2017 № 4 (23)

CATTLE DISEASES

UDC 619:616.98:578.835.2:616-036.22(517)

А. М. Timina, N. G. Zinyakov, A. V. Scherbakov, D. A. Lozovoy

PHYLOGENETIC ANALYSIS OF FMDV ISOLATES RECOVERED IN POST-SOVIET STATES AND MONGOLIA IN 2016

P. 7–8

Federal Centre for Animal Health (FGBI “ARRIAH”, Vladimir, Russia) is an OIE Regional Reference Laboratory for Foot-and-Mouth Disease for Eastern Europe, Central Asia and Transcaucasia and FAO Reference Centre for Foot-and-Mouth Disease for Central Asia and Western Eurasia. The FGBI “ARRIAH’s” objectives as a Regional Reference Laboratory/Centre include continuous FMDV monitoring in post-Soviet states, detection of new genetic virus variants and matching their antigenic properties with antigenic properties of available vaccine virus strains. The paper demonstrates results of phylogenetic analysis of foot-and-mouth disease virus (FMDV) that caused the disease in Armenia, Zabaikalsky Krai (Russia), Middle Asia and Mongolia in 2016. The test results are indicative of the fact that FMD outbreaks in Armenia were caused by the virus belonging to A/G-VII genetic lineage that was widely spread in the Near East in 2015 and had been never reported in post-Soviet states before. The FMD outbreaks in the Zabaikalsky Krai in November – December 2016 were caused by the exotic for Russia FMD virus of O/Ind-2001d genetic lineage. In Middle Asia FMD was caused by O/ME-SA/PanAsia-2 virus belonging to the O/ME-SA/PanAsia-2 genetic lineage and endemic for the region. In Mongolia the FMD outbreak was caused by the virus belonging to A/Sea-97 genetic lineage that had been previously reported in this country in 2013. Results of the virus genotyping are of particular importance for adjustment of the disease control measures.

Key words: foot-and-mouth disease virus (FMDV), phylogenetic analysis.

REFERENCES

1. Scherbakov A. V., Perevozchikova N. A., Drygin V. V. Methodical instructions on FMDV genome indication and strain differentiation using polymerase chain reaction and VP1 gene sequencing // Methodical guidelines for livestock disease diagnosis using polymerase chain reaction. – Vladimir, 1998. – P. 31–41.

2. Application of RT-PCR and nucleotide sequencing in foot-and-mouth disease diagnosis / A. Sherbakov, N. Lomakina, V. Drygin, A. Gusev // Vet. Quart. – 1998. – Vol. 20, No. 2. – P. 32–34.

3. Foot-and-mouth disease. – URL: http: www.foot-andmouth.org (accessed date: 02.02.17).

4. Foot-and-mouth disease, China. – URL: http://oie.int/wahis_2/public/wahid.php/Reviewreport/Review?page_refer=M... (accessed date: 29.09.17).

5. Incursions of foot-and-mouth disease virus into Europe between 1985 and 2006 / J. F. Valarcher, Y. Leforban, M. Rweyemamu [et al.] // Transbound. Emerg. Dis. – 2008. – Vol. 55, No. 1. – P. 14–34.

6. Knowles N. J., Samuel A. R. Molecular epidemiology of foot-and-mouth disease virus // Virus Res. – 2003. – Vol. 91, No. 1. – P. 65–80.

7. Molecular epidemiology reports produced by the FAO World Reference Laboratory for FMD. – URL: http://www.wrlfmd.org/fmd_genotyping (accessed date: 02.02.17).

8. Samuel A. R., Knowles N. J. Foot-and-mouth disease type O viruses exhibit genetically and geographically distinct evolutionary lineages (topotypes) // J. Gen. Virol. – 2001. – Vol. 82, No. 3. – P. 609–621.

 

UDC 619:616.98:578.824.91:616-036.22:616-097

Ye. А. Konstantinova, S. К. Starov, А. V. Konstantinov, V. I. Diev, А. К. Karaulov

EFFECTIVENESS OF SEROLOGICAL TESTS FOR DIAGNOSTICS OF BOVINE EPHEMERAL FEVER

P. 9–13

Review of publications devoted to spread of bovine ephemeral fever (BEF) in the world is presented; epizootic aspect of this problem is highlighted. The disease is not registered in Americas, Island, New Zealand and Europe. African continent, Australia and Asian countries including those neighboring the Russian Federation are considered BEF infected, that is why there is a risk of BEF introduction into our country. Serological tests for BEFV antibodies in the Russian Federation were performed for the first time. In total 13,911 samples, collected in 2008–2010 on farms in 26 regions of the Russian Federation were tested. Testing of cattle sera using prolonged complement fixation test revealed 333 BEFV seropositive samples, submitted from 9 regions in 2008, percentage by region was 2.6–25.9%, average percentage was 9.7% out of the total number of samples tested during the abovementioned period. Sera specific activity was determined when they were used non-diluted or diluted at 1:10–1:20. Sera with positive reactions in prolonged CFT showed specific activity in ELISA at 1:10–1:1,280 dilutions, but if diffusion precipitation test with non-diluted sera was used antibodies were detected only in few samples due to insufficient sensitivity. Test results proved the possibility of serological monitoring using prolonged CFT and ELISA. Due to insufficient sensitivity of diffusion precipitation test, its use for this purpose is not feasible.

Key words: bovine ephemeral fever, prolonged CFT, ELISA, diffusion precipitation test, antibodies, diagnostics.

REFERENCES

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2. Makarov V. V. Sovremennye dannye ob jefemernoj lihoradke krupnogo rogatogo skota // Sel'skoe hozjajstvo za rubezhom. – 1974. – № 11. – S. 34–36.

3. Mirzaev D. Profilaktika i mery bor'by s jefemernoj lihoradkoj // Sel'skoe hozjajstvo Tadzhikistana. – 1984. – № 12. – S. 46.

4. Monitoring rasprostranenija i predotvrashhenija osobo opasnyh boleznej zhivotnyh: sb. materialov konf., Samarkand, 26–27 okt. 2004 g. – Samarkand, 2004. – S. 107–110.

5. Jeksperimental'nye issledovanija po izucheniju kliniki jefemernoj lihoradki krupnogo rogatogo skota i polucheniju diagnosticheskih preparatov / V. I. Diev [I dr.] // Virusnye i mikrobnye bolezni zhivotnyh: sb. nauch. tr. – Vladimir, 1995. – S. 128–131.

6. A large-scale outbreak of bovine ephemeral fever in Turkey, 2012 / S. Tonbak [et al.] // J. Vet. Med. Sci. – 2013. – Vol. 75, No. 11. – P. 1511–1514.

