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1.
Environ Pollut ; 292(Pt A): 118212, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34582921

RESUMO

Varroa mite is one of the major adverse factors causing honey bee population decline. In this study, Varroa destructor-infested and uninfested honey bee colonies were established by selective applying miticide (Apivar® amitraz). Mite population was monitored monthly (April-October 2016), and deformed wing virus (DWV) loading was detected seasonally (April, July, and October). Four immunity- and two physiology-related gene expressions, natural mortality, and susceptibility to five insecticides were comparatively and seasonally examined in field-collected honey bee workers. Results showed that Apivar-treated bee colonies had minor or undetectable mite and DWV (using RT-qPCR) infestations in whole bee season, while untreated colonies had substantially higher mite and DWV infestations. In untreated colonies, Varroa mite population irregularly fluctuated over the bee season with higher mite counts in Jun (318 ± 89 mites dropped in 48 h) or August (302) than that (25 ± 4 or 34) in October, and mite population density was not dynamically or closely correlated with the seasonal shift of honey bee natural mortality (regression slope = -0.5212). Unlike mite, DWV titer in untreated colonies progressively increased over the bee season, and it was highly correlated (R2 = 1) with the seasonal increase of honey bee natural mortality. Significantly lower gene expressions of dor, PPO, mfe, potentially PPOa and eat as well, in untreated colonies also indicated an association of increased DWV infestation with decreased physiological and immunity-related functions in late-season honey bees. Furthermore, bees with lower mite/DWV infestations exhibited generally consistently lower susceptibilities (contact and oral toxicities) to five representative insecticides than the bees without Apivar treatment. All of these data from this study consistently indicated an interaction of Varroa/viral infestations with insecticide susceptibilities in honey bees, potentially through impairing bee's physiology and immunity, emphasizing the importance of mite control in order to minimize honey bee decline.


Assuntos
Acaricidas , Inseticidas , Vírus de RNA , Varroidae , Acaricidas/toxicidade , Animais , Abelhas , Inseticidas/toxicidade
2.
J Gen Virol ; 102(10)2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34704919

RESUMO

The endosymbiont bacteria of the genus Wolbachia are associated with multiple mutualistic effects on insect biology, including nutritional and antiviral properties. Members of the genus Wolbachia naturally occur in fly species of the genus Drosophila, providing an operational model host for studying how virome composition may be affected by its presence. Drosophila simulans populations can carry a variety of strains of members of the genus Wolbachia, with the wAu strain associated with strong antiviral protection under experimental conditions. We used D. simulans sampled from the Perth Hills, Western Australia, to investigate the potential virus protective effect of the wAu strain of Wolbachia on individual wild-caught flies. Our data revealed no appreciable variation in virus composition and abundance between individuals infected or uninfected with Wolbachia associated with the presence or absence of wAu. However, it remains unclear whether wAu might affect viral infection and host survival by increasing tolerance rather than inducing complete resistance. These data also provide new insights into the natural virome diversity of D. simulans. Despite the small number of individuals sampled, we identified a repertoire of RNA viruses, including nora virus, galbut virus, thika virus and La Jolla virus, that have been identified in other species of the genus Drosophila. Chaq virus-like sequences associated with galbut virus were also detected. In addition, we identified five novel viruses from the families Reoviridae, Tombusviridae, Mitoviridae and Bunyaviridae. Overall, this study highlights the complex interaction between Wolbachia and RNA virus infections and provides a baseline description of the natural virome of D. simulans.


Assuntos
Drosophila simulans/microbiologia , Vírus de RNA/fisiologia , Viroma/fisiologia , Wolbachia/fisiologia , Animais , Drosophila simulans/virologia , Feminino , Filogenia , Vírus de RNA/classificação , Vírus de RNA/genética , Vírus de RNA/isolamento & purificação , Simbiose , Viroma/genética , Wolbachia/isolamento & purificação
3.
PLoS Pathog ; 17(10): e1009841, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34648591

