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Bacteriophage mediated control of necrotic enteritis caused by C. perfringens in broiler chickens.

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Abstract

In Egypt, little attention has been paid to the isolation and application of C. perfringens phages for treating necrotic enteritis at the farm level. This study aims to evaluate the efficiency of the podovirus C. perfringens phage in treating necrotic enteritis in broiler chickens. Accordingly, C. perfringens phage was isolated from cecal samples of apparently healthy chickens and characterized by transmission electron microscopy, thermal stability test, and pH stability test. Commercial 14-day-old Arbor Acres broiler chickens were allocated to three groups: group Ӏ received BHI broth and assigned as a negative control, group П served as a positive control group that was challenged with C. perfringens via oral gavage for four successive days, and group Ш was administrated six phage doses on several occasions after oral gavage challenge with C. perfringens. Daily clinical symptoms and mortality were recorded. At three-time intervals, necrotic enteritis lesions were scored. Cecal samples were examined for re-isolation and counting of C. perfringens. The isolated C. perfringens phage was a podovirus with an icosahedral head diameter of 78.7 nm and a short non-contractile tail length of 22.2 nm. It remained stable for 60 min at 30 °C and 50 °C at pH values of 2, 4, 8, and 10. The phage-treated group (Ш) showed mild gross lesions with a lower mortality rate and reduced colony-forming units than the positive control group (П). The findings revealed that the isolated C. perfringens phage effectively treated experimental necrotic enteritis in broiler chickens.

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Acknowledgments

The authors would like to thank Dr. Ahmed Erfan for his help during the examination of the molecular toxinotypes of the selected C. perfringens isolate using conventional polymerase chain reaction.

Availability of data and material

The authors confirm that the data supporting the findings of this study are available within the article and its supplementary information.

Code availability

Non-applicable.

Funding

This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

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Authors and Affiliations

  1. Reference Laboratory for Veterinary Quality Control on Poultry Production, Animal Health Research Institute, Agriculture Research Center, Giza, 264, Egypt

    Reham A. Hosny, Ahmed F. Gaber & Hend K. Sorour

Authors
  1. Reham A. Hosny
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  2. Ahmed F. Gaber
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  3. Hend K. Sorour
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Contributions

Reham A. Hosny and Hend K. Sorour isolated C. perfringens and C. perfringens phage. Reham A. Hosny, Ahmed F. Gaber, and Hend K. Sorour designed and carried out the experiment, performed laboratory and pathological examination, interpreted the results. Reham A. Hosny performed all statistical analyses in the manuscript. Reham A. Hosny, Ahmed F. Gaber, and Hend K Sorour wrote and revised the manuscript. All authors read and approved the manuscript.

Corresponding author

Correspondence to Reham A. Hosny.

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Ethical approval

All animal handling procedures were done according to an approved protocol [AHRI (23) 27/3/2019] by the Ethical Committee of the Animal Health Research Institute, Ministry of Agriculture, Giza, Egypt.

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The authors declare that they have no competing interests.

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Supplementary Information

Fig 1

Toxinotyping of C. perfringens host strain using a polymerase chain reaction. Lane 1: Examined sample, Lane 2: C. perfringens positive control ATCC 12917, Lane 3: C. perfringens positive control ATCC 27324, Lane 4: C. perfringens positive control ATCC 3626, and lane 5: ladder 100–600 bp. (PNG 47 kb)

High resolution image (TIF 2642 kb)

Fig 2

The thermal stability of C. perfringens phage in different dilutions at different temperatures of 30 °C, 50 °C, 70 °C, and 90 °C following a 60-min incubation using the plaque assay test. The phage (109 PFU/ml) was stable to heat at 30 °C with a log titer of 9.9 PFU/ml, with only a slight reduction in log titer of 9.8 PFU/ml at 50 °C, beyond which they were undetectable. (PNG 24 kb)

High resolution image (TIF 5031 kb)

Fig 3

The pH stability of C. perfringens phage in different dilutions at different pH values of 2, 4, 8, and 10 using the plaque assay test. The phage (109 PFU/ml) retained viability across the different pH values in the plaque assay method with log titers of 9.7, 9.8, 9.9, and 9.7 PFU/ml at pH values of 2, 4, and 8, and 10, respectively. (PNG 54 kb)

High resolution image (TIF 6031 kb)

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Hosny, R.A., Gaber, A.F. & Sorour, H.K. Bacteriophage mediated control of necrotic enteritis caused by C. perfringens in broiler chickens.. Vet Res Commun 45, 409–421 (2021). https://doi.org/10.1007/s11259-021-09821-3

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