A Combination of Real-Time PCR and High-Resolution Melting Analysis to Detect and Identify CpGV Genotypes Involved in Type I Resistance

Hinsberger, Aurélie; Germain, Stéphane; Guerrero, P.; Blachère-López, Christine; López‐Ferber, M.; Bayle, Sandrine

doi:10.3390/v11080723

Cited by 6 publications

(5 citation statements)

References 31 publications

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“…Kumar et al ( 2017) have pointed out that type I GVs are characterized by having a slow speed of kill in their hosts compared to type II GVs, even suggesting the need for a higher LD 50 (up to 100 times more) in type I GVs. Some insects can also delay the infection of Type 1 GVs as a rst defense mechanism, blocking viral replication in cells during the early stages of infection (Hinsberger et al 2019;Pauli et al 2018;Wang et al 2008). The larvae infected with the SfGV-CH28 isolate required more time to reach the pupal stage and showed a slower weight gain rate than those of the other experimental groups.…”

Section: Discussionmentioning

confidence: 99%

Morphological, biological, and molecular characterization of Type I granuloviruses of Spodoptera frugiperda

Ordóñez-García,

Bustillos-Rodríguez,

Ornelas-Paz

et al. 2024

Preprint

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The granuloviruses or GVs (Betabaculovirus) associated with the fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), especially those of Type I, have scarcely been studied but they might represent an alternative for the biocontrol of this insect. In this study, the native granuloviruses SfGV-CH13 and SfGV-CH28 isolated from FAW larvae were characterized for morphology, molecular traits, and insecticidal activity. The elapsed time between symptomatic infection of larvae and stop feeding as well as the weight of larvae before death or prior to pupation were also evaluated. Both granuloviruses isolates showed ovoid shape with a length of 0.4 µm. They showed the same DNA restriction profiles and their genome sizes were about 126 kb. The symptomatic infection with tested GVs mainly caused flaccidity of larva body and discoloration of integument. The integument lysis was only observed in 8% of infected larvae. Infected larvae gradually stopped feeding. Overall, these symptoms are characteristic of infections caused by Type I granuloviruses, which are known as monoorganotropic or slow-killing. The median lethal doses (LD50) values for SfGV-CH13 and SfGV-CH28 isolates were 5.4 × 102 and 1.1 × 103 OBs/larva, respectively. The median lethal time (LT50) ranged from 17 to 24 d. LT50 values decreased as the viral dose was increased. The elapsed time since symptomatic infection until pupation (LD50) and body weight of larvae (third instar) were higher with SfGV-CH28 than SfGV-CH13. Both granulovirus isolates were able to kill the FAW larvae from the 12th day.

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Section: Discussionmentioning

confidence: 99%

Morphological, biological, and molecular characterization of Type I granuloviruses of Spodoptera frugiperda

Ordóñez-García,

Bustillos-Rodríguez,

Ornelas-Paz

et al. 2024

Preprint

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“…R GV neonate larvae were first allowed to feed on CpGV-R5 treated medium for 30 min (time necessary for ingestion of sufficient virus to result in double infection [ 36 ]). Larvae were then transferred to non-treated medium and left for various durations (Xi), then retransferred to the medium inoculated with CpGV-M for 30 min, and finally transferred back to non-treated medium for 3 days.…”

Section: Methodsmentioning

confidence: 99%

CpGV-M Replication in Type I Resistant Insects: Helper Virus and Order of Ingestion Are Important

Hinsberger

Blachère-López

Knox

et al. 2021

Viruses

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The genetic diversity of baculoviruses provides a sustainable agronomic solution when resistance to biopesticides seems to be on the rise. This genetic diversity promotes insect infection by several genotypes (i.e., multiple infections) that are more likely to kill the host. However, the mechanism and regulation of these virus interactions are still poorly understood. In this article, we focused on baculoviruses infecting the codling moth, Cydia pomonella: two Cydia pomonella granulovirus genotypes, CpGV-M and CpGV-R5, and Cryptophlebia peltastica nucleopolyhedrovirus (CrpeNPV). The influence of the order of ingestion of the virus genotypes, the existence of an ingestion delay between the genotypes and the specificity of each genotype involved in the success of multiple infection were studied in the case of Cydia pomonella resistance. To obtain a multiple infection in resistant insects, the order of ingestion is a key factor, but the delay for ingestion of the second virus is not. CrpeNPV cannot substitute CpGV-R5 to allow replication of CpGV-M.

