Special Issue "Insect Viruses and Pest Management"

A special issue of Viruses (ISSN 1999-4915). This special issue belongs to the section "Insect Viruses".

Deadline for manuscript submissions: 30 November 2019.

Special Issue Editor

Guest Editor
Prof. Miguel López-Ferber Website E-Mail
Laboratory of Industrial Environmental Engineering, IMT Mines, Centre LGEI, Occitanie, France
Interests: baculovirus genetic diversity; virus–host interactions; biological control with viruses

Special Issue Information

Dear Colleagues,

Increasing public concern about the environmental consequences of the massive use of chemical pesticides and the development of resistances to these products by insects have increased scientists’ interest in finding alternatives for controlling insect pests. Insects are major pests, not only to agricultural crops but also to domestic animals and humans. Viruses offer alternatives for safe and environmentally friendly insect pest control, using various strategies. Baculoviruses have been used as biological control agents with success against various insect pests. Viruses belonging to other families have been proposed for use and are under evaluation for safety, specificity, and efficacy on the control of their insect hosts. Viruses has published a Special Issue on this topic (2014). Since then, our knowledge on viruses infecting insects, and their potential to control pests, has greatly improved, and new questions have emerged. This Issue presents the state-of-the-art on the actual use of viruses in pest control and the new approaches under development.

Prof. Miguel López-Ferber
Guest Editor

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Published Papers (16 papers)

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Research

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Open AccessArticle
Identification of Loci Associated with Enhanced Virulence in Spodoptera litura Nucleopolyhedrovirus Isolates Using Deep Sequencing
Viruses 2019, 11(9), 872; https://doi.org/10.3390/v11090872 - 17 Sep 2019
Abstract
Spodoptera litura is an emerging pest insect in cotton and arable crops in Central Asia. To explore the possibility of using baculoviruses as biological control agents instead of chemical pesticides, in a previous study we characterized a number of S. litura nucleopolyhedrovirus (SpltNPV) [...] Read more.
Spodoptera litura is an emerging pest insect in cotton and arable crops in Central Asia. To explore the possibility of using baculoviruses as biological control agents instead of chemical pesticides, in a previous study we characterized a number of S. litura nucleopolyhedrovirus (SpltNPV) isolates from Pakistan. We found significant differences in speed of kill, an important property of a biological control agent. Here we set out to understand the genetic basis of these differences in speed of kill, by comparing the genome of the fast-killing SpltNPV-Pak-TAX1 isolate with that of the slow-killing SpltNPV-Pak-BNG isolate. These two isolates and the SpltNPV-G2 reference strain from China were deep sequenced with Illumina. As expected, the two Pakistani isolates were closely related with >99% sequence identity, whereas the Chinese isolate was more distantly related. We identified two loci that may be associated with the fast action of the SpltNPV-Pak-TAX1 isolate. First, an analysis of rates of synonymous and non-synonymous mutations identified neutral to positive selection on open reading frame (ORF) 122, encoding a viral fibroblast growth factor (vFGF) that is known to affect virulence in other baculoviruses. Second, the homologous repeat region hr17, a putative enhancer of transcription and origin of replication, is absent in SpltNPV-Pak-TAX1 suggesting it may also affect virulence. Additionally, we found there is little genetic variation within both Pakistani isolates, and we identified four genes under positive selection in both isolates that may have played a role in adaptation of SpltNPV to conditions in Central Asia. Our results contribute to the understanding of the enhanced activity of SpltNPV-Pak-TAX1, and may help to select better SpltNPV isolates for the control of S. litura in Pakistan and elsewhere. Full article
(This article belongs to the Special Issue Insect Viruses and Pest Management)
Open AccessArticle
Generation of Virus- and dsRNA-Derived siRNAs with Species-Dependent Length in Insects
Viruses 2019, 11(8), 738; https://doi.org/10.3390/v11080738 - 11 Aug 2019
Abstract
Double-stranded RNA (dsRNA) molecules of viral origin trigger a post-transcriptional gene-silencing mechanism called RNA interference (RNAi). Specifically, virally derived dsRNA is recognized and cleaved by the enzyme Dicer2 into short interfering RNAs (siRNAs), which further direct sequence-specific RNA silencing, ultimately silencing replication of [...] Read more.
