Alguns dados sobre a Fauna entomológica da ilha das Flores - Açores

IV Expedição Científica do Departamento de Biologia - Flores 1989Com este trabalho, realizado em Julho de 1989 nas Flores - a ilha mais ocidental do Arquipélago dos Açores -, acrescentaram-se onze espécies de Lepidópteros à lista referenciada para aquela ilha, pertencendo uma à família Lycaenidae (Lampides boeticus L.), oito a familia Noctuidae (Agrotis ipsilon HFN., Brotolomia meticulosa L., Chrysodeixis chalcites ESPER., Heliothis armigera HBN., Noctua atlantica WARREN, Noctua pronuba L., Peridroma saucia HBN., Sesamia nonagrioides LEF.), uma à família Nymphalidae (Vanessa atalanta L.) e uma a família Pyralidae (Glyphodes unionalis HBN.). Entre os demais insectos, foram identificadas cerca de duas dezenas e meia de espécies, distribuídas pelas Ordens Dermaptera, Orthoptera, Dictyoptera, Heteroptera, Homoptera, Coleoptera, Neuroptera, Diptera, Hymenoptera e Collembola. Salienta-se ainda a importância, do ponto de vista agronómico, das pragas Mythimna unipuncta (HAWORTH) e Xestia c-nigrun L. naquela ilha.RÉSUMÉ: Avec ce travail, réalisé en Juillet 1989 a Flores - l'île plus occidental de l'archipel des Açores, onze espèces de Lépidoptères ont été ajoutées à la liste des espèces connus pour cette île, dont une appartient a la famille Lycaenidae (Lampides boelicus L.), huit à la famille Noctuidae (Agrotis ipsilon HFN., Brotolomia meticulosa L. Chrysodeicis chalcites ESPER., Heliothis armigera HBN., Noctua atlantica WARREN, Noctua pronuba L., Peridroma saucia HBN., Sesamia nonagrioides LEF.), une à la famille Nymphalidae (Vanessa atalanta L.) et une à la famille Pyralidae (Glyphodes unionalis HBN.). Parmi les autres insects ont été identifiés environ deux dizaines et demie d'espèces, lesquelles sont réparties par les Ordres Dermaptera, Orthoptera, Dictyoptera, Heteroptera, Homoptera, Coleoptera, Neuroptera, Diptera, Hymenoptera et Collembola. On remarque I'importance, du point de vue agronomique, des ravageurs Mythimna unipuncra (HAWORTH) et Xestia c-nigrum L. dans cette île

Characteristics of Corruption in African States : by comparing with Asian states

<div><p>Chikungunya virus (CHIKV) is a mosquito-borne alphavirus that re-emerged in 2004 and has caused massive outbreaks in recent years. The lack of a licensed vaccine or treatment options emphasize the need to obtain more insight into the viral life cycle and CHIKV-host interactions. Infectious cDNA clones are important tools for such studies, and for mechanism of action studies on antiviral compounds. Existing CHIKV cDNA clones are based on a single genome from an individual clinical isolate, which is expected to have evolved specific characteristics in response to the host environment, and possibly also during subsequent cell culture passaging. To obtain a virus expected to have the general characteristics of the recent E1-226V CHIKV isolates, we have constructed a new CHIKV full-length cDNA clone, CHIKV LS3, based on the consensus sequence of their aligned genomes. Here we report the characterization of this synthetic virus and a green fluorescent protein-expressing variant (CHIKV LS3-GFP). Their characteristics were compared to those of natural strain ITA07-RA1, which was isolated during the 2007 outbreak in Italy. In cell culture the synthetic viruses displayed phenotypes comparable to the natural isolate, and in a mouse model they caused lethal infections that were indistinguishable from infections with a natural strain. Compared to ITA07-RA1 and clinical isolate NL10/152, the synthetic viruses displayed similar sensitivities to several antiviral compounds. 3-deaza-adenosine was identified as a new inhibitor of CHIKV replication. Cyclosporin A had no effect on CHIKV replication, suggesting that cyclophilins -opposite to what was found for other +RNA viruses- do not play an essential role in CHIKV replication. The characterization of the consensus sequence-based synthetic viruses and their comparison to natural isolates demonstrated that CHIKV LS3 and LS3-GFP are suitable and representative tools to study CHIKV-host interactions, screen for antiviral compounds and unravel their mode of action.</p></div

Development of RABV-specific antibodies over time in three- and eight-week old mice.

<p>BALB/c mice were inoculated i.m or s.c with (a) 10² TCID₅₀, (b) 10⁴ TCID₅₀ (c) 10⁶ TCID₅₀ of DUVV-NL07 or DUVV-NL07 BPL control. Dotted lines indicate threshold level of the assay.</p

Effect of antiviral compounds on the replication of various CHIKV strains.

