Jovanović, Tanja

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Detection and molecular characterization of equine herpesviruses 1, 2, and 5 in horses in the Republic of Serbia

Radalj, Andrea; Nišavić, Jakov; Krnjaić, Dejan; Valčić, Miroslav; Jovanović, Tanja; Veljović, Ljubiša; Milić, Nenad

(Veterinarni A Farmaceuticka Univerzita Brno, Brno, 2018)

TY  - JOUR
AU  - Radalj, Andrea
AU  - Nišavić, Jakov
AU  - Krnjaić, Dejan
AU  - Valčić, Miroslav
AU  - Jovanović, Tanja
AU  - Veljović, Ljubiša
AU  - Milić, Nenad
PY  - 2018
UR  - https://vet-erinar.vet.bg.ac.rs/handle/123456789/1626
AB  - 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.
PB  - Veterinarni A Farmaceuticka Univerzita Brno, Brno
T2  - Acta Veterinaria - Brno
T1  - Detection and molecular characterization of equine herpesviruses 1, 2, and 5 in horses in the Republic of Serbia
VL  - 87
IS  - 1
SP  - 27
DO  - 10.2754/avb201887010027
ER  - 
@article{
author = "Radalj, Andrea and Nišavić, Jakov and Krnjaić, Dejan and Valčić, Miroslav and Jovanović, Tanja and Veljović, Ljubiša and Milić, Nenad",
year = "2018",
abstract = "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.",
publisher = "Veterinarni A Farmaceuticka Univerzita Brno, Brno",
journal = "Acta Veterinaria - Brno",
title = "Detection and molecular characterization of equine herpesviruses 1, 2, and 5 in horses in the Republic of Serbia",
volume = "87",
number = "1",
pages = "27",
doi = "10.2754/avb201887010027"
}
Radalj, A., Nišavić, J., Krnjaić, D., Valčić, M., Jovanović, T., Veljović, L.,& Milić, N.. (2018). Detection and molecular characterization of equine herpesviruses 1, 2, and 5 in horses in the Republic of Serbia. in Acta Veterinaria - Brno
Veterinarni A Farmaceuticka Univerzita Brno, Brno., 87(1), 27.
https://doi.org/10.2754/avb201887010027
Radalj A, Nišavić J, Krnjaić D, Valčić M, Jovanović T, Veljović L, Milić N. Detection and molecular characterization of equine herpesviruses 1, 2, and 5 in horses in the Republic of Serbia. in Acta Veterinaria - Brno. 2018;87(1):27.
doi:10.2754/avb201887010027 .
Radalj, Andrea, Nišavić, Jakov, Krnjaić, Dejan, Valčić, Miroslav, Jovanović, Tanja, Veljović, Ljubiša, Milić, Nenad, "Detection and molecular characterization of equine herpesviruses 1, 2, and 5 in horses in the Republic of Serbia" in Acta Veterinaria - Brno, 87, no. 1 (2018):27,
https://doi.org/10.2754/avb201887010027 . .
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The application of polymerase chain reaction in detection of bovine herpesvirus 1 in clinical samples

Nišavić, Jakov; Milić, Nenad; Knežević, Aleksandra; Jovanović, Tanja

(Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd, 2010)

