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Updated: Jun 10, 2026

Influenza Virus Propagation in Embryonated Chicken Eggs
Published on: March 19, 2015
Experimental avian paramyxovirus serotype-3 infection in chickens and turkeys
Sachin Kumar1, Flavia Militino Dias, Baibaswata Nayak
1Virginia-Maryland Regional College of Veterinary Medicine, University of Maryland, College Park, MD 20742, USA.
Abstract:
Avian paramyxoviruses (APMV) are divided into nine serotypes. Newcastle disease virus (APMV-1) is the most extensively characterized, while relatively little information is available for the other APMV serotypes. In the present study, we examined the pathogenicity of two divergent strains of APMV-3, Netherlands and Wisconsin, in (i) 9-day-old embryonated chicken eggs, (ii) 1-day-old specific pathogen free (SPF) chicks and turkeys, and (iii) 2-week-old SPF chickens and turkeys. The mean death time in 9-day-old embryonated chicken eggs was 112 h for APMV-3 strain Netherlands and > 168 h for strain Wisconsin. The intracerebral pathogenicity index in 1-day-old chicks for strain Netherlands was 0.39 and for strain Wisconsin was zero. Thus, both strains are lentogenic. Both the strains replicated well in brain tissue when inoculated intracerebrally in 1-day-old SPF chicks, but without causing death. Mild respiratory disease signs were observed in 1-day-old chickens and turkeys when inoculated through oculonasal route with either strain. There were no overt signs of illness in 2-weeks-old chickens and turkeys by either strain, although all the birds seroconverted after infection. The viruses were isolated predominantly from brain, lungs, spleens, trachea, pancreas and kidney. Immunohistochemistry studies also showed the presence of large amount of viral antigens in both epithelial and sub-epithelial lining of respiratory and alimentary tracts. Our result suggests systemic spread of APMV-3 even though the viral fusion glycoprotein does not contain the canonical furin proteases cleavage site. Furthermore, there was little or no disease despite systemic viral spread and abundant viral replication in all the tissues tested.
Insights
Avian paramyxoviruses (APMV-3) showed systemic spread in chickens and turkeys, despite causing mild or no disease. These lentogenic strains replicated widely in multiple tissues, indicating potential for avian health concerns.
Area of Science:
- Veterinary Virology
- Avian Pathology
- Molecular Virology
Background:
- Avian paramyxoviruses (APMV) encompass nine serotypes, with APMV-1 (Newcastle disease virus) being well-studied.
- Limited information exists on the pathogenicity of APMV serotypes beyond APMV-1, necessitating further research into their impact on avian health.
Purpose of the Study:
- To evaluate the pathogenicity of two distinct APMV-3 strains (Netherlands and Wisconsin) in various avian models.
- To investigate the replication and dissemination of APMV-3 within different avian tissues and age groups.
Main Methods:
- Inoculation of embryonated chicken eggs and specific pathogen-free (SPF) chicks and turkeys of different ages (1-day-old and 2-week-old).
- Assessment of pathogenicity through mean death time, intracerebral pathogenicity index (ICPI), and observation of clinical signs.
- Viral isolation and immunohistochemistry to determine tissue tropism and antigen distribution.
Main Results:
- Both APMV-3 strains were classified as lentogenic based on low ICPI values and prolonged mean death times in eggs.
- Mild respiratory signs were observed in 1-day-old birds, with no overt illness in 2-week-old birds, despite seroconversion.
- Systemic viral spread was confirmed by isolation from multiple organs (brain, lungs, spleen, etc.) and presence of viral antigens, even without a canonical furin cleavage site.
Conclusions:
- APMV-3 strains can spread systemically in chickens and turkeys with minimal or no apparent disease.
- The absence of a canonical furin cleavage site does not preclude systemic dissemination of APMV-3.
- Further investigation is warranted to understand the full implications of APMV-3 systemic spread in poultry.
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