accomplished in non-vaccinated and non-challenged birds and birds vaccinated with the VAXXITEK HVT-IBDv vaccine. The recombinant VAXXITEK HVT-IBDv vaccine has provided protection for commercial chicks against challenge with the vvIBDv strain in this experiment. Under field conditions, additional vaccination is possibly needed with supplementary application of live attenuated vaccines. However, the recombinant vector vaccines are providing significant aid against clinical signs and immunosupression caused by the vvIBDv.

The presence of equine herpesviruses 1, 2 and 5 (EHV-1, EHV-2 and EHV-5) was examined in 66 samples of spinal cord, submandibular lymph nodes and spleen of healthy, non-vaccinated abattoir horses from different locations in the Republic of Serbia. Virus isolation was conducted on RK-13 cell line with the confirmation of isolated viral strains by multiplex nested polymerase chain reaction. The cytopathic effect was observed 48-72 h after the first inoculation in 28 (42.4%) organ samples, and after 5 days in 11 other samples (16.7%) that were all confirmed as EHV-1. Four other samples (6.1%) that showed cytopathic effects on day 5 of the third passage were all positive for EHV-5. Additionally, EHV-1, EHV-2, and EHV-5 were directly detected in all organs by multiplex nested PCR in 46 (69.7%), 3 (4.5%), and 7 (10.6%) samples, respectively. The molecular characterization based on nucleotide sequencing of the part of the gB gene showed that Serbian EHV-1 isolates were 100% homogenous and clustered with EHV-1 strains from Turkey, the United Kingdom, the United States, and Japan. The EHV-2 strain from Serbia branched together with Turkish EHV-2 isolates with homogeneity from 96% to 98%. Serbian EHV-5 strains can be separated in one distinct cluster with isolates from Turkey and the United States with homogeneity from 98 to 99%. These data represent the first report of the molecular characterization of EHV-1, EHV-2, and EHV-5 in the horse population of the Republic of Serbia and document the first successful isolation of Serbian EHV-5 strains.

The scientific community has known about canine brucellosis for over four decades, even so, there are no standardized diagnostic protocols, nor a general agreement on the most appropriate diagnostic test. Each laboratory defines its own criteria. This variety of tests and the lack of clearly defined protocols lead to difficulties in interpreting the results of serological tests in different laboratories. For this reason, the goal of this doctoral dissertation has been to improve diagnostics using recommended and new, self-prepared tests. One of the tasks of this dissertation was to examine the usability of the Bruce-ladder multiplex PCR method for testing clinical samples, i.e. dog uterine and testicular tissues. The material (blood, testicles and uteruses) was collected from 225 dogs without owners, 145 female and 80 male dogs. The results obtained showed that from a total of 225 tested samples, 33 or 14,67% of blood sera had measurable antibody titer in 2-ME TAT. 13 or 5,78% blood sera had the lowest tested titer of 1/50, in 8 or 3,55% blood sera a titer of 1/100 was determined, while 12 serum samples or 5,33% had the titer of 1/200. By applying classic bacteriological methods, B. canis was isolated from three samples of homogenized reproductive organ tissue (1,33%), 2 from samples originating from males and one specimen originating from a female. One isolate originated from a serologically negative dog. Of the 225 samples assayed using the Bruce-ladder multiplex PCR method, a positive reaction was established in two (0,88%). Protein concentrations were determined prior to the formulation of the indirect ELISA test. The electrophoretic analysis of antigens retrieved using heat and ultrasound, as well as the densitometric quantification, showed that the antigen retrieved using heat had the most prevalent molecular weight fraction of 10,95 kDa with a participation of as much as 43,12% that corresponds to Brucella R LPS. The same fraction was represented in the antig, Bruceloza pasa je bolest poznata preko četiri decenije, ali i pored toga ne postoje standardizovani dijagnostički protokoli, kao ni generalni dogovor o najprikladnijem dijagnostičkom testu. Svaka laboratorija definiše sopstvene kriterijume. Ovakva raznovrsnost testova i nedostatak jasno definisanih protokola dovodi do teškoća u interpretaciji rezultata seroloških testova u različitim laboratorijama. Iz tog razloga, cilj ove