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20 pages, 7204 KiB  
Article
Structural Features and In Vitro Antiviral Activities of Fungal Metabolites Sphaeropsidins A and B Against Bovine Coronavirus
by Luca Del Sorbo, Maria Michela Salvatore, Clementina Acconcia, Rosa Giugliano, Giovanna Fusco, Massimiliano Galdiero, Violetta Iris Vasinioti, Maria Stella Lucente, Paolo Capozza, Annamaria Pratelli, Luigi Russo, Rosa Iacovino, Anna Andolfi and Filomena Fiorito
Int. J. Mol. Sci. 2025, 26(15), 7045; https://doi.org/10.3390/ijms26157045 - 22 Jul 2025
Viewed by 223
Abstract
The scientific community’s interest in natural compounds with antiviral properties has considerably increased after the emergence of the severe acute respiratory syndrome coronavirus (SARS-CoV-2), especially for their potential use in the treatment of the COVID-19 infection. From this perspective, bovine coronavirus (BCoV), member [...] Read more.
The scientific community’s interest in natural compounds with antiviral properties has considerably increased after the emergence of the severe acute respiratory syndrome coronavirus (SARS-CoV-2), especially for their potential use in the treatment of the COVID-19 infection. From this perspective, bovine coronavirus (BCoV), member of the genus β-CoV, represents a valuable virus model to study human β-CoVs, bypassing the risks of handling highly pathogenic and contagious viruses. Pimarane diterpenes are a significant group of secondary metabolites produced by phytopathogenic fungi, including several Diplodia species. Among the members of this class of natural products, sphaeropsidin A (SphA) and its analog sphaeropsidin B (SphB) are well known for their bioactivities, such as antimicrobial, insecticidal, herbicidal, and anticancer. In this study, the antiviral effects of SphA and SphB were evaluated for the first time on bovine (MDBK) cells infected with BCoV. Our findings showed that both sphaeropsidins significantly increased cell viability in infected cells. These substances also caused substantial declines in the virus yield and in the levels of the viral spike S protein. Interestingly, during the treatment, a cellular defense mechanism was detected in the downregulation of the aryl hydrocarbon receptor (AhR) signaling, which is affected by BCoV infection. We also observed that the presence of SphA and SphB determined the deacidification of the lysosomal environment in infected cells, which may be related to their antiviral activities. In addition, in silico investigations have been performed to elucidate the molecular mechanism governing the recognition of bovine AhR (bAhR) by Sphs. Molecular docking studies revealed significant insights into the structural determinants driving the bAhR binding by the examined compounds. Hence, in vitro and in silico results demonstrated that SphA and SphB are promising drug candidates for the development of efficient therapies able to fight a β-CoV-like BCoV during infection. Full article
(This article belongs to the Special Issue Structure, Function and Dynamics in Proteins: 3rd Edition)
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16 pages, 2821 KiB  
Article
Metabolomic Analysis Uncovers the Presence of Pimarenyl Cation-Derived Diterpenes as Insecticidal Constituents of Sphagneticola trilobata
by Lilia Chérigo, Juan Fernández, Ramy Martínez and Sergio Martínez-Luis
Plants 2025, 14(14), 2219; https://doi.org/10.3390/plants14142219 - 17 Jul 2025
Viewed by 404
Abstract
Aphis gossypii is a significant global pest that impacts numerous agricultural crops and vegetables, causing direct damage to food plants and indirect damage through the transmission of phytopathogenic viruses, primarily begomoviruses. In Panama, particularly in the Azuero region, viral infections transmitted by this [...] Read more.
