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.
277 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.
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)
The purpose of the work was to carry out comparative analysis of the positive and negative on leukemia cattle blood sera in ELISA kits of different constructions. Research was carried out using “DIA®-BLV-Ab” kit, in which the reaction had been performed in the indirect ELISA, and “ID Screen® BLV Competition” kit in a competitive format. There were used 15 cattle blood sera for testing, in which antibodies to BLV were confirmed in the ID and the ELISA “Bovine leukemia virus antibody test kit” (IDEXX), as well as 10 positive cattle blood sera confirmed in ID, 10 weak positive sera tested in ID and 10 sera with a weak line of precipitate in ID, 34 negative for leukemia blood sera tested in ID, from which 24 were also tested in the ELISA “Bovine leukemia virus antibody test kit”. The “DIA®-BLV-Ab” kit and “ID Screen® BLV Competition” kit determined positive 25 blood sera with antibodies to BLV, which were positive in ID, and 15 samples were also confirmed in IDEXX test kit. When analyzing 10 sera, that were weak positive in ID, the “DIA®-BLV-Ab” kit determined 8 sera as positive and 2 samples as negative. The “ID Screen® BLV Competition” kit detected specific antibodies to all sera. When analyzing 14 sera with a weak precipitate line in ID, the “DIA®-BLV-Ab” kit determined 9 samples as positive and 5 as negative. The “ID Screen® BLV Competition” determined specific antibodies in 11 samples When analyzing 3 sera, the test result was negative in both ELISA kits. The “DIA®-BLV-Ab” kit determined as negative all 34 sera, which were negative in ID, 24 samples from them were negative in IDEXX test kit. In the “ID Screen® BLV Competition” kit 5 false positive results were received. Studies have shown that both test kits have a high diagnostic capacity and detect antibodies to BLV at different concentrations in all positive sera. The “DIA®-BLV-Ab” kit determined 34 sera as negative, in which specific antibodies were absent, and the “ID Screen® BLV Competition” kit identified 5 samples with a false positive result
Veterinary drug AmixinR, active substance dihydrochloride 2,7-bis[2-(diethylamine)ethoxy]fluorene-9-one (AMX), was tested for its antiviral activity with using of the epizootic relevance for Ukraine the infectious agents of Pseudorabies (PR), Teschovirus encephalomyelitis (TEM), classical swine fever (CSF), porcine reproductive and respiratory syndrome pigs (PRRS), 2nd type of porcine circovirus (PCV-2) and parvovirus infections (PPVI), swine (SIV) and avian influenza viruses (AIV). The influence of this drug on agents of the avian mycoplasmosis (Myc) and pasteurellosis (Past) was also learned. AMX action was tested in the concentrations of 0.5–15 mg/ml, expositions for 0.25–12 hours and at the room temperature. Under these conditions, the inactivation of 20–60% (the highest concentrations more likely) the 1000 infectious units (TCID, ELD or PFU50/ml, respectively) of all viruses was resulted. The 5000 ELD50/ml of the SIV and AIV viruses were inactivated for 5 hours almost totally. Moderate toxicity of AMX was registered in it doses ≥1 mg/ml (P≤0.01) for tube cultures of PK-15 cell line and Marc-145. At the same time AMX doses ≥0.125 mg/ml inhibited by 20–75% of infective activities of agents of the PR, PRRS, PCV-2, PPVI (n=42, P≤0.01) in cell cultures Marc-145 and PK-15, respectively. The toxicity of the drug for 7–9-days-old embryos chickens began to emerge with a concentration of 1.5 mg/ml (n=18, P≤0.01). Its virostatic effect on SIV, AIV and PR agents was manifested in doses ≥0.25 mg/ml (n=24, P≤0.01). AMX was administrated in a single dose of 540 mg ana partes with sunflower oil and showed no toxicity for suckling piglets (n=4, P≤0.01) and 10-days-old chickens (n=10, P≤0.005). At the same time the antibiotic resistant Myc and Past from the blood of chicken infected by natural mixes of these agents, acquire the sensitivity to commercial food antibiotic after 5-days course of treatment by AMX. These data is interpreted the mechanism of therapeutic and preventive action of AMX through direct antivirus activity. The hypothesis of acquires the antibiotic sensitivity by pathogenic bacteria throughout its bacteriophages inhibition is proposed.
The article presents data on the use of GIS technologies to visualize the spread of Marek’s disease virus in Ukraine since 2011. The ArcGis v. 10.4.1 was used as a geographic information system. Three vector maps were designed, with different data on Marek’s disease virus outbreaks, and a map showing the density of the poultry population in Ukraine, to better understand the possible risks associated with the spread of the virus and to predict the epizootic situation regarding Marek’s disease
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