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Published on: September 13, 2018
Comparative activity of Choristoneura fumiferana nucleopolyhedrovirus propagated in different hosts
1Natural Resources Canada, Great Lakes Forestry Centre, 1219 Queen St E., Sault Ste. Marie, Ontario, Canada P6A 2E5. pebling@nrcan.gc.ca
Abstract:
The biological activity of the Ireland strain of Choristoneura fumiferana (Clem) nucleopolyhedrovirus (CfMNPV) propagated in different hosts was determined to provide the basis upon which genetically modified CfMNPV, or other naturally occurring isolates, should be compared. Occlusion bodies (OB) derived from CF-203 cells were significantly larger and more pathogenic than those propagated in vivo when tested against the fifth larval instar of C fumiferana (Clem) and C occidentalis Freeman. The dose-responses (LD50 and LD95, expressed as occlusion bodies per larva) of C fumiferana larvae to in vitro-propagated OBs were 274 and 5785, respectively. The values of LD50 and LD95 to C occidentalis larvae were 19 and 118, respectively. There were no significant differences in pathogenicity or size when OBs propagated in C fumiferana larvae were tested against either insect species, nor were there significant differences for OBs propagated in C occidentalis larvae. The LD50 and LD95 of in vivo-produced OBs to C fumiferana were 925 and 61988, respectively. The LD50 and LD95 to C occidentalis were 50 and 453, respectively. OBs propagated in vitro had a mean volume of 13.13 microm3, whereas those propagated in vivo ranged from 0.84 to 1.41 microm3. The median survival time-responses (ST50) of fifth-instar C fumiferana or C occidentalis larvae to OBs propagated in vivo were not significantly different from those propagated in vitro at the dosage levels tested. Values of ST50 of C fumiferana larvae to in vitro- and in vivo-produced OBs at dosages causing less than 50% mortality rangedfrom 9.6 to 9.8 days post-inoculation (dpi), whereas a LD95 dose resulted in ST50 values ranging from 7.3 to 7.7 days. ST50 values of C occidentalis larvae at dosages causing less than 50% mortality ranged from 9.8 to 10.2 dpi, whereas a LD95 dose resulted in ST50 values ranging from 9.5 to 9.8 dpi. The median feeding cessation time-response (FT50) of fifth-instar C fumiferana larvae to OBs propagated in vitro (5.7 days) was not significantly different from the FT50 of those propagated in vivo in either insect species (5.3 and 5.7 days) at the dosage level tested (LD95). No significant differences in FT50 values were observed between OBs propagated in either larval host. The FT50 of C occidentalis larvae to OBs propagated in vitro (7.7 days) was not significantly different from that to those propagated in vivo in C occidentalis larvae (7.6days), but somewhat different (7.2 days) from that to those propagated in C fumiferana larvae. Results indicate that CfMNPV can be propagated in vivo in either C fumiferana or C occidentalis larvae (or sequentially through both) without alteration in infectivity, although the use of the CF-203 cell line yields the most biologically active OBs.
Insights
In vitro propagation of Choristoneura fumiferana nucleopolyhedrovirus (CfMNPV) in CF-203 cells produced larger and more pathogenic occlusion bodies (OBs) than in vivo propagation. However, in vivo propagation in either host species did not alter infectivity.
Area of Science:
- Entomology
- Virology
- Pest Management
Background:
- Choristoneura fumiferana nucleopolyhedrovirus (CfMNPV) is a pathogen used for insect biological control.
- Understanding the biological activity of different CfMNPV strains is crucial for developing effective pest management strategies.
- Propagation methods can influence the characteristics and efficacy of viral occlusion bodies (OBs).
Purpose of the Study:
- To determine the biological activity of the Ireland strain of CfMNPV propagated in different hosts.
- To establish a baseline for comparing genetically modified or naturally occurring CfMNPV isolates.
- To evaluate the impact of propagation method (in vitro vs. in vivo) on OB size, pathogenicity, and infectivity.
Main Methods:
- Propagation of CfMNPV in CF-203 cells (in vitro) and in Choristoneura fumiferana and Choristoneura occidentalis larvae (in vivo).
- Determination of occlusion body (OB) size and pathogenicity through dose-response bioassays (LD50 and LD95) against fifth-instar larvae of C. fumiferana and C. occidentalis.
- Assessment of median survival time (ST50) and median feeding cessation time (FT50) post-inoculation.
Main Results:
- OBs propagated in vitro from CF-203 cells were significantly larger (13.13 µm³) and more pathogenic than those propagated in vivo (0.84–1.41 µm³).
- In vitro-propagated OBs showed lower LD50/LD95 values against C. fumiferana and C. occidentalis larvae compared to in vivo-propagated OBs.
- In vivo propagation in either C. fumiferana or C. occidentalis larvae did not significantly alter OB pathogenicity or size when tested against either host species.
- Median survival time (ST50) and feeding cessation time (FT50) responses were not significantly different between in vitro- and in vivo-propagated OBs at tested dosages.
Conclusions:
- The CF-203 cell line yields the most biologically active CfMNPV OBs in terms of size and pathogenicity.
- In vivo propagation of CfMNPV in C. fumiferana or C. occidentalis larvae does not alter its infectivity.
- Results provide a basis for comparing different CfMNPV isolates and inform strategies for optimizing viral insecticide production.
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