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1

Replication of equine herpesvirus type 1 in equine dermal cells transfected with Bam HI[G] restriction fragment of EHV-2 genome

100%
Dzieciatkowski T., Chmielewska A., Turowska A., Tucholska A., Banbura M. W.
Polish Journal of Veterinary Sciences
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2009
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tom 12
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nr 1
In previous experiments, we have demonstrated that the presence of equine herpesvirus 2 (EHV-2) enhanced plaque formation in cell cultures infected with equine herpesvirus type 1. To determine whether a specific region of the EHV-2 genome is responsible for this effect, we have constructed a library of Bam HI fragments of the EHV-2 genome ligated into pcDNA plasmid. Equine dermal (ED) cell cultures were subsequently transfected with the constructs, passaged 5 times, tested for the presence of the plasmids and infected with EHV-1 at MOI=0.01. Only in cultures transfected with the pcDNA/Bam HI[G]construct, designated Δ2/4, the mean number of plaques at 24 hrs p.i. was approximately 10 times higher than in non-transfected controls. Virus titers in culture supernatants as well as in freeze-thawed cells were 4- and 5-fold higher, respectively, than in non-transfected cultures. These differences were observed only at 24 hrs p.i. At 48 hrs p.i. cultures were completely destroyed and, surprisingly, the virus titer was slightly lower in the supernatant of transfected cells. However, the titer of EHV-1 in freeze-thawed culture was exactly the same as in the control. These results suggest that the presence of Bam HI[G] fragment of the EHV-2 genome stimulates (accelerates?) plaque formation only at earlier stages of infection but does not influence the total yield of EHV-1 at 48 hrs p.i. The exact mechanism of this stimulation remains unclear and further experiments are necessary to determine the role of putative EHV-2 proteins encoded by Bam HI [G] fragment of the EHV-genome.
2

Equine herpesvirus type 1 quantification in different types of samples by a real-time PCR

84%
Dzieciatkowski T., Przybylski M., Cymerys J., Turowska A., Chmielewska A., Tucholska A., Banbura M. W.
Polish Journal of Veterinary Sciences
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2009
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tom 12
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nr 3
Equine herpesvirus type 1 (EHV-1) is one of the major viral agents causing diseases in horses common worldwide. A variety of techniques, including PCR, have been used to diagnose EHV-1 infections. In this paper, an attempt of real-time PCR has been described, which uses specific fluorochrome-labeled TaqMan probes for detection of viral DNA. This method does not require post-amplification manipulations, thereby reducing the risk of cross-contamination. The assay was sensitive enough to detect EHV-1 sequences in different clinical samples, as well in mice neuronal cell cultures. The technique was also very specific – there was no cross reaction with other human and equine herpesviruses. Compared to previously used nested PCR technique, the test was more sensitive and should be useful for the common diagnosis based on its specificity and rapidity.
3
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Equine herpesvirus type 1 (EHV-1) replication in primary murine neurons culture

84%
Cymerys J., Dzieciatkowski T., Slonska A., Bierla J., Tucholska A., Chmielewska A., Golke A., Banbura M. W.
Polish Journal of Veterinary Sciences
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2010
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tom 13
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nr 4
Equine herpesvirus-1 (EHV-1) infections cause significant economic losses for equine industries worldwide as a result of abortion, respiratory illness, and neurologic disease in all breeds of horses. The occurrence of abortions caused by EHV-1 has repeatedly been confirmed in Poland, but neurological manifestations of the infection have not been described yet. Also it is unknown how the infection of neurons with non-neuropathogenic strains is regulated. To further understand the virus- neuron interaction we studied two strains of EHV-1 in murine primary neuron cell cultures. Both strains were isolated from aborted fetuses: Rac-H, a reference strain isolated by Woyciechowska in 1959 (Woyciechowska 1960) and Jan-E isolated by Bańbura et al. (Bańbura et al. 2000). Upon infection of primary murine neuronal cell cultures with Jan-E or Rac-H strains, a cytopathic effect was observed, manifested by a changed morphology and disintegration of the cell monolayer. Positive results of immunofluorescence, nPCR and real-time PCR tests indicated high virus concentration in neurons, meaning that both EHV-1 strains were likely to replicate in mouse neurons in vitro without the need for adaptation. Moreover, we demonstrated that some neurons may survive (limited) virus replication during primary infection, and these neurons (eight weeks p.i.) harbour EHV-1 and were still able to transmit infection to other cells.
4
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The xCELLigence system for real-time and label-free analysis of neuronal and dermal cell response to Equine Herpesvirus type 1 infection

67%
Golke A., Cymerys J., Slonska A., Dzieciatkowski T., Chmielewska A., Tucholska A., Banbura M. W.
Polish Journal of Veterinary Sciences
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2012
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tom 15
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nr 1
Real-time cell electronic sensing (RT-CES) based on impedance measurements is an emerging technology for analyzing the status of cells in vitro. It allows label-free, real time monitoring of the biological status of cells. The present study was designed to assess dynamic data on the cell processes during equine herpesvirus type 1 (EHV-1) infection of ED (equine dermal) cells and primary murine neuronal cell culture. We have demonstrated that the xCELLigence system with dynamic monitoring can be used as a rapid diagnostic tool both to analyze cellular behavior and to investigate the effect of viral infection.
5
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Restriction enzyme analysis of EHV1 strains responsible for abortion outbreak in mares

67%
Rola J.
Bulletin of the Veterinary Institute in Puławy
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1997
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tom 41
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nr 2
In the present study field strains of equine herpesvirus 1 (EHV 1) were compared by restriction enzyme analysis. The strains were isolated from aborted fetuses in a horse stud during an abortion outbreak in 1993/1994. We found that all isolates had the same electrophoretic patterns of EHV 1 type, very similar to the pattern of the reference Ab-4 strain of EHV 1. We confirmed also the usefulness of DNA fingerprinting method for differentiation of EHV 1 and EHV4 viruses.
6

Postępowanie w przypadku wystąpienia postaci nerwowej zakażenia wywołanego przez herpeswirus koni typu 1

51%
Rola J., Stasiak K.
Lecznica Dużych Zwierząt. Ogólnopolski Kwartalnik dla Lekarzy Weterynarii
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2018
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tom 13
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nr 2
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