RESUMO

In general, in mammalian cells, cytosolic DNA viruses are sensed by cyclic GMP-AMP synthase (cGAS), and RNA viruses are recognized by retinoic acid-inducible gene I (RIG-I)-like receptors, triggering a series of downstream innate antiviral signaling steps in the host. We previously reported that measles virus (MeV), which possesses an RNA genome, induces rapid antiviral responses, followed by comprehensive downregulation of host gene expression in epithelial cells. Interestingly, gene ontology analysis indicated that genes encoding mitochondrial proteins are enriched among the list of downregulated genes. To evaluate mitochondrial stress after MeV infection, we first observed the mitochondrial morphology of infected cells and found that significantly elongated mitochondrial networks with a hyperfused phenotype were formed. In addition, an increased amount of mitochondrial DNA (mtDNA) in the cytosol was detected during progression of infection. Based on these results, we show that cytosolic mtDNA released from hyperfused mitochondria during MeV infection is captured by cGAS and causes consequent priming of the DNA sensing pathway in addition to canonical RNA sensing. We also ascertained the contribution of cGAS to the in vivo pathogenicity of MeV. In addition, we found that other viruses that induce downregulation of mitochondrial biogenesis as seen for MeV cause similar mitochondrial hyperfusion and cytosolic mtDNA-priming antiviral responses. These findings indicate that the mtDNA-activated cGAS pathway is critical for full innate control of certain viruses, including RNA viruses that cause mitochondrial stress.


Assuntos
Imunidade Inata/imunologia , Sarampo/metabolismo , Mitocôndrias/metabolismo , Nucleotidiltransferases/metabolismo , Animais , Regulação para Baixo , Humanos , Vírus do Sarampo , Camundongos , Camundongos Endogâmicos C57BL , Mitocôndrias/virologia , Biogênese de Organelas , Infecções por Vírus de RNA/metabolismo , Vírus de RNA
4.
Genome Biol Evol ; 13(11)2021 11 05.
Artigo em Inglês | MEDLINE | ID: mdl-34694399

RESUMO

Adenosine Deaminases that Act on RNA (ADARs) are RNA editing enzymes that play a dynamic and nuanced role in regulating transcriptome and proteome diversity. This editing can be highly selective, affecting a specific site within a transcript, or nonselective, resulting in hyperediting. ADAR editing is important for regulating neural functions and autoimmunity, and has a key role in the innate immune response to viral infections, where editing can have a range of pro- or antiviral effects and can contribute to viral evolution. Here we examine the role of ADAR editing across a broad range of viral groups. We propose that the effect of ADAR editing on viral replication, whether pro- or antiviral, is better viewed as an axis rather than a binary, and that the specific position of a given virus on this axis is highly dependent on virus- and host-specific factors, and can change over the course of infection. However, more research needs to be devoted to understanding these dynamic factors and how they affect virus-ADAR interactions and viral evolution. Another area that warrants significant attention is the effect of virus-ADAR interactions on host-ADAR interactions, particularly in light of the crucial role of ADAR in regulating neural functions. Answering these questions will be essential to developing our understanding of the relationship between ADAR editing and viral infection. In turn, this will further our understanding of the effects of viruses such as SARS-CoV-2, as well as many others, and thereby influence our approach to treating these deadly diseases.


Assuntos
Adenosina Desaminase/metabolismo , Edição de RNA , Vírus de RNA/genética , Adenosina Desaminase/genética , Animais , Evolução Molecular , Interações Hospedeiro-Patógeno/imunologia , Humanos , Imunidade , Vírus de RNA/classificação , Vírus de RNA/fisiologia , RNA Viral/genética , RNA Viral/metabolismo , Replicação Viral/genética
5.
BMC Genomics ; 22(1): 720, 2021 Oct 06.
Artigo em Inglês | MEDLINE | ID: mdl-34610790

RESUMO

BACKGROUND: Varroa destructor mites, and the numerous viruses they vector to their honey bee hosts, are among the most serious threats to honey bee populations, causing mortality and morbidity to both the individual honey bee and colony, the negative effects of which convey to the pollination services provided by honey bees worldwide. Here we use a combination of targeted assays and deep RNA sequencing to determine host and microbial changes in resistant and susceptible honey bee lineages. We focus on three study sets. The first involves field sampling of sympatric western bees, some derived from resistant stock and some from stock susceptible to mites. The second experiment contrasts three colonies more deeply, two from susceptible stock from the southeastern U.S. and one from mite-resistant bee stock from Eastern Texas. Finally, to decouple the effects of mites from those of the viruses they vector, we experimentally expose honey bees to DWV in the laboratory, measuring viral growth and host responses. RESULTS: We find strong differences between resistant and susceptible bees in terms of both viral loads and bee gene expression. Interestingly, lineages of bees with naturally low levels of the mite-vectored Deformed wing virus, also carried lower levels of viruses not vectored by mites. By mapping gene expression results against current ontologies and other studies, we describe the impacts of mite parasitism, as well as viruses on bee health against two genetic backgrounds. We identify numerous genes and processes seen in other studies of stress and disease in honey bee colonies, alongside novel genes and new patterns of expression. CONCLUSIONS: We provide evidence that honey bees surviving in the face of parasitic mites do so through their abilities to resist the presence of devastating viruses vectored by these mites. In all cases, the most divergence between stocks was seen when bees were exposed to live mites or viruses, suggesting that gene activation, rather than constitutive expression, is key for these interactions. By revealing responses to viral infection and mite parasitism in different lineages, our data identify candidate proteins for the evolution of mite tolerance and virus resistance.