Aphis gossypii is a significant global pest that impacts numerous agricultural crops and vegetables, causing direct damage to food plants and indirect damage through the transmission of phytopathogenic viruses, primarily begomoviruses. In Panama, particularly in the Azuero region, viral infections transmitted by this aphid can affect a substantial share of tomato crops cultivated for industrial use. A traditional alternative to synthetic pesticides involves exploring plant extracts with insecticidal properties derived from wild plants found in our tropical forests, which can be easily prepared and applied by farmers. In this context, the present research aimed to evaluate the insecticidal activity of ethanolic extracts from the stems and leaves of Sphagneticola trilobata on both nymphs and adults of A. gossypii. Mortality was assessed at 24, 48, and 72 h after applying three doses of each extract (25, 50, and 100 µg/L). A standard phytochemical analysis to determine insecticidal activity revealed that both extracts exhibited significant efficacy at the highest concentration tested; however, the leaf extract demonstrated greater effectiveness at lower concentrations. A comprehensive metabolomic study indicated that the active compounds are diterpenes derived from the pimarenyl cation. These compounds have been extensively documented for their insecticidal potential against various insect species, suggesting that ethanolic extracts from this plant could serve as viable candidates for agricultural insecticides to combat aphid infestations. Full article
(This article belongs to the Special Issue Sustainable Strategies for Managing Plant Diseases)
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17 pages, 449 KiB  
Article
Immunotoxicity Studies on the Insecticide 2-((1-(4-Phenoxyphenoxy)propan-2-yl)oxy)pyridine (MPEP) in Hsd:Harlan Sprague Dawley SD® Rats
by Victor J. Johnson, Stefanie C. M. Burleson, Michael I. Luster, Gary R. Burleson, Barry McIntyre, Veronica G. Robinson, Reshan A. Fernando, James Blake, Donna Browning, Stephen Cooper, Shawn Harris and Dori R. Germolec
Toxics 2025, 13(7), 600; https://doi.org/10.3390/toxics13070600 - 17 Jul 2025
Viewed by 570
Abstract
The broad-spectrum insect growth regulator (IGR) and insecticide 2-((1-(4-Phenoxyphenoxy)propan-2-yl)oxy)pyridine (MPEP; also known as pyriproxyfen) is increasingly being used to address public health programs for vector control, initiated by the spread of Zika virus in 2015–2016. While considered relatively safe for humans under normal [...] Read more.
The broad-spectrum insect growth regulator (IGR) and insecticide 2-((1-(4-Phenoxyphenoxy)propan-2-yl)oxy)pyridine (MPEP; also known as pyriproxyfen) is increasingly being used to address public health programs for vector control, initiated by the spread of Zika virus in 2015–2016. While considered relatively safe for humans under normal conditions, limited toxicology data are available. Current studies were undertaken to address the data gap regarding potential immunotoxicity of MPEP, with particular emphasis on host resistance to viral infection. Hsd:Harlan Sprague Dawley SD® rats were treated for 28 days by oral gavage with doses of 0, 62.5, 125, 250 or 500 mg/kg/day of MPEP in corn oil. There was a dose-dependent increase in liver weights which is consistent with the liver playing a dominant role in MPEP metabolism. However, no histological correlates were observed. Following treatment, rats were subjected to a battery of immune tests as well as an established rat model of influenza virus infection to provide a comprehensive assessment of immune function and host resistance. While several of the immune tests showed minor exposure-related changes, evidenced by negative dose–response trends, most did not show significant differences in any of the MPEP treatment groups relative to vehicle control. Most notable was a negative trend in pulmonary mononuclear cell phagocytosis with increases in dose of MPEP. There was also a positive trend in early humoral immune response (5 days after immunization) to keyhole limpet hemocyanin (KLH) as evidenced by increased serum anti-KLH IgM antibodies which was followed later (14 days following immunization) by decreasing trends in anti-KLH IgM and IgG antibody levels. However, MPEP treatment had no effect on the ability of rats to clear the influenza virus nor the T-dependent IgM and IgG antibody response to the virus. The lack of effects of MPEP on host resistance to influenza suggests the immune effects were minimal and unlikely to present a hazard with respect to susceptibility to respiratory viral infection. Full article
(This article belongs to the Special Issue Environmental Contaminants and Human Health—2nd Edition)
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16 pages, 964 KiB  
Review
Fecal Transmission of Nucleopolyhedroviruses: A Neglected Route to Disease?
by Trevor Williams
Insects 2025, 16(6), 562; https://doi.org/10.3390/insects16060562 - 26 May 2025
Viewed by 542
Abstract
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 [...] Read more.