<p>Dose dependent reduction of CHIKV-induced CPE by (A) 3-deaza-adenosine, (B) 6-aza-uridine, (C) chloroquine and (D) mycophenolic acid in Vero E6 cells infected with CHIKV strains ITA07-RA1, LS3 and LS3-GFP (MOI 0.005). (E) Antiviral effect of ribavirin on CHIKV replication in BHK-21 (black lines) and Vero E6 cells (gray lines). Cell viability was normalized to untreated uninfected cells (100%). The 50% cytotoxic concentration (CC₅₀) of the compounds is indicated in the top left of each panel. (F) Dose-response curves showing the effect of five antiviral compounds on the eGFP expression in Vero E6 cells infected with CHIKV LS3-GFP (MOI 0.05) at 42 h p.i. Values were normalized to eGFP fluorescence in untreated infected cells (100%).</p

Transcriptional and translational shut-off induced by CHIKV.

<p>293/ACE2 cells were infected with CHIKV LS3 or ITA07-RA1 at an MOI of 5, and at the indicated time points post infection metabolic labeling with ³H-uridine (A, B) or ³⁵S-Cys and ³⁵S-Met (C) was performed to analyze total RNA and protein synthesis, respectively. (A) Incorporation of ³H-uridine into viral and cellular RNA as measured by liquid scintillation counting of total RNA samples taken at various time points post infection. (B) ³H-uridine incorporation into viral and cellular RNA during CHIKV LS3 infection as detected by denaturing gel electrophoresis and fluorography. In control samples (ActD) 5 ug/ml Actinomycin D was added 30 min. prior to metabolic labeling to inhibit cellular transcription. (C) Synthesis of ³⁵S-labeled viral and cellular proteins during CHIKV LS3 infection. The control lane labeled CHX contains proteins from cells treated with the translation inhibitor cycloheximide prior to metabolic labeling. CHIKV-specific proteins are indicated with a *.</p

Replication of synthetic CHIKV strains <i>in vivo</i>.

<p>(A) Survival of mice after intraperitoneal injection with 100 TCID₅₀ of CHIKV LS3, LS3-GFP or S27 (5 mice per group). (B) CHIKV titers (in TCID₅₀ equivalents per mg of tissue) in the brains of the mice infected with the 3 CHIKV strains.</p

In vivo replication of DUVV-NL07.

<p>Viral RNA detected in the brain of animals (mean values expressed in copies/ml) infected with high, intermediate or low dose of DUVV-NL07 via peripheral route. The number of DUVV-NL07 RNA positive animals per total number of animals inoculated is indicated within brackets. Numbers in superscript indicate the amount of animals that developed paralysis in the respective groups.</p

Comparison of the amino acid sequences of CHIKV LS3 and various natural isolates.

<p>The CHIKV LS3 amino acid sequences were aligned with those of several highly similar natural isolates, and clinical isolate NL10/152. Only amino acid differences are indicated and identity is represented by dots. The numbering is based on the sequence of LS3, which is also equal to that of LR2006_OPY1. It is important to note that - like all other CHIKV strains in this table - clinical isolate LR2006-OPY1 (Genbank DQ443544) also contains an opal stop codon in the 3′-end of the nsP3 coding region, while its infectious clone with Genbank accession number EU224268 lacks this stop codon.</p

Western blot analysis of CHIKV nsP1 and E2 expression at different time points post infection.

<p>293/ACE2 cells were infected with CHIKV ITA07, LS3 or LS3-GFP at an MOI of 5. At the indicated time points cells were lysed, proteins were separated by SDS-PAGE and viral proteins were detected by Western blotting. The anti-E2 antiserum also recognized the E3E2 (p62) precursor of E2. Actin and the transferrin receptor were used as loading controls.</p

Accumulation of negative- and positive-strand CHIKV RNA in infected cells.

<p>(A) 293/ACE2 cells were infected with CHIKV LS3, LS3-GFP or ITA07-RA1 at an MOI of 5, total RNA was isolated at different time points post infection and strand-specific detection was performed with radioactively labeled oligonucleotides complementary to the 3′ end of either negative- (top panel) or positive-strand (middle panel) CHIKV RNA. Cellular 18S ribosomal RNA was probed as a loading control (lower panel). The positions of genomic RNA, the 26S sgRNA and the second eGFP-encoding sgRNA are indicated to the right of the middle panel. (B) Plot representing the kinetics of CHIKV negative-strand RNA accumulation, based on quantification of data from panel A. After correction for variations in sample loading based on the 18S rRNA signal, the relative abundance of the RNAs was determined by normalizing to the highest value observed (CHIKV LS3-GFP, 8 h p.i.). (C) Kinetics of CHIKV positive-strand RNA accumulation. The relative abundance of RNA was calculated as before, except that data were normalized to the value measured for LS3 sgRNA at 12 h p.i (100%). Genomic RNA levels are indicated with black lines, sgRNA levels with gray lines. The total level of both sgRNAs expressed by LS3-GFP is indicated with the gray dotted line.</p