Assuntos
Vírus de RNA , Varroidae , Viroses , Animais , Abelhas , Vírus de RNA/genética , Carga Viral
6.
PLoS Pathog ; 17(10): e1009726, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34695163

RESUMO

The zinc finger antiviral protein (ZAP) is a broad inhibitor of virus replication. Its best-characterized function is to bind CpG dinucleotides present in viral RNAs and, through the recruitment of TRIM25, KHNYN and other cofactors, target them for degradation or prevent their translation. The long and short isoforms of ZAP (ZAP-L and ZAP-S) have different intracellular localization and it is unclear how this regulates their antiviral activity against viruses with different sites of replication. Using ZAP-sensitive and ZAP-insensitive human immunodeficiency virus type I (HIV-1), which transcribe the viral RNA in the nucleus and assemble virions at the plasma membrane, we show that the catalytically inactive poly-ADP-ribose polymerase (PARP) domain in ZAP-L is essential for CpG-specific viral restriction. Mutation of a crucial cysteine in the C-terminal CaaX box that mediates S-farnesylation and, to a lesser extent, the residues in place of the catalytic site triad within the PARP domain, disrupted the activity of ZAP-L. Addition of the CaaX box to ZAP-S partly restored antiviral activity, explaining why ZAP-S lacks antiviral activity for CpG-enriched HIV-1 despite conservation of the RNA-binding domain. Confocal microscopy confirmed the CaaX motif mediated localization of ZAP-L to vesicular structures and enhanced physical association with intracellular membranes. Importantly, the PARP domain and CaaX box together jointly modulate the interaction between ZAP-L and its cofactors TRIM25 and KHNYN, implying that its proper subcellular localisation is required to establish an antiviral complex. The essential contribution of the PARP domain and CaaX box to ZAP-L antiviral activity was further confirmed by inhibition of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) replication, which replicates in double-membrane vesicles derived from the endoplasmic reticulum. Thus, compartmentalization of ZAP-L on intracellular membranes provides an essential effector function in ZAP-L-mediated antiviral activity against divergent viruses with different subcellular replication sites.


Assuntos
Prenilação/fisiologia , Vírus de RNA/efeitos dos fármacos , Proteínas de Ligação a RNA/farmacologia , Replicação Viral/fisiologia , Ilhas de CpG/fisiologia , Células HEK293 , HIV-1/fisiologia , Células HeLa , Humanos , Vírus de RNA/fisiologia , RNA Viral/química , RNA Viral/metabolismo , Motivos de Ligação ao RNA/fisiologia , Proteínas de Ligação a RNA/química , Proteínas de Ligação a RNA/metabolismo , SARS-CoV-2/fisiologia , Transfecção , Replicação Viral/efeitos dos fármacos
7.
Enzymes ; 49: 83-113, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34696840

RESUMO

DNA and RNA viruses depend on one or more enzymes to copy and transcribe their genome, such as a polymerase, helicase, or exonuclease. Because of the important role of these enzymes in the virus replication cycle, they are key targets for antiviral development. To better understand the function of these enzymes and their interactions with host and viral factors, biochemical, structural and single-molecule approaches have been used to study them. Each of these techniques has its own strengths, and single-molecule methods have proved particularly powerful in providing insight into the step-sizes of motor proteins, heterogeneity of enzymatic activities, transient conformational changes, and force-sensitivity of reactions. Here we will discuss how single-molecule FRET, magnetic tweezers, optical tweezers, atomic force microscopy and flow stretching approaches have revealed novel insights into polymerase fidelity, the mechanism of action of antivirals, and the protein choreography within replication complexes.