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
(This article belongs to the Section Insect Behavior and Pathology)
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13 pages, 1104 KiB  
Article
The Individual and Combined Entomopathogenic Activity of a Spodoptera frugiperda Multiple Nucleopolyhedrovirus and a Type I Spodoptera frugiperda Granulovirus on S. frugiperda Larvae
by Magali Ordóñez-García, Juan Carlos Bustillos-Rodríguez, José de Jesús Ornelas-Paz, Miguel Ángel Salas-Marina, Octavio Jhonathan Cambero-Campos, Carlos Horacio Acosta-Muñiz, David Ignacio Berlanga-Reyes and Claudio Rios-Velasco
Viruses 2025, 17(5), 674; https://doi.org/10.3390/v17050674 - 5 May 2025
Viewed by 583
Abstract
The bioinsecticidal activity of several doses of a Spodoptera frugiperda multiple nucleopolyhedrovirus (SfMNPV-CH-32; LD10, LD50, and LD90) and a Type I Spodoptera frugiperda granulovirus (SfGV-CH13; LD50 and LD90), alone and in co-infection, was evaluated [...] Read more.
The bioinsecticidal activity of several doses of a Spodoptera frugiperda multiple nucleopolyhedrovirus (SfMNPV-CH-32; LD10, LD50, and LD90) and a Type I Spodoptera frugiperda granulovirus (SfGV-CH13; LD50 and LD90), alone and in co-infection, was evaluated on S. frugiperda larvae. In the co-infection assays, one virus was applied at 0 h, and then the second virus was supplied at different times (0, 12, and 24 h) in order to test the effect of the co-infection time on the insecticidal activity of the viruses. The symptoms observed in the co-infected larvae depended on the viral dose supplied at 0 h. The larvae treated with the highest dose (LD90) of SfMNPV-CH32 and co-infected with SfGV-CH13 at LD50 showed symptoms of nucleopolyhedrovirus infection at 14 days post-infection. The larvae initially infected with the highest dose of SfGV-CH13 (LD90) and subsequently co-infected with SfMNPV-CH32 (LD50 and LD10) showed infection symptoms characteristic of both viruses. The insecticidal activity of SfGV-CH13 and SfMNPV-CH32 alone or in combination depended on the viral doses and the time elapsed between the first and second inoculation. An antagonistic effect was observed for most of the treatments tested. A synergistic effect was observed only in treatment 10, where the larvae were first infected with SfMNPV-CH32 at a high dose (LD90) and inoculated 24 h later with SfGV-CH13 (LD50). Full article
(This article belongs to the Special Issue Insect Viruses and Pest Management, the Third Edition)
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17 pages, 3408 KiB  
Article
Evaluation of Yellow Fever Virus Infection in Aedes aegypti Mosquitoes from Pakistan with Distinct Knockdown Resistance Genotypes
by Carlucio Rocha dos Santos, Caleb Guedes Miranda dos Santos, Dinair Couto-Lima, Bárbara Silva Souza, Rafi Ur Rahman, Marcos Dornelas Ribeiro, José Bento Pereira Lima and Ademir Jesus Martins
Insects 2025, 16(1), 33; https://doi.org/10.3390/insects16010033 - 31 Dec 2024
Cited by 1 | Viewed by 1375
Abstract
Background: Yellow fever (YF) is an acute hemorrhagic disease endemic to Africa and Latin America; however, no cases have been reported in Asian regions with high Aedes aegypti infestation. Factors such as environmental conditions and genetic variations in the yellow fever virus (YFV) [...] Read more.
