Systemic spread of an RNA insect virus in plants expressing plant viral movement protein genes

Flock house virus (FHV), a single-stranded RNA insect virus, has previously been reported to cross the kingdom barrier and replicate in barley protoplasts and in inoculated leaves of several plant species [Selling, B. H., Allison, R. F. & Kaesberg, P. (1990) Proc. Natl. Acad. Sci. USA 87, 434–438]. There was no systemic movement of FHV in plants. We tested the ability of movement proteins (MPs) of plant viruses to provide movement functions and cause systemic spread of FHV in plants. We compared the growth of FHV in leaves of nontransgenic and transgenic plants expressing the MP of tobacco mosaic virus or red clover necrotic mosaic virus (RCNMV). Both MPs mobilized cell-to-cell and systemic movement of FHV in Nicotiana benthamiana plants. The yield of FHV was more than 100-fold higher in the inoculated leaves of transgenic plants than in the inoculated leaves of nontransgenic plants. In addition, FHV accumulated in the noninoculated upper leaves of both MP-transgenic plants. RCNMV MP was more efficient in mobilizing FHV to noninoculated upper leaves. We also report here that FHV replicates in inoculated leaves of six additional plant species: alfalfa, Arabidopsis, Brassica, cucumber, maize, and rice. Our results demonstrate that plant viral MPs cause cell-to-cell and long-distance movement of an animal virus in plants and offer approaches to the study of the evolution of viruses and mechanisms governing mRNA trafficking in plants as well as to the development of promising vectors for transient expression of foreign genes in plants.

Complete replication of an animal virus and maintenance of expression vectors derived from it in Saccharomyces cerevisiae.

Here we describe the first instances to our knowledge of animal virus genome replication, and of de novo synthesis of infectious virions by a nonendogenous virus, in the yeast Saccharomyces cerevisiae, whose versatile genetics offers significant advantages for studying viral replication and virus-host interactions. Flock house virus (FHV) is the most extensively studied member of the Nodaviridae family of (+) strand RNA animal viruses. Transfection of yeast with FHV genomic RNA induced viral RNA replication, transcription, and assembly of infectious virions. Genome replication and virus synthesis were robust: all replicating FHV RNA species were readily detected in yeast by Northern blot analysis and yields of virions per cell were similar to those from Drosophila cells. We also describe in vivo expression and maintenance of a selectable yeast marker gene from an engineered FHV RNA derivative dependent on FHV-directed RNA replication. Use of these approaches with FHV and their possible extension to other viruses should facilitate identification and characterization of host factors required for genomic replication, gene expression, and virion assembly.

Impacto socioeconômico das políticas sanitárias sobre os estabelecimentos avícolas comerciais de postura da regional agropecuária de Limeira, estado de São Paulo

É reconhecida a importância da avicultura para o agronegócio e para o desenvolvimento do Brasil. Mas o País tem enfrentado grande concorrência internacional, que se traduz em barreiras sanitárias e exigências cada vez maiores de controle de seu rebanho por parte dos importadores. Neste sentido, o Ministério da Agricultura, Pecuária e Abastecimento (MAPA) publicou uma série de atos legais para viabilizar a organização dos programas de saúde animal, entre eles as Instruções Normativas n° 56/2007, n° 59/2009, n°36/2012 e n°10/2013, para estabelecer os procedimentos para registro e fiscalização de estabelecimentos avícolas comerciais e de reprodução, voltados à biosseguridade do sistema avícola. Como todos os setores produtivos, a avicultura é fortemente influenciada pela sua estrutura de custos, de modo que há um certo sentimento de que a adequação às medidas de biosseguridade, preconizadas pelas normativas, pode impactar a atividade de produção de ovos comerciais de modo a levar, eventualmente, alguns avicultores ao abandono da mesma. O objetivo deste trabalho foi analisar o impacto socioeconômico das políticas sanitárias para estabelecimentos avícolas comerciais de postura da regional agropecuária de Limeira, Estado de São Paulo, analisando o perfil socioeconômico da propriedade e do produtor e, estimando o custo de produção e seu impacto em atendimento às medidas de biosseguridade preconizadas pelas normativas. Para a análise do perfil das propriedades foram utilizados os dados dos 28 estabelecimentos avícolas, levantados por meio de documentação para registro. Para a análise do perfil do produtor e para estimar os custos de produção foram considerados dez estabelecimentos, voluntários à pesquisa, visitados entre os meses de junho e julho de 2013. O resultado do estudo sugere que as adequações à biosseguridade podem ser factíveis de serem realizadas economicamente, sendo o custo relativamente pequeno frente aos possíveis riscos de enfermidades, representando entre 1,61% e 2,09% do custo total de produção. No entanto, as sucessivas alterações nas legislações podem fazer com que o programa de sanidade avícola perca a credibilidade diante da sociedade e dos produtores que demonstram resistência às mudanças nos paradigmas zoosanitários

