The ORF virus causes contagious ecthyma in sheep and goats, which is an economically significant viral skin disease. The goal of this research was to observe the influence of ORFV on small ruminants’ hematological parameters as well as serum biochemical profiles across various districts of Punjab, Pakistan. In 58/441 animals, ORFV was confirmed by molecular analysis. ORFV lineages were classified into five separate groups based on phylogenetic analysis of the GIF/IL2 gene of Parapoxviruses. ORFV sequences from Pakistan isolates were grouped with viruses from China and India in Cluster I. In ORFV infected animals, hematological tests manifested mild to moderate leukocytosis, granulocytosis, severe neutrophilia and severe lymphopenia. Eosinophils, basophils, monocytes, thrombocytes, red blood cells (RBC), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) were not influenced statistically (P>0.05). Infected animals had significantly higher (P<0.05) alanine transaminase (ALT) and albumin to globulin (A/G) ratios when serum biochemical parameters were assessed. Total protein, albumin, globulin, creatinine and urea all indicated a significant decline (P<0.05) in the analysis of data. Bilirubin, aspartate aminotransferase (AST), and alkaline phosphatase (ALP), on the other hand, did not differ significantly (P>0.05). On the lateral commissure of the lips, muzzle, and nostrils of ORFV infected animals, diffused papular and pustular lesions as well as scab formation were visible. Microscopic investigations revealed acanthosis, parakeratosis with micro abscess zones, hyperkeratosis, spongiosis, epithelial cell degeneration, necrotic foci with patches of inflammatory cell aggregation and intracytoplasmic eosinophilic inclusion body in keratinocytes. It may be inferred that the ORFV had a considerable impact on sheep and goats hematological and serum biochemical parameters. The findings helped in the comprehension of disease mechanisms and controlling the infection.