Background: Yellow fever (YF) is an acute hemorrhagic disease endemic to Africa and Latin America; however, no cases have been reported in Asian regions with high Aedes aegypti infestation. Factors such as environmental conditions and genetic variations in the yellow fever virus (YFV) strains and mosquito populations may explain this absence. Mosquito populations have undergone strong selective pressure owing to the excessive use of insecticides. This pressure has led to the spread of alterations, such as knockdown-resistant mutations (kdr), which, while conferring resistance to pyrethroids, also induce various physiological side effects in the insect. Therefore, it is important to investigate whether the presence of kdr mutations influences the infectivity of YFV mosquitoes. This study evaluated the susceptibility of Ae. aegypti from Pakistan with distinct kdr genotypes to different YFV strains under laboratory conditions. Methods: Ae. aegypti from a Pakistani colony were exposed to YFV strains (PR4408/2008 and ES504/2017) along with the Rockefeller strain. After 14 days, RNA and DNA were extracted for viral RNA detection (qPCR) and kdr genotyping (TaqMan qPCR and HRM for T1520I and F1534C SNPs). Results: Pakistani Ae. aegypti were orally susceptible to YFV, with infection rates of 83.7% (PR4408/2008) and 61.3% (ES504), respectively, similar to Rockefeller. Two kdr genotypes (II + CC and TI + FC) were identified, with no significant differences in viral infection or dissemination rates. Conclusions: The Ae. aegypti population from Asia is capable of YFV infection and dissemination, regardless of kdr genotype. Full article
(This article belongs to the Section Medical and Livestock Entomology)
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30 pages, 3508 KiB  
Review
The Global Burden of Emerging and Re-Emerging Orbiviruses in Livestock: An Emphasis on Bluetongue Virus and Epizootic Hemorrhagic Disease Virus
by Shanta Barua, Eaftekhar Ahmed Rana, M. Asaduzzaman Prodhan, Syeda Hasina Akter, Jully Gogoi-Tiwari, Subir Sarker, Henry Annandale, Debbie Eagles, Sam Abraham and Jasim M. Uddin
Viruses 2025, 17(1), 20; https://doi.org/10.3390/v17010020 - 26 Dec 2024
Cited by 5 | Viewed by 2562
Abstract
Bluetongue virus (BTV) and epizootic hemorrhagic disease virus (EHDV) are vector-borne orbiviruses that pose an emerging threat to livestock, including cattle and sheep. This review summarizes the global distribution, genetic diversity, and key factors driving their spread along with the existing knowledge gaps [...] Read more.
Bluetongue virus (BTV) and epizootic hemorrhagic disease virus (EHDV) are vector-borne orbiviruses that pose an emerging threat to livestock, including cattle and sheep. This review summarizes the global distribution, genetic diversity, and key factors driving their spread along with the existing knowledge gaps and recommendations to mitigate their impact. Both viruses cause hemorrhagic disease in susceptible ruminants and are commonly reported in tropical and subtropical regions including North America, Asia, Africa, Oceania, and some parts of Europe. The geographical distribution of these viruses, encompassing 27 BTV and 7 EHDV serotypes, has shifted, particularly with the recent invasion of BTV-3, 4, and 8 and EHDV-8 serotypes in Europe. Several factors contribute to the recent spread of these viruses such as the distribution of virulent strains by the movement of temperature-dependent Culicoides vectors into new areas due to rapid climate change, the reassortment of viral strains during mixed infections, and unrestricted global trade. These diseases cause significant economic impacts including morbidity, mortality, reduced production, high management costs, and the disruption of international trade. Effective prevention and control strategies are paramount and rely on vaccination, vector control using insecticides, and the destruction of breeding sites, husbandry practices including the isolation and quarantine of infected hosts, restriction of animal movement, prompt diagnosis and identification of circulating strains, and effective surveillance and monitoring plans such as the pre-export and post-import screening of semen used for artificial insemination. However, challenges remain with intercontinental virus spread, live vaccines, and the failure of inactivated vaccines to produce protective immunity against dissimilar strains. Significant knowledge gaps highlight the need for a better scientific understanding and a strategic plan to ensure healthy livestock and global food security. Full article
(This article belongs to the Section Animal Viruses)
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21 pages, 6259 KiB  
Article
Rapid and Visual Screening of Virus Infection in Sugar Beets Through Polerovirus-Induced Gene Silencing
by Heemee Devi Bunwaree, Elodie Klein, Guillaume Saubeau, Bruno Desprez, Véronique Ziegler-Graff and David Gilmer
Viruses 2024, 16(12), 1823; https://doi.org/10.3390/v16121823 - 23 Nov 2024
Viewed by 1396
Abstract
Since the ban of neonicotinoid insecticides in the European Union, sugar beet production is threatened by outbreaks of virus yellows (VY) disease, caused by several aphid-transmitted viruses, including the polerovirus beet mild yellowing virus (BMYV). As the symptoms induced may vary depending on [...] Read more.