Assuntos
Vírus de DNA , Vírus de RNA , Replicação Viral , Antivirais , DNA Helicases , Vírus de DNA/enzimologia , Vírus de DNA/fisiologia , Pinças Ópticas , Vírus de RNA/enzimologia , Vírus de RNA/fisiologia
8.
J Anim Ecol ; 90(10): 2230-2233, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34609752

RESUMO

Research Highlight: Norton, A. M., Remnant, E. J., Tom, J., Buchmann, G., Blacquiere, T., & Beekman, M. (2021). Adaptation to vector-based transmission in a honeybee virus. Journal of Animal Ecology, 90, https://besjournals.onlinelibrary.wiley.com/doi/10.1111/1365-2656.13493. In their paper on the adaptation to vector-based transmission via the mite Varroa destructor in a honeybee virus, Norton et al. study how high versus low levels of a viral vector affect viral load and potential competition between two strains of Deformed Wing Virus, an important highly virulent bee virus with the potential to spill-over into other pollinators and bee-associated insect species. This paper addresses two very timely issues, on the one hand on viral evolutionary ecology in response to vector-borne transmission, and on the other hand providing much needed information on an important honey bee pathogen. Using a complex natural system, this study shows that vector-borne transmission, and the control of the vector, can select for complex host-pathogen-vector interactions and that adaptations to changing transmission landscapes in fast evolving pathogens can create conditions for emerging pathogens to transition to endemic diseases.


Assuntos
Vírus de RNA , Varroidae , Aclimatação , Animais , Abelhas , Interações Hospedeiro-Patógeno
9.
Enzymes ; 49: 149-193, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34696831

RESUMO

All RNA viruses encode an RNA-dependent RNA polymerase (RdRp) responsible for genome replication. It is now recognized that enzymes in general, and RdRps specifically, are dynamic macromolecular machines such that their moving parts, including active site loops, play direct functional roles. While X-ray crystallography has provided deep insight into structural elements important for RdRp function, this methodology generally provides only static snapshots, and so is limited in its ability to report on dynamic fluctuations away from the lowest energy conformation. Nuclear magnetic resonance (NMR), molecular dynamics (MD) simulations and other biophysical techniques have brought new insight into RdRp function by their ability to characterize the trajectories, kinetics and thermodynamics of conformational motions. In particular, these methodologies have identified coordinated motions among conserved structural motifs necessary for nucleotide selection and incorporation. Disruption of these motions through amino acid substitutions or inhibitor binding impairs RdRp function. Understanding and re-engineering these motions thus provides exciting new avenues for anti-viral strategies. This chapter outlines the basics of these methodologies, summarizes the dynamic motions observed in different RdRps important for nucleotide selection and incorporation, and illustrates how this information can be leveraged towards rational vaccine strain development and anti-viral drug design.


Assuntos
Vírus de RNA , RNA Polimerase Dependente de RNA , Domínio Catalítico , Cristalografia por Raios X , Simulação de Dinâmica Molecular , RNA Polimerase Dependente de RNA/genética
10.
Artigo em Alemão | MEDLINE | ID: mdl-34666368

RESUMO

OBJECTIVE: Chronic bee paralysis virus (CBPV), a so far unclassified RNA virus that may cause neurological signs and hairless black syndrome in honey bees, has become increasingly prevalent in various European countries in recent years. The disease occasionally leads to immense losses of worker bees and a significant drop in performance, especially in strong colonies. The aim of this retrospective study was to analyse the spread of CBPV in Bavaria from 2018 to 2020 as well as to evaluate therapeutic measures. MATERIAL AND METHODS: In part 1 of the study, analysis results of the Tiergesundheitsdienst Bayern e. V. (Bavarian Animal Health Service) from 302 bee colonies were examined with regard to virus diagnostics and clinical signs with a focus on CBPV. In part 2, data collected with the help of a questionnaire regarding 105 CBPV-positive and symptomatic colonies were evaluated. RESULTS: In part 1, a significant (p = 0.004) increase in CBPV-positive samples from 2018 to 2020 was detected within Bavaria. In addition, the number of cases with clinical signs also increased during the last 2 years. Part 2 of the study showed a clustering of reports of initial onset of CBPV symptoms during springtime. The colonies mostly with a combination of symptoms recovered from the disease in 57 % of the cases. Therapeutic intervention was one decisive factor. A large number of different combinations of measures led to survival, and a lack of intervention mostly resulted in the death of the colony. In 62 % of the diseased and isolated colonies, isolation prevented further spread in the apiary. CONCLUSION UND CLINICAL RELEVANCE: Based on the results, CBPV occurred more frequently in connection with clinical signs in Bavaria in recent years. Due to the large number of applied combinations of measures, no evidence-based therapy recommendation may be derived based on the current evaluations. However, it has been shown that any therapeutic intervention is superior to withholding treatment. For precise therapy recommendations, trials with treatment under standard conditions are necessary.


