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1

Zastosowanie cytometrii przeplywowej do oceny wrazliwosci wybranych linii komorkowych na zakazenie wirusem zapalenia tetnic koni

100%

Larska M., Rola J.

Medycyna Weterynaryjna

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2008

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tom 64

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nr 01

105-109

The purpose of the study was to attempt the application of flow cytometry to evaluate the sensitivity of various cell lines to EAV infections, according to their type and passage number. Monolayers of RK13, Vero, BHK-21 and MDBK cells were infected with reference EAV strain Bucyrus. First of all the susceptibility of each cell line to different titers of the virus was tested. The second step was to establish a time course kinetic of viral infection. Sequentially, starting from 2 to 72 hours post infection the cells were fixed, permeabilized and stained with FITC monoclonal EAV-specific antibodies. The analysis carried out in Coulter Epics XL (Beckman Coulter) flow cytometer considered the percentage of EAV infected cells determined in a time range by gating FITC/Count histograms. Significant differences in the sensitivity to EAV infection in particular cell lines were found. After 24 h. p.i. most RK13 cells infected with Bucyrus strain showed signs of infection in the titer of 10 TCID₅₀, whilst in Vero and BHK-21 a similar histogram was not obtained until 100 and 1000 TCID₅₀ respectively. As early as 10 and 12 h post inoculation significant level of infected cells (37.9 and 18.6%) were detected in line RK13 passage 36 and 11 respectively. Cell line Vero passage 115 indicated a higher sensitivity to EAV infection comparing to Vero passage 153. More than 70% of cells from that line were EAV infected after 24 hours post inoculation. In Vero passage 153 the infected cells (15.4%) were not detected until 36 h.p.i. The presence of infected cells was found also after 36 h post inoculation in BHK-21 cell line passage 10. The obtained results indicate that differences in cell line susceptibility to EAV infection depend on their type and decrease with their passage.

2

Zmiennosc genetyczna wirusa a diagnostyka zakazen EAV

100%

Larska M., Rola J.

Medycyna Weterynaryjna

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2006

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tom 62

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nr 12

1348-1351

Equine arteritis virus (EAV) belongs to the family Arteriviridae of the order Nidovirales. The viral genome composed by single-stranded positive sense RNA is enclosed in a icosahedral nucleocapsid and surrounded by a proteolipid envelope. The genome consists of nine open reading frames (ORFs) coding both structural and non-structural proteins. Although only one serotype of EAV is distinguished, field isolates differ in virulence and pathogenicity. The EAV infection is usually subclinical. 30-60% of stallions after infection become persistent carriers of the virus and can shed EAV with their semen during next several weeks, months or even years. Mares covered by EAV shedding stallions can result in abortions, fetus resorptions, infertility and even death of the newborn foals that lead to large losses in horse breeding. Seropositive stallions sheding the virus in their semen are the main reservoir of the virus; the control of the disease, therefore, should be based on their identification and elimination from breeding. Genetic variability of EAV can lead to the increase of virulence of the isolates and to changes in viral properties having an impact on the results of laboratory testing. Researching genetic modification of the viral genome provides important information about the changes in the nucleotide sequence of currently circulating strains and about the direction of EAV evolution. The purpose of the review is to present current data concerning molecular biology and diagnostics of equine arteritis virus infections.

3

Zastosowanie testu RT-PCR do wykrywania obecnosci wirusa zapalenia tetnic koni w nasieniu ogierow

100%

Rola J., Larska M., Zmudzinski J. F.

Medycyna Weterynaryjna

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2004

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tom 60

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nr 03

289-291

4

Zastosowanie testu immunoperoksydazowego posredniego do potwierdzania tozsamosci izolatow EAV

100%

Larska M., Rola J.

Medycyna Weterynaryjna

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2007

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tom 63

|

nr 06

712-716

The purpose of the study was the elaboration and introduction of an effective method to confirm the identity of EAV isolates obtained in cell culture from the serum of persistently infected stallions. In view of the low cost, simplicity of preparation, and its high specificity, an indirect immunoperoxidase test based on rabbit polyclonal sera was introduced. After immunization of the rabbits with the EAV reference strain Bucyrus, purified by ultracentrifugation, a specific polyclonal serum in the titer of SN antibodies from 1:32 to 1:128 was obtained. Peroxidase-conjugated goat IgG antibodies against horse immunoglobulins were used as a secondary antibody and DAB as a substrate. After the optimalization of the test conditions, its sensitivity and specificity was defined. The sensitivity estimated in reference to the Bucyrus strain was equal to 1 TCID₅₀. The specificity was defined at 100%. After the preliminary validation, the indirect immunoperoxidase test based on the obtained polyclonal sera was affirmed as a method to identify EAV isolates. All 68 EAV field isolates reacted positively in the immunoperoxidase test.

5

Spontaniczne zakazenia wirusem zapalenia tetnic koni u ogierow

100%

Golnik W., Sordyl B.

Medycyna Weterynaryjna

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2004

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tom 60

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nr 06

630-633

6

Artykuł dostępny w postaci pełnego tekstu - kliknij by otworzyć plik

84%

Golnik W., Cierpisz J.

Medycyna Weterynaryjna

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1994

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tom 50

|

nr 10

482-483

Attempts of using stallions naturally infected with Equine arteritis virus in reproduction The objective of the studies was to check both in vitro and vivo the use in reproduction of convalescent stallions of semen free from the Equine arteritis virus (EAV). The examinations were done on 92 stallions from the stud К in which sera of 83.2% of animals were free from antibodies to EAV. Three stallions used for mating in 1993 were seropositive. Attempts to isolate the EAV from semen were performed on RK-13 and Vero cell cultures. From the semen of one stallion a virus identified as AEV by the direct immunofluorescence test was isolated. As for the two remaining convalescent stallions with negative results of the laboratory test for EAV, semen was examined for the EAV transfer on four fully susceptible mares.