Double-stranded RNA (dsRNA) molecules of viral origin trigger a post-transcriptional gene-silencing mechanism called RNA interference (RNAi). Specifically, virally derived dsRNA is recognized and cleaved by the enzyme Dicer2 into short interfering RNAs (siRNAs), which further direct sequence-specific RNA silencing, ultimately silencing replication of the virus. Notably, RNAi can also be artificially triggered by the delivery of gene-specific dsRNA, thereby leading to endogenous gene silencing. This is a widely used technology that holds great potential to contribute to novel pest control strategies. In this regard, research efforts have been set to find methods to efficiently trigger RNAi in the field. In this article, we demonstrate the generation of dsRNA- and/or virus-derived siRNAs—the main RNAi effectors—in six insect species belonging to five economically important orders (Lepidoptera, Orthoptera, Hymenoptera, Coleoptera, and Diptera). In addition, we describe that the siRNA length distribution is species-dependent. Taken together, our results reveal interspecies variability in the (antiviral) RNAi mechanism in insects and show promise to contribute to future research on (viral-based) RNAi-triggering mechanisms in this class of animals. Full article
(This article belongs to the Special Issue Insect Viruses and Pest Management)
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Open AccessArticle
A Combination of Real-Time PCR and High-Resolution Melting Analysis to Detect and Identify CpGV Genotypes Involved in Type I Resistance
Viruses 2019, 11(8), 723; https://doi.org/10.3390/v11080723 - 06 Aug 2019
Abstract
Cydia pomonella granulovirus, in particular CpGV-M isolate, is used as a biological control against the codling moth (CM), Cydia pomonella. As a result of intensive control over the years, codling moth populations have developed resistance against this isolate. This resistance is now [...] Read more.
Cydia pomonella granulovirus, in particular CpGV-M isolate, is used as a biological control against the codling moth (CM), Cydia pomonella. As a result of intensive control over the years, codling moth populations have developed resistance against this isolate. This resistance is now called type I resistance. Isolates, among them, CpGV-R5, have been found that are able to overcome type I resistance. Both CpGV-M and CpGV-R5 are used in orchards to control the codling moth. High resolution melting (HRM) has been adapted to differentiate between CpGV-M and CpGV-R5 isolates. Specific PCR primers have been designed for the CpGV p38 gene, encompassing the variable region responsible for the ability to overcome resistance. Because each amplicon has a specific melting point, it is possible to identify the CpGV-M and CpGV-R5 genotypes and to quantify their relative proportion. This method has been validated using mixtures of occlusion bodies of each isolate at various proportions. Then, the HRM has been used to estimate the proportion of each genotype in infected larvae or in occlusion bodies (OBs) extracted from dead larvae. This method allows a rapid detection of genotype replication and enables the assessment of either success or failure of the infection in field conditions. Full article
(This article belongs to the Special Issue Insect Viruses and Pest Management)
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Open AccessArticle
Bio-Insecticidal Potential of Nucleopolyhedrovirus and Granulovirus Mixtures to Control the Fall Armyworm Spodoptera frugiperda (J.E. Smith, 1797) (Lepidoptera: Noctuidae)
Viruses 2019, 11(8), 684; https://doi.org/10.3390/v11080684 - 26 Jul 2019
Abstract
The ability of the isolate VG008 of S. frugiperda granulovirus (SpfrGV) to enhance the infectivity of the isolate SfCOL of S. frugiperda multiple nucleopolyhedrovirus (SpfrMNPV) was evaluated on S. frugiperda larvae. Bioassays were performed with mixtures by using different proportions 90%:10% (M1), 95%:5% [...] Read more.