Assuntos
Vírus de RNA , Animais , Abelhas , Incidência , Paralisia/veterinária , Estudos Retrospectivos
11.
Braz J Microbiol ; 52(4): 2475-2482, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34562234

RESUMO

Bufotenine, an alkaloid that can be found in plant extracts and skin secretions of amphibians, is reported to have potential antiviral activity. The present study evaluated the antiviral activity of bufotenine against different genetic lineages of rabies virus (RABV, a single-stranded, negative-sense RNA virus), canine coronavirus (CCoV, a positive-sense RNA virus) and two double-stranded DNA viruses (two strains of herpes simplex virus type 1/HSV-1 [KOS and the acyclovir-resistant HSV-1 strain 29R] and canine adenovirus 2, CAV-2). The maximal non-toxic bufotenine concentrations in Vero and BHK-21 cells were determined by MTT assays. The antiviral activity of bufotenine against each virus was assessed by examination of reductions in infectious virus titres and plaque assays. All experiments were performed with and without bufotenine, and the results were compared. Bufotenine demonstrated significant RABV inhibitory activity. No antiviral action was observed against CCoV, CAV-2 or HSV-1. These findings indicate that the antiviral activity of bufotenine is somewhat linked to the particular infectious dose used and the genetic lineage of the virus, although the mechanisms of its effects remain undetermined.


Assuntos
Antivirais , Bufotenina , Vírus de DNA/efeitos dos fármacos , Vírus de RNA/efeitos dos fármacos , Animais , Antivirais/farmacologia , Bufotenina/farmacologia , Chlorocebus aethiops , Cricetinae , Células Vero
12.
PLoS Pathog ; 17(9): e1009901, 2021 09.
Artigo em Inglês | MEDLINE | ID: mdl-34506605

RESUMO

Neddylation, an important type of post-translational modification, has been implicated in innate and adapted immunity. But the role of neddylation in innate immune response against RNA viruses remains elusive. Here we report that neddylation promotes RNA virus-induced type I IFN production, especially IFN-α. More importantly, myeloid deficiency of UBA3 or NEDD8 renders mice less resistant to RNA virus infection. Neddylation is essential for RNA virus-triggered activation of Ifna gene promoters. Further exploration has revealed that mammalian IRF7undergoes neddylation, which is enhanced after RNA virus infection. Even though neddylation blockade does not hinder RNA virus-triggered IRF7 expression, IRF7 mutant defective in neddylation exhibits reduced ability to activate Ifna gene promoters. Neddylation blockade impedes RNA virus-induced IRF7 nuclear translocation without hindering its phosphorylation and dimerization with IRF3. By contrast, IRF7 mutant defective in neddylation shows enhanced dimerization with IRF5, an Ifna repressor when interacting with IRF7. In conclusion, our data demonstrate that myeloid neddylation contributes to host anti-viral innate immunity through targeting IRF7 and promoting its transcriptional activity.


Assuntos
Imunidade Inata/imunologia , Fator Regulador 7 de Interferon/imunologia , Células Mieloides/imunologia , Infecções por Vírus de RNA/imunologia , Vírus de RNA/imunologia , Animais , Fator Regulador 7 de Interferon/biossíntese , Camundongos , Células Mieloides/metabolismo , Proteína NEDD8/deficiência , Processamento de Proteína Pós-Traducional , Ubiquitinas/deficiência
14.
J Fish Dis ; 44(12): 2097-2109, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34477227