Since the ban of neonicotinoid insecticides in the European Union, sugar beet production is threatened by outbreaks of virus yellows (VY) disease, caused by several aphid-transmitted viruses, including the polerovirus beet mild yellowing virus (BMYV). As the symptoms induced may vary depending on multiple infections and other stresses, there is an urgent need for fast screening tests to evaluate resistance/tolerance traits in sugar beet accessions. To address this issue, we exploited the virus-induced gene silencing (VIGS) system, by introducing a fragment of a Beta vulgaris gene involved in chlorophyll synthesis in the BMYV genome. This recombinant virus was able to generate early clear vein chlorosis symptoms in infected sugar beets, allowing easy and rapid visual discernment of infected plants across five sugar beet lines. The recombinant virus displayed similar infectivity as the wild-type, and the insert remained stable within the viral progeny. We demonstrated that the percentage of VIGS-symptomatic plants was representative of the infection rate of each evaluated line, and depending on the susceptibility of the line to BMYV infection, VIGS symptoms may last over months. Our work provides a polerovirus-based VIGS system adapted to sugar beet crop allowing visual and rapid large-scale screens for resistance or functional genomic studies. Full article
(This article belongs to the Special Issue Application of Genetically Engineered Plant Viruses)
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10 pages, 1430 KiB  
Article
Antiviral Efficacy of Coridothymus capitatus Essential Oil Against HSV-1 and HSV-2
by Virginia Fuochi, Pio Maria Furneri, Salvatore Furnari and Adriana Garozzo
Life 2024, 14(8), 1023; https://doi.org/10.3390/life14081023 - 18 Aug 2024
Cited by 1 | Viewed by 1646
Abstract
Coridothymus capitatus is a perennial herb with aromatic leaves and flowers, distinct from Thymus vulgaris in its chemical composition, resulting in a unique Thymus Essential Oil (TEO). A main component of TEO, carvacrol, is known for its antimicrobial and insecticidal activity. Carvacrol has [...] Read more.
Coridothymus capitatus is a perennial herb with aromatic leaves and flowers, distinct from Thymus vulgaris in its chemical composition, resulting in a unique Thymus Essential Oil (TEO). A main component of TEO, carvacrol, is known for its antimicrobial and insecticidal activity. Carvacrol has potent antibacterial, antioxidant, anti-inflammatory, and antifungal properties, generating interest in traditional medicine. However, studies on its antiviral activity are limited. Given the rise in viral infections and limitations of synthetic antiviral drugs, natural antiviral agents are promising due to their efficacy, lower resistance development, and reduced side effects. This study assessed the antiviral efficacy of TEO compared to that of pure carvacrol. We tested various viruses, revealing significant inhibitory effects of TEO on the replication of only Simplexvirus humanalpha1 (HSV-1) and Simplexvirus humanalpha2 (HSV-2), with specific interference during the early stages of the viral replication cycle after the adsorption period. TEO exhibited inhibitory effects at doses below the cytotoxic threshold, with IC50 values of 47 μg/mL for HSV-1 and 40 μg/mL for HSV-2. Maximum virus inhibition was achieved when TEO was added within 90 min post-infection, indicating interference with early viral replication steps. These findings highlight the potential of TEO as a natural antiviral agent and suggest further research into its mechanisms and clinical applications. Full article
(This article belongs to the Special Issue Development and Application of Antimicrobials)
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17 pages, 4117 KiB  
Article
Evaluation of Integrated Pest and Disease Management Combinations against Major Insect Pests and Diseases of Tomato in Tamil Nadu, India
by Sankaran Pagalahalli Shanmugam, Marimuthu Murugan, Mookiah Shanthi, Thiyagarajan Elaiyabharathi, Kathithachalam Angappan, Gandhi Karthikeyan, Gopal Arulkumar, Palanisamy Manjari, Manickam Ravishankar, Paola Sotelo-Cardona, Ricardo Oliva and Ramasamy Srinivasan
Horticulturae 2024, 10(7), 766; https://doi.org/10.3390/horticulturae10070766 - 19 Jul 2024
Cited by 9 | Viewed by 4353
Abstract
Tomatoes are one of the predominant vegetable crops grown throughout the year in Tamil Nadu, India. Their perishable nature and resource-intensive cultivation make them susceptible to biotic stress. The damage caused by invasive insect pests, bacterial wilt during the rainy season, and viral [...] Read more.