doktorske disertacije obuhvatio je unapređenje dijagnostike primenom preporučenih i novih, sopstveno pripremljenih testova. Jedan od zadataka ove disertacije je bilo i ispitivanje upotrebljivosti Bruce-ladder multiplex PCR metode za ispitivanje kliničkih uzoraka odnosno tkiva uterusa i testisa pasa. Sakupljen je materijal (krv, testisi i materice) od 225 nevlasničkih pasa i to 145 ženki i 80 mužjaka. Dobijeni rezultati su pokazali da je od ukupno 225 ispitanih uzoraka, 33 ili 14,67% krvnih seruma imalo merljiv titar antitela u 2-ME TAT. Najniži ispitivani titar od 1/50 imalo je 13 krvnih seruma ili 5,78%, kod 8 krvnih seruma ustanovljen je titar od 1/100 ili 3,55%, dok je titar od 1/200 imalo 12 uzoraka seruma ili 5,33%. Primenom klasičnih bakterioloških metoda, B. canis je izolovana iz tri uzorka homogenizata tkiva reproduktivnih organa (1,33%) i to iz 2 uzorka poreklom od mužjaka i jednog uzorka poreklom od ženke. Jedan izolat je poticao od serološki negativnog psa. Od 225 uzoraka ispitanih Bruce-ladder multiplex PCR metodom, pozitivna reakcija je ustanovljena kod dva (0,88%). Pre formulacije indirektnog ELISA testa, određivane su koncentracije proteina. Elektroforetska analiza antigena dobijenih toplotom i ultrazvukom kao i denzitometrijska kvantifikacija su pokazale da je u antigenu dobijenom toplotom, najzastupljenija frakcija molekulske mase 10,95 kDa sa učešćem od čak 43,12% koja odgovara R-LPS-u brucela. Ista frakcija je u antigenu dobijenom ultrazvukom bila zastupljena sa 11,56%, odnosno u količini koja je bila 3,7x manja...

The aim of this PhD thesis was the identification and molecular characterization of equine herpesviruses from horses originating from the Republic of Serbia and The Republic of Srpska (Bosnia and Herzegovina), as well as archivated EHV-1 strains in the possession of The Department of Microbiology, Faculty of Veterinary Medicine, University of Belgrade. Archivated EHV-1 strains were isolated during abortion storms that occured on „Ljubičevo“ stud farm during the 1980s. Samples originating from 137 unvaccinated horses from stud farms and private breeders, i.e. 112 nasal swab samples and 100 organ samples from 25 horses (submandibular lymph nodes, spleen, spinal cord and medulla) were examined for the presence of equine herpesviruses 1, 4, 2 and 5 (EHV-1, EHV-4, EHV-2 and EHV-5) using standard and molecular virological methods. Archivated EHV-1 samples were examined by molecular methods in order to perform their genetic characterization. Equine herpesvirus 1 was isolated and identified by virus neutralization test, direct immunofluorescence and Nested multiplex PCR in 72 samples. Equine herpesvirus 5 was isolated from 7 examined samples and identified by Nested multiplex PCR, whilst the isolation of EHV-4 and EHV-2 was not successful. The direct examination of the presence of equine herpesviruses 1, 4, 2 and 5 was performed by Nested multiplex PCR directly from the samples of organs and nasal swabs and confirmed 162 positive samples of which 16.67% accounted for mixed infections with multiple equine herpesviruses. Equine herpesvirus 1 was confirmed in 153 samples, EHV-4 in 11, EHV-2 in 4 and EHV-5 in 23 samples, whilst EHV-2 and EHV-4 were only detected in mixed infections with EHV-1 and/or EHV-5. Partial gB gene nucleotide sequences of identified EHV-1 strains were 98 to 100% homologous amongst each other and with sequences from GenBank whilst their phylogenetic analysis showed grouping with strains from Turkey, UK, USA and Japan..., Cilj ove doktorske disertacije je identifikacija i molekularna karakterizacija sojeva herpesvirusa konja poreklom iz uzoraka od konja sa teritorije Republike Srbije i Republike Srpske (BiH) kao i arhiviranih sojeva EHV-1 Katedre za mikrobiologiju Fakulteta veterinarske medicine Univerziteta u Beogradu. Arhivirani sojevi EHV-1 su liofilizati pomenutog virusa izolovani osamdesetih godina prošlog veka iz uzoraka pobačenih fetusa konja, kao i iz uginule novorođene ždrebadi sa ergele „Ljubičevo“. Ispitivani su uzorci poreklom od ukupno 137 nevakcinisanih konja sa ergela i iz privatnog sektora koji su obuhvatali 112 uzoraka nosnih briseva i 100 uzoraka organa - submandibularnih limfnih čvorova, slezine, produžene moždine i kičmene moždine prikupljenih od 25 konja. Prikupljeni uzorci nosnih briseva i organa konja su primenom klasičnih i molekularnih metoda virusološke dijagnostike ispitivani na prisustvo konjskih herpesvirusa 1, 4, 2 i 5 (EHV-1, EHV-4, EHV-2 i EHV-5), dok su