Search Results (40)

The mosquito-associated virome may modulate host biology and influence vector competence, highlighting the importance of understanding its composition. Here, a metagenomic analysis was conducted to characterize the virome of the sea-rock pool mosquito Aedes mariae across sexes and developmental stages, together with water from its sea-rock pool breeding site in San Felice Circeo (Italy). A total of 51 viral taxa were identified, including viruses associated with bacteria and archaea (39%), plants, algae, fungi, and protists (35%), vertebrates (8%), and invertebrates (18%), including insect-specific viruses such as Mesoniviridae, Baculoviridae, Nudiviridae, Iridoviridae and Totiviridae. Twenty-five percent of the taxa were shared across samples, suggesting acquisition from breeding-site water and persistence across stages during development. Interestingly, the need for host genome filtering highlights the potential sequence similarity between viral and mosquito genomes, which may reflect the presence of endogenous viral elements or historical virus–host interactions. These findings represent the first characterization of the virome of Aedes mariae and highlight the role of aquatic breeding sites in shaping mosquito virome. Finally, we argue the importance of adequate sequencing depth and host genome filtering to capture the diversity of the mosquito virome. Full article

The Spodoptera frugiperda multiple nucleopolyhedrovirus (SfMNPV) is an important pathogen of the fall armyworm and is used as the basis for biological insecticides. In this study, we examined the relationship between the size–frequency distribution of SfMNPV occlusion bodies (OBs) and their insecticidal characteristics when collected at the end of the replication cycle. Exposure of OBs to 40%, 70%, and 90% (wt/wt) glycerol had no effect on OB pathogenicity. Glycerol density gradient (50–100%) centrifugation was used to separate OBs into two fractions. OBs recovered from the upper fraction of the gradient had a significantly smaller median cross-sectional area than those harvested from the lower fraction. These fractions also differed significantly in their size–frequency distributions. The OB concentration–mortality response of S. frugiperda second instars did not differ significantly between the two fractions or with non-centrifuged OBs. The median survival time was similar for insects inoculated with OBs from the upper and lower fractions but was significantly shorter in insects inoculated with non-centrifuged OBs. The proportion of mature OBs (67–71%) and the number of viral genome copies (1.33–1.40 × 10⁸ copies/µL) did not differ significantly between the upper and lower OB fractions. These findings suggest that altering the size–frequency distribution by density gradient centrifugation is not a useful technique for selecting large OBs with high insecticidal activity as part of the baculovirus insecticide production process. Future studies should evaluate a range of OB size separation techniques to determine their effects on OB insecticidal characteristics. Full article

Glyphosate is a broad-spectrum, systemic herbicide that has attracted concern over its non-target effects, environmental persistence, and the presence of residues in food. The fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae), is a major invasive pest of maize that can be controlled by application of its homologous nucleopolyhedrovirus (SfMNPV), an occluded virus in the family Baculoviridae. We examined the effects of a glyphosate-based herbicide on S. frugiperda growth and survival and on virus occlusion bodies (OBs) exposed to product label-recommended concentrations of the herbicide. Larval growth, time to pupation, pupal weight, duration of the pupal stage and sex ratio were not affected by exposure to the herbicide (1% v/v solution) applied to the surface of semi-synthetic diet. Exposure to 1–2% herbicide solution had no effect on the median lethal concentration (LC₅₀) of OBs, the susceptibility of second instar larvae to virus infection, or the production of OBs in virus-killed larvae. Virus acquisition did not vary significantly when larvae fed on virus-sprayed maize plants at 1 and 6 days after they had been treated with herbicide, compared to healthy plants. Finally, the presence of 2% herbicide solution did not influence the persistence of OBs in non-sterilized soil samples over a 6-week greenhouse experiment. Although the laboratory and greenhouse experiments indicated that the glyphosate-based herbicide tested was unlikely to influence the transmission or persistence of SfMNPV OBs, future studies should verify these findings across a range of field conditions, soil types and different herbicide formulations. Full article