7. Application of a real-time polymerase chain reaction assay to the diagnosis of bovine ephemeral fever during an outbreak in New South Wales and northern Victoria in 2009-10 / D. S. Finlaison [et al.] // Aust. Vet. J. – 2014. – Vol. 92, No. 1–2. – P. 24–27.

8. Circulation of bovine ephemeral fever in the Middle East – strong evidence for transmission by winds and animal transport / O. Aziz-Boaron [et al.] // Vet. Microbiol. – 2012. – Vol. 158, No. 3–4. – P. 300–307.

9. Epidemiological analysis of bovine ephemeral fever in 2012–2013 in the subtropical islands of Japan / Y. Hayama [et al.] // BMC Vet. Res. – 2016. – URL: https://bmcvetres.biomedcentral.com/articles/10.1186/s12917-016-0673-0 (дата обращения: 03.08.16).

10. Epidemiological investigation of bovine ephemeral fever outbreaks in Israel / I. Yeruham [et al.] // Vet. Med. Int. – 2010. – URL: http://www.hindawi.com/journals/vmi/2010/290541/ (дата обращения: 03.08.16).

11. Epizootics of bowine ephemeral fewer on dairy farms in Saudi Arabia / M. A. Farag [et al.] // Rev. Sci. Tech. OIE. – 1998. – Vol. 17, No. 3. – P. 713–722.

12. Finlaison D. S., Read A. J., Kirkland P. D. An epizootic of bovine ephemeral fever in New South Wales in 2008 associated with long-distance dispersal of vectors // Aust. Vet. J. – 2010. – Vol. 88, No. 8. – P. 301–306.

13. Large-scale serological survey of bovine ephemeral fever in China / Z. Li [et al.] // Vet. Microbiol. – 2015. – Vol. 176, No. 1–2. – P. 155–160.

14. Molecular epidemiology of Bovine ephemeral fever virus in cattle and buffaloes in Iran / H. Momtaz [et al.] // Revue Méd. Vét. – 2012. – Vol. 163, No. 8–9. – P. 415–418.

15. Occurrence of bovine ephemeral fever in Okinawa Prefecture, Japan, in 2012 and development of a reverse-transcription polymerase chain reaction assay to detect bovine ephemeral fever virus gene / T. Niwa [et al.] // J. Vet. Med. Sci. – 2015. – Vol. 77, No. 4. – P. 455–460.

16. Preliminary studies of the complement fixation test to confirm the diagnosis of bovine ephemeral fever / Bai Wenbin [et al.] // Aust. J. Biol. Sci. – 1987. – Vol. 40, No. 2. – P. 137–141.

17. Relationships of bovine ephemeral fever epizootics to population immunity and virus variation / L.-J. Ting [et al.] // Vet. Microbiol. – 2014. –Vol. 173, No. 3–4. – P. 241–248.

18. Resurgence of bovine ephemeral fever in mainland Japan in 2015 after a 23-year absence / Y. Hirashima [et al.] // J. Vet. Med. Sci. – 2017. – Vol. 79, No. 5. – P. 904–911.

19. Sero-survey on Aino, Akabane, Chuzan, bovine ephemeral fever and Japanese encephalitis virus of cattle and swine in Korea / S. In Lim [et al.] // J. Vet. Sci. – 2007. – Vol. 8, No. 1. – P. 45–49.

20. The follow up study after massive outbreak of Akabane and bovine ephemeral fever viruses in Korea / D. K. Yang [et al.] // Korean J. Vet. Serv. – 2013. – Vol. 36, No. 3. – P. 151–155.

21. The geographical distribution and first molecular analysis of Culicoides Latreille (Diptera: Ceratopogonidae) species in the Southern and Southeastern Turkey during the 2012 outbreak of bovine ephemeral fever / B. Dik [et al.] // Parasitol. Res. – 2014. – Vol. 113, No. 11. – P. 4225–4232.

22. The history of bluetongue, Akabane and ephemeral fever viruses and their vectors in Australia 1975–1999 / T. D. St. George [et al.]. – Canberra, Australia, 2001. – 101 p. – URL: https://espace.library.uq.edu.au/view/UQ:290721/TSG_2_34.pdf (дата обращения: 05.08.16).

23. Zheng F., Qiu C. Phylogenetic relationships of the glycoprotein gene of bovine ephemeral fever virus isolated from mainland China, Taiwan, Japan, Turkey, Israel and Australia // Virol. J. – 2012. – Vol. 9. – URLhttp://virologyj.biomedcentral.com/articles/10.1186/1743-422X-9-268 (дата обращения: 05.08.16).

 

 

 

UDC 619:613.11:636.2

М. S. Mansurova

BLOOD MORPHOBIOCHEMICAL PARAMETERS OF HEREFORD COWS IMPORTED FROM AUSTRALIA IN SPRING

P. 14–16

The results of morphological and biochemical analysis of Hereford cows imported in 2012 from Australia to the Amur Oblast are presented in the paper. Blood parameter assessment of heifers obtained from the abovementioned cows, cross mated with local Hereford bulls, is given here. All animals were managed under identical conditions: on natural pastures in summer and at stables during cold seasons. Deterioration in some types of metabolic processes was noted in mother cows, in particular based on sera test results, the decrease in albumen synthesis resulting from the increase in blood urea nitrogen in protein metabolism, reduction of glucose content in carbohydrate metabolism, low contents of calcium and magnesium in the mineral metabolism. Fat metabolism parameters (lipid, cholesterol and triglyceride contents) were within the normal range. Biochemical parameters of heifer sera as compared to mother cows were closer to the normal range; i. e. albumen concentration and glucose content were higher, BUN level was lower, calcium and magnesium contents were identical to adult animals’ ones. It was established that during winter time and after it young animals are more demanding to feeding and management conditions.

Key words: imported cattle, adaptation, Hereford, blood analysis.

REFERENCES

1. Ahmetov A. M. Kartina krovi krupnogo rogatogo skota gerefordskoj porody shvedskoj selekcii v processe akklimatizacii v uslovijah Severnogo Zaural'ja // Perspektivy razvitija APK v rabotah molodyh uchenyh: sb. materialov region. nauch.-prakt. konf. molodyh uchenyh / GAU Severnogo Zaural'ja. – 2014. – S. 7–10.

2. Biohimicheskij sostav krovi matochnogo pogolov'ja aberdin-angusskoj porody v processe adaptacii / S. V. Sidunov, I. S. Petrushko, S. A. Petrushko [i dr.] // Zootehnicheskaja nauka Belarusi. – 2013. – T. 48, № 2. – S. 224–231.

3. Bol'shakov D. S., Nikeshina T. B. Biohimicheskie pokazateli syvorotki krovi sel'skohozjajstvennyh zhivotnyh // Veterinarija segodnja. – 2015. – № 4 (15). – S. 51–55.