The ability of the isolate VG008 of S. frugiperda granulovirus (SpfrGV) to enhance the infectivity of the isolate SfCOL of S. frugiperda multiple nucleopolyhedrovirus (SpfrMNPV) was evaluated on S. frugiperda larvae. Bioassays were performed with mixtures by using different proportions 90%:10% (M1), 95%:5% (M2) and 97.5%:2.5% (M3) of SfCOL:VG008, respectively. All mixtures showed higher insecticidal activity that SfCOL. The mixture M3 showed the highest enhancement of SfCOL reducing 11.40 times the Mean Lethal Concentration and 96 h in the Mean Time to Death. The enhancer activity of proteins derived from VG008 (GVPs) were also evaluated in mixture with SfCOL. The GVPs increased 27% larval mortality caused by SfCOL and damaged the peritrophic membrane of S. litura larvae, suggesting that the key point in this enhancing activity is the initial step of the larva colonization, the midgut infection. M3 was formulated and evaluated under greenhouse conditions in maize plants using different doses. The highest efficacy was obtained with the highest dose of M3 (8 × 1011 OBs/ha), which was similar to that found when formulated SfCOL was applied using an approximately twofold higher dose. The viral mixture M3 was selected as the active ingredient for developing a new biopesticide for a more efficient management of the pest in the field. Full article
(This article belongs to the Special Issue Insect Viruses and Pest Management)
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Open AccessArticle
Genetic Variation and Biological Activity of Two Closely Related Alphabaculoviruses during Serial Passage in Permissive and Semi-Permissive Heterologous Hosts
Viruses 2019, 11(7), 660; https://doi.org/10.3390/v11070660 - 18 Jul 2019
Abstract
Phylogenetic analyses suggest that Mamestra brassicae multiple nucleopolyhedrovirus (MbMNPV) and Helicoverpa armigera multiple nucleopolyhedrovirus (HearMNPV) may be strains of the same virus species. Most of the studies comparing their biological activities have been performed in their homologous hosts. A comparison of host range [...] Read more.
Phylogenetic analyses suggest that Mamestra brassicae multiple nucleopolyhedrovirus (MbMNPV) and Helicoverpa armigera multiple nucleopolyhedrovirus (HearMNPV) may be strains of the same virus species. Most of the studies comparing their biological activities have been performed in their homologous hosts. A comparison of host range and stability in alternative hosts was performed. The host range of these viruses was compared using high concentrations of inoculum to inoculate second instars of six species of Lepidoptera. One semi-permissive host (Spodoptera littoralis) and one permissive host (S. exigua) were then selected and used to perform six serial passages involving a concentration corresponding to the ~25% lethal concentration for both viruses. Restriction endonuclease analysis showed fragment length polymorphisms in every host-virus system studied. In S. littoralis, serial passage of MbMNPV resulted in decreased pathogenicity and an increase in speed-of-kill, whereas no significant changes were detected for HearMNPV with respect to the initial inoculum. In contrast, both viruses showed a similar trend in S. exigua. These results highlight the low genetic diversity and a high phenotypic stability of HearMNPV with respect to the original inoculum after six successive passages in both insect hosts. This study concludes that host-baculovirus interactions during serial passage are complex and the process of adaptation to a novel semi-permissive host is far from predictable. Full article
(This article belongs to the Special Issue Insect Viruses and Pest Management)
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Open AccessCommunication
Genome Analysis of A Novel South African Cydia pomonella granulovirus (CpGV-SA) with Resistance-Breaking Potential
Viruses 2019, 11(7), 658; https://doi.org/10.3390/v11070658 - 18 Jul 2019
Cited by 1
Abstract
The complete genome of an endemic South African Cydia pomonella granulovirus isolate was sequenced and analyzed. Several missing or truncated open reading frames (ORFs) were identified, including a 24 bp deletion in the pe38 gene which is reported to be associated with type [...] Read more.
The complete genome of an endemic South African Cydia pomonella granulovirus isolate was sequenced and analyzed. Several missing or truncated open reading frames (ORFs) were identified, including a 24 bp deletion in the pe38 gene which is reported to be associated with type I resistance-breaking potential. Comparison of single nucleotide polymorphisms (SNPs) with five other fully sequenced CpGV isolates identified 67 unique events, 47 of which occurred within ORFs, leading to several amino acid changes. Further analysis of single nucleotide variations (SNVs) within CpGV-SA revealed that this isolate consists of mixed genotypes. Phylogenetic analysis using complete genome sequences placed CpGV-SA basal to M, I12 and E2 and distal to S and I07 but with no distinct classification into any of the previously defined CpGV genogroups. These results suggest that CpGV-SA is a novel and genetically distinct isolate with significant potential as a biopesticide for management of codling moth (CM), not only in South Africa, but potentially in other pome fruit producing countries, particularly where CM resistance to CpGV has been reported. Full article
(This article belongs to the Special Issue Insect Viruses and Pest Management)
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Open AccessArticle
Identification of Multiple Replication Stages and Origins in the Nucleopolyhedrovirus of Anticarsia gemmatalis
Viruses 2019, 11(7), 648; https://doi.org/10.3390/v11070648 - 15 Jul 2019
Abstract
To understand the mechanism of replication used by baculoviruses, it is essential to describe all the factors involved, including virus and host proteins and the sequences where DNA synthesis starts. A lot of work on this topic has been done, but there is [...] Read more.