12

Zroznicowanie genetyczne szczepow EAV wsrod trwale zakazonych ogierow w stadach ogierow w Polsce

84%

Larska M., Rola J.

Medycyna Weterynaryjna

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2007

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tom 63

|

nr 11

1381-1386

The purpose of the study was to analyse the genetic diversity of Polish EAV isolates. Genetic variability can lead to increased virulence of isolates and to significant changes in EAV antigen properties influencing the results of laboratory testing. Studies on genetic modifications of viral genomes as well as on the phylogenetic affinity of strains have facilitated the investigation of viral evolution. Phylogenetic analysis was performed on 32 isolates that were isolated from the semen of asymptomatic virus-shedding stallions originating from 8 national studs. These isolates were compared with 15 reference EAV strains commonly used in phylogenesis. On the basis of the nucleotide sequence analysis of ORF5 gene encoding GP5 glycoprotein it was shown that Polish EAV isolates belonged to two subgroups and demonstrated the closest relationship to the European strains. None of these strains had any relationship to the first Polish strain Wroclaw-2 isolated in 1976. The homology of ORF5 nucleotide and predicted GP5 amino-acid sequences of Polish isolates attained a level of 81.2-99.0% and 90.1-99.4% respectively. Analyzing the genetic diversity of ORF5 facilitated the conducting of retrospective epizootic investigations.

13

Wystepowanie zakazen wirusem zapalenia tetnic koni u zrebiat w okresie przed odsadzeniem

84%

Golnik W., Paweska J.

Medycyna Weterynaryjna

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1997

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tom 53

|

nr 11

654-656

14

Wykrywanie zakazen wirusem zapalenia tetnic koni w nasieniu ogierow metoda cytometrii przeplywowej

67%

Cierpisz J., Golnik W., Laskowska A., Ugorski M.

Medycyna Weterynaryjna

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1998

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tom 54

|

nr 09

626-627

In the study flow cytometry was attempted to identify EAV infection in stallions’ semen. Material for the study consisted of 8 semen samples taken from EAV-seropositive stallions. The samples were put on RK-13 and Vero cells. After a 24-hour incubation period the cells were treated with a specific conjugate and then analyzed in FACS aparatus. In the case of two samples positive fluorescence was observed. The above positive result was confirmed by virus isolation in both cell cultures used.

15

Epidemiologia zakazen wirusem zapalenia tetnic koni u ogierow. Proba dekontaminacji nasienia z zanieczyszczen wirusowych

67%

Sordyl B.

Acta Scientiarum Polonorum. Medicina Veterinaria

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2005

|

tom 04

|

nr 1

15-26

16

Wirusowe choroby układu oddechowego koni. Cz.II

67%

Zietek-Barszcz A., Kalinowski M., Pukaluk K., Gradzki Z., Adaszek L.

Magazyn Weterynaryjny

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2011

|

tom 20

|

nr 01

17

67%

Golnik W.

Magazyn Weterynaryjny

|

2000

|

tom Supl.Konie

24-26

Ograniczanie wyników

Zastosowanie cytometrii przeplywowej do oceny wrazliwosci wybranych linii komorkowych na zakazenie wirusem zapalenia tetnic koni

Zmiennosc genetyczna wirusa a diagnostyka zakazen EAV

Zastosowanie testu RT-PCR do wykrywania obecnosci wirusa zapalenia tetnic koni w nasieniu ogierow

Zastosowanie testu immunoperoksydazowego posredniego do potwierdzania tozsamosci izolatow EAV

Spontaniczne zakazenia wirusem zapalenia tetnic koni u ogierow

Wirusowe zapalenie tętnic koni (EVA)

Odsetek zakazen naturalnych wirusem zapalenia tetnic koni u ogierow roznych ras

Zastosowanie preparatu EquiPure do pozyskiwania morfologicznie prawidlowych plemnikow ogiera oraz dekontaminacji spermy z zanieczyszczen wirusem zapalenia tetnic koni

Zakazne zapalenie tetnic koni - choroba, o ktora pytaja hodowcy

Porownanie przydatnosci metody RT-PCR i badania wirusologicznego w diagnostyce wirusowego zapalenia tetnic koni

proby uzytkowania ogierow czolowych zakazonych spontanicznie wirusem zapalenia tetnic koni

Zroznicowanie genetyczne szczepow EAV wsrod trwale zakazonych ogierow w stadach ogierow w Polsce

Wystepowanie zakazen wirusem zapalenia tetnic koni u zrebiat w okresie przed odsadzeniem

Wykrywanie zakazen wirusem zapalenia tetnic koni w nasieniu ogierow metoda cytometrii przeplywowej

Epidemiologia zakazen wirusem zapalenia tetnic koni u ogierow. Proba dekontaminacji nasienia z zanieczyszczen wirusowych

Wirusowe choroby układu oddechowego koni. Cz.II

Analiza molekularna fragmentu genu nukleoproteiny wirusa zapalenia tętnic koni izolowanego z nasienia trwałych siewców wirusa

Zakazenia wirusowe drog oddechowych u koni

Wirusowe zapalenie tetnic koni - choroba wciaz aktualna

Wirusowe zapalenie tetnic koni