TY  - JOUR
AU  - Nišavić, Jakov
AU  - Milić, Nenad
AU  - Knežević, Aleksandra
AU  - Jovanović, Tanja
PY  - 2010
UR  - https://vet-erinar.vet.bg.ac.rs/handle/123456789/662
AB  - The objective of this study was the application of polymerase chain reaction in detection of bovine herpesvirus 1 in clinical samples. Twenty samples of bovine nasal swabs were separately inoculated in Vero cells. Laboratory strain TN 41 of bovine herpesvirus 1 served as a control in the experiment. The cytopathic effects were recovered in the cell line previously inoculated with a sample of bovine nasal swab, as well as in cells inoculated with the laboratory strain TN 41 of the bovine herpesvirus 1 after a period of incubation of 24h, 36h and 48h. Identification of the isolated strains of the virus was done by the virus - neutralization test with specific immune sera against BHV 1, with a titre of 1:16. Comparative analysis of DNA fragments of the laboratory strain of BHV1 and isolated strain obtained by polymerase chain reaction with primers for viral gB glycoprotein and thymidine-kinase coding region, confirmed that the isolated strain of the virus belongs to bovine herpesvirus 1 (BHV 1).
AB  - Cilj nàših istraživanja je bio primena lančane reakcije polimeraze u dokazivanju prisustva goveđeg herpesvirusa 1 u kliničkim uzorcima materijala. Dvadeset uzoraka nosnih briseva goveda pojedinačno je inokulisano u ćelijsku liniju Vero. Referentni soj TN 41 virusa BHV 1 služio je kao pozitivna kontrola u ogledu. Posle 24h, 36h i 48h od inokulacije, utvrđena je pojava citopatogenog efekta u ćelijskoj liniji inokulisanoj uzorkom poreklom od jednog nosnog brisa goveda. Iste promene su ustanovljene i posle inokulacije referentnog soja TN 41 virusa BHV 1 u ćelijsku liniju Vero. Identifikacija izolovanog soja virusa poreklom iz uzorka nosnog brisa goveda, vršena je primenom testa virus - neutralizacije uz korišćenje specifičnog imunog seruma protiv virusa BHV 1, titra od 1:16. Uporednom analizom DNK fragmenata referentnog soja virusa BHV 1 i izolovanog soja virusa, dobijenih primenom lančane reakcije polimeraze uz korišćenje dva para prajmera koji su amplifikovali gene na molekulu DNK koji kodiraju sintezu glikoproteina B spoljašnjeg omotača virusa BHV 1 i timidin - kinaze, potvrđena je pripadnost izolovanog soja virusa goveđem herpesvirusu 1.
PB  - Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd
T2  - Acta Veterinaria-Beograd
T1  - The application of polymerase chain reaction in detection of bovine herpesvirus 1 in clinical samples
T1  - Primena lančane reakcije polimeraze u dokazivanju prisustva goveđeg herpesvirusa 1 u kliničkim uzorcima materijala
VL  - 60
IS  - 1
SP  - 39
EP  - 48
DO  - 10.2298/AVB1001039N
ER  - 
@article{
author = "Nišavić, Jakov and Milić, Nenad and Knežević, Aleksandra and Jovanović, Tanja",
year = "2010",
abstract = "The objective of this study was the application of polymerase chain reaction in detection of bovine herpesvirus 1 in clinical samples. Twenty samples of bovine nasal swabs were separately inoculated in Vero cells. Laboratory strain TN 41 of bovine herpesvirus 1 served as a control in the experiment. The cytopathic effects were recovered in the cell line previously inoculated with a sample of bovine nasal swab, as well as in cells inoculated with the laboratory strain TN 41 of the bovine herpesvirus 1 after a period of incubation of 24h, 36h and 48h. Identification of the isolated strains of the virus was done by the virus - neutralization test with specific immune sera against BHV 1, with a titre of 1:16. Comparative analysis of DNA fragments of the laboratory strain of BHV1 and isolated strain obtained by polymerase chain reaction with primers for viral gB glycoprotein and thymidine-kinase coding region, confirmed that the isolated strain of the virus belongs to bovine herpesvirus 1 (BHV 1)., Cilj nàših istraživanja je bio primena lančane reakcije polimeraze u dokazivanju prisustva goveđeg herpesvirusa 1 u kliničkim uzorcima materijala. Dvadeset uzoraka nosnih briseva goveda pojedinačno je inokulisano u ćelijsku liniju Vero. Referentni soj TN 41 virusa BHV 1 služio je kao pozitivna kontrola u ogledu. Posle 24h, 36h i 48h od inokulacije, utvrđena je pojava citopatogenog efekta u ćelijskoj liniji inokulisanoj uzorkom poreklom od jednog nosnog brisa goveda. Iste promene su ustanovljene i posle inokulacije referentnog soja TN 41 virusa BHV 1 u ćelijsku liniju Vero. Identifikacija izolovanog soja virusa poreklom iz uzorka nosnog brisa goveda, vršena je primenom testa virus - neutralizacije uz korišćenje specifičnog imunog seruma protiv virusa BHV 1, titra od 1:16. Uporednom analizom DNK fragmenata referentnog soja virusa BHV 1 i izolovanog soja virusa, dobijenih primenom lančane reakcije polimeraze uz korišćenje dva para prajmera koji su amplifikovali gene na molekulu DNK koji kodiraju sintezu glikoproteina B spoljašnjeg omotača virusa BHV 1 i timidin - kinaze, potvrđena je pripadnost izolovanog soja virusa goveđem herpesvirusu 1.",
publisher = "Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd",
journal = "Acta Veterinaria-Beograd",
title = "The application of polymerase chain reaction in detection of bovine herpesvirus 1 in clinical samples, Primena lančane reakcije polimeraze u dokazivanju prisustva goveđeg herpesvirusa 1 u kliničkim uzorcima materijala",
volume = "60",
number = "1",
pages = "39-48",
doi = "10.2298/AVB1001039N"
}
Nišavić, J., Milić, N., Knežević, A.,& Jovanović, T.. (2010). The application of polymerase chain reaction in detection of bovine herpesvirus 1 in clinical samples. in Acta Veterinaria-Beograd
Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd., 60(1), 39-48.
https://doi.org/10.2298/AVB1001039N
Nišavić J, Milić N, Knežević A, Jovanović T. The application of polymerase chain reaction in detection of bovine herpesvirus 1 in clinical samples. in Acta Veterinaria-Beograd. 2010;60(1):39-48.
doi:10.2298/AVB1001039N .
Nišavić, Jakov, Milić, Nenad, Knežević, Aleksandra, Jovanović, Tanja, "The application of polymerase chain reaction in detection of bovine herpesvirus 1 in clinical samples" in Acta Veterinaria-Beograd, 60, no. 1 (2010):39-48,
https://doi.org/10.2298/AVB1001039N . .
5
3
3

Examination of presence of specific antibodies against avian influenza virus in some species of wild birds

Šekler, Milanko; Ašanin, Ružica; Krnjaić, Dejan; Palić, T.; Milić, Nenad; Jovanović, Tanja; Kovačević, Dragana; Plavšić, B.; Stojanović, Dragica; Vidanović, Dejan; Ašanin, N.

(Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd, 2009)

TY  - JOUR
AU  - Šekler, Milanko
AU  - Ašanin, Ružica
AU  - Krnjaić, Dejan
AU  - Palić, T.
AU  - Milić, Nenad
AU  - Jovanović, Tanja
AU  - Kovačević, Dragana
AU  - Plavšić, B.
AU  - Stojanović, Dragica
AU  - Vidanović, Dejan
AU  - Ašanin, N.
PY  - 2009
UR  - https://vet-erinar.vet.bg.ac.rs/handle/123456789/652
AB  - Infections caused by the avian influenza virus have been known for a long time and they are present, to a smaller or greater extent, in both extensive and intensive poultry production in many parts of the world. Epidemiological investigations have established a definite significance of the population of wild birds in maintaining and spreading this infection. Avian influenza is a zoonosis, and the virus has a great potential for causing mortality in humans, in particular its subtypes H5 and H7, which is why it has lately been provoking much attention among scientists and experts, as well as the general public. The objective of the work was to catch a certain number of wild birds in several locations in the Republic of Serbia, to identify them, and to collect samples of their blood serum for the determination of specific antibodies against the avian influenza virus. Birds were caught in ten locations in a manner that was safe for the birds themselves, as well as for the staff that did the catching. The birds were caught in especially produced nets, and in some cases in special traps. The caught wild birds were identified using the methods described in reference literature. All the names of the wild birds were coordinated with the valid Serbian nomenclature of European birds, prepared by prominent ornithologist and bird lover Milan Ružić. Following catching and identification, blood samples were taken from the birds from the wing vein (in bigger birds) or from the leg vein (in smaller birds). In taking blood samples from wild birds, all the principles of asepsis and antisepsis were followed in order to prevent any possibility of infection. After that, the birds were returned to their natural habitat, to the same locations in which they were caught. Serums were isolated from the taken blood samples and they were stored at -20ºC until the final examinations. A total of 46 species of wild birds were identified among a total of 259 birds from which 259 samples of blood serum were isolated. The following were used for the detection of the presence of specific antibodies against the avian influenza virus in blood serum samples of wild birds: agar gel precipitation (AGP), the hemagglutination inhibition test (HI) for subtypes H5 and H7, the cELISA test with antigen for the A type avian infleunza virus, and the cELISA test with antigen for subtype H5 of the avian influenza virus. Due to the fact that about 360 different species of wild birds live in the Republic of Serbia, the number of 46 identified species covered by these investigations account for 12.77% of the total number of bird species present in Serbia, which is considered a good sample. Specific antibodies against the A type avian influenza virus were established in serum samples of only 9 of the 259 birds covered by examinations using the cELISA test. Of the 46 identified wild bird species, 6 belonged to birds that live exclusively in water habitats and are considered a reservoir of the avian influenza virus (white stork, mallard, mute swan, common pochard, common goldeneye, and Eurasian coot). Among the listed species, particular attention was drawn to 4 species of wild birds of the order Anseriformes and the family Anatidae (mallard, mute swan, common pochard, common goldeneye) of which there were 30 birds among the total of 259 examined. In the 30 blood serum samples of the listed bird species, specific antibodies against the A type avian influenza virus were established in 9 (30%) serum samples using cELISA. Specific antibodies against the avian influenza virus subtype H5 were established in 3 serum samples of mute swans (one serum sample originated from a mute swan which was tagged in Poland) and in one blood serum sample of a common pochard, or a total of 4 (13.33%) serum samples, using the hemagglutination inhibition test. Specific antibodies against the avian inluenza virus subtype H7 were established in 3 (10%) blood serum samples, in two serum samples from mallards and one sample from a mute swan, using the hemagglutination inhibition test. Specific antibodies against the avian influenza virus type A were not established in any examined bird species using the AGP test. In the population of wild bird species in the Republic of Serbia covered by these investigations, specific antibodies against the avian influenza virus were established only in serum samples of birds of the family Anatidae. Specific antibodies against the avian influenza virus type A established in 3 (6.52%) species of wild birds, and against subtypes H5 and H7 in 2 (4.34%) of the total of 46 examined species. The sensitivity of the cELISA test for the avian influenza virus subtype H5 and the hemagglutination inhibition test for subtype H5 amounted to 100%.
