DETERMINATION OF GROWTH CHARACTERISTICS OF THE FOOT AND MOUTH VIRUSES (A, O, ASIA-1) IN BHK-21 AN30 AND BHK-21 AN73 CELL CULTURES

Year 2017, Volume: 2 Issue: 1, 1 - 5, 30.04.2017

Neslihan Taşçene Banu Bayri Özbilge Sadık Onur Karaçam Veli Gülyaz Mustafa Hasöksüz

Abstract

Bu çalışmada, BHK-21 An₇₃ hücre kültürü ile
BHK-21 An₃₀ hücre kültürlerinin üreme hızı, miktarı ve virus titresi
üzerine etkilerinin araştırılması amaçlandı. Bu amaçla BHK-21 An₃₀ ve
BHK-21 An₇₃ süspanse hücre kültürlerinde, hücrelerin üreme
kinetikleri belirlendi. Bu hücre kültürlerinde A TUR 11, O TUR 07 ve Asia-1/11
şap aşı suşlarının üretimi gerçekleştirildi. BHK-21 An₃₀ hücre
kültürlerinin 7 günlük inkubasyon periyodu esnasında monolayer formda 4. günde
1,3x10⁶/ml hücre sayısıyla, süspanse formda ise 4. günde 4,1x10⁵
hücre sayısı ile pik noktaya ulaşırken BHK-21 An₇₃  kültürlerinde ise monolayer formda 6. günde
2,2x10⁶/ml hücre sayısına ulaşılırken, süspanse formda 3. günde 2,8
x10⁵ hücre sayısına ulaştığı belirlendi.  Süspanse BHK-21 An₃₀ hücre
kültürlerinde A, O ve Asia-1 aşı virusunun üretimi sonrası 146S değerleri
sırasıyla ortalama; 0,51 μg/ml, 0,18 μg/ml ve 0,16 μg/ml olarak saptanırken
enfektif titreleri ortalama 6,87 pfu/ml, 6,22 pfu/ml ve 6,49 pfu/ml olarak
belirlendi.

Süspanse BHK-21 An₇₃ hücre kültürlerinde A,
O ve Asia-1 aşı virusunun üretimi sonrası değerleri sırasıyla ortalama; 2,11
μg/ml, 2,59 μg/ml ve 0,53 μg/ml olarak saptanırken enfektif titreleri ortalama
6,99 pfu/ml, 7,82 pfu/ml ve 6,37 pfu/ml olarak tespit edildi.

Sonuç olarak, aşı üretiminde kullanılan A, O ve Asia-1
aşı viruslarının BHK-21 An₇₃ hücre kültüründe daha yüksek 146S ve
enfektif titrede üreme gösterdiği ve bundan dolayı şap aşısı üretiminde BHK-21
An₇₃ hücre kültürlerinin kullanılmasının gerek üretim zamanı gerekse
maliyet olarak BHK-21 An₃₀ hücre kültürlerinden daha uygun olduğu
kanaatine varılmıştır. 

Keywords

Şap Hastalığı, Şap Virusu suşları, BHK-21 hücre kültürü

DETERMINATION OF GROWTH CHARACTERISTICS OF THE FOOT AND MOUTH VIRUSES (A, O, ASIA-1) IN BHK-21 AN30 AND BHK-21 AN73 CELL CULTURES

