ISSN: 0973-7510

E-ISSN: 2581-690X

Research Article | Open Access
Gopal Sarkar1, Katherukamem Rajukumar1 , Dhanapal Senthilkumar1, Fateh Singh1, Govindarajulu Venkatesh1, Jaswant Patel1, Rohit Sahu1, Nourin Khan1, Amandeep2, Vijendra Pal Singh1 and Aniket Sanyal1
1ICAR-National Institute of High Security Animal Diseases (ICAR-NIHSAD), Bhopal, Madhya Pradesh, India.
2Northern Regional Disease Diagnostic Laboratory (NRDDL), Jalandhar, Punjab, India.
Article Number: 11692 | © The Author(s). 2026
J Pure Appl Microbiol. 2026;20(2):1737-1745. https://doi.org/10.22207/JPAM.20.2.64
Received: 21 April 2026 | Accepted: 18 May 2026 | Published online: 05 June 2026
Issue online: June 2026
Abstract

African swine fever (ASF), a deadly disease caused by African swine fever virus (ASFV), emerged in north-eastern India in 2020 and subsequently spread to several states, devastating swine herds due to the lack of commercial vaccines. We report the isolation, genetic characterization and growth kinetics of ASFV isolated from Punjab, India in primary porcine pulmonary alveolar macrophages (PAM). Partial sequencing of the B646L gene categorized the virus as genotype II, showing complete nucleotide identity with previously reported Indian genotype II isolates. Phylogenetic analysis placed the isolate amongst genotype II ASFVs circulating in Europe and the Asia-Pacific region since 2007. Multistep growth curves were constructed based on HAD50 titres and viral DNA copy numbers after infection of PAM cultures at 1.0 and 0.1 multiplicity of infection (MOI) observed up to 168 hrs post-infection (hpi). At 1.0 MOI, viral titres and genome copy numbers in the cell associated (CA) fraction peaked earlier (120-144 hpi) before declining, whereas the culture supernatant (CS) fraction showed a delayed but sustained increase, reaching maximum levels at 168 hpi. At 0.1 MOI, both HAD50 titres and viral genome copies increased progressively in CA and CS fractions, with higher peak levels observed in the CS fraction at later time points. By observing up to 168 hpi, this study reveals higher extracellular virus production with lower DNA copy to HAD ratio in PAM cultures infected at 0.1 MOI, demonstrating optimal duration for higher virus yield in cell culture supernatants, which would support efficient ASFV propagation, pathogenesis and vaccine development studies.

Keywords

African Swine Fever, African Swine Fever Virus, ASFV, Growth Kinetics, PAM

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© The Author(s) 2026. Open Access. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License which permits unrestricted use, sharing, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.