4. Ermishin A. S. K voprosu o celesoobraznosti ispol'zovanija importnogo skota v Jaroslavskoj oblasti // Agrarnyj vestnik Verhnevolzh'ja. – 2016. – № 4. – S. 58–65.

5. Kadraliev D. S., Hismetov I. H., Evstigneev V. V. Problemy adaptacii i akklimatizacii krupnogo rogatogo skota zarubezhnoj selekcii v uslovijah Astrahanskoj oblasti // Formirovanie i razvitie sel'skohozjajstvennoj nauki v XXI veke: sb. nauch. statej / FGBNU «PNIIAZ». – 2016. – S. 460–471.

6. Klinicheskaja laboratornaja diagnostika v veterinarii / I. P. Kondrahin, N. V. Kurilov, A. G. Malahov [i dr.]. – M.: Agropromizdat, 1985. – 287 s.

7. Koksharova T. E., Cydypov C. C. Metodicheskie ukazanija po matematicheskoj obrabotke rezul'tatov issledovanija s ispol'zovaniem tablichnogo processora Excel. – Ulan-Udje: VSGTU. – 2002. – 40 s.

8. Problemy i prinimaemye mery po adaptacii zavoznogo krupnogo rogatogo skota v Uzbekistane / Sh. A. Akmal'hanov, B. H. Zhumadullaev, A. P. Bezverhov [i dr.] // Sovremennoe jekologicheskoe sostojanie prirodnoj sredy i nauchno-prakticheskie aspekty racional'nogo prirodopol'zovanija: sb. nauch. statej. – 2016. – S. 3155–3162.

9. Salihov A. R., Sedyh T. A. Hozjajstvenno-biologicheskie osobennosti gerefordskoj porody avstralijskoj selekcii pri chistoporodnom razvedenii v uslovijah Juzhnogo Urala // Fundamental'nye issledovanija. – 2013. – № 4–5. – S. 1161–1163.

10. Sedyh T. A. Kliniko-fiziologicheskie pokazateli gerefordskogo skota zarubezhnoj selekcii pri akklimatizacii v uslovijah Bashkortostana // Izvestija OGAU. – 2017. – № 1 (63). – S. 97–100.

11. Tuzov I. N., Usenkov I. S. Biohimicheskaja harakteristika syvorotki krovi golshtinskih zhivotnyh, zavezennyh iz Kanady neteljami // KubGAU – 2013. – № 88 (04). – S. 1–21.

12. Chugunov A. V., Zaharova L. N. K probleme akklimatizacii porod // Evrazijskij sojuz uchenyh (ESU). – 2015. – № 6 (15). – S. 35–37.

 

 

 

SMALL RUMINANTS DISEASES

UDС 619:616.98:578.831.2:636.3

S. V. Scherbinin, А. К. Karaulov, V. М. Zakharov

RISK ANALYSIS OF PESTE DES PETITS RUMINANTS INTRODUCTION INTO THE TERRITORY OF THE RUSSIAN FEDERATION

P. 21–22

Peste des petits ruminants (PPR) has never been registered in the Russian Federation, but the epidemic situation in the neighboring countries indicates that it is a typical transboundary disease. Due to significant social and economic losses, negative effect on food safety of many countries of the world, peste des petits ruminants has been included in the List of priority diseases of the FAO/OIE Global Framework for the Progressive Control of Transboundary Animal Diseases (GF-TADs) A Five-Year Action Plan. The disease has already spread in the countries which share borders with Russia, therefore there is an urgent need to assess the risk of introduction and subsequent spread of peste des petits ruminants in the territory of the Russian Federation. The performed analysis of PPR epidemic situation and disease spread tendencies in the Russian Federation neighboring countries over the last years using map documents and government sources has demonstrated that there is a high risk of PPR penetration into the territory of our country. The epidemic situation in Mongolia and China, in Georgia, Turkey and Iran is of primary concern.

Key words: peste des petits ruminants (PPR), PPR Global Eradication Programme, Global Strategy, Roadmap Meeting, ways of PPR penetration in the Russian Federation.

REFERENCES

1. Presentations on the Second PPR Regional Roadmap Meeting. Dushanbe, Tadjikistan. 28.02–2.03.2017. – URL: htps://www.dropbox.com/sh/8tdj0hdnu1xk82z/AADhRlKKQssJEo09JuRFir-5a?dl=0 (accessed date: 12.03.17).

2. Animal disease risk forecasts in the Russian Federation for 2017 / FGBI “ARRIAH”. – Vladimir, 2016. – P. 191–199.

3. Peste des petits ruminants / А. V. Mischenko, V. А. Mischenko, V. P. Melnikov, О. Yu. Chernykh // Veterinariya. – 2016. – No. 9. – P. 3–9.

4. Peste des petits ruminants (current status, epizootology, specific prevention and control measures) / N. I. Zakutsky, V. М. Balyshev, А. V. Knize [et al.] // Scientific journal KubGAU. – 2012. – No. 83. – P. 10–24.

5. Disease outbreak maps: Peste des petits ruminants. – URL: http://www.oie.int/wahis_2/public/wahid.php/Diseaseinformation/Diseaseoutbreakmaps?disease_type_hidden=&disease_id_hidden=&selected_disease_name_hidden=&disease_type=0&disease_id_terrestrial=5&disease_id_aquatic=-999&selected_start_day=1&selected_start_month=1&selected_start_year=2014&selected_end_day=1&selected_end_month=12&selected_end_year=2015&submit2=OK.

6. Disease outbreak maps: Peste des petits ruminants. – URL: http://www.oie.int/wahis_2/public/wahid.php/Diseaseinformation/Diseaseoutbreakmaps?disease_type_hidden=&disease_id_hidden=&selected_disease_name_hidden=&disease_type=0&disease_id_terrestrial=5&disease_id_aquatic=-999&selected_start_day=1&selected_start_month=1&selected_start_year=2015&selected_end_day=1&selected_end_month=12&selected_end_year=2017&submit2=OK.

7. Disease outbreak maps: Peste des petits ruminants. – URL: http://www.oie.int/wahis_2/public/wahid.php/Diseaseinformation/Diseaseoutbreakmaps?disease_type_hidden=&disease_id_hidden=&selected_disease_name_hidden=&disease_type=0&disease_id_terrestrial=5&disease_id_aquatic=-999&selected_start_day=1&selected_start_month=1&selected_start_year=2015&selected_end_day=1&selected_end_month=12&selected_end_year=2016&submit2=OK.

8. Event summary: Peste des petits ruminants, Mongolia. – URL: http://www.oie.int/wahis_2/public/wahid.php/Reviewreport/Review/viewsummary?fupser=&dothis=&reportid=22395.

