Objective. To research the molecular characteristics of two HPAI strains – A/Ch/Syvash/02/05/H5N1 and A/Ch/Krasnogvardeysk/58/08/H5N1, which were identifi ed as representatives of the highly pathogenic H5N1 viruses. Methods. RNA extraction, real-time polymerase chain reaction (PCR). Results. The phylogenetic studies revealed that the above mentioned strains belong to two various genetic lineages originated from the Eastern European strains isolated in 2005, but differ from the viruses introduced to the Central and Western Europe in 2005/2006, and also the lineages consisting of H5N1 viruses isolated in the Europe and Middle East in late 2007. Conclusions. Relying on experimental studies, it can be concluded that the strains of A/Ch/Syvash/02/05/H5N1 and A/Ch/Krasnogvardeysk/58/08/H5N1 are highly pathogenic.
Aim. This study was focused on (i) detection of specifi c BVDV-antibodies within selected cattle farms, (ii) identifi cation of persistently infected (PI) animals and (iii) genetic typing of selected BVDV isolates. Methods. RNA extraction, real-time polymerase chain reaction, ELISA technique, sequencing. Results. Specifi c BVDV- antibodies were detected in 713 of 1,059 analyzed samples (67.3 per cent). This number is in agreement with fi ndings in many cattle herds around the world. However, the number of positive samples differed in the herds. While 57 samples out of 283 (20.1 per cent) were identifi ed in the fi rst herd, 400 out of 475 (84.2 per cent) and 256 out of 301 (85 per cent) animals were positive in the second and third herd. High number of animals with BVDV RNA was detected in all herds. The real-time PCR assay detected BVDV RNA in 5 of 1068 samples analyzed (0.5 per cent). 4 positive samples out of 490 (0.8 per cent) and 1 out of 301 (0.33 per cent) were found in the second and third herd. The
genetic materials of BVDV were not found in the fi rst herd. Data on the number of PI animals were in accord with serological fi ndings in the cattle herds involved in our study. The genetic typing of viral isolates revealed that only BVDV, Type 1 viruses were present. The phylogenetic analysis confi rmed two BVDV-1 subtypes, namely b and f and revealed that all 4 viruses from the second farm were typed as BVDV-1b and all of them were absolutely identical in 5’-UTR, but virus from the third farm was typed as BVDV-1f. Conclusion. Our results indicated that the BVDV infection is widespread in cattle herds in the eastern Ukraine, that requires further research and development of new approaches to improve the current situation.
Isolation of the virus from biological material from a two-month-old calf with pathology of the respiratory system from a herd with a morbidity rate of 48% was performed. After detection the presence of IRT antigens in the lungs of the dead animal, the pathogen was isolated on a continuous culture of calf kidney cells, where a characteristic cytopathic effect was observed. The genetic material of the bovine herpesvirus type 1 (Bovine herpesvirus-1, BHV-1) was identified by polymerase chain reaction in the test sample. The virus isolate was adapted to continuous cell cultures of calf kidneys, sheep kidney, cow embryo lung and calf trachea, and the most suitable biological system was determined, where adsorption and reproductive properties of the virus were more pronounced. It was found that the highest titer of infectious activity of BHV-1 isolate (6.1 lg TCD50/cm3) was obtained on continuous culture of lung embryonic cells of a cow embryo after its reproduction during three consecutive passages (observation period)
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Conclusions and prospects for further research:It has been established that in the case of double subcutaneous administration, the inactivated vaccine against contagious agalactia of sheep and goats (NSC "IECVM") provides 100% protection for susceptible animals from clinical manifestations of the disease and is harmless, areactogenic, characterized by immunogenic and protective properties equal to the vaccine, produced in Romania, and can be recommended for further implementation. УДК 57.043;578.71 СТЕГНІЙ М.Ю., канд. біол. наук, доцент СТЕГНІЙ Б.Т., док. вет. наук, професор, академік НААН Національний науковий центр «Інститут експериментальної і клінічної ветеринарної медицини» м. Харків ВИКОРИСТАННЯ КРІОКОНСЕРВУВАННЯ ВІРУСУ ЛЕЙКОЗУ ВЕЛИКОЇ РОГАТОЇ ХУДОБИ У БІОТЕХНОЛОГІЇ ВИРОБНИЦТВА ЛЕЙКОЗНОГО ДІАГНОСТИКУМУ Розроблено спосіб кріоконсервування (FLK-SBBL), який включає поетапне заморожування, на першому етапі впродовж години за температури плюс чотири градуси за Цельсієм, на другому етапі: в парах рідкого азоту впродовж 18 годин з наступним занурюванням у рідкий азот; суміш поживних середовищ DMEM і 199 (1:1) (70 %), з сироваткою крові ВРХ (20 %) та дімексиду (10 %). Розроблений спосіб кріоконсервування (FLK-SBBL) дозволяє зберігати на вихідному рівні антигенпродукуючу та віруспродукуючу активність штамів FLK-BLV за довготривалого зберігання в умовах кріобанку ННЦ «ІЕКВМ». За допомогою електронно-мікроскопічних досліджень встановлено, що максимальна кількість продукції вірусу лейкозу відбувалася з п'ятої по сьому доби культивування після розморожування (FLK-SBBL) Ключові слова. лейкозний антиген, FLK-BLV, кріоконсервування.
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