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“…The non-template control was added in triplicate with nuclease-free water as a template. The qPCR amplification was conducted in CFX96™ Real-Time PCR Detection System (Bio-Rad) [31]. The C T of all replicates was obtained and the mean of each sample was determined.…”

Section: Quantitative Polymerase Chain Reaction Amplification Of Extr...mentioning

confidence: 99%

Efficacy, humoral, and cell-mediated immune response of inactivated fowl adenovirus 8b propagated in chicken embryo liver cells using bioreactor in broiler chickens

et al. 2022

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Background and Aim: Fowl adenovirus (FAdV) 8b causes inclusion body hepatitis, resulting in major economic losses globally among chickens. The objectives were to inactivate FAdV 8b isolate propagated in chicken embryo liver (CEL) cells using a stirred tank bioreactor (UPM08136P5B1) and determine the humoral and cell-mediated immune response, efficacy, and virus shedding in broiler chickens. Materials and Methods: The FAdV 8b isolate UPM08136P5B1 was inactivated using binary ethyleneimine, adjuvanted with Montanide 71VG, inoculated into day-old broiler chickens in a booster group (BG) and non-booster group (NBG), and challenged with a pathogenic FAdV 8b strain. Clinical signs, gross lesions, body weight (BW), liver: body weight ratio, FAdV antibody titer using enzyme-linked immunosorbent assay, and histopathological changes were recorded. The CD3+, CD4+, and CD8+ T-lymphocyte profiles of the liver, spleen, and thymus using flow cytometry, and viral load in liver and cloacal shedding using quantitative polymerase chain reaction were evaluated. Results: Chickens in the challenged control group (CCG) exhibited mild clinical signs, gross lesions, and histopathological changes, which were absent in the inoculated groups, and had lower BW and higher liver BW ratio than chickens in the unchallenged control group (UCG); BG and NBG on 35- and 42-days post-inoculation (DPI). Chickens in NBG and BG had higher antibodies than UCG on 7, 21, 35, and 42 DPI. The challenged BG and NBG produced higher antibodies than the CCG on 35 DPI. T-lymphocytes were higher among the inoculated groups than UCG in the liver, spleen, and thymus. Inoculated challenged groups recorded higher CD3+, CD4+, and CD8+ T-lymphocytes on 35 and 42 DPI than CCG. The challenged control group had a significantly higher viral load in the liver than challenged that in BG on 35 DPI and BG and NBG on 42 DPI. The challenged control group had significantly higher challenge FAdV shedding than challenged inoculated groups on 35 and NBG on 42 DPI. Conclusion: UPM08136P5B1 was successfully inactivated and mixed with Montanide 71VG. The inactivated vaccine candidate that induced humoral and cellular immunity was effective, reduced FAdV load in the liver, and shedding in the cloaca, and could be useful against FAdV 8b infections in chickens.

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A Combination of Real-Time PCR and High-Resolution Melting Analysis to Detect and Identify CpGV Genotypes Involved in Type I Resistance

Cited by 6 publications

References 31 publications

Morphological, biological, and molecular characterization of Type I granuloviruses of Spodoptera frugiperda

Morphological, biological, and molecular characterization of Type I granuloviruses of Spodoptera frugiperda

CpGV-M Replication in Type I Resistant Insects: Helper Virus and Order of Ingestion Are Important

Efficacy, humoral, and cell-mediated immune response of inactivated fowl adenovirus 8b propagated in chicken embryo liver cells using bioreactor in broiler chickens

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