To understand the mechanism of replication used by baculoviruses, it is essential to describe all the factors involved, including virus and host proteins and the sequences where DNA synthesis starts. A lot of work on this topic has been done, but there is still confusion in defining what sequence/s act in such functions, and the mechanism of replication is not very well understood. In this work, we performed an AgMNPV replication kinetics into the susceptible UFL-Ag-286 cells to estimate viral genome synthesis rates. We found that the viral DNA exponentially increases in two different phases that are temporally separated by an interval of 5 h, probably suggesting the occurrence of two different mechanisms of replication. Then, we prepared a plasmid library containing virus fragments (0.5–2 kbp), which were transfected and infected with AgMNPV in UFL-Ag-286 cells. We identified 12 virus fragments which acted as origins of replication (ORI). Those fragments are in close proximity to core genes. This association to the core genome would ensure vertical transmission of ORIs. We also predict the presence of common structures on those fragments that probably recruit the replication machinery, a structure also present in previously reported ORIs in baculoviruses. Full article
(This article belongs to the Special Issue Insect Viruses and Pest Management)
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Open AccessArticle
Importance of the Host Phenotype on the Preservation of the Genetic Diversity in Codling Moth Granulovirus
Viruses 2019, 11(7), 621; https://doi.org/10.3390/v11070621 - 05 Jul 2019
Abstract
To test the importance of the host genotype in maintaining virus genetic diversity, five experimental populations were constructed by mixing two Cydia pomonella granulovirus isolates, the Mexican isolate CpGV-M and the CpGV-R5, in ratios of 99% M + 1% R, 95% M + [...] Read more.
To test the importance of the host genotype in maintaining virus genetic diversity, five experimental populations were constructed by mixing two Cydia pomonella granulovirus isolates, the Mexican isolate CpGV-M and the CpGV-R5, in ratios of 99% M + 1% R, 95% M + 5% R, 90% M + 10% R, 50% M + 50% R, and 10% M + 90% R. CpGV-M and CpGV-R5 differ in their ability to replicate in codling moth larvae carrying the type I resistance. This ability is associated with a genetic marker located in the virus pe38 gene. Six successive cycles of replication were carried out with each virus population on a fully-permissive codling moth colony (CpNPP), as well as on a host colony (RGV) that carries the type I resistance, and thus blocks CpGV-M replication. The infectivity of offspring viruses was tested on both hosts. Replication on the CpNPP leads to virus lineages preserving the pe38 markers characteristic of both isolates, while replication on the RGV colony drastically reduces the frequency of the CpGV-M pe38 marker. Virus progeny obtained after replication on CpNPP show consistently higher pathogenicity than that of progeny viruses obtained by replication on RGV, independently of the host used for testing. Full article
(This article belongs to the Special Issue Insect Viruses and Pest Management)
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Open AccessArticle
Genetic Variability of Chrysodeixis Includens Nucleopolyhedrovirus (ChinNPV) and the Insecticidal Characteristics of Selected Genotypic Variants
Viruses 2019, 11(7), 581; https://doi.org/10.3390/v11070581 - 26 Jun 2019
Cited by 1
Abstract
Genetic variation in baculoviruses is recognized as a key factor, not only due to the influence of such variation on pathogen transmission and virulence traits, but also because genetic variants can form the basis for novel biological insecticides. In this study, we examined [...] Read more.
