Biosafety of recombinant and wild type nucleopolyhedroviruses as bioinsecticides

Mohamed Bassem Ashour, Didair A. Ragheb, El Sayed A El-Sheikh, El Adarosy A Gomaa, Shizuo G. Kamita, Bruce D. Hammock

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

The entomopathogenic Autographa californica (Speyer) nucleopolyhedrovirus (AcMNPV) has been genetically modified to increase its speed of kill. The potential adverse effects of a recombinant AcMNPV (AcAaIT) as well as wild type AcMNPV and wild type Spodoptera littoralis NPV (SlNPV) were studied. Cotton plants were treated with these viruses at concentrations that were adjusted to resemble the recommended field application rate (4 × 1012 PIBs/feddan, feddan = 4,200 m2) and 3rd instar larvae of S. littoralis were allowed to feed on the contaminated plants. SDS-PAGE, ELISA, and DNA analyses were used to confirm that larvae that fed on these plants were virus-infected. Polyhedra that were purified from the infected larvae were subjected to structural protein analysis. A 32 KDa protein was found in polyhedra that were isolated from all of the viruses. Subtle differences were found in the size and abundance of ODV proteins. Antisera against polyhedral proteins isolated from AcAaIT polyhedra were raised in rabbits. The terminal bleeds from rabbits were screened against four coating antigens (i.e., polyhedral proteins from AcAaIT, AcAaIT from field-infected larvae (AcAaIT-field), AcMNPV, and SlNPV) using a two-dimensional titration method with the coated antigen format. Competitive inhibition experiments were conducted in parallel to optimize antibody and coating antigen concentrations for ELISA. The IC50 values for each combination ranged from 1.42 to 163 μg/ml. AcAaIT-derived polyhedrin gave the lowest IC50 value, followed by those of SlNPV, AcAaIT-field, and AcMNPV. The optimized ELISA system showed low cross reactivity for AcMNPV (0.87%), AcAaIT-field (1.2%), and SlNPV (4.0%). Genomic DNAs isolated from AcAaIT that were passaged in larvae of S. littoralis that were reared in the laboratory or field did not show any detectable differences. Albino rats (male and female) that were treated with AcAaIT, AcMNPV or SlNPV (either orally or by intraperitoneal injection at doses of 1 × 108 or 1 × 107 PIBs/rat, respectively) appeared to be healthy and showed increased body weight at 21 days posttreatment. The effect of virus administration on hematological, serum biochemical, and histopathological parameters were determined. Slight to moderate differences were observed in most of the hematological parameters. Specifically, serum proteins were decreased markedly in female rats treated orally with SlNPV, and in male rats injected with AcAaIT. SDS-PAGE analysis also showed some changes in serum protein profiles. No marked changes in acetylcholine esterase (AChE) activity were found. Changes in serum glucose, alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), bilirubin, creatinin, and urea were also observed. Immunohistochemical observation of tissues from stomach, intestine, liver, kidney, brain, spleen, and lung also showed slight changes. Fish (Tilapia nilotica) were also exposed to AcAaIT, AcMNPV or SlNPV by incorporating each of the viruses into diet (1 × 109 PIBs/group). No mortality was found in treated or untreated fish during the experimental period (28 days). Macrophage phagocytic activity of fish head kidney cells increased with time, reaching maximum values at 180 min for both treated and control fish.

Original languageEnglish (US)
Pages (from-to)111-125
Number of pages15
JournalInternational Journal of Environmental Research and Public Health
Volume4
Issue number2
DOIs
StatePublished - Jun 2007

Fingerprint

biosafety
Nucleopolyhedrovirus
Spodoptera
Viruses
Larva
Fish
Rats
larva
protein
virus
serum
Fishes
antigen
Proteins
Antigens
Blood Proteins
Enzyme-Linked Immunosorbent Assay
fish
Inhibitory Concentration 50
Coatings

Keywords

  • Bioinsecticide
  • Biosaftey
  • Histopathology
  • Immunotoxicity
  • Macrophage phagocytosis
  • Nucleopolyhedrovirus
  • Recombinant baculovirus
  • Spodoptera littoralis
  • Toxicity

ASJC Scopus subject areas

  • Public Health, Environmental and Occupational Health
  • Health, Toxicology and Mutagenesis
  • Environmental Chemistry

Cite this

Ashour, M. B., Ragheb, D. A., El-Sheikh, E. S. A., Gomaa, E. A. A., Kamita, S. G., & Hammock, B. D. (2007). Biosafety of recombinant and wild type nucleopolyhedroviruses as bioinsecticides. International Journal of Environmental Research and Public Health, 4(2), 111-125. https://doi.org/10.3390/ijerph2007040005

Biosafety of recombinant and wild type nucleopolyhedroviruses as bioinsecticides. / Ashour, Mohamed Bassem; Ragheb, Didair A.; El-Sheikh, El Sayed A; Gomaa, El Adarosy A; Kamita, Shizuo G.; Hammock, Bruce D.

In: International Journal of Environmental Research and Public Health, Vol. 4, No. 2, 06.2007, p. 111-125.

Research output: Contribution to journalArticle

Ashour, Mohamed Bassem ; Ragheb, Didair A. ; El-Sheikh, El Sayed A ; Gomaa, El Adarosy A ; Kamita, Shizuo G. ; Hammock, Bruce D. / Biosafety of recombinant and wild type nucleopolyhedroviruses as bioinsecticides. In: International Journal of Environmental Research and Public Health. 2007 ; Vol. 4, No. 2. pp. 111-125.
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KW - Recombinant baculovirus

KW - Spodoptera littoralis

KW - Toxicity

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