Attenue mavidil serotip-4 aşısının farklı stabilizatörler ile üretilmesi

Öz

Bu çalışma ile attenue mavidil serotip-4 aşısının halihazırda kullanılmakta olan embriyo ekstraktı ve yumurta sarısından oluşan stabilizatörünün yerine farklı stabilizatörlerin kullanılması ve bunların liyofilizasyon ve raf ömrüne etkileri araştırıldı. Bu amaçla Ankara Etlik Merkez Veteriner Kontrol ve Araştırma Enstitüsü Viral Aşı Üretim Laboratuarı bünyesinde bulunan ve Vero hücrelerinde üretilen SA BT/4 attenüe suşu kullanıldı. Hazırlanan ham aşı sırasıyla; Lactalbumin hydrolysate %5 -sucrose %10 (LH), Weybridge Medium (%2.5 lactalbümin hyrolysate , %5 sucrose ve %1 Sodium glutamate) (WBM), Lactalbumin hydrolysate %5 -Mannitol %10 (LM), Jelatin sorbitol (hidrolize jelatin %3.5- D-sorbitol %3.5) (JS), distile suda %5 oranında Trehalose hidrate (TD) ile hazırlanan değişik stabilizatörlerle karıştırılıp liyofilizasyona tabi tutuldu. Liyofilizasyon sonucunda ortaya çıkan final ürünün kalite kontrol testleri ve raf ömrü çalışmaları gerçekleştirildi. Bu testler sonucunda; liyofilizasyon sonucunda aşı virusunda meydan gelen infektif titredeki kayıp, denenen bütün stabilizatörler için kabul edilir düzeydeydi. Denemesi yapılan stabilizatörlerle hazırlanan aşılar zararsızdı. Raf ömrü çalışmalarında en iyi performansı Lactalbumin hydrolysate %5 -sucrose %10 (LH) ile hazırlanan aşı gösterdi.

Anahtar Kelimeler

• Aşı
• bağışıklık
• mavidil serotip 4
• raf ömrü
• stabilizatör
• Ahmet Burak GÜNGÖR
• Özden KABAKLI
• Elvin ÇALIŞKAN
• 2011
• Etlik Veteriner Mikrobiyoloji Dergisi

Production of attenuated live bluetongue serotype-4 vaccine with different stabilizers

Öz

In this study, effects of different stabilizers other than embryo extract and egg yolk used at present on lyophilisation and shelf life of attenuated lyophilized Bluetongue serotype-4 vaccine was investigated. For this purpose, SA BT/4 attenuated vaccine strain present at Etlik Central Veterinary Control and Research Institude Viral Vaccine Production Laboratory produced in Vero cell line is used. Bulk vaccine was mixed with 5% lactalbumin hydrolysate10% sucrose (LS), Weybridge Medium (2.5% lactalbümin hyrolysate , 5% sucrose and 1% Sodium glutamate) (WBM), 5% lactalbumin hydrolysate -10% Mannitol (LM), Gelatin sorbitole (3.5% hidrolised gelatine - 3.5% D-sorbitole) (JS) and 5% Trehalose hidrate (TD) in distilled water respectively as stabilizer. At the end of lyophilisation, quality control tests and shelf life studies of the vaccines were carried out. Results indicated that; the loss of infective titre of the vaccine virus after lyophilisation procedure was acceptable for all stabilizers, all the vaccines prepeared with mentioned stabilizers were harmless, the vaccine, prepeared with 5% lactalbumin hydrolysate-10% sucrose (LS) stabilizer had the best performance amongst in shelf life studies.

Anahtar Kelimeler

• Bluetongue serotype 4
• vaccine
• stabilizer
• immunity
• shelf life.

