Determination of Anti-phage Antibodies in Calf Sera Following Application of Escherichia Coli and Mannheimia Haemolytica-specific Bacteriophages

应用大肠杆菌和溶血曼氏菌特异性噬菌体后小牛血清中抗噬菌体抗体的测定

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作者:Renata Urban-Chmiel, Anna Nowaczek, Paulina Chmiel, Jacek Wawrzykowski, Ewelina Pyzik, Agnieszka Marek, Dagmara Stępien-Pyśniak, Mohammed M M Alomari, Marta Dec

Conclusion

Despite the induction of an anti-phage response, no significant negative effect on the antibacterial activity of phages was observed in vitro.

Material and methods

The study was conducted using phage components of experimental preparations and sera from calves treated and not treated with phages. Levels of G, M and A immunoglobulins were analysed by ELISA. The assay plates were coated with whole Escherichia coli and Mannheimia haemolytica phages and selected phage proteins obtained in sodium dodecyl sulphate-polyacrylamide gel electrophoresis and two-dimensional electrophoresis. Neutralisation of phages by immunoglobulins was assessed by determining phage titres using double-layer plates.

Methods

The study was conducted using phage components of experimental preparations and sera from calves treated and not treated with phages. Levels of G, M and A immunoglobulins were analysed by ELISA. The assay plates were coated with whole Escherichia coli and Mannheimia haemolytica phages and selected phage proteins obtained in sodium dodecyl sulphate-polyacrylamide gel electrophoresis and two-dimensional electrophoresis. Neutralisation of phages by immunoglobulins was assessed by determining phage titres using double-layer plates.

Results

The results confirmed an increased anti-phage response affecting all immunoglobulin classes in the calf sera. The highest significant (P ≤ 0.05) level of antibodies was observed for IgG in the sera of calves receiving phages. The phage neutralisation test showed a significant differences (P ≤ 0.05) in the reduction of phage titres in comparison to untreated calves.

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