arhivirani sojevi ispitivani primenom molekularnih metoda u cilju molekularne karakterizacije. Posle izolacije i identifikacije virusa na kulturi tkiva primenom testa virus neutralizacije, direktne imunofluorescencije i metode Nested multiplex PCR od ukupno 212 ispitivanih uzoraka organa i nosnih briseva konja prisustvo konjskog herpesvirusa 1 je utvrđeno u 72 uzorka, dok je metodom izolacije virusa na kulturi ćelija sa identifikacijom primenom metode Nested multiplex PCR iz prethodno navedenih uzoraka konjski herpesvirus 5 detektovan u 7 uzoraka, a konjski herpesvirusi 4 i 2 nisu izolovani ni iz jednog uzorka nosnih briseva i organa konja. Primenom metode Nested multiplex PCR za ispitivanje prisustva nukleinskih kiselina konjskih herpesvirusa 1, 4, 2 i 5 direktno u uzorcima ispitivanog materijala (organa i nosnih briseva konja) detektovano je ukupno 162 uzorka pri čemu je EHV-1 detektovan u 153 uzorka, EHV-4 u 11, EHV-2 u 4, a EHV-5 u 23 uzorka ispitivanog materijala. Od ukupnog broja pozitivnih uzora

Факултет ветеринарске медицине представља један од факултета који пружају практична знања и вештине, а планови и програми појединих предмета дају солидну основу да по дипломирању лекар ветеринарске медицине зна да постави сумњу на неку заразну и паразитску болест, обави хируршку операцију, анализира резултате дијагностичких испитивања телесних флуида, уведе животињу у анестезију, обави телење, анализира резултате испитивања квалитета сточарских производа, апликује терапију у зависности од дијагнозе и потреба животиња, саветује сточара у вези технологије узгоја и биосиурносних мера итд. Кроз сваки од наведених (и ненаведених) послова, провлачи се један заједнички именитељ. Наиме, појединачни случајеви тј. пацијенти и најчешће позитиван исход третмана, не би значили пуно ако се не би анализовали са становишта правилности и учесталости појављивања болести и успеха спроведених мера. Као лекари ветеринарске медицине, често у помоћ позивамо статистичаре који нам са мање или више успеха, користећи статистичке методе, објасне како се појединачни случајеви болести уклапају у целу слику односа епизоотиолошких детерминанти: узрока (микроорганизма), пријемчиве врсте животиња (макроорганизма) и спољашњег фактора. Међутим, биолошки закони који представљају основу ветеринарске медицине често или нису до краја познати или по природи ствари нису елементи које статистичари уче. Отуда је од великог значаја да сваки лекар ветеринарске медицине буде и епизоотиолог и да на основу својих искустава (рада), покуша да пронађе закономерности поремећаја здравља, производних карактеристика и добробити у популацијама животињских врста као и да установи који су то фактори који утичу на појаву ових поремећаја, а са циљем изналажења мера за њихово уклањање и/или смањењe штета. Имајући у виду професионални профил епизоотиолога, може да се каже да епизоотиолошка служба обједињује податке о поремећајима здравља, производних карактеристика и добробити животиња и има задатак да анализује добијене информ, The Faculty of Veterinary Medicine is one of the „hands on“ faculties whose curriculum offers a sound base for a graduate to diagnose an infective or parasitic disease, perform a surgical intervention, analyze results of body fluids tests, assist delivery, assess the quality of livestock products, apply treatment according to the diagnosis and need of the animal, offer council to the farmers, etc. In all the listed (and not listed) duties there is a common denominator, as individual cases would not be of relevance if not observed within the framework of disease regularity and incidence, as well as success of the performed measures. Doctors of veterinary medicine very often appeal to the help of statisticians which to a higher or lesser extent, with the aid of statistical methods, explain how individual cases fit within the whole picture of epizootiological factors: cause (microorganisms), susceptible animal (macro organism), and environmental factors. However, biological rules which represent the core of veterinary medicine very often are not elucidated, or are not taken into account by the statisticians. Thus, the importance for every veterinarian to find the epizootiologist within, and according to his/her knowledge and experience try to define the rules according to which a disease develops or production and welfare are affected. At the same time the veterinarian should be able to define which factors affect the presence of these disorders, all with the aim of prevention and/or damage mitigation. Bearing in mind the professional profile of an epizootiologist, it can be said that the epizootiological service encompasses data on health disorders, productive characteristics and animal welfare and within the Ministry of Agriculture has a role to analyze the obtained data, suggest and carry out control measures, combat and eradicate above all infectious diseases. Also, the Directorate for Veterinary Affairs forwards data to the International Office thus collaborating

