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Microbiol Resour Announc ; 10(26): e0132120, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34197198


Eight isolates of Streptococcus equi subsp. zooepidemicus were isolated from mares with clinical cases of endometritis. S. equi subsp. zooepidemicus strains were chosen for sequencing based on differing levels of biofilm production in vitro. Using Illumina short-read sequencing in conjunction with MinION sequencing, we report the genomes of eight isolates.

Vaccines (Basel) ; 9(4)2021 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-33916180


The COVID-19 pandemic has generated intense interest in the rapid development and evaluation of vaccine candidates for this disease and other emerging diseases. Several novel methods for preparing vaccine candidates are currently undergoing clinical evaluation in response to the urgent need to prevent the spread of COVID-19. In many cases, these methods rely on new approaches for vaccine production and immune stimulation. We report on the use of a novel method (SolaVAX) for production of an inactivated vaccine candidate and the testing of that candidate in a hamster animal model for its ability to prevent infection upon challenge with SARS-CoV-2 virus. The studies employed in this work included an evaluation of the levels of neutralizing antibody produced post-vaccination, levels of specific antibody sub-types to RBD and spike protein that were generated, evaluation of viral shedding post-challenge, flow cytometric and single cell sequencing data on cellular fractions and histopathological evaluation of tissues post-challenge. The results from this preliminary evaluation provide insight into the immunological responses occurring as a result of vaccination with the proposed vaccine candidate and the impact that adjuvant formulations, specifically developed to promote Th1 type immune responses, have on vaccine efficacy and protection against infection following challenge with live SARS-CoV-2. This data may have utility in the development of effective vaccine candidates broadly. Furthermore, the results of this preliminary evaluation suggest that preparation of a whole virion vaccine for COVID-19 using this specific photochemical method may have potential utility in the preparation of one such vaccine candidate.

PLoS Pathog ; 17(3): e1009315, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33647063


Bunyaviruses (Negarnaviricota: Bunyavirales) are a large and diverse group of viruses that include important human, veterinary, and plant pathogens. The rapid characterization of known and new emerging pathogens depends on the availability of comprehensive reference sequence databases that can be used to match unknowns, infer evolutionary relationships and pathogenic potential, and make response decisions in an evidence-based manner. In this study, we determined the coding-complete genome sequences of 99 bunyaviruses in the Centers for Disease Control and Prevention's Arbovirus Reference Collection, focusing on orthonairoviruses (family Nairoviridae), orthobunyaviruses (Peribunyaviridae), and phleboviruses (Phenuiviridae) that either completely or partially lacked genome sequences. These viruses had been collected over 66 years from 27 countries from vertebrates and arthropods representing 37 genera. Many of the viruses had been characterized serologically and through experimental infection of animals but were isolated in the pre-sequencing era. We took advantage of our unusually large sample size to systematically evaluate genomic characteristics of these viruses, including reassortment, and co-infection. We corroborated our findings using several independent molecular and virologic approaches, including Sanger sequencing of 197 genome segments, and plaque isolation of viruses from putative co-infected virus stocks. This study contributes to the described genetic diversity of bunyaviruses and will enhance the capacity to characterize emerging human pathogenic bunyaviruses.

Genoma Viral/genética , Nairovirus/genética , Orthobunyavirus/genética , Vírus de RNA/genética , Animais , Arbovírus/genética , Artrópodes/genética , Sequência de Bases , Humanos , Filogenia
Viruses ; 12(6)2020 06 02.
Artigo em Inglês | MEDLINE | ID: mdl-32498297