RESUMO

Tilapia tilapinevirus (also known as tilapia lake virus, TiLV) is considered to be a new threat to the global tilapia industry. The objective of this study was to develop simple cell culture-based heat-killed (HKV) and formalin-killed (FKV) vaccines for the prevention of disease caused by TiLV. The fish were immunized with 100 µl of either HKV or FKV by intraperitoneal injection with each vaccine containing 1.8 × 106 TCID50- inactivated virus. A booster vaccination was carried out at 21-day post-vaccination (dpv) using the same protocol. The fish were then challenged with a lethal dose of TiLV at 28 dpv. The expression of five immune genes (IgM, IgD, IgT, CD4 and CD8) in the head kidney and spleen of experimental fish was assessed at 14 and 21 dpv and again after the booster vaccination at 28 dpv. TiLV-specific IgM responses were measured by ELISA at the same time points. The results showed that both vaccines conferred significant protection, with relative percentage survival of 71.3% and 79.6% for HKV and FKV, respectively. Significant up-regulation of IgM and IgT was observed in the head kidney of fish vaccinated with HKV at 21 dpv, while IgM, IgD and CD4 expression increased in the head kidney of fish receiving FKV at the same time point. After booster vaccination, IgT and CD8 transcripts were significantly increased in the spleen of fish vaccinated with the HKV, but not with FKV. Both vaccines induced a specific IgM response in both serum and mucus. In summary, this study showed that both HKV and FKV are promising injectable vaccines for the prevention of disease caused by TiLV in Nile tilapia.


Assuntos
Doenças dos Peixes/prevenção & controle , Infecções por Vírus de RNA/prevenção & controle , Vírus de RNA/imunologia , Vacinas Virais/imunologia , Animais , Ciclídeos/genética , Ciclídeos/imunologia , Doenças dos Peixes/imunologia , Doenças dos Peixes/virologia , Injeções Intraperitoneais , Vacinas de Produtos Inativados/administração & dosagem , Vacinas de Produtos Inativados/imunologia , Vacinas Virais/administração & dosagem
15.
Appl Environ Microbiol ; 87(23): e0144821, 2021 11 10.
Artigo em Inglês | MEDLINE | ID: mdl-34550753

RESUMO

Municipal wastewater provides an integrated sample of a diversity of human-associated microbes across a sewershed, including viruses. Wastewater-based epidemiology (WBE) is a promising strategy to detect pathogens and may serve as an early warning system for disease outbreaks. Notably, WBE has garnered substantial interest during the coronavirus disease 2019 (COVID-19) pandemic to track disease burden through analyses of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA. Throughout the COVID-19 outbreak, tracking SARS-CoV-2 in wastewater has been an important tool for understanding the spread of the virus. Unlike traditional sequencing of SARS-CoV-2 isolated from clinical samples, which adds testing burden to the health care system, in this study, metatranscriptomics was used to sequence virus directly from wastewater. Here, we present a study in which we explored RNA viral diversity through sequencing 94 wastewater influent samples across seven wastewater treatment plants (WTPs), collected from August 2020 to January 2021, representing approximately 16 million people in Southern California. Enriched viral libraries identified a wide diversity of RNA viruses that differed between WTPs and over time, with detected viruses including coronaviruses, influenza A, and noroviruses. Furthermore, single-nucleotide variants (SNVs) of SARS-CoV-2 were identified in wastewater, and we measured proportions of overall virus and SNVs across several months. We detected several SNVs that are markers for clinically important SARS-CoV-2 variants along with SNVs of unknown function, prevalence, or epidemiological consequence. Our study shows the potential of WBE to detect viruses in wastewater and to track the diversity and spread of viral variants in urban and suburban locations, which may aid public health efforts to monitor disease outbreaks. IMPORTANCE Wastewater-based epidemiology (WBE) can detect pathogens across sewersheds, which represents the collective waste of human populations. As there is a wide diversity of RNA viruses in wastewater, monitoring the presence of these viruses is useful for public health, industry, and ecological studies. Specific to public health, WBE has proven valuable during the coronavirus disease 2019 (COVID-19) pandemic to track the spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) without adding burden to health care systems. In this study, we used metatranscriptomics and reverse transcription-droplet digital PCR (RT-ddPCR) to assay RNA viruses across Southern California wastewater from August 2020 to January 2021, representing approximately 16 million people from Los Angeles, Orange, and San Diego counties. We found that SARS-CoV-2 quantification in wastewater correlates well with county-wide COVID-19 case data, and that we can detect SARS-CoV-2 single-nucleotide variants through sequencing. Likewise, wastewater treatment plants (WTPs) harbored different viromes, and we detected other human pathogens, such as noroviruses and adenoviruses, furthering our understanding of wastewater viral ecology.