9. Geographical distribution of PPR. – URL: http://www.oie.int/animal-health-in-the-world/ppr-portal/distribution/.

10. Global Strategy for the Control and Eradication of PPR. – URL: http://www.fao.org/emergencies/resources/documents/resources-detail/it/c/282777/.

11. Peste des Petits Ruminants Global Eradication Programme / FAO/OIE. – Rome, 2016. – URL: http://www.fao.org/3/a-i6316e.pdf.

 

 

PORCINE DISEASES

UDC 619:616.98:578.842.1:338.14

V. M. Gulyonkin, I. M. Klinovitskaya, O. N. Petrova, A. K. Karaulov, D. A. Lozovoy

AFRICAN SWINE FEVER IN RUSSIAN FEDERATION: ECONOMIC IMPACT

P. 23–27

Following the results of the earlier studies the economic losses due to African swine fever amounted to about 26.56 bln RUB (5.4% inflation-adjusted). In 2016 ASF further spread in the population of domestic pigs and wild boars in the 27 Subjects of the Russian Federation. All disease outbreaks were reported in the European part of the Russian Federation. Total of 26 infected regions were reported where ASF was diagnosed in domestic pigs and 15 infected regions were reported where ASF was detected in wild boars. In the Ivanovo Oblast the ASF was diagnosed only in wild boars (one case). In 15 RF Subjects the disease was reported both in domestic pigs and wild boars. Estimated data demonstrated that economic losses due to ASF outbreaks in domestic pigs and wild boars will amount to 7.89 bln RUB this year. Total economic losses due to ASF spread in the country in 2008–2016 will amount to about 34.45 bln RUB in monetary terms.

Key words: African swine fever, economic losses, anti-epidemic measures, pig industry.

REFERENCES

1. Veterinarnye pravila osushhestvlenija profilakticheskih, diagnosticheskih, ogranichitel'nyh i inyh meroprijatij, ustanovlenija i otmeny karantina i inyh ogranichenij, napravlennyh na predotvrashhenie rasprostranenija i likvidaciju ochagov afrikanskoj chumy svinej / MSH RF. – 2016. – 16 s.

2. Instrukcija o meroprijatijah po preduprezhdeniju i likvidacii afrikanskoj chumy svinej / MSH SSSR. – 1980. – 10 s.

3. Klinovickaja I. M., Gulenkin V. M., Karaulov A. K. Ocenka jekonomicheskih poter' pri likvidacii afrikanskoj chumy svinej v regionah RF v 2015 g. // BIO. – 2016. – № 10. – S. 24–30.

4. Metodika ischislenija razmera vreda, prichinennogo ohotnich'im resursam: utv. prikazom Minprirody Rossii ot 8 dekabrja 2011 g. № 948 (v red. prikaza Minprirody Rossii ot 22 ijulja 2013 g. № 252).

5. Metodika opredelenija jekonomicheskoj jeffektivnosti veterinarnyh meroprijatij / P. A. Chulkov, I. N. Nikitin [i dr.]. – M., 1982. – 55 s.

6. Metodicheskoe rukovodstvo po opredeleniju jekonomicheskogo ushherba i jeffektivnosti protivojepizooticheskih meroprijatij pri osobo opasnyh boleznjah zhivotnyh (gruppa A) / V. M. Gulenkin [i dr.]; FGU «VNIIZZh». – Vladimir, 1996. – 86 s.

7. O tendencijah rasprostranenija AChS v RF i Evrope v 2016 godu. – URL: http://fsvps.ru/fsvps/news/19990.html.

8. Federal'naja sluzhba gosudarstvennoj statistiki. – URL: http://www.gks.ru/.

9. Jekonomicheskie posledstvija rasprostranenija afrikanskoj chumy svinej v Rossijskoj Federacii / V. M. Gulenkin, I. M. Klinovickaja, A. K. Karaulov, A. S. Igolkin // Veterinarija. – 2015. – № 12. – S. 7–11.

10. Prevention and control of animal diseases worldwide. Part I: Economic analysis: Prevention versus outbreak costs / C. Caspari [et al.]; OIE; Agra CEAS Consulting. – 2007. – 257 p. – URL: http://www.oie.int/fileadmin/Home/eng/Support_to_OIE_Members/docs/pdf/OIE_-_Cost-Benefit_Analysis__Part_I_.pdf.

11. World Organisation for Animal Health (OIE). – URL: http://www.oie.int.

 

 

 

 

 

 

AVIAN DISEASES

UDC 619:616.98:579.943.94

V. А. Evgrafova, А. V. Potekhin

VIRULENCE OF INFECTIOUS CORYZA CAUSATIVE AGENT ISOLATES

P. 28–32

Virulence of 10 isolates of infectious coryza agent recovered from diseased 38-211-day-old chicken on poultry farms in the Vladimir, Moscow, Kostroma, Yaroslavl, Orenburg and Ulyanovsk Oblasts, as well as the Republic of Mordovia and Tatarstan was determined. One isolate was recovered from poultry in the Republic of Belarus. The criteria for virulence determination of Avibacterium paragallinarum were assessed according to a point system based on severity of inflammatory processes in upper respiratory airways of experimentally infected chicken. Nine out of ten isolates were found pathogenic for poultry. Chicken infected with different isolates of A. paragallinarum demonstrated the same duration of the disease stages. Diseased poultry showed similar clinical signs: rhinitis, sinusitis and conjunctivitis. In spite of the typical dynamic of the disease in infected poultry, the virulence of the isolates was different. The most virulent were isolates recovered from poultry farms in the Kostroma and Orenburg Oblasts, as well as the Republic of Tatarstan and Mordovia. The diseased chicken demonstrated abundant nasal discharge with marked swelling of the infraorbital sinus. A. paragallinarum isolates recovered in the farms of the Yaroslavl and Ulyanovsk Oblasts, as well as the Republic of Belarus showed medium virulence. The infection was characterized by moderate nasal discharges with swelling in the area of the infraorbital sinuses. Isolates recovered from the farms in the Moscow Oblast' had low virulence. Infected poultry demonstrated minor discharge from nasal passages with insignificant swelling in infraorbital sinuses. The isolate recovered in the Vladimir Oblast was found avirulent, the clinical signs were absent. Based on the suggested differentiation scheme the inhomogenuity of virulent properties of A. paragallinarum isolates was shown.

Key words: infectious coryza (Haemophilus infection) in chickens, isolates, Avibacterium paragallinarum, virulence.

REFERENCES

1. Gemofilez ptic / T. N. Rozhdestvenskaja, E. V. Kononenko, S. A. Emel'janova [i dr.] // Veterinarnaja medicina. – 2016. – № 4. – S. 50–53.