AB  - Infekcije izazvane virusom avijarne influence, su već odavno poznate i prisutne u manjem ili većem obimu, kako u ekstenzivnoj, tako i u intenzivnoj živinarskoj proizvodnji, u mnogim delovima sveta. Epidemiološkim ispitivanjima je utvrđen nesumnjiv značaj populacije divljih ptica u očuvanju i širenju ove infekcije. Avijarna influenca je zoonoza, a virus ima veliki potencijal da izazove visoku smrtnost kod ljudi, posebno njegovi podtipovi H5 i H7, tako da u novije vreme izaziva veliku pažnju, kako naučne i stručne, tako i najšire javnosti. Cilj ovog rada je bio da se na nekoliko lokacija u Republici Srbiji uhvati određeni broj divljih ptica, izvrši njihova identifikacija i prikupe uzorci krvnog seruma radi otkrivanja specifičnih antitela protiv virusa avijarne influence. Hvatanje ptica vršeno je na deset lokacija na bezbedan način, kako za same ptice, tako i za osoblje koje ih je hvatalo. Hvatanje ptica obavljano je posebnim za te svrhe proizvedenim mrežama, a u nekim slučajevima i posebnim zamkama (klopkama). Za identifikaciju uhvaćenih divljih ptica korišćene su metode koje su opisane u stručnoj literaturi. Svi nazivi divljih ptica usklađeni su sa važećom srpskom nomenklaturom ptica Evrope. Nakon hvatanja i identifikacije, pticama je uzimana krv iz krilne vene (kod većih ptica) ili iz nožne vene (kod malih ptica). Prilikom uzimanja krvi od divljih ptica poštovani su svi principi asepse i antisepse, kako bi se sprečila svaka mogućnost infekcije. Nakon toga, ptice su vraćane u prirodu, na iste lokacije na kojima su i uhvaćene. Od uzetih uzoraka krvi izdvojeni su serumi koji su ostavljani na - 20ºC i čuvani do konačnog ispitivanja. Identifikovano je 46 vrsta divljih ptica sa ukupno 259 jedinki od kojih je izdvojeno 259 uzoraka krvnog seruma. Za otkrivanje prisustva specifičnih antitela protiv virusa avijarne influence u uzorcima krvnog seruma divljih ptica korišćeni su agar gel precipitacija (AGP), test inhibicije hemaglutinacije (IH) za podtipove H5 i H7, cELISA test sa antigenom A tipa virusa avijarne influence i cELISA sa antigenom podtipa H5 virusa avijarne influence. S obzirom na činjenicu da na teritoriji Republike Srbije živi oko 360 različitih vrsta divljih ptica, broj od 46 identifikovanih vrsta obuhvaćenih ispitivanjem, činio je 12,77% od ukupnog broja prisutnih vrsta ptica u Srbiji, što se smatra dobrim uzorkom. Specifična antitela protiv A tipa virusa avijarne influence ustanovljena su u uzorcima seruma samo 9 od 259 jedinki koje su bile obuhvaćene ispitivanjem primenom cELISA testa. U identifikovanih 46 vrsta divljih ptica 6 je pripadalo pticama koje žive isključivo u vodenim staništima i smatraju se rezervoarom virusa avijarne influence (bela roda, patka gluvara, labud grbac, riđoglava patka, patka dupljašica i liska). Od navedenih vrsta posebnu pažnju privukle su 4 vrste divljih ptica iz reda Anseriformes i familije Anatidae (patka gluvara, labud grbac, riđoglava patka, patka dupljašica) kojima je od ukupno 259 ptica pripadalo 30 jedinki. U 30 uzoraka krvnog seruma navedenih vrsta ptica, specifična antitela protiv A tipa virusa avijarne influence utvrđena su u 9 (30%) uzoraka seruma, primenom cELISA. Specifična antitela protiv podtipa H5 virusa avijarne influence su ustanovljena u 3 uzorka seruma labudova grbaca (jedan uzorak seruma je poticao od labuda grbca koji je prstenovan u Poljskoj) i u jednom uzorku krvnog seruma riđoglave patke, ili ukupno u 4 (13,33%) uzorka seruma, primenom testa inhibicije hemaglutinacije. Specifična antitela protiv podtipa H7 virusa avijarne influence utvrđena su u 3 (10%) uzorka krvnog seruma i to u dva seruma pataka gluvara i u jednom serumu labuda grbca, primenom inhibicije hemaglutinacije. Specifična antitela protiv A tipa virusa avijarne influence nisu ustanovljena ni kod jedne ispitivane vrste ptice, primenom AGP testa. U populaciji divljih vrta ptica u Republici Srbiji obuhvaćenih ovim ispitivanjem, specifična antitela protiv virusa avijarne influence ustanovljena su samo u uzorcima seruma ptica iz familije Anatidae. Specifična antitela protiv A tipa virusa avijarne influence su otkrivena kod 3 (6,52%) vrste divljih ptica, a protiv podtipova H5 i H7 kod 2 (4,34%) od ukupno 46 vrsta koje su ispitivane. Senzitivnost cELISA testa za podtip H5 virusa avijarne influence i testa inhibicije hemaglutinacije za isti podtip iznosila je 100%.
PB  - Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd
T2  - Acta Veterinaria-Beograd
T1  - Examination of presence of specific antibodies against avian influenza virus in some species of wild birds
T1  - Ispitivanje prisustva specifičnih antitela protiv virusa avijarne influence kod nekih vrsta divljih ptica
VL  - 59
IS  - 4
SP  - 381
EP  - 403
DO  - 10.2298/AVB0904381S
ER  - 
@article{
author = "Šekler, Milanko and Ašanin, Ružica and Krnjaić, Dejan and Palić, T. and Milić, Nenad and Jovanović, Tanja and Kovačević, Dragana and Plavšić, B. and Stojanović, Dragica and Vidanović, Dejan and Ašanin, N.",
year = "2009",