Assuntos
Vírus de RNA/isolamento & purificação , SARS-CoV-2/isolamento & purificação , Viroma , Águas Residuárias/virologia , Vigilância Epidemiológica Baseada em Águas Residuárias , COVID-19/epidemiologia , California , Perfilação da Expressão Gênica , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Reação em Cadeia da Polimerase , Vírus de RNA/classificação , Vírus de RNA/genética , SARS-CoV-2/classificação , SARS-CoV-2/genética , Análise de Sequência de RNA
16.
J Invertebr Pathol ; 185: 107667, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34560106

RESUMO

Managed and wild bee populations are in decline around the globe due to several biotic and abiotic stressors. Pathogenic viruses associated with the Western honey bee (Apis mellifera) have been identified as key contributors to losses of managed honey bee colonies, and are known to be transmitted to wild bee populations through shared floral resources. However, little is known about the prevalence and intensity of these viruses in wild bee populations, or how bee visitation to flowers impacts viral transmission in agroecosystems. This study surveyed honey bee, bumble bee (Bombus impatiens) and wild squash bee (Eucera (Peponapis) pruinosa) populations in Cucurbita agroecosystems across Pennsylvania (USA) for the prevalence and intensity of five honey bee viruses: acute bee paralysis virus (ABPV), deformed wing virus (DWV), Israeli acute paralysis virus (IAPV), Kashmir bee virus (KBV), and slow bee paralysis virus (SBPV). We investigated the potential role of bee visitation rate to flowers on DWV intensity among species in the pollinator community, with the expectation that increased bee visitation to flowers would increase the opportunity for transmission events between host species. We found that honey bee viruses are highly prevalent but in lower titers in wild E. pruinosa and B. impatiens than in A. mellifera populations throughout Pennsylvania (USA). DWV was detected in 88% of B. impatiens, 48% of E. pruinosa, and 95% of A. mellifera. IAPV was detected in 5% of B. impatiens and 4% of E. pruinosa, compared to 9% in A. mellifera. KBV was detected in 1% of B. impatiens and 5% of E. pruinosa, compared to 32% in A. mellifera. Our results indicate that DWV titers are not correlated with bee visitation in Cucurbita fields. The potential fitness impacts of these low viral titers detected in E. pruinosa remain to be investigated.


Assuntos
Abelhas/virologia , Vírus de Insetos/fisiologia , Vírus de RNA de Cadeia Positiva/fisiologia , Animais , Produtos Agrícolas , Cucurbita , Dicistroviridae/fisiologia , Pennsylvania , Polinização , Vírus de RNA/fisiologia , Especificidade da Espécie
17.
Braz J Biol ; 83: e248975, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34550289

RESUMO

Colletotrichum is one of the most economically important fungal genera, which affects a wide range of hosts, specifically tropical and subtropical crops. Thus far, there have been several records of mycovirus infection in Colletotrichum spp., primarily by viruses of the Partitiviridae family. There have also been records of infections by mycoviruses of the Chrysoviridae family. Mycoviruses are (+)ssRNA and dsRNA genome viruses, which may or may not be enveloped. To date, no mycovirus with a DNA genome has been isolated from Colletotrichum spp. Typically, mycoviruses cause latent infections, although hypo- and hypervirulence have also been reported in Colletotrichum spp. In addition to its effects on pathogenic behavior, mycovirus infection can lead to important physiological changes, such as altered morphological characteristics, reduced vegetative growth, and suppressed conidia production. Therefore, research on mycoviruses infecting phytopathogenic fungi can help develop alternative methods to chemical control, which can cause irreversible damage to humans and the environment. From an agricultural perspective, mycoviruses can contribute to sustainable agriculture as biological control agents via changes in fungal physiology, ultimately resulting in the total loss of or reduction in the virulence of these pathogens.


Assuntos
Colletotrichum , Micovírus , Vírus de RNA , Micovírus/genética , Humanos , Filogenia , Esporos Fúngicos , Virulência
18.
Acta Virol ; 65(3): 320-323, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34565161

RESUMO

The complete genome of a novel virus from Arma chinensis was determined by RNA sequencing and rapid amplification of cDNA ends. This virus has a single-stranded RNA genome of 10,540 nucleotides (nt) excluding the poly(A) tail. Two non-overlapping open reading frames (ORFs) in the sense direction were predicted: one long ORF at the 5' end of the genome (6,219 nt) that encodes a polypeptide of 2,072 amino acids (aa), and one short ORF at the 3' end of the genome (3,033 nt) that encodes a polypeptide of 1,010 aa. Phylogenetic analysis indicated that the virus clusters within a large cluster of currently unidentified picorna-like viruses with a high bootstrap value. We named the virus isolate Arma chinensis picorna-like virus 1 (AcPV-1). The prevalence of AcPV-1 infection in samples of Arma chinensis from the wild was at a low level (5.48%, 8 positives in 146 samples). Keywords: Arma chinensis; genomic characterization; phylogenetic analysis; Arma chinensis picorna-like virus 1; prevalence.