Genetic variation in baculoviruses is recognized as a key factor, not only due to the influence of such variation on pathogen transmission and virulence traits, but also because genetic variants can form the basis for novel biological insecticides. In this study, we examined the genetic variability of Chrysodeixis includens nucleopolyhedrovirus (ChinNPV) present in field isolates obtained from virus-killed larvae. Different ChinNPV strains were identified by restriction endonuclease analysis, from which genetic variants were isolated by plaque assay. Biological characterization studies were based on pathogenicity, median time to death (MTD), and viral occlusion body (OB) production (OBs/larva). Nine different isolates were obtained from eleven virus-killed larvae collected from fields of soybean in Mexico. An equimolar mixture of these isolates, named ChinNPV-Mex1, showed good insecticidal properties and yielded 23 genetic variants by plaque assay, one of which (ChinNPV-R) caused the highest mortality in second instars of C. includens. Five of these variants were selected: ChinNPV-F, ChinNPV-J, ChinNPV-K, ChinNPV-R, and ChinNPV-V. No differences in median time to death were found between them, while ChinNPV-F, ChinNPV-K, ChinNPV-R and ChinNPV-V were more productive than ChinNPV-J and the original mixture of field isolates ChinNPV-Mex1. These results demonstrate the high variability present in natural populations of this virus and support the use of these new genetic variants as promising active substances for baculovirus-based bioinsecticides. Full article
(This article belongs to the Special Issue Insect Viruses and Pest Management)
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Open AccessArticle
A Novel Alphabaculovirus from the Soybean Looper, Chrysodeixis includens, that Produces Tetrahedral Occlusion Bodies and Encodes Two Copies of he65
Viruses 2019, 11(7), 579; https://doi.org/10.3390/v11070579 - 26 Jun 2019
Abstract
Isolates of the alphabaculovirus species, Chrysodeixis includens nucleopolyhedrovirus, have been identified that produce polyhedral occlusion bodies and infect larvae of the soybean looper, Chrysodeixis includens. In this study, we report the discovery and characterization of a novel C. includens-infecting alphabaculovirus, [...] Read more.
Isolates of the alphabaculovirus species, Chrysodeixis includens nucleopolyhedrovirus, have been identified that produce polyhedral occlusion bodies and infect larvae of the soybean looper, Chrysodeixis includens. In this study, we report the discovery and characterization of a novel C. includens-infecting alphabaculovirus, Chrysodeixis includens nucleopolyhedrovirus #1 (ChinNPV#1), that produces tetrahedral occlusion bodies. In bioassays against C. includens larvae, ChinNPV #1 exhibited a degree of pathogenicity that was similar to that of other ChinNPV isolates, but killed larvae more slowly. The host range of ChinNPV#1 was found to be very narrow, with no indication of infection occurring in larvae of Trichoplusia ni and six other noctuid species. The ChinNPV#1 genome sequence was determined to be 130,540 bp, with 126 open reading frames (ORFs) annotated but containing no homologous repeat (hr) regions. Phylogenetic analysis placed ChinNPV#1 in a clade with other Group II alphabaculoviruses from hosts of lepidopteran subfamily Plusiinae, including Chrysodeixis chalcites nucleopolyhedrovirus and Trichoplusia ni single nucleopolyhedrovirus. A unique feature of the ChinNPV#1 genome was the presence of two full-length copies of the he65 ORF. The results indicate that ChinNPV#1 is related to, but distinct from, other ChinNPV isolates. Full article
(This article belongs to the Special Issue Insect Viruses and Pest Management)
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Open AccessArticle
Partial Loss of Inheritable Type I Resistance of Codling Moth to Cydia pomonella granulovirus
Viruses 2019, 11(6), 570; https://doi.org/10.3390/v11060570 - 20 Jun 2019
Abstract
Current knowledge of the field resistance of codling moth (CM, Cydia pomonella, L) against Cydia pomonella granulovirus (CpGV) is based mainly on the interaction between the Mexican isolate CpGV-M and CpRR1, a genetically homogeneous CM inbreed line carrying type I resistance. The [...] Read more.
Current knowledge of the field resistance of codling moth (CM, Cydia pomonella, L) against Cydia pomonella granulovirus (CpGV) is based mainly on the interaction between the Mexican isolate CpGV-M and CpRR1, a genetically homogeneous CM inbreed line carrying type I resistance. The resistance level of laboratory-reared CpRR1 to CpGV-M was recently found to have decreased considerably, compared to the initially high resistance. To understand the background of this phenomenon, CpRR1 larvae were exposed over several generations to CpGV-M for re-selection of the original resistance level. After five and seven generations of selection, new CpRR1_F5 and CpRR1_F7 lines were established. The resistance ratio of these selected lines was determined by full range bioassays. The CpRR1_F5 strain regained a higher level of resistance against CpGV up to 104-fold based on LC50 values compared to susceptible larvae (CpS), which indicated that the absence of virus selection had resulted in a reduction of resistance under laboratory rearing conditions. In addition, some fitness costs of fecundity were observed in CpRR1_F5. Single-pair crossings between CpRR1_F5 or CpRR1_F7 with susceptible CpS moths revealed a dominant but not fully sex-linked inheritance, which suggests a partial loss of previous resistance traits in CpRR1. Full article
(This article belongs to the Special Issue Insect Viruses and Pest Management)
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Open AccessArticle
DWV-A Lethal to Honey Bees (Apis mellifera): A Colony Level Survey of DWV Variants (A, B, and C) in England, Wales, and 32 States across the US
Viruses 2019, 11(5), 426; https://doi.org/10.3390/v11050426 - 09 May 2019
Cited by 2
Abstract
The strong association between Varroa destructor, deformed wing virus (DWV), and high overwintering colony losses (OCL) of honey bees is well established. Three DWV master variants (DWV-A, -B, and -C) have been described, and their role in colony mortality remains an open [...] Read more.