2. Danilova V. A., Potehin A. V., Stepanova I. A. Osobennosti vydelenija i identifikacii vozbuditelja infekcionnogo rinita (gemofileza) kur // Veterinarija segodnja. – 2016. – № 3 (18). – S. 65–70.

3. Blackall P. J., Matsumoto M. Infectious coryza // Diseases of Poultry / ed. Y. M. Saif [et al.]. – 11th ed. – Ames, Iowa, 2003. – P. 691–703.

4. Bragg R. R. Effects of differences in virulence of different serovars of Haemophilus paragallinarum on perceived vaccine efficacy // Onderstepoort J. Vet. Res. – 2005. – Vol. 72, No. 1. – P. 1–6.

5., Evaluation of two experimental infection models for Avibacterium paragallinarum / Q. Zhao, Y. N. Sun, X. X. Zhang [et al.] // Vet. Microbiol. – 2010. – Vol. 141, No. 1–2. – P. 68–72.

6. Reclassification of Pasteurella gallinarum, [Haemophilus] paragallinarum, Pasteurella avium and Pasteurella volantium as Avibacterium gallinarum gen. nov., comb. nov., Avibacterium paragallinarum comb. nov., Avibacterium avium comb. nov. and Avibacterium volantium comb. nov. / P. J. Blackall, H. Christensen, T. Beckenham [et al.] // Int. J. Syst. Evol. Microbiol. – 2005. – Vol. 55, No. 1. – P. 353–362.

7. Soriano-Vargas E., Terzolo H. R. Haemophilus paragallinarum: Etiology of infectious coryza // Vet. Méx. – 2004. – Vol. 35, No. 3. – P. 245–259.

8. The vaccination-challenge trial: the gold standard test to evaluate the protective efficacy of infectious coryza vaccines / A. García, F. Romo, A. M. Ortiz [et al.] // Avian Pathol. – 2008. – Vol. 37, No. 2. – P. 183–186.

9. Virulence characterization of Avibacterium paragallinarum isolates from Uganda / D. K. Byarugaba, U. M. Minga, P. S. Gwakisa [et al.] // Avian Pathol. – 2007. – Vol. 36, No. 1. – P. 35–42.

10. Virulence of South African isolates of Haemophilus paragallinarum. Part 3: Experimentally produced NAD-independant isolate / M. Taole, J. Albertyn, E. Van Heerden [et. al.] // Onderstepoort J. Vet. Res. – 2002. – Vol. 69, No. 3. – P. 189–196.

11. Virulence of the nine serovar reference strains of Haemophilus paragallinarum / V. E. Soriano, G. M. Longinos, R. P. Fernández [et al.] // Avian Dis. – 2004. – Vol. 48, No. 4. – P. 886–889.

12. Yamamoto R. Infectious coryza // Diseases of Poultry / ed. B. W. Calnek [et al.]. – 9th ed. – Ames, Iowa, 1991. – P. 186–195.

 

 

 

GENERAL ISSUES

UDC 619:616.98:578.824.11:616-036.22(470.311)

А. V. Paroshin, S. V. Voskresensky, А. А. Mukovnin, А. V. Yenshin,

А. Ye. Metlin, К. N. Gruzdev

RETROSPECTIVE ANALYSIS OF RABIES EPIZOOTIC SITUATION IN MOSCOW REGION IN 2010–2016

P. 33–37

Rabies is a viral disease of animals and humans that is characterized by signs of polioencephalomyelitis and is in most cases lethal. This is one of the oldest and most dangerous infectious diseases. As estimated by the WHO this disease is in the top five zoonoses that cause major economic losses; it poses a constant threat to animal and human health and is included in the OIE list of notifiable diseases. Rabies in animals (especially wild carnivores) is one of the most important international criteria (WHO, FAO, OIE) for assessing the biological and environmental safety of human habitat. The Moscow Oblast' is a unique region in Russia, which is an urbanized megalopolis where rabies is reported in animals. Occupying a large territory, it borders on seven Subjects of the Russian Federation with the unfavourable epidemic situation with regard to rabies. This paper presents statistical data on the epizootic situation in the Moscow Oblast, Moscow and neighboring regions in 2010–2016. The dynamics of the epizootic process over the years and the need for its continuous monitoring are shown. In the territory of the Moscow Oblast and neighbouring regions, oral vaccination is one of the main ways to control rabies in animals in the wild. In order to prevent introduction, as well as to ensure prevention and eradication of this transboundary disease, a deeper scientific rationale is required for carrying out a complex of veterinary activities taking into account epizootic processes occurring in the border areas of neighbouring territories.

Key words: rabies virus, retrospective analysis, epizootic situation.

REFERENCES

1. Analiticheskij obzor meroprijatij po profilaktike i bor'be s beshenstvom zhivotnyh v Rossijskoj Federacii / V. A. Vedernikov, A. A. Shabejkin, A. M. Guljukin [i dr.]. – M., 2014. – 23 s.

2. Baldina I. V. Jepizootologicheskie osnovy sovershenstvovanija profilaktiki beshenstva v Moskovskom regione: dis. … kand. vet. nauk. – M., 2004. – 142 s.

3. Bel'chihina A. V., Karaulov A. K. Retrospektivnyj analiz jepizooticheskoj situacii po beshenstvu zhivotnyh na territorii Rossijskoj Federacii // Veterinarija segodnja. – 2016. – № 1 (16). – S. 64–70.

4. Beshenstvo v Rossijskoj Federacii: inform.-analit. bjulleten' / E. M. Poleshhuk, G. N. Sidorov, D. G. Sidorova, N. M. Kolychev. – Omsk, 2009. – 46 s.

5. Guljukin A. M. Beshenstvo. Sovremennaja sistema analiza i kontrolja jepizooticheskogo processa na territorii Rossijskoj Federacii: avtoref. dis. … d-ra vet. nauk. – SPb., 2017. – 49 s.

6. Kontrol' jeffektivnosti vakcinoprofilaktiki beshenstva dikoj fauny na territorii Smolenskoj oblasti / N. A. Hismatullina, A. M. Guljukin, V. V. Sabirova [i dr.] // Tr. VIJeV. – 2013. – T. 77. – S. 197–200.

7. Makarov V. V. Sostojanie i vozmozhnye napravlenija razvitija central'no-evropejskogo superareala beshenstva // Vet. konsul'tant. – 2004. – № 6. – S. 6–8.

8. Makarov V. V., Vorob'ev A. A. Aktual'nye problemy beshenstva: prirodnaja ochagovost', metodologija issledovanij i kontrolja v centre Rossii // Vet. patologija. – 2004. – № 3. – S. 102–116.

9. Poljushkina G. S., Gorkunov A. A. Jepizooticheskaja situacija po beshenstvu v Moskovskoj oblasti i puti ee uluchshenija // Voprosy virusologii. – 1998. – № 1. – S. 47–48.

