Experimental pathology of two highly pathogenic H5N1 viruses isolated from crows in BALB/c mice.

Subbiah Kombiah1 Manoj Kumar2 Harshad Vinayakrao Murugkar3 Shanmugasundaram Nagarajan3 Chakradhar Tosh3 Dhanapal Senthil Kumar3 Katherukamem Rajukumar3 Siddharth Gautam1 Rajendra Singh1 Mathesh Karikalan1 Anil Kumar Sharma1 Vijendra Pal Singh3
Affiliations 3 institutions
  1. ICAR- Indian Veterinary and Research Institute, Izatnagar, Uttar Pradesh, India.
  2. ICAR- National Institute of High Security Animal Diseases, Bhopal, Madhya Pradesh, India. Electronic address: [email protected].
  3. ICAR- National Institute of High Security Animal Diseases, Bhopal, Madhya Pradesh, India.
Active Last checked 2026-10-06 03:30:27 Source: NCBI PubMed

Abstract

In this study, we assessed the pathogenicity of two H5N1 viruses isolated from crows in mice. Eighteen 6-8 weeks BALB/c mice each were intranasally inoculated with 106 EID50/ml of H5N1 viruses A/crow/India/03CA04/2015 (H9N2-PB2 reassortant H5N1) and A/crow/India/02CA01/2012 (Non-reassortant H5N1). The infected mice showed dullness, weight loss and ruffled fur coat. Histopathological examination of lungs showed severe congestion, haemorrhage, thrombus, fibrinous exudate in perivascular area, interstitial septal thickening, bronchiolitis and alveolitis leading to severe pneumonic changes and these lesions were less pronounced in reassortant virus infected mice. Viral replication was demonstrated in nasal mucosa, lungs, trachea and brain in both the groups. Brain, lung, nasal mucosa and trachea showed significantly higher viral RNA copies and presence of antigen in immunohistochemistry in both the groups. This study concludes that both the crow viruses caused morbidity and mortality in mice and the viruses were phenotypically highly virulent in mice. The H5N1 viruses isolated from synanthropes pose a serious public health concern and should be monitored continuously for their human spill-over.

Supporting text Virus Host Location
Avian influenza 58 BALB/c mice 2 Crow isolates 1 H9N2-PB2 reassortant H5N1 1 Animals 1981 Biopsy 2 Crows 4 Disease Susceptibility 21 Histocytochemistry 1 Influenza A Virus, H5N1 Subtype 308 Influenza in Birds 352 Mice 256 Mice, Inbred BALB C 73 Orthomyxoviridae Infections 235 Reassortant Viruses 109 RNA, Viral 194 Viral Load 36 Virus Replication 195

Evidence records

6 total
Zoonotic Surveillance
3 records · 2 evidence types
Evidence type
2 records
OVE3611
Key finding

The H5N1 virus isolate A/crow/India/03CA04/2015 was recovered from crow samples in India.

Virus
Host
Location
Not specified
Supporting text

two H5N1 viruses isolated from crows in mice

Sample type
crow samples
OVE3612
Key finding

The H5N1 virus isolate A/crow/India/02CA01/2012 was recovered from crow samples in India.

Virus
Host
Location
Not specified
Supporting text

two H5N1 viruses isolated from crows in mice

Sample type
crow samples
Evidence type
1 records
OVE3615
Key finding

H5N1 viral RNA and antigen were detected in the nasal mucosa, lungs, trachea, and brain of infected BALB/c mice.

Virus
Host
Location
Not specified
Supporting text

Viral replication was demonstrated in nasal mucosa, lungs, trachea and brain in both the groups. Brain, lung, nasal mucosa and trachea showed significantly higher viral RNA copies and presence of antigen in immunohistochemistry in both the groups.

Method
viral RNA quantification | immunohistochemistry
Sample type
nasal mucosa | lungs | trachea | brain
Experimental Infection
2 records · 1 evidence types
Evidence type
2 records
OVE3613
Key finding

Experimental infection of BALB/c mice with A/crow/India/03CA04/2015 (H9N2-PB2 reassortant H5N1) caused morbidity, mortality, and severe lung pathology, indicating high virulence.

Virus
Host
Location
Not specified
Supporting text

Eighteen 6-8 weeks BALB/c mice were intranasally inoculated with 10^6 EID50/ml of H5N1 virus A/crow/India/03CA04/2015. The infected mice showed dullness, weight loss and ruffled fur coat. Histopathological examination of lungs showed severe congestion, haemorrhage, thrombus, fibrinous exudate leading to severe pneumonic changes, and the virus was phenotypically highly virulent in mice.

Method
intranasal inoculation | histopathological examination of lungs | observation of clinical signs
Experimental system
Mouse infection model
OVE3614
Key finding

Experimental infection of BALB/c mice with A/crow/India/02CA01/2012 (non-reassortant H5N1) caused morbidity, mortality, and severe pneumonic lesions similar to the reassortant strain, demonstrating high virulence.

Virus
Host
Location
Not specified
Supporting text

Eighteen 6-8 weeks BALB/c mice were intranasally inoculated with 10^6 EID50/ml of H5N1 virus A/crow/India/02CA01/2012. The infected mice showed dullness, weight loss and ruffled fur coat. Histopathological examination of lungs showed severe congestion, haemorrhage, thrombus, fibrinous exudate leading to severe pneumonic changes; both the crow viruses caused morbidity and mortality in mice and were phenotypically highly virulent.

Method
intranasal inoculation | histopathological examination of lungs | observation of clinical signs
Experimental system
Mouse infection model
Genomic Evolution
1 records · 1 evidence types
Evidence type
1 records
OVE3616
Key finding

The H5N1 virus isolate A/crow/India/03CA04/2015 was identified as an H9N2-PB2 reassortant, indicating genome-segment reassortment between H5N1 and H9N2 viruses.

Virus
Host
Not specified
Location
Not specified
Supporting text

A/crow/India/03CA04/2015 (H9N2-PB2 reassortant H5N1)

Event type
reassortment
Genes or segments
PB2