abstract = "Infections caused by the avian influenza virus have been known for a long time and they are present, to a smaller or greater extent, in both extensive and intensive poultry production in many parts of the world. Epidemiological investigations have established a definite significance of the population of wild birds in maintaining and spreading this infection. Avian influenza is a zoonosis, and the virus has a great potential for causing mortality in humans, in particular its subtypes H5 and H7, which is why it has lately been provoking much attention among scientists and experts, as well as the general public. The objective of the work was to catch a certain number of wild birds in several locations in the Republic of Serbia, to identify them, and to collect samples of their blood serum for the determination of specific antibodies against the avian influenza virus. Birds were caught in ten locations in a manner that was safe for the birds themselves, as well as for the staff that did the catching. The birds were caught in especially produced nets, and in some cases in special traps. The caught wild birds were identified using the methods described in reference literature. All the names of the wild birds were coordinated with the valid Serbian nomenclature of European birds, prepared by prominent ornithologist and bird lover Milan Ružić. Following catching and identification, blood samples were taken from the birds from the wing vein (in bigger birds) or from the leg vein (in smaller birds). In taking blood samples from wild birds, all the principles of asepsis and antisepsis were followed in order to prevent any possibility of infection. After that, the birds were returned to their natural habitat, to the same locations in which they were caught. Serums were isolated from the taken blood samples and they were stored at -20ºC until the final examinations. A total of 46 species of wild birds were identified among a total of 259 birds from which 259 samples of blood serum were isolated. The following were used for the detection of the presence of specific antibodies against the avian influenza virus in blood serum samples of wild birds: agar gel precipitation (AGP), the hemagglutination inhibition test (HI) for subtypes H5 and H7, the cELISA test with antigen for the A type avian infleunza virus, and the cELISA test with antigen for subtype H5 of the avian influenza virus. Due to the fact that about 360 different species of wild birds live in the Republic of Serbia, the number of 46 identified species covered by these investigations account for 12.77% of the total number of bird species present in Serbia, which is considered a good sample. Specific antibodies against the A type avian influenza virus were established in serum samples of only 9 of the 259 birds covered by examinations using the cELISA test. Of the 46 identified wild bird species, 6 belonged to birds that live exclusively in water habitats and are considered a reservoir of the avian influenza virus (white stork, mallard, mute swan, common pochard, common goldeneye, and Eurasian coot). Among the listed species, particular attention was drawn to 4 species of wild birds of the order Anseriformes and the family Anatidae (mallard, mute swan, common pochard, common goldeneye) of which there were 30 birds among the total of 259 examined. In the 30 blood serum samples of the listed bird species, specific antibodies against the A type avian influenza virus were established in 9 (30%) serum samples using cELISA. Specific antibodies against the avian influenza virus subtype H5 were established in 3 serum samples of mute swans (one serum sample originated from a mute swan which was tagged in Poland) and in one blood serum sample of a common pochard, or a total of 4 (13.33%) serum samples, using the hemagglutination inhibition test. Specific antibodies against the avian inluenza virus subtype H7 were established in 3 (10%) blood serum samples, in two serum samples from mallards and one sample from a mute swan, using the hemagglutination inhibition test. Specific antibodies against the avian influenza virus type A were not established in any examined bird species using the AGP test. In the population of wild bird species in the Republic of Serbia covered by these investigations, specific antibodies against the avian influenza virus were established only in serum samples of birds of the family Anatidae. Specific antibodies against the avian influenza virus type A established in 3 (6.52%) species of wild birds, and against subtypes H5 and H7 in 2 (4.34%) of the total of 46 examined species. The sensitivity of the cELISA test for the avian influenza virus subtype H5 and the hemagglutination inhibition test for subtype H5 amounted to 100%., Infekcije izazvane virusom avijarne influence, su već odavno poznate i prisutne u manjem ili većem obimu, kako u ekstenzivnoj, tako i u intenzivnoj živinarskoj proizvodnji, u mnogim delovima sveta. Epidemiološkim ispitivanjima je utvrđen nesumnjiv značaj populacije divljih ptica u očuvanju i širenju ove infekcije. Avijarna influenca je zoonoza, a virus ima veliki potencijal da izazove visoku smrtnost kod ljudi, posebno njegovi podtipovi H5 i H7, tako da u novije vreme izaziva veliku pažnju, kako naučne i stručne, tako i najšire javnosti. Cilj ovog rada je bio da se na nekoliko lokacija u Republici Srbiji uhvati određeni broj divljih ptica, izvrši njihova