Infection by Spodoptera frugiperda multiple nucleopolyhedrovirus (SfMNPV) profoundly alters the physiology of S. frugiperda (Lepidoptera: Noctuidae), promoting molecular responses that activate genes related to cellular defense. These responses demand substantial energy and lead to feeding dysfunction. Knowledge about the effects of SfMNPV on weight gain and leaf consumption rate is still incipient; in this context, we evaluated the survival rate, weight gain, leaf consumption rate, and body size of S. frugiperda exposed to different concentrations of SfMNPV. A completely randomized design was used in the laboratory. Treatments consisted of SfMNPV from the commercial product Cartugen^®, diluted at different concentrations and applied on a Petri dish: 9.00 × 10³, 1.80 × 10⁴, 3.75 × 10⁴, 7.50 × 10⁴, 1.35 × 10⁵, 2.25 × 10⁵, and 3.75 × 10⁵ occlusion bodies (OBs)/mL. Fifty first-instar larvae were used per treatment. The median lethal concentration was estimated at 1.32 × 10⁵ OBs/mL (95% confidence interval = 1.22 × 10⁵–1.43 × 10⁵ OBs/mL). Nonlinear regression analysis of sublethal effects showed that the expected weight of the control (W₀), the angular parameter (B), and the effective concentration capable of reducing larval weight by 50% (EC₅₀) were 47.40 mg, 1.42, and 1.16 × 10⁴ OBs/mL, respectively. Leaf consumption was inhibited at lower concentrations and increased at higher concentrations among surviving larvae, and larval growth (measured by head diameter, body length, and body width) was lower when larvae were exposed to SfMNPV than in the control. Our data suggest that SfMNPV at low concentrations causes dysfunctions that prevent normal development in surviving individuals, resulting in reduced consumption rate, body growth, and weight gain. Overall, our analysis indicates that the impact of SfMNPV extends beyond mortality; low concentrations can affect larval growth and feeding consumption rate in S. frugiperda. Full article

Nile tilapia (Oreochromis niloticus) is an indispensable source of high-quality protein worldwide. Along with the exponential expansion of tilapia aquaculture, several novel pathogenic viruses have emerged, and some cause significant economic losses. Unfortunately, there is scarce information on the biology and epidemiology of these viruses. This exploratory metagenomic study used Oxford Nanopore Technology (ONT) sequencing to profile the virome compositions of both wild and farmed Nile tilapia across five regions in Egypt. The Nile tilapia virome was dominated by two double-stranded DNA bacteriophages, Muvirus mu and M. sfmu, which constituted 79.8% of the detected sequences. Eukaryotic viruses, including members of the families Amnoonviridae, Peribunyaviridae, and Baculoviridae, were also identified. Two giant DNA viruses known to infect Acanthamoeba spp., Mollivirus sp., and Pandoravirus sp. were identified in the spleen virome of tilapia from a single sampling site. The diversity analysis showed no significant differences among tissue types or sampling sites. Phylogenetic analyses were performed on a single virus detected of potential pathogenicity, an amnoonvirus. The analyses demonstrated that the detected virus is a member of the family Amnoonviridae and placed it alongside members of the Tilapinevirus genus. The virus, however, was distinct from the other two members in the genus: T. tilapae and T. poikilos. This study underscores the usefulness of ONT in providing a foundational understanding of the Nile tilapia virome. Full article

Nucleopolyhedroviruses of lepidopteran larvae (Alphabaculovirus, Baculoviridae) form the basis for effective and highly selective biological insecticides for the control of caterpillar pests of greenhouse and field crops and forests. Horizontal transmission is usually achieved following the release of large quantities of viral occlusion bodies (OBs) from virus-killed insects. In the present review, I examine the evidence for productive midgut infection in different host species and the resulting transmission through the release of OBs in the feces (frass) of the host. This has been a neglected aspect of virus transmission since it was initially studied over six decades ago. The different host–virus pathosystems vary markedly in the quantity of OBs released in feces and in their ability to contaminate the host’s food plant. The release of fecal OBs tends to increase over time as the infection progresses. Although based on a small number of studies, the prevalence of transmission of fecal inoculum is comparable with that of recognized alternative routes for transmission and dissemination, such as cannibalism and interactions with predators and parasitoids. Finally, I outline a series of predictions that would affect the importance of OBs in feces as a source of inoculum in the environment and which could form the basis for future lines of research. Full article

