Determining genetic diversity of prevalent G and P genotype of Bovine Rotavirus A from neonatal calves of Gujarat, India

Fig. 1. Flow diagram of present research work for detection, isolation, and molecular characterization of Bovine rotavirus A.

Fig. 2. (A) Non-infected/Uninoculated monolayer of MA-104 cells three days after. (B) Characteristic ballooning (black arrow) and spindle formation (white arrow) on passage 3 in infected MA-104 cells after 24 h. (C) Characteristic granulation (black arrow), rounding, and clumping occur singly or in clusters in MA-104 cells infected with rotavirus 36 h after infection (Scale bar: 50 µm; phase-contrast microscopy). (D) Characteristic rounding, necrosis (black arrow), and sloughing off (white arrow) of infected MA-104 monolayer cells after 48 h. of infection (Scale bar: 50 µm). (E) Later stage of infection with extensive cell degeneration, i.e., necrosis and sloughing off (black arrow) in the MA-104 monolayer cells infected with Bovine rotavirus A after 60 h. of infection. Scale bars: (A) 50 µm, without staining; (B-E) 50 µm.

Fig. 3. Comparison of RNA-PAGE performed directly from fecal samples (left side) and RNA-PAGE after the isolation and propagation of Bovine rotavirus A in cell culture (right side) (Sample code: S-3, S-10, S-21, A-26, A-28).

Fig. 4. Agarose gel shows amplified G and P genotyping products of Bovine rotavirus A isolates. L: DNA Molecular weight ladder – 100 bp, 1–2 lane: G10 (714 bp), 3–4 lane: G8 (273 bp), 5 lane: Mixed infection (G10 + G8), 6–7 lane: P[5] (659 bp) and 8–9 lane: P[11] (335 bp) (Sample code: A-9, S-14, A-25, S-18, A-16, A-9, A-30, A-16, S-23; lane: left to right side).