The strong association between Varroa destructor, deformed wing virus (DWV), and high overwintering colony losses (OCL) of honey bees is well established. Three DWV master variants (DWV-A, -B, and -C) have been described, and their role in colony mortality remains an open question. Therefore, the aim of this study is to investigate the seasonal prevalence, viral load, and changing distribution of the three DWV master variants within honey bee colonies from England, Wales, and 32 states across the United States. Here, we report that in 2016, DWV-B was prevalent (100%, n = 249) and dominant (95%) in England and Wales, compared to the US. (56%, n = 217 and 23%, respectively), where DWV-A was prevalent (83%, n = 217) and dominant (63%). DWV-C was regularly detected in low viral loads (<1 × 107 genome equivalents per bee) and at lower prevalence (58% in England and Wales, n = 203, and 14% across the United States, n = 124) compared to DWV-A and -B. DWV-B prevalence and dominance in England and Wales coincided with low OCL (6%). Meanwhile, a 60% loss was reported by participating U.S. beekeepers. In the United States, DWV-A prevalence (89%, n = 18) and viral load were significantly (p = 0.002) higher (1 × 10 8–1 × 1011) in colonies that died when compared to the surviving colonies (49% (n = 27), 1 × 106–1 × 1010). DWV-B had low prevalence (56%, n = 18) in the colonies that died with viral loads of <1 × 1010. However, DWV-B was routinely detected in high viral loads (>1 × 1010) in surviving colonies from all sample locations, providing further supporting evidence of DWV-A exhibiting increased virulence over DWV-B at the colony level. Full article
(This article belongs to the Special Issue Insect Viruses and Pest Management)
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Open AccessArticle
Effects of a Covert Infection with Phthorimaea operculella granulovirus in Insect Populations of Phthorimaea operculella
Viruses 2019, 11(4), 337; https://doi.org/10.3390/v11040337 - 09 Apr 2019
Abstract
Virus infections of insects can easily stay undetected, neither showing typical signs of a disease, nor being lethal. Such a stable and most of the time covert infection with Phthorimaea operculella granulovirus (PhopGV) was detected in a Phthorimaea operculella laboratory colony, which originated [...] Read more.
Virus infections of insects can easily stay undetected, neither showing typical signs of a disease, nor being lethal. Such a stable and most of the time covert infection with Phthorimaea operculella granulovirus (PhopGV) was detected in a Phthorimaea operculella laboratory colony, which originated from Italy (Phop-IT). This covert virus (named PhopGV-R) was isolated, purified and characterized at the genetic level by full genome sequencing. Furthermore, the insect colony Phop-IT was used to study the crowding effect, double infection with other PhopGV isolates (CR3 and GR1), and co-infection exclusion. An infection with a second homologous virus (PhopGV-CR3) activated the covert virus, while a co-infection with another virus isolate (PhopGV-GR1) led to its suppression. This study shows that stable virus infections can be common for insect populations and have an impact on population dynamics because they can suppress or enable co-infection with another virus isolate of the same species. Full article
(This article belongs to the Special Issue Insect Viruses and Pest Management)
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Open AccessArticle
Multiple Virus Infections in Western Honeybee (Apis mellifera L.) Ejaculate Used for Instrumental Insemination
Viruses 2019, 11(4), 306; https://doi.org/10.3390/v11040306 - 29 Mar 2019
Abstract
Instrumental insemination of Apis mellifera L. queens is a widely employed technique used in honeybee breeding that enables the effective control of mating. However, drone semen represents a potential source of honeybee viruses. In this study, 43 semen doses collected from apparently healthy [...] Read more.