The production of viable nucleopolyhedrovirus in the feces of infected lepidopteran larvae represents a poorly understood route for virus transmission prior to host death. In the present study, we examined the presence of viable virus in the feces of fourth-instar Spodoptera frugiperda larvae infected with the Nicaraguan isolate of Spodoptera frugiperda multiple nucleopolyhedrovirus (SfMNPV-NIC). Feces production increased in samples taken at 2 to 6 days post-inoculation but was significantly lower in infected insects compared to controls. Second instars experienced 3.9 to 68.3% of polyhedrosis disease following consumption of feces collected at 2–5 days post-inoculation, which subsequently fell to 29.1% in the 6-day sample. Calibration of the insect bioassay using OB-spiked samples of feces indicated that the concentration of OBs varied between 5.4 × 10² and 4.4 × 10⁵ OBs/100 mg of feces in infected fourth instars. Quantitative PCR analysis of fecal samples indicated the presence of 0 to 7629 copies/mg feces following amplification targeted at the polyhedrin gene. However, no correlation was detected between qPCR estimates of virus concentration and time of sample collection or the quantity of feces collected. The qPCR estimates were positively correlated with the prevalence of lethal infection observed in the insect bioassay, but the correlation was weak and several samples did not amplify. Calibration of the qPCR assay using OB-spiked samples of feces provided estimates that were 5- to 10-fold lower than the insect bioassay, indicating inhibition of the amplification reaction or loss of material during processing. In a greenhouse experiment, 2.5–48.3% of second-instar larvae acquired lethal infection following a 24 h period of feeding on maize plants on which fourth instar larvae had deposited their feces at 3 days and 4 days post-infection, respectively. These findings highlight the potential of OB-contaminated feces as a source of biologically significant quantities of inoculum for virus transmission prior to the death of infected insects and represent an additional contribution to the biological control of lepidopteran pests by these pathogens. Full article

The use of CpGV strains as the basis for bioinsecticides is an effective and safe way to control Cydia pomonella. The research is aimed at the identification and study of new CpGV strains. Objects of identification and bioinformatic analysis: 18 CpGV strains. Sequencing was carried out on a NextSeq550. Genome assembly and annotation were carried out using Spades, Samtools 1.9, MinYS, Pilon, Gfinisher, Quast, and Prokka. Comparative genomic analysis was carried out in relation to the reference genome present in the «Madex Tween» strain-producer (biological standard) according to the average nucleotide identity (ANI) criterion. The presence/absence of IAP, cathepsin, MMP, and chitinase in the genetic sequences of the strains was determined using simply phylogeny. Entomopathogenic activity was assessed against C. pomonella according to the criterion of biological efficacy. Thus, molecular genetic identification revealed that 18 CpGV strains belong to a genus of Betabaculovirus. For all the strains under study ANI values of 99% or more were obtained, and the presence of the cathepsin, chitinase, IAP, and MMP genes was noted. The strains BZR GV 1, BZR GV 3, BZR GV 7, BZR GV 10, and BZR GV L-8 showed the maximum biological efficacy: 100% on the 15th day of observation. Strains BZR GV 4, BZR GV 8, and BZR GV 13 showed efficacy at the level of the «Madex Tween» preparation: 89.5% on the 15th day of observation. The strains with the highest mortality rate of the host insect were identified: BZR GV 9, BZR GV 10, BZR GV L-6, and BZR GV L-8. Full article