Kada se bruceloza ustanovi u jednoj zemlji, međunarodni veterinarski propisi nameću ograničenja kretanja životinja i trgovine, što dovodi do velikih ekonomskih gubitaka. Izolacija Brucella spp. konvencionalnim bakteriološkim tehnikama je dugotrajna, rizična za laboratorijske radnike i niske osetljivosti zbog česte kontaminacije materijala. Negativna izolacija ne isključuje postojanje bruceloze. Najbolji rezultati do sada su dobijeni kombinovanjem metoda izolacije i PCR metode na kliničkim uzorcima. Nedostatak PCR metoda baziranih na razlici među sojevima unutar vrste stimulisao je razvoj novih tehnika „otisaka prstiju”. Podaci o sekvenci celog genoma brucela omogućili su identifikaciju i razlikovanje brucela na nivou vrste, biovara i upoređivanje sojeva što je olakšalo pronalaženje izvora infekcije. Indirektni dijagnostički testovi su zasnovani na detekciji imunskog odgovora izazvanog infekcijom. Ovi testovi pokazuju različit stepen osetljivosti i specifičnosti zavisno od brojnih varijabli, poput stepena i načina inficiranja, prisustva takozvanih „unakrsno reaktivnih bakterija” antigenski sličnih Brucella spp, kinetike indukovanog imunskog odgovora i prethodne vakcinacije. Imajući u vidu kinetiku nastajanja imunskog odgovora indukovanog nakon infekcije, vreme kada će se različiti testovi izvoditi ima veliki uticaj na rezultate., When brucellosis is detected in a country, international veterinary regulations impose restrictions on animal movements and trade, which result in huge economic losses. The isolation of Brucella spp. through conventional bacteriological techniques takes a long time, it’s risky for the laboratory workers, and has low sensitivity due to the frequent contamination of materials. Negative cultures do not rule out the disease. The best results have so far been obtained by combining culture and PCR detection on clinical samples. The lack of PCR-based methods for differentiation among strains within a species stimulate the development of new techniques-”fingerprinting methods”. Data on the sequence of the genome of Brucella enabled the identification and differentiation of Brucella at the level of species and biovar and comparison strains which is easier to find the source of infection. Indirect diagnostic tests are based on the detection of immune responses induced by infection. These tests show different sensitivities and specificities depending on numerous variables, such as the infection dose and route, the presence of so-called “cross-reactive bacteria” antigenically similar to Brucella spp., the kinetics of the induced immune response, and previous vaccination. Bearing in mind kinetics of the immune response induced after infection, the time when the different tests carried out has a major impact on the results.

Canine Monocytic Ehrlichiosis is a zoonotic bacterial disease with worldwide distribution. With regards to the population of stray dogs, the disease is facilitated due to their lifestyle and the lack of anti-parasitic protection. The aim of this study was to provide serological data on the presence of a specific Ehrlichia canis IgG antibodies in stray dogs, originating from 7 municipalities in Serbia. During the period from April 2013 to June 2014, 217 canine sera were submitted to the laboratory of the Department of Infectious Diseases of Animals and Bees, Faculty of Veterinary Medicine in Belgrade. An immunofluorescent antibody test (IFAT) was performed to detect antibodies to Ehrlichia canis (cut off, 1: 50). Seropositive dogs were found in 5 out of 7 counties with a seroprevalence varying from 3.57% to 20% and an overall seroprevalence of 11.06% (24/217). There was no statistically significant difference between the prevalence of infection and the host age or gender. Results showed that stray dogs contribute to maintaining and spreading of Ehrlichia canis in Serbia. Due to the close relationship between people and dogs, it is of great importance to constantly monitor and improve prevention of this disease.