Determination of a novel parvovirus pathogen associated with massive mortality in adult tilapia
Fig 5
Characterization of the TiPV genome, PCR detection of TiPV in different samples and cumulative mortality of artificial infected tilapia.
(A) The genome organization of TiPV. The NS1, NP, VP1 proteins, the alternative ORF1 and ORF2 are showed in different colors; (B) Alignment of conservative domains of NS1 proteins from different parvoviruses by Muscle package. The HuH (u indicates hydrophobic residues) and the walker loop motif, including A, B, B′ and C Walker box of helicase domains are marked; (C) Lane M: DL1000 bp DNA ladder; Lane 1: positive control; Lane 2: negative control; Lanes 3: the kidney tissues sample from natural infected tilapia; Lanes 4: TiB cell culture 3rd-passage viral supernatant; Lane 5: The artificial infected tilapia peritoneal injection with 0.5 ml of 0.22μm filtrate of diseased fish tissue homogenates; Lanes 6: the mock-infected tilapia; (D) Fish in the test group 1 (▲) were challenged by intraperitoneal injection with the 2nd-passage of viral supernatant (104.0TCID50/ml) from cell culture; fish in test group 2 (■) were challenged by intraperitoneal injection with 0.5 ml filtrated supernatant homogenate from diseased fish tissues; and fish in the control group (●) were injected intraperitoneally with Dulbecco’s PBS.