Kaynakça

1. Alexander RA, Haig DA, (1951). The use of egg attenuated bluetongue virus in the production of a polyvalent vaccine for sheep. Onderstepoort J Vet Res, 26, 3-15.
2. Anonim, Bluetongue vaccine, method of producing same and method of immunizing rumminants therewith. Erişim adresi: http://www.wipo.int/pctdb/en/wo.jsp?IA=US1983000052&DISPLAY=DESC, Erişim tarihi: 03.03.2011
3. Anonim, European Commision Health Consumer Protection Directorate General (Sanco/C3/AH/R19/2000) Possible use of vaccination against Bluetongue in Europe. Scientific Committee on animal health and animal welfare. Adopte 27 june 2000.
4. Bréard E, Sailleau C, Coupier H, Mure-Ravaud K, Hammoumi S, Gicquel B, Hamblin C, Dubourget P, Zientara S, (2003). Comparison of genome segments 2, 7 and 10 of bluetongue viruses serotype 2 for differentiation between field isolates and the vaccine strain. Vet Res. 34, 777-789.
5. Erasmus BJ, (1990). Bluetongue virus. Dinter Z, Morein B. eds. Virus Infections of Ruminants. Elsevier Science Publishers. Chapter 21, p. 227-237.
6. Ertürk A, Tatar N, Kabaklı O, İncoğlu S, Çizmeci SG, Barut FM, (2004). The current situation of bluetongue in Turkey. Vet Ital. 40 (3), 137-140.
7. Hammoumi S, Breard E, Sailleau C, Russo P, Grillet C, Cetre-Sossah C, Albina E, Sanchis R, Pepin M, Guibert J, Zientra S, (2003). Studies on the Safety and Immunogenicity of the South African BluetongueVirus Serotype 2 Monovalent Vaccine: Specific Detection of the Vaccine Strain Genome by RT-PCR. J Vet Med. 50, 316-21.
8. Kirkland PD, Hawkes RA, (2004). A comparisation of laboratory and wild strains of bluetongue virus- is there any difference and dose it matter? Vet Ital. 40 (4), 448-455.

9. Mann N, Mann S, Singh KP, Samuel AR, Mertens P, (2004). Development of reverse transcriptase-polymerase chain reaction-basedassays and sequencing for typing European strains of bluetongue virusand differential diagnosis of field and vaccine strains. Vet Ital. 40 (4), 552-561.
10. Mellor PS, Wittmann EJ, (2002). Bluetongue virus in the mediterranean basin 1998-2001. Vet J. 164(1), 20-37.
11. Mariner JC, House AE, Sollod C, Mebus CA, (1990). Comparison of the effect of various stabilizers and lyophilization cycleson the thermostability of a Vero Cell-adapted rinderpest vaccine. Vet Microbiol. 21(3), 195-209.
12. Monaco F, Camma C, Serini, Savini G, (2006). Differentiation between field and vaccine strain of bluetongue virus serotype 16. Vet Microb. 166, 45-52.
13. Murphy FA, Gibbs JEP, Horzineck CM, Studdent MJ, eds., (1999). Veterinary Virology. Third Edition. Raven Press Ltd. New York.
14. Murray PK, Eaton BT, (1996). Vaccines for bluetongue. Aust Vet J. 73(6), 207-210.
15. OIE, (2000). Manual of Standarts for Diagnostic Tests and Vaccines Bluetongue. Erişim adresi: http://www.oie.int.norms.MMANUAL/A_00026.htm. Erişim tarihi: 03.03.2011.
16. Ozawa Y, Bahrami S, (1968). Effects of Freezing on African Horse Sickness Virus. Arc Virol. 26(2), 201-210.
17. Rashid A, Asım M, Chaudhary H, (2008). Effect of various stabilizers on titre of lyophilized live-attenuated Peste Des Petits Rumminants (PPR) vaccine. Pakistan Vet J. 28(4), 203-204.
18. Sarkar J, Sareenivasa BP, Singh RP, Dhar P, Bandyopadhyay SK, (2003). Comparative efficiency of various chemical stabilizers on the thermostability of a live-attenuated Peste de Petits Rumminants (PPR) vaccine. 21, 4728-4735.
19. Sparkes JD, Fenje P, (1972). The effect of residual moisture in lyophilized smallpox vaccine on its stability at different temperatures. Bull World Health Organ. 46 (6), 729-734.
20. Wittmann E, Mellr P, Baylis M, (2001). Using climate data to map the potential distribution of Culicoides imicola (Diptera: Ceratopogonidae) in Europe. Rev Sci Tech. 20 (3): 731-740.
21. Yonguç AD, Taylor WP, Csontos L, Worrall E, (1982). Bluetongue in Western Turkey. Vet Rec Aug. 111, 144-146.