Tomatoes are one of the predominant vegetable crops grown throughout the year in Tamil Nadu, India. Their perishable nature and resource-intensive cultivation make them susceptible to biotic stress. The damage caused by invasive insect pests, bacterial wilt during the rainy season, and viral diseases are major yield-limiting factors, and the farmers mostly depend on calendar-based insecticide applications for insect pest and disease management in tomatoes. The desired tomato hybrids grafted onto bacterial wilt-resistant eggplant rootstocks offer protection against bacterial wilt during the rainy season. The integrated pest and disease management (IPDM) practices consist of resistant grafted tomato seedlings (wild eggplant rootstocks EG 203 and TS 03), bioinoculants (Bacillus subtilis + Trichoderma asperellum + Purpureocillium lilacinum), pheromone traps (Phthorimaea absoluta and Helicoverpa armigera), botanicals (azadirachtin), microbial pesticides (Bacillus thuringiensis, Metarhizium anisopliae, and Beauveria bassiana), and bio-rationals, which were evaluated in four locations in two major tomato-growing tracts of Tamil Nadu. The results revealed that the treatment EG 203 eggplant rootstock-grafted tomato along with IPDM practices performed better across all experimental locations than the other treatment combinations viz., TS 03 eggplant rootstock-grafted tomato + IPDM, tomato + IPDM, grafted tomato + farmers’ practice and tomato + farmers’ practice. The EG 203-grafted tomato recorded a higher yield than the farmers’ practice with significantly superior biometric parameters. The treatment of EG 203-grafted tomato and IPDM practices can be adopted for safer tomato production by enabling a reduction in pesticide applications while enhancing productivity. Full article
(This article belongs to the Section Insect Pest Management)
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13 pages, 800 KiB  
Article
A qPCR Assay for the Quantification of Selected Genotypic Variants of Spodoptera frugiperda Multiple Nucleopolyhedrovirus (Baculoviridae)
by Cindy S. Molina-Ruiz, Jesús Alejandro Zamora-Briseño, Oihane Simón, Rodrigo Lasa and Trevor Williams
Viruses 2024, 16(6), 881; https://doi.org/10.3390/v16060881 - 30 May 2024
Cited by 2 | Viewed by 1444
Abstract
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 [...] Read more.
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
(This article belongs to the Section Invertebrate Viruses)
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13 pages, 2079 KiB  
Article
High Frequencies of kdr Mutation and Chikungunya Infection in Aedes aegypti Population from Minas Gerais, Brazil
by Pedro Augusto Almeida-Souza, Cirilo Henrique de Oliveira, Luiz Paulo Brito, Thaynara de Jesus Teixeira, Iago Alves Celestino, Gabriele Barbosa Penha, Ronaldo Medeiros dos Santos, Wexley Miranda Mendes, Bergmann Morais Ribeiro, Fabrício Souza Campos, Paulo Michel Roehe, Natalia Rocha Guimarães, Felipe C. M. Iani, Ademir Jesus Martins and Filipe Vieira Santos de Abreu
Pathogens 2024, 13(6), 457; https://doi.org/10.3390/pathogens13060457 - 28 May 2024
Cited by 3 | Viewed by 3378
Abstract
The Chikungunya virus (CHIKV) presents global health challenges, with Brazil experiencing outbreaks since its introduction in 2014. In 2023, following a CHIKV outbreak in Minas Gerais (MG), social media was used to optimize an entomological survey aimed at identifying vectors and viral lineages [...] Read more.