Abstract

In
this study, it was aimed to investigate BHK-21An₇₃ and BHK-21An₃₀
growth rate of cell cultures and the effects on foot and mouth disease (FMD)
vaccine virus strains titers. For this purpose, growth rate of suspended cell
cultures of BHK-21An₇₃ and BHK-21An₃₀ were determined. A
TUR 11, O TUR 07 and Asia-1/11 strains of foot and mouth disease (FMD) vaccine
virus strains were produced separately on BHK-21An₃₀ and BHK-21An_73.
BHK-21An₃₀ cell amounts during seven incubation days of
reached to peak level in monolayer form at the 4th day with 1,3x10⁶/ml
cell numbers, in suspension form at the 4th day with 4,1x10⁵ cell
numbers. BHK-21An₇₃ reached to peak level in monolayer form at the
6th day 2,2x10⁶/ml cell numbers while in suspension form at the 3rd
day with 2,8 x10⁵ cell numbers. After the production of A, O and
Asia-1 vaccine viruses, in suspension BHK-21An₃₀ cell cultures
average of 146S values respectively; 0,51 μg/ml, 0,18 μg/ml and 0,16 μg/ml were
detected while infective titers were determined as average 6,87 (Plaque Forming
Units) pfu/ml, 6,22 pfu/ml and 6,49 pfu/ml. After the production of A, O and
Asia-1 vaccine viruses, in suspended BHK-21An₇₃ cell cultures
average of 146S values respectively; 2,11 μg/ml, 2,59 μg/ml and 0,53 μg/ml were
detected while infective titers were determined as average 6,99 pfu/ml, 7,82
pfu/ml and 6,37 pfu/ml. As a result, in the production of A, O and Asia-1
vaccine viruses in BHK-21An_73, resulted in higher 146S values and
higher infective titers than BHK-21An_30. It is concluded that for
FMD vaccine production the using of BHK-21An₇₃ cell culture is more
appropriate for both of cost of manufacture and productive time.

Keywords

Foot and Mouth Disease, FMDV vaccine strains, BHK-21 cell culture

References

• 1. Abbas, F., Khan, F, A., Ahmad, F., Hussain, A., Ahmad, M., Awan, M.A., Tariq, M.,M., Kakar, M. A., Wadood, A., Ali, M. (2011). Production of Foot and Mouth Disease Virus Vaccine (O Type) on BHK-21 Cell Line, Iğdır Univ. J. Ins. Sci. Tech. 1(2), 155-159.
• 2. Ali, S.,M., Ismail, A.,H., Soliman, E.,M., Hanaa, A., M. (2013). Studies on Growth Kinetics of the FMDV Serotype SAT-2 Egyptian Strain in Cell Culture, J.Vet.Adv. 3(2), 92-97
• 3. Alexandersen, S., Zhang, Z., Donaldson, A., I., Garland, A., J., M. (2003). The Pathogenesis and Diagnosis of Foot and Mouth Disease. J.Comp Path. 129, 1-36.
• 4. Bartelling, S., J., Vreeswijk, J. (1991). Developments in foot- and-mouth disease vaccines. Central Veterinary Institute, Lelystad, The Netherlands. Vaccine. Feb, 9(2),5-88.
• 5. Bartelling, S., J. (2002). Development and performance of inactivated vaccines against foot and mouth disease. Rev. Sci. Tech. Off.int. Epiz, 21, 577-588.
• 6. Grubman, M.,J., Baxt, B. (2004). Foot-and-Mouth Disease. Clin. Microbiol. Rev., 17 (2), 465–493
• 7. Harmsen, M.,M., Fitjen, H.,P.,D., Westra, D.,F., Coco-martin, J., M. (2011). Effect of thiomersal on dissociation of intact (146S) foot-and-mouth disease virions into 12S particles as assessed by novel ELISAs specific for either 146S or 12S particles. Vaccine, 29, 2682-2690.
• 8. Institute of Foot and Mouth Disease Protocol, 2010.
• 9. Radwan MEI, Khalifa NO, Fahmy HA (2016) Molecular epidemiology of foot and mouth disease virus during 2014 with references to biochemical changes in Egyptian buffaloes . World Journal of Biology and Medical Sciences. 3(1)68-81.
• 10. Rahman, S., U., Rabbani, M., Sahidullah, K., Muhammed, Z. (2007). Studies on In Vitro Culture Characteristics of Adherent Baby Hamster Kidney-21(BHK-21) Cell Line. Int. J. of Agri-Biol. 1560-8530, 821-826.
• 11. Rweyeamu, M.,M., Umehara, O., Giorci, W., Medeiros, R., Lucca, D.,N., Balzatar, M. (1989), Effect of formaldehyde and binary ethyleneimine (BEI) on the Integrity of Foot and Mouth Disease Virus Capsid. Rev. Sci. Tech. Off. Int. Epiz., 8(3), 747-764.
• 12.Shirai, J., Chatchawanchonteera, A., Sinsuwongwat, W., Makarasen, P., Sugimura,T. (1990), Estimation of 146S Paticles in Foot and Mouth Disease Virus (FMDV) Vaccine by Using Computer Analysing System. Jpn.J.Vet., Sci.52(3):621-630.