Insecticides based on baculoviruses have become an alternative for pest control in different agricultural crops. The aim of this study was to assess the biological activity of the bioinsecticide Cartugen^™ (SfMNPV: Baculoviridae: Alphabaculovirus) on larvae of Spodoptera frugiperda J. E. Smith (Lepidoptera: Noctuidae) resistant to Bt corn expressing the insecticidal proteins Cry1A.105+Cry2Ab2+Cry1F. In addition, we assessed the efficiency of SfMNPV on S. frugiperda control in the field from natural infestation of the pest during two agricultural seasons. The results showed that no larvae survived 10 days after being inoculated with Bt and non-Bt corn leaves contaminated with 1.50 × 10¹⁰ occlusion bodies (OBs)/L (equivalent to the recommended dose of the product). However, when using doses equivalent to 50% (7.50 × 10⁹ OBs/L) and 25% (3.75 × 10⁹ OBs/L), the larval mortality ranged from 21.12% to 46.55%, respectively. Although larvae resistant to the Cry1A.105+Cry2Ab2+Cry1F proteins, when exposed to 50% of the SfMNPV dose (7.50 × 10⁹ OBs/L), showed reductions in larval weight (52 to 67% reduction), pupal weight (32 to 59% reduction), and total fecundity (67 to 86% reduction) compared to the control. Furthermore, doses above 25% (3.75 × 10⁹ OBs/L⁻¹) caused a population decrease in the growth of the species in both Bt and non-Bt corn according to the fertility life table. In the field, at 7 and 10 DAA (days after application), corn plants sprayed with SfMNPV (1.50 × 10¹⁰ OBs/L) showed reductions in leaf damage according to the Davis scale. However, from 14 to 21 DAA, there was an increase in leaf damage in corn leaves from both treatments, with or without the application of SfMNPV. This shows that SfMNPV may be an important strategy in the integrated management and resistance management of S. frugiperda. Full article

Alphabaculoviruses are lethal dsDNA viruses of Lepidoptera that have high genetic diversity and are transmitted in aggregates within proteinaceous occlusion bodies. This mode of transmission has implications for their efficacy as biological insecticides. A Nicaraguan isolate of Spodoptera frugiperda multiple nucleopolyhedrovirus (SfMNPV-NIC) comprising nine genotypic variants has been the subject of considerable study due to the influence of variant interactions on the insecticidal properties of mixed-variant occlusion bodies. As part of a systematic study on the replication and transmission of variant mixtures, a tool for the accurate quantification of a selection of genotypic variants was developed based on the quantitative PCR technique (qPCR). First, primer pairs were designed around a region of high variability in four variants named SfNic-A, SfNic-B, SfNic-C and SfNic-E to produce amplicons of 103–150 bp. Then, using cloned purified amplicons as standards, amplification was demonstrated over a dynamic range of 10⁸–10¹ copies of each target. The assay was efficient (mean ± SD: 98.5 ± 0.8%), reproducible, as shown by low inter- and intra-assay coefficients of variation (<5%), and specific to the target variants (99.7–100% specificity across variants). The quantification method was validated on mixtures of genotype-specific amplicons and demonstrated accurate quantification. Finally, mixtures of the four variants were quantified based on mixtures of budded virions and mixtures of DNA extracted from occlusion-derived virions. In both cases, mixed-variant preparations compared favorably to total viral genome numbers by quantification of the polyhedrin (polh) gene that is present in all variants. This technique should prove invaluable in elucidating the influence of variant diversity on the transmission and insecticidal characteristics of this pathogen. Full article

Baculoviridae, a virus family characterized by a single large double stranded DNA, encompasses the majority of viral bioinsecticides, representing a highly promising and environmentally friendly pesticide approach to insect control. This study focuses on the characterization of a baculovirus isolated from larvae of Calliteara abietis (Erebidae, Lymantriidae) collected in Mongolian pinaceae forests. This new isolate was called Calliteara abietis nucleopolyhedrovirus (CaabNPV). CaabNPV exhibits an irregular polyhedron shape, and significant variation in the diameter of its occlusion bodies (OBs) was observed. Nucleotide distance calculations confirmed CaabNPV as a novel baculovirus. The CaabNPV genome spans 177,161 bp with a G+C content of 45.12% and harbors 150 potential open reading frames (ORFs), including 38 core genes. A comprehensive genomic analysis categorizes CaabNPV within Group II alphabaculovirus, revealing a close phylogenetic relationship with Alphabaculovirus orleucostigmae (OrleNPV). Additionally, repeat sequence analysis identified three highly repetitive sequences consisting of 112 bp repeat units, known as homologous regions (hrs). This research contributes valuable insights into CaabNPV’s phylogenetic placement, genomic structure, and its potential applications in insect biocontrol. Full article