Instrumental insemination of Apis mellifera L. queens is a widely employed technique used in honeybee breeding that enables the effective control of mating. However, drone semen represents a potential source of honeybee viruses. In this study, 43 semen doses collected from apparently healthy drones, and consequently used in instrumental insemination, were analysed using PCR or RT-PCR to detect the presence of viral genome of 11 honeybee viruses. In 91% of samples, viral infection was detected. The survey revealed genomes of five viruses, namely Deformed wing virus (DWV), Acute bee paralysis virus (ABPV), Black queen cell virus (BQCV), Sacbrood virus (SBV), and A. mellifera filamentous virus (AmFV) in 84%, 19%, 14%, 2%, and 67% of samples, respectively. Single infection (30% of samples) as well as multiple infection (61% of samples) of two, three or four pathogens were also evaluated. To the best of our knowledge, this is the first study describing the presence of the BQCV and SBV genome sequence in drone ejaculate. Phylogenetic analysis of BQCV partial helicase gene sequence revealed the high similarity of nucleotide sequence of described Czech strains, which varied from 91.4% to 99.6%. The findings of our study indicate the possibility of venereal transmission of BQCV and SBV. Full article
(This article belongs to the Special Issue Insect Viruses and Pest Management)
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Open AccessArticle
New Method for Differentiation of Granuloviruses (Betabaculoviruses) Based on Real-Time Polymerase Chain Reaction (Real-Time PCR)
Viruses 2019, 11(2), 115; https://doi.org/10.3390/v11020115 - 29 Jan 2019
Cited by 1
Abstract
Baculoviridae is a highly diverse family of rod-shaped viruses with double-stranded DNA. To date, almost 100 species have had their complete genomic sequences deposited in the GenBank database, a quarter of which comprises granuloviruses (GVs). Many of the genomes are sequenced using next-generation [...] Read more.
Baculoviridae is a highly diverse family of rod-shaped viruses with double-stranded DNA. To date, almost 100 species have had their complete genomic sequences deposited in the GenBank database, a quarter of which comprises granuloviruses (GVs). Many of the genomes are sequenced using next-generation sequencing, which is currently considered the best method for characterizing new species, but it is time-consuming and expensive. Baculoviruses form a safe alternative to overused chemical pesticides and therefore there is a constant need for identifying new species that can be active components of novel biological insecticides. In this study, we have described a fast and reliable method for the detection of new and differentiation of previously analyzed granulovirus species based on a real-time polymerase chain reaction (PCR) technique with melting point curve analysis. The sequences of highly conserved baculovirus genes, such as granulin and late expression factors 8 and 9 (lef-8 and lef-9), derived from GVs available to date have been analyzed and used for degenerate primer design. The developed method was tested on a representative group of eight betabaculoviruses with comparisons of melting temperatures to allow for quick and preliminary granulovirus detection. The proposed real-time PCR procedure may be a very useful tool as an easily accessible screening method in a majority of laboratories. Full article
(This article belongs to the Special Issue Insect Viruses and Pest Management)
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Review

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Open AccessReview
Nucleocapsid Assembly of Baculoviruses
Viruses 2019, 11(7), 595; https://doi.org/10.3390/v11070595 - 01 Jul 2019
Abstract
The baculovirus nucleocapsid is formed through a rod-like capsid encapsulating a genomic DNA molecule of 80~180 kbp. The viral capsid is a large oligomer composed of many copies of various protein subunits. The assembly of viral capsids is a complex oligomerization process. The [...] Read more.
The baculovirus nucleocapsid is formed through a rod-like capsid encapsulating a genomic DNA molecule of 80~180 kbp. The viral capsid is a large oligomer composed of many copies of various protein subunits. The assembly of viral capsids is a complex oligomerization process. The timing of expression of nucleocapsid-related proteins, transport pathways, and their interactions can affect the assembly process of preformed capsids. In addition, the selection of viral DNA and the injection of the viral genome into empty capsids are the critical steps in nucleocapsid assembly. This paper reviews the replication and recombination of baculovirus DNA, expression and transport of capsid proteins, formation of preformed capsids, DNA encapsulation, and nucleocapsid formation. This review will provide a basis for further study of the nucleocapsid assembly mechanism of baculovirus. Full article
(This article belongs to the Special Issue Insect Viruses and Pest Management)
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