identifikacija i prikupe uzorci krvnog seruma radi otkrivanja specifičnih antitela protiv virusa avijarne influence. Hvatanje ptica vršeno je na deset lokacija na bezbedan način, kako za same ptice, tako i za osoblje koje ih je hvatalo. Hvatanje ptica obavljano je posebnim za te svrhe proizvedenim mrežama, a u nekim slučajevima i posebnim zamkama (klopkama). Za identifikaciju uhvaćenih divljih ptica korišćene su metode koje su opisane u stručnoj literaturi. Svi nazivi divljih ptica usklađeni su sa važećom srpskom nomenklaturom ptica Evrope. Nakon hvatanja i identifikacije, pticama je uzimana krv iz krilne vene (kod većih ptica) ili iz nožne vene (kod malih ptica). Prilikom uzimanja krvi od divljih ptica poštovani su svi principi asepse i antisepse, kako bi se sprečila svaka mogućnost infekcije. Nakon toga, ptice su vraćane u prirodu, na iste lokacije na kojima su i uhvaćene. Od uzetih uzoraka krvi izdvojeni su serumi koji su ostavljani na - 20ºC i čuvani do konačnog ispitivanja. Identifikovano je 46 vrsta divljih ptica sa ukupno 259 jedinki od kojih je izdvojeno 259 uzoraka krvnog seruma. Za otkrivanje prisustva specifičnih antitela protiv virusa avijarne influence u uzorcima krvnog seruma divljih ptica korišćeni su agar gel precipitacija (AGP), test inhibicije hemaglutinacije (IH) za podtipove H5 i H7, cELISA test sa antigenom A tipa virusa avijarne influence i cELISA sa antigenom podtipa H5 virusa avijarne influence. S obzirom na činjenicu da na teritoriji Republike Srbije živi oko 360 različitih vrsta divljih ptica, broj od 46 identifikovanih vrsta obuhvaćenih ispitivanjem, činio je 12,77% od ukupnog broja prisutnih vrsta ptica u Srbiji, što se smatra dobrim uzorkom. Specifična antitela protiv A tipa virusa avijarne influence ustanovljena su u uzorcima seruma samo 9 od 259 jedinki koje su bile obuhvaćene ispitivanjem primenom cELISA testa. U identifikovanih 46 vrsta divljih ptica 6 je pripadalo pticama koje žive isključivo u vodenim staništima i smatraju se rezervoarom virusa avijarne influence (bela roda, patka gluvara, labud grbac, riđoglava patka, patka dupljašica i liska). Od navedenih vrsta posebnu pažnju privukle su 4 vrste divljih ptica iz reda Anseriformes i familije Anatidae (patka gluvara, labud grbac, riđoglava patka, patka dupljašica) kojima je od ukupno 259 ptica pripadalo 30 jedinki. U 30 uzoraka krvnog seruma navedenih vrsta ptica, specifična antitela protiv A tipa virusa avijarne influence utvrđena su u 9 (30%) uzoraka seruma, primenom cELISA. Specifična antitela protiv podtipa H5 virusa avijarne influence su ustanovljena u 3 uzorka seruma labudova grbaca (jedan uzorak seruma je poticao od labuda grbca koji je prstenovan u Poljskoj) i u jednom uzorku krvnog seruma riđoglave patke, ili ukupno u 4 (13,33%) uzorka seruma, primenom testa inhibicije hemaglutinacije. Specifična antitela protiv podtipa H7 virusa avijarne influence utvrđena su u 3 (10%) uzorka krvnog seruma i to u dva seruma pataka gluvara i u jednom serumu labuda grbca, primenom inhibicije hemaglutinacije. Specifična antitela protiv A tipa virusa avijarne influence nisu ustanovljena ni kod jedne ispitivane vrste ptice, primenom AGP testa. U populaciji divljih vrta ptica u Republici Srbiji obuhvaćenih ovim ispitivanjem, specifična antitela protiv virusa avijarne influence ustanovljena su samo u uzorcima seruma ptica iz familije Anatidae. Specifična antitela protiv A tipa virusa avijarne influence su otkrivena kod 3 (6,52%) vrste divljih ptica, a protiv podtipova H5 i H7 kod 2 (4,34%) od ukupno 46 vrsta koje su ispitivane. Senzitivnost cELISA testa za podtip H5 virusa avijarne influence i testa inhibicije hemaglutinacije za isti podtip iznosila je 100%.",
publisher = "Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd",
journal = "Acta Veterinaria-Beograd",
title = "Examination of presence of specific antibodies against avian influenza virus in some species of wild birds, Ispitivanje prisustva specifičnih antitela protiv virusa avijarne influence kod nekih vrsta divljih ptica",
volume = "59",
number = "4",
pages = "381-403",
doi = "10.2298/AVB0904381S"
}
Šekler, M., Ašanin, R., Krnjaić, D., Palić, T., Milić, N., Jovanović, T., Kovačević, D., Plavšić, B., Stojanović, D., Vidanović, D.,& Ašanin, N.. (2009). Examination of presence of specific antibodies against avian influenza virus in some species of wild birds. in Acta Veterinaria-Beograd
Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd., 59(4), 381-403.
https://doi.org/10.2298/AVB0904381S
Šekler M, Ašanin R, Krnjaić D, Palić T, Milić N, Jovanović T, Kovačević D, Plavšić B, Stojanović D, Vidanović D, Ašanin N. Examination of presence of specific antibodies against avian influenza virus in some species of wild birds. in Acta Veterinaria-Beograd. 2009;59(4):381-403.
doi:10.2298/AVB0904381S .
Šekler, Milanko, Ašanin, Ružica, Krnjaić, Dejan, Palić, T., Milić, Nenad, Jovanović, Tanja, Kovačević, Dragana, Plavšić, B., Stojanović, Dragica, Vidanović, Dejan, Ašanin, N., "Examination of presence of specific antibodies against avian influenza virus in some species of wild birds" in Acta Veterinaria-Beograd, 59, no. 4 (2009):381-403,
https://doi.org/10.2298/AVB0904381S . .
2
1
2

