Antiviral Properties of Streptomyces tuirus DBZ39 Mediated Gold Nanoparticles Against Bluetongue virus.

<b>Background and Objective:</b> The proposed study involves the approach from the point of anti-viral activity of gold nanoparticles against the <i>Bluetongue virus</i>. Among viral diseases, Bluetongue is regarded as an economically scouring disease. Neither a vaccine nor an antiviral drug is available for the prevention or treatment of this disease. The antiviral activity of gold nanoparticles synthesized by a novel isolate of <i>Streptomyces tuirus</i> DBZ39 is the breakthrough of the study. <i>Streptomyces tuirus </i>DBZ39, a novel isolate obtained from alkaline soil was proved to be efficient actinomycetes, for the extracellular synthesis of gold nanoparticles. <b>Materials and Methods:</b> An upstream bioprocess was optimized and developed for the synthesis of controlled size gold nanoparticles with solitary mono dispersal pattern in aurum chloride solution. The characterization and confirmation of gold nanoparticles were illustrated by Scanning Electron Microscopy (SEM), Transmission Electron Microscopy (TEM), Energy Dispersive X-ray Analysis (EDAX) and Fourier Transmission Infrared Radiation Analysis (FTIR). <b>Results:</b> Biomass size of 3 g, substrate concentration of 1 mM, pH of 8.5 and temperature of 45°C were observed as optimum conditions for the synthesis of 15-24 nm size gold nanoparticles. The <i>Bluetongue virus</i> (BTV) which belongs to the genus Orbivirus in the family Reoviridae with 26 serotypes is an etiological agent of infectious and non-contagious Bluetongue disease of main sheep and several other domestic animals. <b>Conclusion:</b> Gold nanoparticles for the 1st time, at a higher concentration of 1:64 dilutions revealed a very promising and novel antiviral property against the <i>Bluetongue virus</i>.

Bluetongue virus infection alters the impedance of monolayers of bovine endothelial cells as a result of cell death.

Bluetongue virus (BTV) is the cause of bluetongue, an emerging, arthropod-transmitted disease of ungulates. Bluetongue is characterized by vascular injury with hemorrhage, tissue infarction and widespread edema, lesions that are consistent with those of the so-called viral hemorrhagic fevers. To further investigate the pathogenesis of vascular injury in bluetongue, we utilized an electrical impedance assay and immunofluorescence staining to compare the effects of BTV infection on cultured bovine endothelial cells (bPAEC) with those of inducers of cell death (Triton X-100) and interendothelial gap formation (tissue necrosis factor [TNF]). The data confirm that the adherens junctions of BTV-infected bPAECs remained intact until 24h post-infection, and that loss of monolayer impedance precisely coincided with onset of virus-induced cell death. In contrast, recombinant bovine TNF-alpha caused rapid loss of bPAEC monolayer impedance that was associated with interendothelial gap formation and redistribution of VE-cadherin, but without early cell death. The data from these in vitro studies are consistent with a pathogenesis of bluetongue that involves virus-induced vascular injury leading to thrombosis, hemorrhage and tissue necrosis. However, the contribution of cytokine-induced interendothelial gap formation with subsequent edema and hypovolemic shock contributes to the pathogenesis of bluetongue remains to be fully characterized.

The role of endothelial cell-derived inflammatory and vasoactive mediators in the pathogenesis of bluetongue.

Bluetongue is an insect-transmitted disease of sheep and wild ruminants that is caused by bluetongue virus (BTV). Cattle are asymptomatic reservoir hosts of BTV. Infection of lung microvascular endothelial cells (ECs) is central to the pathogenesis of BTV infection of both sheep and cattle, but it is uncertain as to why sheep are highly susceptible to BTV-induced microvascular injury, whereas cattle are not. Thus, to better characterize the pathogenesis of bluetongue, the transcription of genes encoding a variety of vasoactive and inflammatory mediators was quantitated in primary ovine lung microvascular ECs (OLmVECs) exposed to BTV and/or inflammatory mediators. BTV infection of OLmVECs increased the transcription of genes encoding interleukin- (IL) 1 and IL-8, but less so IL-6, cyclooxygenase-2, and inducible nitric oxide synthase. In contrast, we previously have shown that transcription of genes encoding all of these same mediators is markedly increased in BTV-infected bovine lung microvascular ECs and that BTV-infected bovine ECs produce substantially greater quantities of prostacyclin than do sheep ECs. Thus, sheep and cattle were experimentally infected with BTV to further investigate the role of EC-derived vasoactive mediators in the pathogenesis of bluetongue. The ratio of thromboxane to prostacyclin increased during BTV infection of both sheep and cattle, but was significantly greater in sheep (P = 0.001). Increases in the ratio of thromboxane to prostacyclin, indicative of enhanced coagulation, coincided with the occurrence of clinical manifestations of bluetongue in BTV-infected sheep. The data suggest that inherent species-specific differences in the production and activities of EC-derived mediators contribute to the sensitivity of sheep to BTV-induced microvascular injury.

Aspectos epidemiológicos y entomólogicos de la enfermedad de lengua azul en ganado bovino / Epidemiologic and entomologic aspects of bluetongue in cattle in the Isthmus of Tehuantepec, Oaxaca, Mexico

Se realizó un estudio prospectivo de la enfermedad de lengua azul en 2 hatos de ganado bovino en la región del Istmo de Tehuantepec. El estudio incluyó la detección menstrual de anticuerpos grupo-específicos contra el virus de lengua azul (VLA) y la captura de moscas culicoides. En enero de 1988, el total de ganado bovino en los 2 hatos (600) fue clasificado seropositivo al VLA, utilizando un método ELISA. Con base en este resultado, 40 bovinos adultos y 35 becerros nacidos entre octubre de 1987 y enero de 1988 fueron evaluados mensualmente (enero a diciembre de 1988) para detectar la producción de anticuerpos grupo-específicos VLA. Se detectó un patrón positivo de seroconversión contra el VLA en becerros, durante la temporada de lluvia en los meses del verano (junio-septiembre). Sin embargo, en algunos becerros se determinó serológicamente actividad de VLA en la temporada seca, durante marzo y abril. Debido a la ausencia de anticuerpos maternos, la mayoría de los becerros fueron susceptibles de infección con el VLA a los 6 meses de edad. La evidencia serológica de la actividad del VLA en becerros se relacionó con la abundancia de moscas Culicoides insignis capturadas durante el estudio

Experimental infection of pregnant cattle with bluetongue virus serotype 11 between postbreeding days 21 and 48.

Four bluetongue virus (BTV)-seronegative heifers and 2 BTV-seropositive heifers were inoculated with the virulent strain UC-8 of BTV-11 between postbreeding days (PBD) 21 and 30. The heifers were observed for 10-18 days after inoculation for clinical signs, and pregnancy was monitored by ultrasound examination of the uterus and by plasma progesterone levels. Blood samples were collected daily after inoculation and processed for virus isolation and titration. Heifers were euthanized between PBD 31 and PBD 48, and tissues were collected for virologic and pathologic examination. All but 1 heifer inoculated on PBD 21 remained pregnant after BTV inoculation. A cystic corpus luteum was found in the ovary of the nonpregnant heifer, but BTV was not isolated from the reproductive tract of this heifer. Three of the inoculated heifers that remained pregnant showed mild multifocal areas of perivascular lymphocytic infiltration in the ovary. BTV was reisolated from spleen and prescapular and peribronchial lymph nodes 10 days after inoculation from 3 of the 4 BTV-seronegative heifers. BTV was also reisolated from the uterus of 1 of the heifers that remained pregnant, but microscopic lesions were not found in this organ.