Publication: Tavuk yumurtasından immunoglobulin y (ıgy) eldesi ve etkinliğinin gösterilmesi
Abstract
1. ÖZET Dünya Sağlık Örgütü tarafından antimikrobiyel direnç mekanizmaları nedeniyle yeni nesil antibiyotiklerin hayvanlarda, bitkilerde ve akuakültürde kullanılmaması, mevcut antibiyotiklerin kullanımının sınırlandırılması önerilmektedir. Antibiyotiklerin amacı dışında kullanımı; gıda zincirinde antibiyotik rezidü sorunu oluşturmakta, duyarlı bireylerde sağlık problemlerini artırmakta ve kamunun kıt kaynaklarının tüketilmesine neden olmaktadır. Pasif immunoterapi yoluyla antimikrobiyel kalıntı ve direnç gelişim mekanizmalarının bertarafı sayesinde özellikle riskli dönemlerde antibiyoterapiye alternatif olarak kullanılabilecek bir mücadele yöntemidir. Araştırmamızda Salmonella-free olarak temin edilen Ross 308 damızlık tavuklar kullanıldı. Salmonella gallinarum, Salmonella pullorum, Salmonella gallinarum + Salmonella pullorum şeklinde Freund’un Adjuvanı kullanılarak aşı hazırlandı ve 21 gün arayla 2 aşı uygulaması yapıldı. Aşılanan tavuklardan yumurtalar günlük olarak toplandı ve +4°C’de muhafaza edildi. Amonyum sülfat çöktürme ve diyaliz ile IgY saflaştırıldı. Toplam protein miktarı Bradford yöntemi ile tayin edildi. Elde edilen proteinler SDS-PAGE ile tanımlandı. Daha sonra yapılan in vitro etkinlik denemelerinde tüm fraksiyonların mikrobiyel büyümeyi inhibe ettiği görüldü. Gruplardan özellikle Salmonella gallinarum + Salmonella pullorum antikorlarının etkinliğinin yüksek olduğu gözlendi. Araştırmamız sonucunda, yumurta sarısı antikorlarının kısa süre içerisinde elde edilebileceği, Salmonella dışında da etken spesifik antikor üretilebileceği ve mikrobiyel büyümeyi inhibe edici yönleriyle antibiyotiklere alternatif olarak kullanılabileceği bir yöntem ve süreç geliştirildi. Antikor üretimi, Bakteriyel Büyüme İnhibisyonu, Immunoglobulin Y, Immunoterapi, Salmonella. 2.
Isolation of Immunoglobulin Y from Hen Egg and Determination of Its Efficiency The World Health Organization recommends not to use the new age antibiotics on animals, plants and aquaculture because of their antimicrobial resistance mechanisms. It further recommends to restrict the use of existing antibiotics. The misuse of antibiotics causes a problem of antibiotic residual in the food chain. It also causes an increase in health problems of individuals, a decrease of effectiveness and reduces low public budgetary. Passive immunotherapy that counteracts pathogens helps destroying antimicrobial residual and resistance mechanisms. It can be used even in the presence of infection so it is a good alternative for antibiotic therapy, especially in hazardous periods. In our research a Salmonella-free Ross 308 parent stock was used. Immunization was achieved using Salmonella gallinarum, Salmonella pullorum, Salmonella gallinarum+Salmonella pullorum and Freund's Adjuvant. Two doses of immunization was given for 21 days. Eggs were collected daily and saved. The amount of total protein was measured by Bradford method. Ammonium sulfate precipitation and dialysis methods were used for IgY purification.Proteins were identified by SDS-PAGE. Subsequently in vitro activity of all fractions in inhibiting microbial growth was determined. Especially Salmonella gallinarum + Salmonella pullorum antibodies were observed to have high activity. In conclusion, we have developed a method and process whereby egg yolk antibodies can be obtained in a short time and can be produced as specific antibodies against other antigens than Salmonella. Key Words: Antibody Generation, Bacterial Growth Inhibition, Immunoglobulin Y, Immunotherapy, Salmonella.
Isolation of Immunoglobulin Y from Hen Egg and Determination of Its Efficiency The World Health Organization recommends not to use the new age antibiotics on animals, plants and aquaculture because of their antimicrobial resistance mechanisms. It further recommends to restrict the use of existing antibiotics. The misuse of antibiotics causes a problem of antibiotic residual in the food chain. It also causes an increase in health problems of individuals, a decrease of effectiveness and reduces low public budgetary. Passive immunotherapy that counteracts pathogens helps destroying antimicrobial residual and resistance mechanisms. It can be used even in the presence of infection so it is a good alternative for antibiotic therapy, especially in hazardous periods. In our research a Salmonella-free Ross 308 parent stock was used. Immunization was achieved using Salmonella gallinarum, Salmonella pullorum, Salmonella gallinarum+Salmonella pullorum and Freund's Adjuvant. Two doses of immunization was given for 21 days. Eggs were collected daily and saved. The amount of total protein was measured by Bradford method. Ammonium sulfate precipitation and dialysis methods were used for IgY purification.Proteins were identified by SDS-PAGE. Subsequently in vitro activity of all fractions in inhibiting microbial growth was determined. Especially Salmonella gallinarum + Salmonella pullorum antibodies were observed to have high activity. In conclusion, we have developed a method and process whereby egg yolk antibodies can be obtained in a short time and can be produced as specific antibodies against other antigens than Salmonella. Key Words: Antibody Generation, Bacterial Growth Inhibition, Immunoglobulin Y, Immunotherapy, Salmonella.