Genomic heterogeneity of small ruminant lentiviruses detected by PCR.

In order to detect a large spectrum of small ruminant lentiviruses, primers for PCR were chosen in conserved parts of the LTR and GAG genes of Icelandic Visna virus 1514 and of the POL gene of caprine arthritis-encephalitis virus. This set of primers was tested in six different caprine arthritis-encephalitis virus (CAEV)- and Maedi-Visna virus isolates of Dutch, American and Swiss origin. The LTR primers allowed the detection of the corresponding fragments of all isolates. The GAG primers allowed amplification of the corresponding fragments of all but the Swiss Maedi-Visna virus strain OLV. Using the POL primers, one Maedi-Visna- and two caprine arthritis-encephalitis virus strains were detected after one round of amplification. Sequencing of the GAG and POL amplification products and comparison to Icelandic Visna virus and CAEV strain CO revealed total heterogeneity of 38% for the GAG- and 28% for the POL fragment. The virus strains studied fall into two groups which are more closely related to one another than to Icelandic Visna virus.     

Cloning of <Emphasis Type="Bold">β</Emphasis>-tubulin and succinate dehydrogenase genes from <Emphasis Type="Italic">Uromyces fabae</Emphasis> and establishing selection conditions for their use in transformation

The obligate biotrophic nature of rust fungi calls for an in planta selection scheme as a means of developing a rust transformation technology. We show that the fungicides benomyl (used as its formulated product benlate) and carboxin suppress morphogenesis of the rust fungus Uromyces fabae in vitro and disease in planta, the latter without affecting the health of the host. The limits of their applicability were determined regarding concentration, method of application and optimal time intervals of treatment. Besides procedures for selection, a stable transformation system will also need to include genetic markers allowing to enrich for transformed cells within a large background of untransformed cells. Since the molecular targets of benlate and carboxin had been identified as -tubulin and succinate dehydrogenase, respectively, the corresponding genes (Uf-TBBIand Uf-SUCDHI) were cloned and characterized. Molecular phylogenies demonstrate that both are typical homologs to those of other Basidiomycota. RT-PCR analysis confirmed that both genes are constitutively expressed in all developmental stages of the mitotic uredospore multiplication cycle. Since homologs of Uf-TBB1and Uf-SUCDH1 have been successfully used as selection markers in other fungal systems, they provide valuable tools to develop additional corner stones of a stable transformation system for rust fungi.     

The effectiveness of conservation measures to enhance nest survival in a meadow bird suffering from anthropogenic nest loss

The main reason for meadow bird declines is supposed to be the intensification of grassland management with earlier first harvest dates and more frequent harvests, resulting in high nest destruction rates. To increase productivity of meadow bird populations in intensified grassland areas a delay of mowing date and individual nest protection measures have been proposed. However, for ground-nesting songbirds such as the whinchat (Saxicola rubetra) the effectiveness of such measures remains widely unknown. In particular, if nest predation rate is high, measures to protect nests from agricultural destruction alone might be questionable. Here, we quantify whinchat nest survival of (1) unprotected nests situated in early mown meadows, (2) protected nests situated in early mown meadows, and (3) nests situated in late mown meadows. Analyses considered the fact that successful and unsuccessful nests are not found with equal probabilities. Periods of reduced nest survival were associated with mowing periods in the different types of meadows. In early mown meadows nest survival rates were low (S < 0.1), and both conservation measures, individual nest protection and delayed mowing, resulted in significantly increased nest survival rates (S > 0.7). Individual nest protection cannot avoid changes in habitat quality of intensively managed meadows, and therefore is only suitable as small-scale and short-term measure to increase nest success until a high portion of late mown meadows is established. Thus, we suggest that a combination of the two measures applied to intensified grassland fields should be provided to maintain viable sizes of endangered meadow bird populations.     

Isolation and identification of Salmonella spp. from red foxes (Vulpes vulpes) and badgers (Meles meles) in northern Italy

Background

Salmonella spp. have been isolated from a wide range of wild animals. Opportunistic wild carnivores such as red foxes (Vulpes vulpes) and badgers (Meles meles) may act as environmental indicators or as potential sources of salmonellosis in humans. The present study characterizes Salmonella spp. isolated from the intestinal contents of hunted or dead red foxes (n = 509) and badgers (n = 17) in northern Italy.

Findings

Thirty-one strains of Salmonella belonging to 3 Salmonella enterica subspecies were isolated. Fourteen different serovars of S. enterica subsp. enterica were identified, among which were serovars often associated with human illness.

Conclusions

Wild opportunistic predators can influence the probability of infection of both domestic animals and humans through active shedding of the pathogen to the environment. The epidemiological role of wild carnivores in the spread of salmonellosis needs to be further studied.     

Antigenic and genetic variations in European and North American equine influenza virus strains (H3N8) isolated from 2006 to 2007

Equine influenza virus (EIV) surveillance is important in the management of equine influenza. It provides data on circulating and newly emerging strains for vaccine strain selection. To this end, antigenic characterisation by haemaggluttination inhibition (HI) assay and phylogenetic analysis was carried out on 28 EIV strains isolated in North America and Europe during 2006 and 2007. In the UK, 20 viruses were isolated from 28 nasopharyngeal swabs that tested positive by enzyme-linked immunosorbent assay. All except two of the UK viruses were characterised as members of the Florida sublineage with similarity to A/eq/Newmarket/5/03 (clade 2). One isolate, A/eq/Cheshire/1/06, was characterised as an American lineage strain similar to viruses isolated up to 10 years earlier. A second isolate, A/eq/Lincolnshire/1/07 was characterised as a member of the Florida sublineage (clade 1) with similarity to A/eq/Wisconsin/03. Furthermore, A/eq/Lincolnshire/1/06 was a member of the Florida sublineage (clade 2) by haemagglutinin (HA) gene sequence, but appeared to be a member of the Eurasian lineage by the non-structural gene (NS) sequence suggesting that reassortment had occurred. A/eq/Switzerland/P112/07 was characterised as a member of the Eurasian lineage, the first time since 2005 that isolation of a virus from this lineage has been reported. Seven viruses from North America were classified as members of the Florida sublineage (clade 1), similar to A/eq/Wisconsin/03. In conclusion, a variety of antigenically distinct EIVs continue to circulate worldwide. Florida sublineage clade 1 viruses appear to predominate in North America, clade 2 viruses in Europe.     

Phagocytosis of Astaxanthin-Loaded Microparticles Modulates TGFβ Production and Intracellular ROS Levels in J774A.1 Macrophages

Radiation-induced fibrosis is a serious long-lasting side effect of radiation therapy. Central to this condition is the role of macrophages that, activated by radiation-induced reactive oxygen species and tissue cell damage, produce pro-inflammatory cytokines, such as transforming growth factor beta (TGFβ). This, in turn, recruits fibroblasts at the site of the lesion that initiates fibrosis. We investigated whether astaxanthin, an antioxidant molecule extracted from marine and freshwater organisms, could help control macrophage activation. To this purpose, we encapsulated food-grade astaxanthin from Haematococcus pluvialis into micrometer-sized whey protein particles to specifically target macrophages that can uptake material within this size range by phagocytosis. The data show that astaxanthin-loaded microparticles are resistant to radiation, are well-tolerated by J774A.1 macrophages, induce in these cells a significant reduction of intracellular reactive oxygen species and inhibit the release of active TGFβ as evaluated in a bioassay with transformed MFB-F11 fibroblasts. Micro-encapsulation of bioactive molecules is a promising strategy to specifically target phagocytic cells and modulate their own functions.