Examination of humoral and cellular immune responses after immunization with subunit vaccine against Herpes simplex virus 1 and 2

Milić, Nenad; Jovanović, Tanja; Gađanski-Omerović, Gordana; Knežević, Ivana; Ašanin, Ružica

(Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd, 2001)

TY  - JOUR
AU  - Milić, Nenad
AU  - Jovanović, Tanja
AU  - Gađanski-Omerović, Gordana
AU  - Knežević, Ivana
AU  - Ašanin, Ružica
PY  - 2001
UR  - https://vet-erinar.vet.bg.ac.rs/handle/123456789/170
AB  - The objective of our study was to examine the humoral and cellular immune response in mice after immunization with bivalent subunit vaccine against Herpes simplex viruses (HSV 1 and HSV2). These examinations were carried out in biological assay on 27 female Swiss albino mice (20 g bm). The subunit vaccine was prepared from purified glycoprotein antigens (subunits) isolated from the external envelopes of HSV 1 and HSV 2 and adsorbed on the adjuvant AI (OH)3 at a final concentration of 5 mg/ml. This vaccine contained 0.14 mg/ml total protein for the HSV 1 subunits and 0.16 mg/ml for HSV2 subunits. Two groups of 9 and 7 experimental mice were immunized with the HSV1/HSV2 subunit vaccine (0.5 ml s/c). A group of 11 healthy non-immunized mice served as the control. The specific humoral immune response of the vaccinated mice was examined by a standard method of indirect immunofluorescence. The specific cellular immune response of the vaccinated animals was tested by measuring the lymphocyte proliferation response to HSV1 and HSV2 antigens with tritiated methyl thymidine (*128.5 Bq/mmol), in vitro. The titers of the specific IgG and IgM class antibodies, in the blood sera of the first group of 9 immunized mice, on day 10 after vaccination were: from 1:8 to 1:64 IgG and 1:32 to 1:128 IgM for HSV1 and 1:32 to 1:128 IgG and IgM antibodies for HSV2. The equivalent titers for the second group of 7 immunized mice, on day 24 after vaccination were: from 1:64 to 1:128 IgG and 1:16 to 1:64 IgM for HSV1; from 1:32 to 1:128 IgG and IgM for HSV2. The mean values for radioactivity in the proliferation test of lymphocytes for HSV1 and HSV2 antigens were 1471 cpm and 1839 cpm respectively on day 10 after vaccination, The mean value for radioactivity in the medium after spontaneous proliferation of lymphocytes for this group of mice was 893 cpm. The mean radioactivity of lymphocytes from the second group of mice, stimulated with HSV1 antigens was 4158 cpm on day 24 after vaccination, with a spontaneous proliferation value of 364 cpm. Thus, the low subunit antigen concentrations in the bivalent subunit vaccine, induced satisfactory humoral and cellular immune responses to HSV 1 and HSV2 in the vaccinated mice.
AB  - Cilj naših istraživanja je bilo ispitivanje humoralnog i ćelijskog imunskog odgovora posle imunizacije miševa eksperimentalnom bivalentnom subjediničnom vakcinom protiv Herpes simplex virusa tipa 1 i 2 (HSV1 i HSV2). Ova ispitivanja su izvršena u biološkom ogledu na 27 ženki Swiss albino miševa, težine 20 g. Subjedinična vakcina je pripremljena od prečišćenih glikoproteinskih antigena (subjedinica) izolovanih iz spoljašnjih omotača HSV1 i HSV2 i adsorbovanih na adjuvans A1(OH)3. Finalna koncentracija A1(OH)3 u vakcini iznosila je 5 mg/ml. Ukupne koncentracije antigena u vakcini iznosile su 0.14 mg/ml za HSV1 i 0.16 mg/ml za HSV2. Dve grupe od 9 i 7 eksperimentalnih miševa imunizovane su supkutano sa istim dozama od 0.5 ml HSV1/HSV2 subjedinične vakcine. dok je grupa od 11 zdravih, neimunizovanih miševa predstavljala kontrolnu grupu. Specifični humoralni imunski odgovor vakcinisanih rniševa ispitivan je standardnom metodom indirektne imunofluorescencije. Specifični celularni imunski odgovor vakcinisanih životinja ispitivan je in vitro standardnom metodom proliferacije limfocita na HSV1 i HSV2 antigene pomoću tricijum metil timidina (128.5 Bq/mmol). Specifična antitela Ig G i Ig M klase ustanovljena su u krvnom serumu prve grupe miševa, desetog dana posle vakcinacije i njihov titar se kretao u opsegu od 1:8 do 1:64 (Ig G) i od 1:32 do 1:128 (Ig M) za HSV1 i od 1:32 do 1:128 (Ig G i Ig M) za HSV2. Titar specifičnih antitela u serumu miševa druge grupe, 24-tog dana posle vakcinacije, kretao se u opsegu od 1:64 do 1:128 (IgG) i 1:16 do 1:64 (Ig M) za HSV1 i od 1:32 do 1:128 (IgG i IgM) za HSV2. Srednje vrednosti testa proliferacije limfocita stimulisanih sa HSV1 i HSV2 antigenima kod prve grupe miševa, 10 dana posle imunizacije, iznosile su 1471 c/min (za HSV1) i 1839 c/min za HSV2. Srednja vrednost radioaktivnosti uzoraka u testu spontane proliferacije limfocita kod ovih miševa u medijumu, bila je 893 c/min. Srednja vrednost radioaktivnosti limfocita stimulisanih sa HSV1 antigenima u drugoj grupi miševa, 24-tog dana od vakcinacije iznosila je 4158 c/min, dok je vrednost spontane proliferacije limfocita ovih miševa bila 364 c/min. Niske koncentracije subjediničnih antigena u vakcini indukovale su zadovoljavajući humoralni i ćelijski imunski odgovor kod vakcinisanih miševa protiv HSV1 i HSV2.
PB  - Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd
T2  - Acta Veterinaria-Beograd
T1  - Examination of humoral and cellular immune responses after immunization with subunit vaccine against Herpes simplex virus 1 and 2
T1  - Ispitivanja humoralnog i ćelijskog imunskog odgovora posle imunizacije sa subjediničnom vakcinom protiv Herpes simplex virusa tipa 1 i 2
VL  - 51
IS  - 1
SP  - 35
EP  - 43
UR  - https://hdl.handle.net/21.15107/rcub_veterinar_170
ER  - 
@article{
author = "Milić, Nenad and Jovanović, Tanja and Gađanski-Omerović, Gordana and Knežević, Ivana and Ašanin, Ružica",
year = "2001",