10. Cvil' L. A., Rodina L. V., Lebedeva I. R. Sostojanie antirabicheskoj pomoshhi i organizacija jepidnadzora za beshenstvom na territorii g. Moskvy // Sovremennye problemy rabiologii: tez. dokl. nauch. konf. – M., 1998. – S. 15–16.

 

 

 

UDC 619:579.843.96:57.082.26

V. S. Rusaleyev, D. A. Vasilyev

LOW TEMPERATURE STORAGE OF ACTINOBACILLUS PLEUROPNEUMONIAE CULTURES

P. 38–41

Basic tasks during storage of any microorganism cultures including Actinobacillus pleuropneumoniae involve maintenance of their vitality and stable taxonomically relevant signs. In different collections the microorganism vitality is mainly maintained using the following methods: repeated passages, lyophilisation and storage with mineral oil at low and ultra low temperatures. Promising method of long-term storage of microorganisms includes cryopreservation that demonstrates a number of advantages. The paper examines approaches to possible improvement of means for long term storage of actinobacilli. Experimental data are reported that were obtained during research of potential effect of liquid nitrogen freezing on vitality and some biological properties of actinobacilli. After one year storage (observation period) at −196 °С the microorganism cultures were demonstrated to preserve vitality and species-specific cultural-morphologic and biochemical properties. Factors relevant for Actinobacillus pleuropneumoniae preparation for cryopreservation and affecting storage results are demonstrated.

Key words: Actinobacillus pleuropneumoniae, cultivation, bacterial vitality, cryopreservation, cryoprotectors.

REFERENCES

1. Braun V. Genetika bakterij / pod red. S. I. Alihanjana. – M.: Nauka, 1968. – 446 s.

2. Zhiznesposobnost' liofilizirovannyh bakterij v zavisimosti ot temperatury ohlazhdenija / M. V. Danilova, I. M. Nadirova, T. V. Emceva, E. V. Kuznecova // Izvestija AN SSSR. Serija biologicheskaja. – 1981. – № 2. – S. 307–309.

3. Kriokonservacija – perspektivnyj metod hranenija promyshlenno cennyh shtammov molochnokislyh bakterij i drozhzhej / O. A. Savkina, G. V. Ternovskoj, M. N. Lokachuk [i dr.] // Sel'skohozjajstvennaja biologija. – 2014. – № 4. – S. 112–119.

4. Osin A. V., Chervjakova N. S., Valova T. V. Liofilizacija shtammov patogennyh mikroorganizmov na sublimacionnyh ustanovkah raznogo tipa i ocenka kachestva poluchennyh preparatov // Problemy osobo opasnyh infekcij. – 2016. – Vyp. 3. – S. 66–70.

5. Cucaeva A. A., Anan'ina A. E. Vlijanie uslovij hranenija na svojstva kriokonservirovannoj i liofilizirovannoj kul'tury Streptomyces fradiae 25Ab12 // Problemy kriobiologii. – 2001. – № 1. – S. 52–59.

6. Ashwood-Smith M. J. Mechanisms of dehydration injury in bacteria // Int. J. Refrig. – 1980. – Vol. 3, No. 4. – P. 205–212.

7. Portner D. C., Leuschner R. G. K., Murray B. S. Optimising the viability during storage of freeze-dried cell preparations of Campylobacter jejuni // Cryobiology. – 2007. – Vol. 54, No. 3. – P. 265–270.

 

 

 

UDC 619:615.371:615.91:665.656

I. V. Pigoleva, A. S. Sharypov, T. N. Shabalina, S. V. Zaglyadova, M. V. Kitova,

S. А. Antonov, V. V. Mikhalishin, V. А. Starikov, D. V. Mikhalishin

TESTING OF EMULSION VACCINES BASED ON WHITE OILS OBTAINED
AS A RESULT OF HYDROCATALYTIC PROCESSING OF OILSTOCK

P. 42–48

The paper demonstrates the possibility to use white mineral oils resulting from hydrocatalytic processing of unconverted vacuum gas oil as a basis for an oil adjuvant used in the production of FMD vaccines intended for livestock. Samples of the developed white oils were tested for their reactogenicity and pyrogenicity, and emulsion vaccines were tested for their reactogenicity, potency, and emulsion stability. Tests results were compared with the ones for a foreign analogue. Blood samples were taken from animals before vaccination and 14 days after it to obtain sera which were further tested for FMD antibodies using VNT. Assessment of the white oil sample reactogenicity demonstrated that sample BM 1.4 obtained during hydroisomerization was the least reactogenic. Sample V-BM 1.4 demonstrated the best result in comparison with other samples within the group as well as in comparison with the control sample during the emulsion stability test. According to the potency test results, the emulsion vaccine based of the white oil sample BM 1.4 demonstrated the greatest difference in antibody titers in 14 days – 4.15 log2, while the one based on the control sample – 3.50 log2. This means that it the use of this vaccine results in 17.7-fold increase in FMD antibodies, while the use of control emulsion vaccine in 11.3-fold increase. Vaccines based on BM 1.3 and BM 2.2 oil samples demonstrated lower performance in comparison with the control vaccine in terms of these parameters.

Key words: white mineral oils, oil adjuvants, emulsion vaccines, hydrocatalytic processes.

REFERENCES

1. Ad"juvanty v sovremennoj vakcinologii / E. Ju. Isaenko, E. M. Babich, I. V. Eliseeva [i dr.] // Annaly Mechnikovskogo instituta. – 2013. – № 4. – S. 5–21.

2. Belye masla – proizvodstvo, trebovanija, tehnologii / S. V. Zagljadova [i dr.] // II Ezhegodnaja mezhdunar. konf. «Masla i topliva SNG», 27–29 maja 2014 g.

3. Bol'shoj medicinskij jenciklopedicheskij slovar' / pod red. V. I. Borodulina. – 4-e izd., ispr. i dop. – M.: Ripol Klassik, 2007. – 960 s.

4. Mamkov N. S. Masljanye ad"juvanty dlja protivojashhurnyh vakcin: dis. … d-ra vet. nauk. – Vladimir, 1999. – 255 s.

5. Medunicyn N. V. Vakcinologija. – 2-e izd., pererab. i dop. – M.: Triada-H, 2004. – 448 s.

6. Mihalishin V. V., Mamkov N. S. Ad"juvanty i ih ispol'zovanie // Tr. Federal'nogo centra ohrany zdorov'ja zhivotnyh: mater. Mezhdunar. nauch. konf. «Infekc. patologija zh-nyh», posvjashh. 50-letiju FGU «VNIIZZh». – 2008. – T. 6. – S. 340–371.

