Infectivity and Effects of Gypsy Moth and Spruce Budworm Nuclear Polyhedrosis Viruses Ingested by Rainbow Trout

Kreutzweiser, David P.; Ebling, Peter M; Holmes, Stephen B.

doi:10.1006/eesa.1997.1562

Cited by 4 publications

(6 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The apparent toxicity of injected viruses to oral exposure was assumed to be related to a general foreign body response. Our findings are in general agreement with previous studies concerning the effect of NPV exposure in non insect cells and tissues NPVs [ 33 – 40 ]. Additionally, there was no correlation between any effects induced by the wild type or recombinant baculoviruses in rats or fish.…”

Section: Discussionsupporting

confidence: 93%

Biosafety of Recombinant and Wild Type Nucleopolyhedroviruses as Bioinsecticides

Ashour

Ragheb

El-Sheikh

et al. 2007

IJERPH

View full text Add to dashboard Cite

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...

show abstract

Section: Discussionsupporting

confidence: 93%

Biosafety of Recombinant and Wild Type Nucleopolyhedroviruses as Bioinsecticides

Ashour

Ragheb

El-Sheikh

et al. 2007

IJERPH

View full text Add to dashboard Cite

show abstract

“…Preparation of virus suspension. Naturally occurring CfMNPV and CfDefNPV were propagated in fifth-instar larvae of C. fumiferana and C. occidentalis using contaminated artificial diet (McMorran 1965), as per Arif and Brown (1975) and Kreutzweiser et al (1997). Ebling (2004) found no difference in the activity of CfMNPV (Ireland strain) when propagated in these two hosts.…”

Section: Methodsmentioning

confidence: 99%

Host-range testing of a mixture of two nucleopolyhedroviruses of Choristoneura fumiferana (Lepidoptera: Tortricidae)

et al. 2011

Self Cite

View full text Add to dashboard Cite

The host range of a mixture of Choristoneura fumiferana (Clemens) nucleopolyhedroviruses (CfMNPV and CfDefNPV) was investigated using a per os bioassay of larvae of 29 species of Lepidoptera and adult males of Megachile rotundata (F.) (Hymenoptera: Megachilidae). Using a whole-genomic DNA probe, positive results were obtained in 8 of 10 Tortricidae: Archips cerasivorana (Fitch), Choristoneura fractivittana (Clemens), C. fumiferana, Choristoneura occidentalis Freeman, Choristoneura pinus pinus Freeman, Choristoneura rosaceana (Harris), Clepsis persicana (Fitch), and Cydia pomonella (L.); one Crambidae: Ostrinia nubilalis (Hübner); one arctiine Erebidae: Estigmene acrea (Drury); and two Noctuidae: Oligia illocata (Walker) and Pyrrhia exprimens (Walker). Mortality rates were highest among C. fumiferana, C. occidentalis, C. pinus pinus, A. cerasivorana, and C. pomonella. Sequenced polymerase chain reaction (PCR) amplicons from infected individuals from several species confirmed that the primer sets amplified the target viruses. CfMNPV was consistently found in virus-fed C. fumiferana; whereas, CfDefNPV was present only occasionally. The presence of CfMNPV and CfDefNPV in A. cerasivorana was confirmed by PCR and DNA sequencing. Significant treatment-mortality rates were induced in the noctuids P. exprimens and Acronicta impleta Walker; PCR determined that both viruses were present in treated P. exprimens but only CfMNPV was present in A. impleta. No virus was detected in M. rotundata.

show abstract

“…Occlusion bodies produced using the diet surface contamination method were separated from larval cadavers by filtration and centrifugation using a procedure modified from Kreutzweiser et al 4 Infected insects were macerated in distilled water and sodium dodecyl sulphate added to a final concentration of 3g litre −1 . This solution was stirred for 2 h at room temperature, filtered through three layers of cheesecloth, one layer of Kleenex®, and the OBs were pelleted out of suspension by centrifugation at 1483 g for 30 min at 15 °C.…”

Section: Experimental Methodsmentioning

confidence: 99%

“…To determine whether the probe would react to uninfected tissues, as reported by some researchers,4 twelve non‐infected laboratory‐reared second‐instar O leucostigma larvae were macerated individually in equal volumes of water and 20‐µl volumes blotted onto the membrane.…”

Section: Experimental Methodsmentioning

confidence: 99%

“…Although NPV has routinely been detected by microscopic examination of either stained or unstained preparations of larval tissue smears, this can be unreliable at low levels or early stages of infection 2. Researchers2–6 have successfully demonstrated the use of horseradish peroxidase (HRP)‐labelled whole genomic DNA probes and enhanced chemiluminescence (ECL) for the detection of baculoviruses in various other types of samples blotted onto nylon membranes. This methodology has previously not been utilized for the detection of NPV in O leucostigma larvae ( Ol NPV) nor have detection thresholds for any NPV ever been quantified to account for the effects of larval tissues and pre‐occluded viral components.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

DNA hybridization assay for detection of nucleopolyhedrovirus in whitemarked tussock moth (Orgyia leucostigma) larvae

2001

View full text Add to dashboard Cite

DNA dot-blot hybridization assays utilizing a horseradish peroxidase-labelled whole genomic DNA probe and enhanced chemiluminescence were conducted to quantify detection thresholds of nucleopolyhedrovirus (NPV) in whitemarked tussock moth (Orgyia leucostigma) larvae. The minimum detection thresholds for an aqueous suspension of occlusion bodies (OBs), OBs added to macerates of non-infected larvae and OBs in macerates of diseased larvae were 7.8 x 10(3), 7.8 x 10(3), and 1.5 x 10(3) OBs, respectively. Purified viral DNA was detected at a concentration of 1.6 x 10(-1) ng in a 20 microliters volume. The presence of pre-occluded viral nucleocapsids and DNA, inherent to infected larvae, improved the detection threshold five-fold compared with OBs alone. Larval tissues did not block the detection system utilized, nor did they bind non-specifically to the probe. Detection thresholds, upon sequential hybridization of the same membrane, on average deteriorated two-fold between the first and second hybridization and an additional six-fold between the second and third hybridization. NPV infection was detected two days post-inoculation (pi) in about one-third of the larvae examined and in almost all larvae three days pi. Microscopic analysis of stained larval smears missed NPV infection in almost all larvae two days pi and about two-thirds of the larvae three days pi. Results from the two methods of analysis were not comparable until four days pi. The detection system utilized is a reliable, efficient and simple method for the early detection of NPV infection in large numbers of larvae and may be used for further studies quantifying the role of this baculovirus in the ecology of whitemarked tussock moth populations.

show abstract

Infectivity and Effects of Gypsy Moth and Spruce Budworm Nuclear Polyhedrosis Viruses Ingested by Rainbow Trout

Cited by 4 publications

References 21 publications

Biosafety of Recombinant and Wild Type Nucleopolyhedroviruses as Bioinsecticides

Biosafety of Recombinant and Wild Type Nucleopolyhedroviruses as Bioinsecticides

Host-range testing of a mixture of two nucleopolyhedroviruses of Choristoneura fumiferana (Lepidoptera: Tortricidae)

DNA hybridization assay for detection of nucleopolyhedrovirus in whitemarked tussock moth (Orgyia leucostigma) larvae

Contact Info

Product

Resources

About