Assuntos
Genoma Viral , Vírus de RNA , Sequência de Aminoácidos , Fases de Leitura Aberta , Filogenia , Vírus de RNA/genética , RNA Viral/genética
19.
Future Microbiol ; 16: 1105-1133, 2021 09.
Artigo em Inglês | MEDLINE | ID: mdl-34468163

RESUMO

SARS-CoV-2 is the etiological agent of the current pandemic worldwide and its associated disease COVID-19. In this review, we have analyzed SARS-CoV-2 characteristics and those ones of other well-known RNA viruses viz. HIV, HCV and Influenza viruses, collecting their historical data, clinical manifestations and pathogenetic mechanisms. The aim of the work is obtaining useful insights and lessons for a better understanding of SARS-CoV-2. These pathogens present a distinct mode of transmission, as SARS-CoV-2 and Influenza viruses are airborne, whereas HIV and HCV are bloodborne. However, these viruses exhibit some potential similar clinical manifestations and pathogenetic mechanisms and their understanding may contribute to establishing preventive measures and new therapies against SARS-CoV-2.


Assuntos
COVID-19/história , Pandemias/história , SARS-CoV-2/fisiologia , SARS-CoV-2/patogenicidade , Antivirais/uso terapêutico , COVID-19/tratamento farmacológico , COVID-19/epidemiologia , COVID-19/transmissão , Clima , Reservatórios de Doenças/virologia , Genoma Viral , História do Século XIX , História do Século XX , História do Século XXI , Humanos , Mutação , Vírus de RNA/patogenicidade , Vírus de RNA/fisiologia , Reinfecção/epidemiologia , Reinfecção/história , Reinfecção/transmissão , Reinfecção/virologia , Infecções Respiratórias/tratamento farmacológico , Infecções Respiratórias/epidemiologia , Infecções Respiratórias/história , Infecções Respiratórias/transmissão , Replicação Viral
20.
Int J Mol Sci ; 22(17)2021 Aug 29.
Artigo em Inglês | MEDLINE | ID: mdl-34502274

RESUMO

Heat shock proteins (HSPs) are a large group of chaperones found in most eukaryotes and bacteria. They are responsible for the correct protein folding, protection of the cell against stressors, presenting immune and inflammatory cytokines; furthermore, they are important factors in regulating cell differentiation, survival and death. Although the biological function of HSPs is to maintain cell homeostasis, some of them can be used by viruses both to fold their proteins and increase the chances of survival in unfavorable host conditions. Folding viral proteins as well as replicating many different viruses are carried out by, among others, proteins from the HSP70 and HSP90 families. In some cases, the HSP70 family proteins directly interact with viral polymerase to enhance viral replication or they can facilitate the formation of a viral replication complex and/or maintain the stability of complex proteins. It is known that HSP90 is important for the expression of viral genes at both the transcriptional and the translational levels. Both of these HSPs can form a complex with HSP90 and, consequently, facilitate the entry of the virus into the cell. Current studies have shown the biological significance of HSPs in the course of infection SARS-CoV-2. A comprehensive understanding of chaperone use during viral infection will provide new insight into viral replication mechanisms and therapeutic potential. The aim of this study is to describe the molecular basis of HSP70 and HSP90 participation in some viral infections and the potential use of these proteins in antiviral therapy.


Assuntos
Proteínas de Choque Térmico HSP70/metabolismo , Proteínas de Choque Térmico HSP90/metabolismo , Viroses/patologia , COVID-19/metabolismo , COVID-19/patologia , COVID-19/virologia , Vírus de DNA/fisiologia , Humanos , Isoformas de Proteínas/metabolismo , Vírus de RNA/fisiologia , SARS-CoV-2/isolamento & purificação , Viroses/metabolismo , Viroses/virologia
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