As part of research and wildlife disease surveillance efforts, we performed necropsy examinations of 125 free-ranging (n = 114) and captive (n = 11) prairie dogs in Colorado from 2009 to 2017. From these cases, we identified three cases of thymic lymphoma in free-ranging Gunnison's prairie dogs (Cynomys gunnisoni), and we identified a novel retroviral sequence associated with these tumors. The viral sequence is 7700 nucleotides in length and exhibits a genetic organization that is consistent with the characteristics of a type D betaretrovirus. The proposed name of this virus is Gunnison's prairie dog retrovirus (GPDRV). We screened all 125 prairie dogs for the presence of GPDRV using PCR with envelope-specific primers and DNA extracted from spleen samples. Samples were from Gunnison's prairie dogs (n = 59), black-tailed prairie dogs (Cynomys ludovicianus) (n = 40), and white-tailed prairie dogs (Cynomys leucurus) (n = 26). We identified GPDRV in a total of 7/125 (5.6%) samples including all three of the prairie dogs with thymic lymphoma, as well as spleen from an additional four Gunnison's prairie dogs with no tumors recognized at necropsy. None of the GPDRV-negative Gunnison's prairie dogs had thymic lymphomas. We also identified a related, apparently endogenous retroviral sequence in all prairie dog samples. These results suggest that GPDRV infection may lead to development of thymic lymphoma in Gunnison's prairie dogs.

Linfoma/veterinária , Retroviridae/isolamento & purificação , Doenças dos Roedores/virologia , Timoma/veterinária , Sequência de Aminoácidos , Animais , Animais Selvagens/virologia , Colorado , Feminino , Genoma Viral , Linfoma/patologia , Linfoma/virologia , Filogenia , Retroviridae/química , Retroviridae/classificação , Retroviridae/genética , Doenças dos Roedores/epidemiologia , Doenças dos Roedores/patologia , Sciuridae/classificação , Sciuridae/virologia , Alinhamento de Sequência , Timoma/patologia , Timoma/virologia , Proteínas Virais/química , Proteínas Virais/genética
Viruses ; 10(7)2018 07 22.
Artigo em Inglês | MEDLINE | ID: mdl-30037148


Ixodes scapularis ticks harbor a variety of microorganisms, including eukaryotes, bacteria and viruses. Some of these can be transmitted to and cause disease in humans and other vertebrates. Others are not pathogenic, but may impact the ability of the tick to harbor and transmit pathogens. A growing number of studies have examined the influence of bacteria on tick vector competence but the influence of the tick virome remains less clear, despite a surge in the discovery of tick-associated viruses. In this study, we performed shotgun RNA sequencing on 112 individual adult I. scapularis collected in Wisconsin, USA. We characterized the abundance, prevalence and co-infection rates of viruses, bacteria and eukaryotic microorganisms. We identified pairs of tick-infecting microorganisms whose observed co-infection rates were higher or lower than would be expected, or whose RNA levels were positively correlated in co-infected ticks. Many of these co-occurrence and correlation relationships involved two bunyaviruses, South Bay virus and blacklegged tick phlebovirus-1. These viruses were also the most prevalent microorganisms in the ticks we sampled, and had the highest average RNA levels. Evidence of associations between microbes included a positive correlation between RNA levels of South Bay virus and Borrelia burgdorferi, the Lyme disease agent. These findings contribute to the rationale for experimental studies on the impact of viruses on tick biology and vector competence.

Coinfecção/microbiologia , Coinfecção/virologia , Ixodes/microbiologia , Ixodes/virologia , Doenças Transmitidas por Carrapatos/epidemiologia , Animais , Bactérias/genética , Bactérias/isolamento & purificação , Borrelia burgdorferi/genética , Borrelia burgdorferi/isolamento & purificação , Coinfecção/epidemiologia , Eucariotos/genética , Eucariotos/isolamento & purificação , Sequenciamento de Nucleotídeos em Larga Escala , Ixodes/genética , Doença de Lyme , Microbiota/genética , Orthobunyavirus/genética , Orthobunyavirus/isolamento & purificação , Phlebovirus/genética , Phlebovirus/isolamento & purificação , Prevalência , Simbiose , Infestações por Carrapato/epidemiologia , Infestações por Carrapato/microbiologia , Infestações por Carrapato/virologia , Doenças Transmitidas por Carrapatos/microbiologia , Doenças Transmitidas por Carrapatos/virologia , Vírus/genética , Vírus/isolamento & purificação , Wisconsin/epidemiologia