The Chikungunya virus (CHIKV) presents global health challenges, with Brazil experiencing outbreaks since its introduction in 2014. In 2023, following a CHIKV outbreak in Minas Gerais (MG), social media was used to optimize an entomological survey aimed at identifying vectors and viral lineages and assessing insecticide resistance. Following Instagram posts, residents with suspected CHIKV infection were able to schedule mosquito aspirations. In total, 421 mosquitoes (165 Aedes aegypti and 256 Culex quinquefasciatus) were captured from 40 households in Salinas city (MG) and tested for the Dengue, Zika, and Chikungunya viruses through RT-qPCR. Twelve of 57 pools (10 Ae. aegypti and two Cx. quinquefasciatus) tested positive for CHIKV RNA. Viral RNA was also detected in the heads of nine Ae. aegypti, indicating viral dissemination but not in Cx. quinquefasciatus. Genome sequencing yielded the first near-complete genome from the 2023 outbreak, unveiling that the CHIKV strain belonged to the East/Central/South African (ECSA) genotype. Additionally, genetic analyses revealed high frequencies of kdr alleles, including in CHIKV-infected mosquitoes, suggesting resistance to pyrethroid insecticides in this Ae. aegypti population. Social media was important for guiding mosquito-capture efforts in CHIKV transmission hotspots, thus optimizing the opportunity for viral detection. These findings emphasize the urgent need for innovative vector studies and control strategies, as well as interdisciplinary approaches in public health interventions. Full article
(This article belongs to the Special Issue Molecular Epidemiology of Emerging Viruses)
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15 pages, 6933 KiB  
Article
Managing Super Pests: Interplay between Pathogens and Symbionts Informs Biocontrol of Whiteflies
by Weili Yan, Saixian Wang, Jialei Liu, Dan Zhai, Hang Lu, Jingjing Li, Rune Bai, Caiyan Lei, Luyang Song, Chenchen Zhao and Fengming Yan
Microorganisms 2024, 12(5), 887; https://doi.org/10.3390/microorganisms12050887 - 28 Apr 2024
Cited by 1 | Viewed by 2027
Abstract
Bemisia tabaci is distributed globally and incurs considerable economic and ecological costs as an agricultural pest and viral vector. The entomopathogenic fungus Metarhizium anisopliae has been known for its insecticidal activity, but its impacts on whiteflies are understudied. We investigated how infection with [...] Read more.
Bemisia tabaci is distributed globally and incurs considerable economic and ecological costs as an agricultural pest and viral vector. The entomopathogenic fungus Metarhizium anisopliae has been known for its insecticidal activity, but its impacts on whiteflies are understudied. We investigated how infection with the semi-persistently transmitted Cucurbit chlorotic yellows virus (CCYV) affects whitefly susceptibility to M. anisopliae exposure. We discovered that viruliferous whiteflies exhibited increased mortality when fungus infection was present compared to non-viruliferous insects. High throughput 16S rRNA sequencing also revealed significant alterations of the whitefly bacterial microbiome diversity and structure due to both CCYV and fungal presence. Specifically, the obligate symbiont Portiera decreased in relative abundance in viruliferous whiteflies exposed to M. anisopliae. Facultative Hamiltonella and Rickettsia symbionts exhibited variability across groups but dominated in fungus-treated non-viruliferous whiteflies. Our results illuminate triangular interplay between pest insects, their pathogens, and symbionts—dynamics which can inform integrated management strategies leveraging biopesticides This work underscores the promise of M. anisopliae for sustainable whitefly control while laying the groundwork for elucidating mechanisms behind microbe-mediated shifts in vector competence. Full article
(This article belongs to the Special Issue Plant Pathogens: Monitoring, Identification and Biological Control)
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26 pages, 1978 KiB  
Systematic Review
Evaluating the Metabolomic Profile and Anti-Pathogenic Properties of Cannabis Species
by Shadrack Monyela, Prudence Ngalula Kayoka, Wonder Ngezimana and Lufuno Ethel Nemadodzi
Metabolites 2024, 14(5), 253; https://doi.org/10.3390/metabo14050253 - 26 Apr 2024
Cited by 5 | Viewed by 3149
Abstract
The Cannabis species is one of the potent ancient medicinal plants acclaimed for its medicinal properties and recreational purposes. The plant parts are used and exploited all over the world for several agricultural and industrial applications. For many years Cannabis spp. has proven [...] Read more.