Baculoviruses are viral pathogens that infect different species of Lepidoptera, Diptera, and Hymenoptera, with a global distribution. Due to their biological characteristics and the biotechnological applications derived from these entities, the Baculoviridae family is an important subject of study and manipulation in the natural sciences. With the advent of RNA interference mechanisms, the presence of baculoviral genes that do not code for proteins but instead generate transcripts similar to microRNAs (miRNAs) has been described. These miRNAs are functionally associated with the regulation of gene expression, both in viral and host sequences. This article provides a comprehensive review of miRNA biogenesis, function, and characterization in general, with a specific focus on those identified in baculoviruses. Furthermore, it delves into the specific roles of baculoviral miRNAs in regulating viral and host genes and presents structural and thermodynamic stability studies that are useful for detecting shared characteristics with predictive utility. This review aims to expand our understanding of the baculoviral miRNAome, contributing to improvements in the production of baculovirus-based biopesticides, management of resistance phenomena in pests, enhancement of recombinant protein production systems, and development of diverse and improved BacMam vectors to meet biomedical demands. Full article

Outbreaks of Anticarsia gemmatalis (Hübner, 1818) (Lepidoptera: Erebidae), a major pest of soybean, can be controlled below economic thresholds with methods that do not involve the application of synthetic insecticides. Formulations based on natural isolates of the Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV) (Baculoviridae: Alphabaculovirus) played a significant role in integrated pest management programs in the early 2000s, but a new generation of chemical insecticides and transgenic soybean have displaced AgMNPV-based products over the past decade. However, the marked genotypic variability present among and within alphabaculovirus isolates suggests that highly insecticidal genotypic variants can be isolated and used to reduce virus production costs or overcome isolate-dependent host resistance. This study aimed to select novel variants of AgMNPV with suitable insecticidal traits that could complement the existing AgMNPV active ingredients. Three distinct AgMNPV isolates were compared using their restriction endonuclease profile and in terms of their occlusion body (OB) pathogenicity. One isolate was selected (AgABB51) from which eighteen genotypic variants were plaque purified and characterized in terms of their insecticidal properties. The five most pathogenic variants varied in OB pathogenicity, although none of them was faster-killing or had higher OB production characteristics than the wild-type isolate. We conclude that the AgABB51 wild-type isolates appear to be genotypically structured for fast speed of kill and high OB production, both of which would favor horizontal transmission. Interactions among the component variants are likely to influence this insecticidal phenotype. Full article

Baculoviruses are entomopathogens that carry large, double-stranded circular DNA genomes and infect insect larvae of Lepidoptera, Hymenoptera and Diptera, with applications in the biological control of agricultural pests, in the production of recombinant proteins and as viral vectors for various purposes in mammals. These viruses have a variable genetic composition that differs between species, with some sequences shared by all known members, and others that are lineage-specific or unique to isolates. Based on the analysis of nearly 300 sequenced genomes, a thorough bioinformatic investigation was conducted on all the baculoviral protein coding sequences, characterizing their orthology and phylogeny. This analysis confirmed the 38 protein coding sequences currently considered as core genes, while also identifying novel coding sequences as candidates to join this set. Accordingly, homology was found among all the major occlusion body proteins, thus proposing that the polyhedrin, granulin and CUN085 genes be considered as the 39th core gene of Baculoviridae. Full article

Cryptophlebia leucotreta granulovirus (CrleGV), a double-stranded DNA virus (genus Betabaculovirus, family Baculoviridae), is highly infective to the citrus insect pest Thaumatotibia leucotreta. The South African isolate CrleGV-SA is formulated into a commercial biopesticide and registered for use in several countries. In South Africa, it is used as a biopesticide in a multi-faceted integrated pest management approach for citrus crops involving chemical and biological control methods. The virus nucleocapsid is surrounded and protected by an occlusion body (OB) composed of granulin protein in a crystalline matrix. Like all other baculoviruses, CrleGV is susceptible to ultraviolet (UV) radiation from sunlight. This reduces its efficacy as a biopesticide in the field and necessitates frequent respraying. UV damage to baculovirus biopesticides is detected by means of functional bioassays. However, bioassays do not give an indication of whether any structural damage has occurred that may contribute to functional loss. In this study, transmission electron microscopy (TEM) was used to observe damage to the OB and nucleocapsid (NC) of CrleGV-SA, following controlled UV irradiation in the laboratory to mimic field conditions. The resultant images were compared with images of non-irradiated CrleGV-SA virus. TEM images of irradiated CrleGV-SA samples revealed changes to the OB crystalline faceting, a reduction in the size of the OBs, and damage to the NC following UV exposure for 72 h. Full article