Madeeha TARIQ, Aft ab Ahmad ANJUM, Ali Ahmad SHEIKH, Ali Raza AWAN, Muhammad Asad ALİ, Mian Muhammad Khubaib SATTAR, Shahid HUSSAIN, Tehreem ALI

Preparation and Evaluation of Alum Precipitate and Oil Adjuvant Multivalent Vaccines Against Clostridium perfringens

Enterotoxaemia is one of the hazardous diseases of the livestock. In Pakistan prophylaxis failure is due to the vaccination with type D monovalentvaccine. There is a need to develop a cost eff ective multivalent vaccine against enterotoxaemia using characterized toxinotypes isolated fromfield. Indigenously (Punjab, Pakistan) characterized Clostridium perfringens toxinotypes A (MW551947.1), B (MW332247.1) and D (MW332258.1)(n=1 each) were used. These toxinotypes were used to produce higher amount of alpha, beta and epsilon toxin units under culture conditions.Colony forming units (CFU) of each bacterium was determined through the standard plate count method and 106CFU/mL bacteria were usedfor vaccine dose. Monovalent, bivalent and multivalent oil adjuvant and alum precipitate vaccines were prepared. Formulated vaccines werepassed the stability, sterility and safety test. Bacterin plus toxoid oil adjuvant vaccine produced higher (868.25±3.54 IU/mL) antibody titer at 28thday post vaccination in rabbits and 100% protection was observed after challenge. Multivalent bacterin plus toxoid oil adjuvant vaccine wasused in field trials. Increased antibody response was detected after 4 months in sheep (1294.81±1.90 IU/mL) and goats (1091.85±2.51 IU/mL).During the experimental and field trials commercial vaccine did not produced higher antibody titer. Multivalent bacterin plus toxoid oil adjuvantvaccine proved as an excellent candidate for vaccination of animals against C. perfringens diseases, and it produced specific and efficient immuneresponse to be used in field.

Clostridium perfringens’e Karşı Alum Presipite ve Yağ Adjuvanlı Multivalan Aşıların Hazırlanması ve Değerlendirilmesi

Enterotoksemi, çiftlik hayvanları için tehlikeli hastalıklardan birisidir. Pakistan’daki profilaksinin başarısızlığı tip D monovalan aşıdan kaynaklanmaktadır. Sahadan izole edilen ve karakterize edilmiş toksinotipler kullanılarak enterotoksemiye karşı uygun maliyetli bir multivalan aşı geliştirmeye ihtiyaç vardır. Yöreye özgü (Punjab, Pakistan) karakterize edilmiş Clostridium perfringens toksinotipleri, A (MW551947.1), B (MW332247.1) ve D (MW332258.1) (n=1 her biri için) kullanıldı. Bu toksinotipler, kültür ortamında yüksek miktarda alfa, beta ve epsilon toksinlerinin üretiminde kullanıldı. Her bakterinin koloni oluşturan birimleri (KOB) standart plak sayım yöntemiyle belirlendi ve aşı dozu olarak 106 CFU/mL kullanıldı. Yağ adjuvanlı ve alum presipite monovalan, bivalan ve multivalan aşılar hazırlandı. Formüle edilen aşılar stabilite, sterilite ve güvenlik testlerinden geçirildi. Bakterin + toksoid yağ adjuvanlı aşı, tavşanlarda aşılamadan sonraki 28. günde yüksek (868.25±3.54 IU/mL) antikor titresine yol açtı ve eprüvasyon sonrası %100 koruma gözlendi. Saha çalışmalarında multivalan bakterin + toksoid yağ adjuvanlı aşı kullanıldı. Aşılamadan 4 ay sonra koyun (1294.81±1.90 IU/mL) ve keçilerde (1091.85±2.51 IU/mL) antikor yanıtında artış saptandı. Deneysel ve saha çalışmaları sırasında ticari aşının daha yüksek antikor titresi üretmediği gözlendi. Hayvanların C. perfringens enfeksiyonlarına karşı aşılanmasında multivalan bakterin + toksoid yağ adjuvanlı aşının mükemmel bir aday olduğu kanıtlandı ve bu aşı sahada kullanılmak üzere spesifik ve etkili bir bağışıklık yanıtı üretti.

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