The Cannabis species is one of the potent ancient medicinal plants acclaimed for its medicinal properties and recreational purposes. The plant parts are used and exploited all over the world for several agricultural and industrial applications. For many years Cannabis spp. has proven to present a highly diverse metabolomic profile with a pool of bioactive metabolites used for numerous pharmacological purposes ranging from anti-inflammatory to antimicrobial. Cannabis sativa has since been an extensive subject of investigation, monopolizing the research. Hence, there are fewer studies with a comprehensive understanding of the composition of bioactive metabolites grown in different environmental conditions, especially C. indica and a few other Cannabis strains. These pharmacological properties are mostly attributed to a few phytocannabinoids and some phytochemicals such as terpenoids or essential oils which have been tested for antimicrobial properties. Many other discovered compounds are yet to be tested for antimicrobial properties. These phytochemicals have a series of useful properties including anti-insecticidal, anti-acaricidal, anti-nematicidal, anti-bacterial, anti-fungal, and anti-viral properties. Research studies have reported excellent antibacterial activity against Gram-positive and Gram-negative multidrug-resistant bacteria as well as methicillin-resistant Staphylococcus aureus (MRSA). Although there has been an extensive investigation on the antimicrobial properties of Cannabis, the antimicrobial properties of Cannabis on phytopathogens and aquatic animal pathogens, mostly those affecting fish, remain under-researched. Therefore, the current review intends to investigate the existing body of research on metabolomic profile and anti-microbial properties whilst trying to expand the scope of the properties of the Cannabis plant to benefit the health of other animal species and plant crops, particularly in agriculture. Full article
(This article belongs to the Special Issue Metabolomics in Plant Natural Products Research)
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13 pages, 1858 KiB  
Article
Nicotinic Acetylcholine Receptor Alpha6 Contributes to Antiviral Immunity via IMD Pathway in Drosophila melanogaster
by Zhiying Wang, Xiaoju Lin, Wangpeng Shi and Chuan Cao
Viruses 2024, 16(4), 562; https://doi.org/10.3390/v16040562 - 3 Apr 2024
Cited by 3 | Viewed by 1896 | Correction
Abstract
Currently, insecticides that target nicotinic acetylcholine receptors (nAChR) are widely used. Studies on the sublethal effects of insecticides have found that they can affect the amount of virus in insects. The mechanism by which insecticides affect insect virus load remain unclear. Here, we [...] Read more.
Currently, insecticides that target nicotinic acetylcholine receptors (nAChR) are widely used. Studies on the sublethal effects of insecticides have found that they can affect the amount of virus in insects. The mechanism by which insecticides affect insect virus load remain unclear. Here, we show that nAChR targeting insecticide can affect viral replication through the immune deficiency (IMD) pathway. We demonstrate that a low dose of spinosad (6.8 ng/mL), acting as an antagonist to Drosophila melanogaster nicotinic acetylcholine receptor α6 (Dα6), significantly elevates Drosophila melanogaster sigmavirus (DMelSV) virus titers in adults of Drosophila melanogaster. Conversely, a high dose of spinosad (50 ng/mL), acting as an agonist to Dα6, substantially decreases viral load. This bidirectional regulation of virus levels is absent in Dα6-knockout flies, signifying the specificity of spinosad’s action through Dα6. Furthermore, the knockdown of Dα6 results in decreased expression of genes in the IMD pathway, including dredd, imd, relish, and downstream antimicrobial peptide genes AttA and AttB, indicating a reduced innate immune response. Subsequent investigations reveal no significant difference in viral titers between relish mutant flies and Dα6-relish double mutants, suggesting that the IMD pathway’s role in antiviral defense is dependent on Dα6. Collectively, our findings shed light on the intricate interplay between nAChR signaling and the IMD pathway in mediating antiviral immunity, highlighting the potential for nAChR-targeting compounds to inadvertently influence viral dynamics in insect hosts. This knowledge may inform the development of integrated pest management strategies that consider the broader ecological impact of insecticide use. Full article
(This article belongs to the Special Issue Molecular Virus-Insect Interactions)
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