abstract = "The objective of our study was to examine the humoral and cellular immune response in mice after immunization with bivalent subunit vaccine against Herpes simplex viruses (HSV 1 and HSV2). These examinations were carried out in biological assay on 27 female Swiss albino mice (20 g bm). The subunit vaccine was prepared from purified glycoprotein antigens (subunits) isolated from the external envelopes of HSV 1 and HSV 2 and adsorbed on the adjuvant AI (OH)3 at a final concentration of 5 mg/ml. This vaccine contained 0.14 mg/ml total protein for the HSV 1 subunits and 0.16 mg/ml for HSV2 subunits. Two groups of 9 and 7 experimental mice were immunized with the HSV1/HSV2 subunit vaccine (0.5 ml s/c). A group of 11 healthy non-immunized mice served as the control. The specific humoral immune response of the vaccinated mice was examined by a standard method of indirect immunofluorescence. The specific cellular immune response of the vaccinated animals was tested by measuring the lymphocyte proliferation response to HSV1 and HSV2 antigens with tritiated methyl thymidine (*128.5 Bq/mmol), in vitro. The titers of the specific IgG and IgM class antibodies, in the blood sera of the first group of 9 immunized mice, on day 10 after vaccination were: from 1:8 to 1:64 IgG and 1:32 to 1:128 IgM for HSV1 and 1:32 to 1:128 IgG and IgM antibodies for HSV2. The equivalent titers for the second group of 7 immunized mice, on day 24 after vaccination were: from 1:64 to 1:128 IgG and 1:16 to 1:64 IgM for HSV1; from 1:32 to 1:128 IgG and IgM for HSV2. The mean values for radioactivity in the proliferation test of lymphocytes for HSV1 and HSV2 antigens were 1471 cpm and 1839 cpm respectively on day 10 after vaccination, The mean value for radioactivity in the medium after spontaneous proliferation of lymphocytes for this group of mice was 893 cpm. The mean radioactivity of lymphocytes from the second group of mice, stimulated with HSV1 antigens was 4158 cpm on day 24 after vaccination, with a spontaneous proliferation value of 364 cpm. Thus, the low subunit antigen concentrations in the bivalent subunit vaccine, induced satisfactory humoral and cellular immune responses to HSV 1 and HSV2 in the vaccinated mice., Cilj naših istraživanja je bilo ispitivanje humoralnog i ćelijskog imunskog odgovora posle imunizacije miševa eksperimentalnom bivalentnom subjediničnom vakcinom protiv Herpes simplex virusa tipa 1 i 2 (HSV1 i HSV2). Ova ispitivanja su izvršena u biološkom ogledu na 27 ženki Swiss albino miševa, težine 20 g. Subjedinična vakcina je pripremljena od prečišćenih glikoproteinskih antigena (subjedinica) izolovanih iz spoljašnjih omotača HSV1 i HSV2 i adsorbovanih na adjuvans A1(OH)3. Finalna koncentracija A1(OH)3 u vakcini iznosila je 5 mg/ml. Ukupne koncentracije antigena u vakcini iznosile su 0.14 mg/ml za HSV1 i 0.16 mg/ml za HSV2. Dve grupe od 9 i 7 eksperimentalnih miševa imunizovane su supkutano sa istim dozama od 0.5 ml HSV1/HSV2 subjedinične vakcine. dok je grupa od 11 zdravih, neimunizovanih miševa predstavljala kontrolnu grupu. Specifični humoralni imunski odgovor vakcinisanih rniševa ispitivan je standardnom metodom indirektne imunofluorescencije. Specifični celularni imunski odgovor vakcinisanih životinja ispitivan je in vitro standardnom metodom proliferacije limfocita na HSV1 i HSV2 antigene pomoću tricijum metil timidina (128.5 Bq/mmol). Specifična antitela Ig G i Ig M klase ustanovljena su u krvnom serumu prve grupe miševa, desetog dana posle vakcinacije i njihov titar se kretao u opsegu od 1:8 do 1:64 (Ig G) i od 1:32 do 1:128 (Ig M) za HSV1 i od 1:32 do 1:128 (Ig G i Ig M) za HSV2. Titar specifičnih antitela u serumu miševa druge grupe, 24-tog dana posle vakcinacije, kretao se u opsegu od 1:64 do 1:128 (IgG) i 1:16 do 1:64 (Ig M) za HSV1 i od 1:32 do 1:128 (IgG i IgM) za HSV2. Srednje vrednosti testa proliferacije limfocita stimulisanih sa HSV1 i HSV2 antigenima kod prve grupe miševa, 10 dana posle imunizacije, iznosile su 1471 c/min (za HSV1) i 1839 c/min za HSV2. Srednja vrednost radioaktivnosti uzoraka u testu spontane proliferacije limfocita kod ovih miševa u medijumu, bila je 893 c/min. Srednja vrednost radioaktivnosti limfocita stimulisanih sa HSV1 antigenima u drugoj grupi miševa, 24-tog dana od vakcinacije iznosila je 4158 c/min, dok je vrednost spontane proliferacije limfocita ovih miševa bila 364 c/min. Niske koncentracije subjediničnih antigena u vakcini indukovale su zadovoljavajući humoralni i ćelijski imunski odgovor kod vakcinisanih miševa protiv HSV1 i HSV2.",
publisher = "Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd",
journal = "Acta Veterinaria-Beograd",
title = "Examination of humoral and cellular immune responses after immunization with subunit vaccine against Herpes simplex virus 1 and 2, Ispitivanja humoralnog i ćelijskog imunskog odgovora posle imunizacije sa subjediničnom vakcinom protiv Herpes simplex virusa tipa 1 i 2",
volume = "51",
number = "1",
pages = "35-43",
url = "https://hdl.handle.net/21.15107/rcub_veterinar_170"
}
Milić, N., Jovanović, T., Gađanski-Omerović, G., Knežević, I.,& Ašanin, R.. (2001). Examination of humoral and cellular immune responses after immunization with subunit vaccine against Herpes simplex virus 1 and 2. in Acta Veterinaria-Beograd
Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd., 51(1), 35-43.
https://hdl.handle.net/21.15107/rcub_veterinar_170
Milić N, Jovanović T, Gađanski-Omerović G, Knežević I, Ašanin R. Examination of humoral and cellular immune responses after immunization with subunit vaccine against Herpes simplex virus 1 and 2. in Acta Veterinaria-Beograd. 2001;51(1):35-43.
https://hdl.handle.net/21.15107/rcub_veterinar_170 .
Milić, Nenad, Jovanović, Tanja, Gađanski-Omerović, Gordana, Knežević, Ivana, Ašanin, Ružica, "Examination of humoral and cellular immune responses after immunization with subunit vaccine against Herpes simplex virus 1 and 2" in Acta Veterinaria-Beograd, 51, no. 1 (2001):35-43,
https://hdl.handle.net/21.15107/rcub_veterinar_170 .