Journal of Oil Palm Research Vol. 28  December 2016 p.  452-462
REPLICATION OF Oryctes NUDIVIRUS (OrNV) IN INSECT CELL LINE DSIR-HA-1179 AND ITS INFECTIVITY ON NEONATES OF RHINOCEROS BEETLE, Oryctes rhinoceros

Author(s): NUR AIN FARHAH ROS SAIDON KHUDRI*; WAHIZATUL AFZAN AZMI *; NORMAN KAMARUDIN**; SITI RAMLAH AHMAD ALI** and RAMLE MOSLIM**

The Oryctes nudi­virus (OrNV) or pre­vi­ously known as Oryctes bac­ulovirus is a bio­con­trol agent for rhi­noc­eros bee­tle, Oryctes rhi­noc­eros. The repli­ca­tion of three types of OrNV, the types A, B, and C in the insect cell line DSIR-HA-1179 was stud­ied. The cells were grown in PS100 media con­tain­ing 10% fetal bovine serum. The signs of infec­tion in cells by all types of OrNV were clearly seen at 10 to 14 days after treat­ment (DAT). The infected cells expe­ri­enced cyto­pathic effects (CPE) involv­ing for­ma­tion of high num­ber of vesi­cle or glob­ule like-structures around the cells. The den­sity of viable cells in flasks inoc­u­lated by the nudi­virus decreased over time. At 16 DAT, the viable cell den­sity in flasks inoc­u­lated with OrNV types A, B, and C was 1.49 × 105, 0.29 × 105, and 4.78 × 105 cells ml-1 respec­tively, sig­nif­i­cantly lower (P<0.05) compared to control flask of 7.98 × 105 cells ml-1. Based on the Tissue Culture Infectious Dose 50% (TCID50), it was found that the OrNV type B was more virulent than the OrNV types A and C as it produced high infectious virus particles. The TCID50 values for the OrNV types A and C were 3.00 × 107 infectious unit (IU ml-1) and 8.03 x 105 IU ml-1 as compared to type B which was 7.09 × 107 IU ml-1. The PCR analysis showed that all types of OrNV had successfully replicated in the cells as early as 4 DAT. Pathogenicity study on neonates using cells-propagated OrNV has again showed that the OrNV type B was more pathogenic than the other types of OrNV. The LT50 value for the OrNV type B was 33.47 days, shorter than the OrNV type A (49.35 days) and the OrNV type C (48.58 days). The DNA profiles of OrNV extracted from dead infected neonates in the bioassay were the same as those from infected gut tissues collected from field population. Thus, it is suggested that the Malaysian OrNV types A, B and C were successfully produced in cell line DSIR-HA-1179.

Keywords: , , ,

Author Information
* Malaysian Palm Oil Board, 6 Persiaran Institusi, Bandar Baru Bangi, 43000 Kajang, Selangor, Malaysia. E-mail: ainfarhah@mpob.gov.my


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