Nucleotides sequence alignment of the RPO30 gene of CaPVs highlighting the snapback tail binding site.

<p>The RPO30 gene sequences of 7 CaPVs representing GTPVs, LSDVs and SPPVs were aligned. A sequence of sixteen nucleotides complementary to the snapback tail in GTPV (100% match), as well as the corresponding positions in SPPV and LSDV are shown in the box. Note the targeted single nucleotide mismatches inside the box: T:A between GTPV and SPPV, and T:G between GTPV and LSDV. Conserved nucleotides are shown as dots.</p

Secondary structure of the expected GTPV PCR amplicon.

<p>The Snapback hairpin with a 18-nucleotide stem and a loop of 55 bases is shown. The predictions were done on the Mfold web server <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0075971#pone.0075971-Zuker1" target="_blank">[28]</a> using the default parameters of the DNA folding form except for the temperature which was set at 45°C and the salt concentration set at 50 mM.</p

Snapback primer genotyping of CaPVs.

<p>The fluorescence melting curve analysis of the PCR products shows two melting peaks for each of the CaPV three genotypes (GTPV, SPPV and LSDV) corresponding to the snapback stem melting peak at lower temperature and the full-length PCR amplicon melting peak at higher temperature (see arrows).</p

Sequences of the snapback and reverse primers.

<p>The snapback tail of 16 bases is shown as underlined; 2 nucleotides mismatch at the 5′end are indicated as lowercase; and mismatch nucleotide as underlined bold (A).</p

High-Resolution melting curve analysis of CaPVs using the Precision Melt Analysis™ software (BioRad).

<p>A: the normalized melt curve of the full-length amplicon; B: the difference curve of full-length amplicon; C: the normalized melt curve of snapback stem; D: The difference curve of the snapback stem. The species are indicated by the arrows: G = GTPV, S = SPPV and L = LSDV.</p

Transovarial passage and transmission of LSDV by Amblyomma hebraeum, Rhipicephalus appendiculatus and Rhipicephalus decoloratus

Lumpy skin disease (LSD), an acute, sub-acute or inapparent disease of cattle, is caused by lumpy skin disease virus (LSDV), a member of the genus Capripoxvirus in the family Poxviridae. LSD is characterised by high fever, formation of circumscribed skin lesions and ulcerative lesions on the mucous membranes of the mouth, respiratory and digestive tracts. It is an economically important disease due to the permanent damage to hides, the reduction in productivity and trade restrictions imposed on affected areas. Transmission has been associated with blood-feeding insects such as stable flies (Stomoxysis calcitrans) and mosquitoes (Aedes aegypti). Mechanical (intrastadial) and transstadial transmission by Amblyomma hebraeum and Rhipicephalus appendiculatus as well as transovarial transmission by R. decoloratus have been reported. In this study transovarial passage of LSDV to larvae and subsequent transmission to recipient animals were demonstrated. The finding of transovarial passage of LSDV in female ticks shows the potential for A. hebraeum, R. appendiculatus and R. decoloratus to be reservoir hosts for LSDV.The Combating Infectious Diseases of Livestock for International Development (CIDLID) research programme, the Department of International Biotechnology and Biological Sciences Research Council (BBSRC), the UK government, the Department for International Development (DFID) and the Scottish Government (CIDLID project number BB/H009361/1)http://link.springer.com/journal/10493hb2014ab201