7. Sposob poluchenija malovjazkih belyh masel: pat. 2549898 Rossijskaja Federacija / E. V. Kashin, S. V. Zagljadova, T. N. Shabalina [i dr.]; OAO «Neftjanaja kompanija «Rosneft'». – № 2014105866/04; zajavl. 18.02.14; opubl. 10.05.2015.

8. FS.2.2.0004.15 Maslo vazelinovoe // Gosudarstvennaja farmakopeja Rossijskoj Federacii. – T. 3. – 13-e izd. – M.: FJeMB, 2015. – 1292 s.

9. Shabalina T. N. Razrabotka tehnologii proizvodstva neftjanyh malovjazkih masel s primeneniem gidrokataliticheskih processov: dis. … d-ra tehn. nauk. – Moskva, 1999. – 361 s.

10. European Pharmacopeia 8.0. Vol. 1–2 / Directorate for the Quality of Medicines & HealthCare of the Council of Europe (EDQM). – 8th ed. – Strasbourg, 2014.

11. FDA 21 CFR 172.878. Food additives permitted for direct addition to food for human consumption. White mineral oil. – URL: https://www.accessdata.fda.gov/SCRIPTs/cdrh/cfdocs/cfcfr/CFRSearch.cfm?fr=172.878.

12. FDA 21 CFR 178.3620. Indirect food additives: adjuvants, production aids and sanitizers. Mineral oil. – URL: https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfcfr/CFRSearch.cfm?FR=178.3620.

13. The United States Pharmacopeia 38 – National Formulary 33. – 38th еd. – 2015.

14. Vaccine Adjuvants: Preparation Methods and Research Protocols / ed. D. T. O’Hagan // Methods in Molecular Medicine / ed. J. M. Walker. – Totowa, NJ: Humana Press, Inc., 2000. – Vol. 42. – 342 р.

 

 

 

UDC 619:00058(470)

M. A. Shibayev, I. M. Klinovitskaya, A. S. Oganesyan, A. V. Belchikhina,

K. N. Gruzdev, A. K. Karaulov, N. V. Lebedev

ARRANGEMENT AND IMPLEMENTATION OF REGIONAL OFFICIAL VETERINARY SURVEILLANCE IN RUSSIAN FEDERATION IN 2016

P. 49–57

The paper is devoted to the arrangement of the official veterinary surveillance in the Russian Federation. Examples of veterinary surveillance arrangement in federal states are demonstrated. Summarized results of quantitative analysis of the scope of the regional official monitoring in 2016 are first published; special properties of the regional distribution of the control and surveillance measures are identified. Analysis of the coverage of the facilities subject to the regional official veterinary surveillance is performed, and following its results relatively low percent of the coverage of the controlled facilities is identified. Analysis of some parameters of veterinary surveillance performance including comparison of routine and random control and surveillance measures was performed. Staff and financial resources available for the RF veterinary surveillance authorities were quantified and analysis aimed at detection of interrelation between the resource volume and different factors and variables was made. It was concluded that for the purpose of adequate public protection the transition from total control to risk-oriented policy is needed and that risk-oriented policy should involve planning of checks depending on the animal health risk level. It is additionally urgent to pay closer attention to the development and revision of unified approaches to determination of inspectors’ staff size and improvement of methodical approaches to the calculation of financial expenses on surveillance implementation. Results of the analytical investigation performed can be used for the development of optimal approaches to addressing practical tasks and for total improvement of veterinary surveillance in the Russian Federation.

Key words: official veterinary surveillance, surveillance measures, veterinary service, controlled facilities, provision of surveillance authorities with resources, checks.

REFERENCES

1. Administrativnyj reglament komiteta gosudarstvennogo veterinarnogo nadzora Nizhegorodskoj oblasti po ispolneniju gosudarstvennoj funkcii «Osushhestvlenie regional'nogo gosudarstvennogo veterinarnogo nadzora». – URL: http://vetnadzor.government-nnov.ru/?id=13837 (data obrashhenija: 17.02.17).

2. Dobroljubova E. I., Juzhakov V. N. Ocenka rezul'tativnosti i jeffektivnosti kontrol'no-nadzornoj dejatel'nosti // Vopr. gos. i municip. upravlenija. – 2015. – № 4. – S. 41–64.

3. Doklad ob osushhestvlenii Rossel'hoznadzorom gosudarstvennogo kontrolja (nadzora), municipal'nogo kontrolja za 2016 god. – URL: http://gasu.gov.ru/infopanel?id=11824 (data obrashhenija: 02.03.17).

4. Kontrol'no-nadzornaja dejatel'nost' v Rossijskoj Federacii: analit. dokl. – 2014. – M.: MAKS Press, 2015. – 120 s.

5. Kontrol'no-nadzornaja i razreshitel'naja dejatel'nost' v Rossijskoj Federacii: analit. dokl. – 2016. – M.: Izd. dom RSPP, 2017. – 141 s.

6. Modernizacija sistemy kontrol'no-nadzornyh polnomochij v Rossijskoj Federacii: analit. dokl. / koll. avt.; Nac. issled. un-t «Vysshaja shkola jekonomiki». – M.: Izd. dom Vysshej shkoly jekonomiki, 2014. – 89 s.

7. O veterinarii: Zakon RF ot 14.05.1993 № 4979-1 (red. ot 03.07.2016). – M.: Konsul'tant Pljus, 2016. – 19 s.

8. O zashhite prav juridicheskih lic i individual'nyh predprinimatelej pri osushhestvlenii gosudarstvennogo kontrolja (nadzora) i municipal'nogo kontrolja: Federal'nyj zakon ot 26.12.2008 № 294-FZ (red. ot 01.05.2017). – M.: Konsul'tant Pljus, 2017. – 41 s.

9. O primenenii risk-orientirovannogo podhoda pri organizacii otdel'nyh vidov gosudarstvennogo kontrolja (nadzora) i vnesenii izmenenij v nekotorye akty Pravitel'stva Rossijskoj Federacii: utv. postanovleniem Pravitel'stva RF ot 17.08.2016 № 806. – M.: Konsul'tant Pljus, 2016. – 23 s.

10. Ob utverzhdenii administrativnogo reglamenta ispolnenija upravleniem veterinarii Altajskogo kraja gosudarstvennoj funkcii po osushhestvleniju regional'nogo gosudarstvennogo veterinarnogo nadzora: prikaz upravlenija veterinarii Altajskogo kraja ot 27.03.2012 № 67-p. – URL: http://base.garant.ru/7341300 (data obrashhenija: 19.02.17).

