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Molecular and genetic detection of infectious laryngeotrachitis disease virus in broiler farms after a disease outbreak in Egypt

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Abstract

Infectious laryngotracheitis (ILT) is a viral respiratory illness in poultry that causes massive financial losses. This research aimed to isolate and identify the ILT virus in suspected outbreaks of broiler flocks in Egypt during 2020–2021, besides investigating its genetic link with other circulating strains. Real-time-PCR was used to test 57 samples taken from unvaccinated broiler farms. Ten samples are positive for ILTV, and the virus is being isolated in SPF chicken embryos. The Sanger sequencing was used to conduct (partial) sequencing of the infected cell protein4 gene (ICP4) for eight isolates. Phylogenetic analysis conducted Maximum Likelihood, comparative sequencing analysis of ICP4 of strains under study with vaccination ILT reference strains reveled that all isolates were clustered into two major groups. The (OM291843and OM291846) clustered together with the chicken embryo origin vaccine strains (IV and V group). The remaining six strains belong to the TCO vaccine(I, II and III group). The total sequence similarity between the strains under study and the various Egyptian strains varied from (97 to 100%) while the similarity with TCO or chicken embryo origin -vaccine strains ranged from (95to 100%). There were no deletions detected in the 272–283-bp region of the ICP4 gene. Detection of arginine to methionine substitutions at position 180 (R180M) and change of Serine to Asparagine at position 227 (S227N) in the (OM291843 and OM291846) which were previously described in chicken embryo origin -vaccine strains. This reveals that field strains may have evolved from vaccine strains, notably identification of non-synonymous substitutions which might be linked to the virulence strains’ attenuation. Finally, independent of geographical distribution, both chicken embryo origin-vaccine-like and TCO-Vaccine-like virus strains were circulating in Egyptian non-vaccinated broiler flocks in 2020 and 2021. Despite their genetic differences, both viruses caused significant illnesses in the field.

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Data Availability

The data presented in this study are available on request from the corresponding authors.

Abbreviations

ILTV:

Infectious laryngotracheitis

Real-Time-PCR:

Real-time-Polymeras Chain Reaction

TCO:

tissue culture origin

CEO:

chicken embryo origin

S:

Serine

N:

Asparagine

R:

arginine

M:

methionine

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Acknowledgements

The authors extend a cordial thanks to all the farms owners, and veterinarians who help in this study.

Funding

This research received no external funding. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

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Authors and Affiliations

  1. Reference Laboratory for Veterinary Quality Control on Poultry Production, Animal Health Research Institute, Agriculture Research Center (ARC), 12618, Giza, Egypt

    Zienab mossad, Saad A. Moussa, M. Saied, Mustafa M. Fathy & Ali Mahmoud Zanaty

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  1. Zienab mossad
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  2. Saad A. Moussa
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  3. M. Saied
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  4. Mustafa M. Fathy
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Contributions

Conceptualization, Zeinab mossad and Ali Mahmoud Zanaty methodology, Moussa,M. Saied, Mustafa M. Fathy.; software, Zeinabmossad.; validation Ali Mahmoud Zanaty.; analysis, Zeinabmossad investigation, Ali Mahmoud Zanaty and J Moussa,M. Saied resources Mustafa M. Fathy data curation, Mustafa M. Fathy, writing—original draft preparation Zeinab mossad and Ali Mahmoud Zanaty writing review and editing, Moussa,M. Saied, Mustafa M. Fathy. funding acquisition, All authors read and agreed to the published version of the manuscript.

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Correspondence to Zienab mossad.

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mossad, Z., Moussa, S.A., Saied, M. et al. Molecular and genetic detection of infectious laryngeotrachitis disease virus in broiler farms after a disease outbreak in Egypt. VirusDis. 33, 404–412 (2022). https://doi.org/10.1007/s13337-022-00792-w

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