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Lorusso A.,OIE Reference Laboratory for Bluetongue | Sghaier S.,Institute Of La Recherche Veterinaire Of Tunisie Irvt | Carvelli A.,OIE Reference Laboratory for Bluetongue | Di Gennaro A.,OIE Reference Laboratory for Bluetongue | And 7 more authors.
Infection, Genetics and Evolution | Year: 2013

Since 2000 several bluetongue virus (BTV) incursions have occurred in Sardinia (Italy) involving serotypes 1, 2, 4, 8 and 16. In October 2012, new BT outbreaks caused by BTV-1 and BTV-4 were reported. Nearly 500 flocks were infected and 9238 sheep died because of the infection. When sequenced, Seg-10 of both strains shared 99% similarity at nucleotide level with BTV strains that have circulated in the Mediterranean basin in the last few years. Similarly, Seg-5 sequences of BTV-1 and BTV-4 newly isolated Sardinian strains are identical and cluster together with recent BTV-1 circulating in the Mediterranean Basin and the BTV-4 strains isolated in Tunisia in 2007 and 2009. These BTV-4 strains differ from the ones that circulated in Europe from 2003 to 2005 and appear to be reassortant strains. © 2013 Elsevier B.V. Source


Monaco F.,OIE Reference Laboratory for Bluetongue | Benedetto L.,OIE Reference Laboratory for Bluetongue | di Marcello V.,OIE Reference Laboratory for Bluetongue | Lelli R.,OIE Reference Laboratory for Bluetongue | Goffredo M.,OIE Reference Laboratory for Bluetongue
Veterinaria Italiana | Year: 2010

A real-time polymerase chain reaction (PCR) using a green fluorescence dye, followed by a comparison of derivative melting curves in the post-amplification phase, was developed to distinguish species of Culicoides within the Obsoletus Complex. The selected target sequence was internal transcribed spacer 2 (ITS 2) of the ribosomal DNA (rDNA). Using the newly developed method, 140 midges were morphologically classified in the Obsoletus Complex and were processed. The results were compared to those obtained by combining the morphological identification with the gel based reverse transcriptase (RT)-PCR. By analysing the species-specific pattern of the dissociation curves, it was possible to identify 52 midges as Culicoides scoticus, 82 midges as C. obsoletus sensu strictu and 6 as C. montanus. These results matched those obtained by the combination of gel-based PCR and morphological identification used on a routine basis. Given its diagnostic flexibility, rapid results, automation capability, high quality result performance and expression, the real-time ITS 2 rDNA PCR appears to be more functional and efficient than the gel-based PCR, especially when dealing with large-scale monitoring of midges belonging to the Obsoletus Complex. © IZS A&M 2010. Source


Lorusso A.,OIE Reference Laboratory for Bluetongue | Sghaier S.,Institute Of La Recherche Veterinaire Of Tunisie Irvt | Ancora M.,OIE Reference Laboratory for Bluetongue | Marcacci M.,OIE Reference Laboratory for Bluetongue | And 9 more authors.
Infection, Genetics and Evolution | Year: 2014

Western BTV-1 emerged in the Mediterranean basin in 2006 and it has since been isolated in southern and northern European countries. Six BTV-1 strains isolated from infected sheep in Italy between 2006 and 2013 and a BTV-1 strain isolated from an infected sheep in Tunisia in 2011 were fully sequenced. The seven strains were shown to be nearly identical in each gene segment. The Seg-2 sequences of the BTV-1 strains group according to the year of isolation reflecting the time of BTV incursions in Italy. Combined results suggest that BTV-1 strains isolated in Sardinia, Sicily and mainland Italy in 2012 and 2013 have a direct northern African origin. The Italian strains originated from a strain closely related to a BTV-1 strain isolated in Tunisia in 2011. Better surveillance programs with northern and sub-Saharan African countries should be implemented making the control of spread of BTV easier and effective. © 2014 Elsevier B.V. Source


Spedicato M.,OIE Reference Laboratory for Bluetongue | Carmine I.,OIE Reference Laboratory for Bluetongue | Teodori L.,OIE Reference Laboratory for Bluetongue | Leone A.,OIE Reference Laboratory for Bluetongue | And 5 more authors.
Vaccine | Year: 2016

Epizootic hemorrhagic disease (EHD) is an arthropod-borne infectious viral disease sustained by the epizootic hemorrhagic disease virus (EHDV). The only commercially available and currently used vaccines are manufactured for EHDV-2 in Japan, either live or inactivated vaccines. In this study we tested the innocuity for fetuses of the live attenuated EHDV-2 vaccine in five late-term pregnant cows. Whole blood and serum samples were collected from dams and screened for the presence of EHDV-2 RNA, infectious virus and antibodies. After calving, whole blood and serum samples collected from calves, before and after colostrum intake, were also tested for antibodies and for virus detection. In dams, neither fever nor clinical signs were observed. All of them seroconverted and a strong humoral response was detected throughout the sampling period. All blood samples tested negative for EHDV-2 except for one sample collected from a dam 11 days post-vaccination which tested positive at virus isolation at the third cell passage following two rounds of blind passages. Although they had free access to colostrum, calves tested serologically negative for EHDV-2 during the entire course of the experiment. Overall, the tested live attenuated vaccine can be safely administered to late-term pregnant cows as it was not demonstrated to cross the placental barrier. The safety of the live-attenuated vaccine is further confirmed by the emergence of Ibaraki virus in 2013 in Japan which is apparently not related to the spread of the vaccine strain currently used in Japan. © 2016 Elsevier Ltd. Source


Lorusso A.,OIE Reference Laboratory for Bluetongue | Baba D.,Center National Delevage Et Of Recherches Veterinaires Cnerv | Spedicato M.,OIE Reference Laboratory for Bluetongue | Teodori L.,OIE Reference Laboratory for Bluetongue | And 11 more authors.
Infection, Genetics and Evolution | Year: 2016

In March 2013, EDTA-blood and serum samples were collected from 119 cattle and 159 dromedaries at the slaughterhouse of Nouakchott, the capital city of the Islamic Republic of Mauritania. Serum samples were screened for the presence of Bluetongue (BT) antibodies by competitive ELISA (cELISA). Positive samples were then tested by serum-neutralization (SN) to determine BTV serotype. RNA from blood samples was first tested by a genus-specific quantitative RT-PCR assay which is able to detect all 27 existing BTV serotypes (RT-qPCR1-27). Positive samples were further screened by a RT-qPCR assay which, instead, is able to detect the classical 24 BTV serotypes only (RT-qPCR1-24). Of the 278 serum samples tested, 177 (mean = 63.7%; 95% CI: 57.9%-69.1%) resulted positive by cELISA. Of these, 69 were from cattle (mean = 58.0%; 95% CI: 49.0%-66.5%) and 108 from dromedaries (mean = 67.9%; 95% CI: 60.3%-74.7%). BTV-26 neutralizing antibodies were by far the most frequently found as they were detected in 146 animals with titres ranging from 1:10 to 1:80. Out of 278 blood samples, 25 (mean = 9.0%; 95% CI: 6.2%-12.9%) were found positive for BTV by RT-qPCR1-27, 20 (mean = 16.8%; 95% CI: 11.2%-24.6%) were from cattle and 5 (mean = 3.1%; 95% CI: 1.4%-7.1%) from dromedaries. When tested by RT-qPCR1-24 the 25 BTV positive samples were negative. Unfortunately, no genetic information by molecular typing or by next generation sequencing has been obtained as for the very low levels of RNA in the blood samples. © 2016 Elsevier B.V. Source

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