11. Ob utverzhdenii statisticheskogo instrumentarija dlja organizacii Minjekonomrazvitija Rossii federal'nogo statisticheskogo nabljudenija za osushhestvleniem gosudarstvennogo kontrolja (nadzora) i municipal'nogo kontrolja: prikaz Rosstata ot 21.12.2011 № 503. – M.: Konsul'tant Pljus, 2016. – 20 s.

12. Osnovnye napravlenija razrabotki i vnedrenija sistemy ocenki rezul'tativnosti i jeffektivnosti kontrol'no-nadzornoj dejatel'nosti: utv. rasporjazheniem Pravitel'stva RF ot 17.05.2016 № 934-r. – M.: Konsul'tant Pljus, 2017. – 58 s.

13. Plan meroprijatij («dorozhnaja karta») po sovershenstvovaniju kontrol'no-nadzornoj dejatel'nosti v Rossijskoj Federacii na 2016–2017 gody: utv. rasporjazheniem Pravitel'stva RF ot 01.04.2016 № 559-r. – M.: Konsul'tant Pljus, 2016. – 32 s.

14. Polozhenie o gosudarstvennom veterinarnom nadzore: utv. postanovleniem Pravitel'stva RF ot 05.06.2013 № 476. – M.: Konsul'tant Pljus, 2017.

15. Polozhenie o regional'nom gosudarstvennom veterinarnom nadzore v Krasnodarskom krae: utv. postanovleniem glavy administracii (gubernatora) Krasnodarskogo kraja ot 15.07.2014 № 701. – URL: http://docs.cntd.ru/document/444741428 (data obrashhenija: 19.02.17).

16. Porjadok organizacii i osushhestvlenija regional'nogo gosudarstvennogo veterinarnogo nadzora v gorode Moskve i Administrativnogo reglamenta ispolnenija Komitetom veterinarii goroda Moskvy gosudarstvennoj funkcii po osushhestvleniju regional'nogo gosudarstvennogo veterinarnogo nadzora: utv. postanovleniem Pravitel'stva Moskvy ot 23.09.2014 № 550-PP. – URL: http://www.garant.ru/products/ipo/prime/doc/70648282 (data obrashhenija: 19.02.17).

17. Porjadok organizacii i osushhestvlenija regional'nogo gosudarstvennogo veterinarnogo nadzora na territorii Respubliki Burjatija: utv. postanovleniem Pravitel'stva Respubliki Burjatija ot 12.01.2012 № 3. – URL: http://docs.cntd.ru/document/473809890 (data obrashhenija: 20.02.17).

18. Pravila podgotovki dokladov ob osushhestvlenii gosudarstvennogo kontrolja (nadzora), municipal'nogo kontrolja v sootvetstvujushhih sferah dejatel'nosti i ob jeffektivnosti takogo kontrolja (nadzora): utv. postanovleniem Pravitel'stva RF ot 05.04.2010 № 215. – M.: Konsul'tant Pljus, 2012. – 11 s.

19. Rekomendacii po organizacii i osushhestvleniju regional'nogo gosudarstvennogo veterinarnogo nadzora na territorii sub''ektov Rossijskoj Federacii: instruktivno-metod. izdanie / S. G. Dresvjannikova [i dr.]. – M.: FGBNU «Rosinformagroteh», 2015. – 64 s.

 

 

UDC 619:576.35:57.082.26

M. A. Shevchenko, M. N. Guseva, D. V. Mikhalishin, B. L. Manin, А. А. Shishkova

STUDIES OF THE BHK-21/2-17 LINEAGE USING FLOW CYTOMETRY

P. 58–62

The paper presents results of the ВНК-21/2-17 lineage cell cycle tests using flow cytometry. This cell line is widely used in biological industry that is why studying the ВНК-21/2-17 cell suspension pattern using the proposed technique and determination of tools and methods to prevent morphological and structural cell modifications in the culture affected by negative factors is of great interest. Flow cytometry allows to determine quantitative parameters of cell cycle phases and perform correlation between these phases and viability of the working cell population. It was mentioned that the general trend of this cell lineage suspension cultivation for 48 hours has a standard dynamics, herewith at all stages of the cell cycle the primary growth stage – G1 prevailed. Under unfavorable conditions of cultivation apoptotic debris could increase up to 31–59%, i. e. part of the cell population could be potentially non-reproductive. Unsuccessful cultivation stages associated with low pH content in the cultivator where pH was not maintained at the constant level were detected. During optimization of cell cultivation physical and chemical parameters proliferation intensity increased. Viable cell yield during cultivation when a specific pH was maintained was higher and at the synthetic stage G2 + M no cycle delay occurred.

Key words: flow cytometry, apoptosis, cell cycle phases.

REFERENCES

1. Vojtkova V. V. Izuchenie apoptoza metodom protochnoj citofluorimetrii (obzor literatury) // Bjulleten' VSNC SO RAMN. – 2010. – № 6 (76), ch. 1. – S. 220–225.

2. Zhivotnaja kletka v kul'ture (metody i primenenie v biotehnologii) / pod red. L. P. D'jakonova. – M.: Sputnik+, 2009. – 656 s.

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UDC 619:616.98:578.823.91:616-078:616-097

О. P. Bjadovskaya, Ye. А. Bukhon, А. А. Nesterov

DEVELOPMENT AND USE OF THE COMPETITIVE ELISA-BASED TEST SYSTEM FOR DETECTION OF ANTIBODIES TO GROUP A ROTAVIRUS IN LIVESTOCK SERA

P. 63–66

Test-system based on competitive enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to Group A rotavirus in sera collected from livestock of various species (cattle, pigs) was developed. Bovine rotavirus TE87 strain of serogroup A adapted to continuous porcine embryo kidney cell culture was selected as an antigen for ELISA. The said strain has the infectivity titre of 7.0–7.5 lg TCID50/cm3. It is group-specific and the most technologically suitable for production antigen scaling-up. Specificity of prepared components, rotavirus antigen and hyperimmune rabbit serum, was confirmed by real-time PCR and immunoblotting. The development of competitive ELISA-based test-system included selection of optimal blocking buffer composition as well as optimal antigen dilution, specific hyperimmune serum and anti-species conjugate. Admissible optical density values of control sera and cut-off values were determined. The test-system was examined for its specificity and sensitivity as compared to those ones of INGENASA test-kit (Spain) by parallel testing of 203 cattle sera. Performed studies resulted in the development of sensitive (92%) and specific (94%) competitive ELISA enabling detection of antibodies against Group A rotavirus in cattle and porcine sera with 92.6% accuracy. A total of 2,259 cattle and porcine sera submitted from 48 farms located in various Russian Federation regions in 2016–2017 were tested with the developed test-system. Average proportion of Group A rotavirus-positive cattle and pigs was 42% and 84%, respectively.

Key words: rotavirus, cell culture, enzyme-linked immunosorbent assay, sera, livestock, pigs.

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