Assessing Activity and Location of Individual Laying Hens in Large Groups Using Modern Technology

Tracking individual animals within large groups is increasingly possible, offering an exciting opportunity to researchers. Whereas previously only relatively indistinguishable groups of individual animals could be observed and combined into pen level data, we can now focus on individual actors within these large groups and track their activities across time and space with minimal intervention and disturbance. The development is particularly relevant to the poultry industry as, due to a shift away from battery cages, flock sizes are increasingly becoming larger and environments more complex. Many efforts have been made to track individual bird behavior and activity in large groups using a variety of methodologies with variable success. Of the technologies in use, each has associated benefits and detriments, which can make the approach more or less suitable for certain environments and experiments. Within this article, we have divided several tracking systems that are currently available into two major categories (radio frequency identification and radio signal strength) and review the strengths and weaknesses of each, as well as environments or conditions for which they may be most suitable. We also describe related topics including types of analysis for the data and concerns with selecting focal birds.

John Wiclif, of Wycliffe, & other essays,/

Partly reprinted from various periodicals.

The French ideal; Pascal, Fénelon and other essays,

Contains bibliographies.

The French ideal; Pascal, Fénelon and other essays,

Contains bibliographies.

Bayesian inferences on the recent island colonization history by the bird Zosterops lateralis lateralis

The founding of new populations by small numbers of colonists has been considered a potentially important mechanism promoting evolutionary change in island populations. Colonizing species, such as members of the avian species complex Zosterops lateralis, have been used to support this idea. A large amount of background information on recent colonization history is available for one Zosterops subspecies, Z. lateralis lateralis, providing the opportunity to reconstruct the population dynamics of its colonization sequence. We used a Bayesian approach to combine historical and demographic information available on Z. l. lateralis with genotypic data from six microsatellite loci, and a rejection algorithm to make simultaneous inferences on the demographic parameters describing the recent colonization history of this subspecies in four southwest Pacific islands. Demographic models assuming mutation–drift equilibrium or a large number of founders were better supported than models assuming founder events for three of four recently colonized island populations. Posterior distributions of demographic parameters supported (i) a large stable effective population size of several thousands individuals with point estimates around 4000–5000; (ii) a founder event of very low intensity with a large effective number of founders around 150–200 individuals for each island in three of four islands, suggesting the colonization of those islands by one flock of large size or several flocks of average size; and (iii) a founder event of higher intensity on Norfolk Island with an effective number of founders around 20 individuals, suggesting colonization by a single flock of moderate size. Our inferences on demographic parameters, especially those on the number of founders, were relatively insensitive to the precise choice of prior distributions for microsatellite mutation processes and demographic parameters, suggesting that our analysis provides a robust description of the recent colonization history of the subspecies.

Heterologous RNA encapsidated in pariacoto virus-like particles forms a dodecahedral cage similar to genomic RNA in wild-type virions

The genome of some icosahedral RNA viruses plays an essential role in capsid assembly and structure. In T=3 particles of the nodavirus Pariacoto virus (PaV), a remarkable 35% of the single-stranded RNA genome is icosahedrally ordered. This ordered RNA can be visualized at high resolution by X-ray crystallography as a dodecahedral cage consisting of 30 24-nucleotide A-form RNA duplex segments that each underlie a twofold icosahedral axis of the virus particle and interact extensively with the basic N-terminal region of 60 subunits of the capsid protein. To examine whether the PaV genome is a specific determinant of the RNA structure, we produced virus-like particles (VLPs) by expressing the wild-type capsid protein open reading frame from a recombinant baculovirus. VLPs produced by this system encapsidated similar total amounts of RNA as authentic virus particles, but only about 6% of this RNA was PaV specific, the rest being of cellular or baculovirus origin. Examination of the VLPs by electron cryomicroscopy and image reconstruction at 15.4-Angstrom resolution showed that the encapsidated RNA formed a dodecahedral cage similar to that of wild-type particles. These results demonstrate that the specific nucleotide sequence of the PaV genome is not required to form the dodecahedral cage of ordered RNA.

Computing with elation groups

We have proved that elation groups of a certain infinite family of Roman generalized quadrangles are not isomorphic to those of associated flock generalized quadrangles. The proof is theoretical, but is based upon detailed computations. Here we elaborate on the explicit computer calculations which inspired the proof.