Ticks from migratory birds as possible route to spread Crimean-Congo Hemorrhagic fever virus : preliminary data about species composition from three Italian islands

Migratory birds have been known to be passive carriers of arthropod vectors of various pathogens. A study to assess the potential role of migratory birds, which are tick-carriers, as a possible route to spread the Crimean-Congo Hemorrhagic Fever Virus (CCHFV) into Europe was initiated in 2017 by the Italian Ministry of Health. Ticks were collected from birds during seasonal bird ringing activities in Mediterranean islands known to be stop-over sites for migratory birds from Africa or from East Europe and Turkey.peer-reviewe

First external quality assessment of molecular and serological detection of rift valley fever in the western Mediterranean region

Pas de clé UTRift Valley fever (RVF) is a mosquito-borne viral zoonosis which affects humans and a wide range of domestic and wild ruminants. The large spread of RVF in Africa and its potential to emerge beyond its geographic range requires the development of surveillance strategies to promptly detect the disease outbreaks in order to implement efficient control measures, which could prevent the widespread of the virus to humans. The Animal Health Mediterranean Network (REMESA) linking some Northern African countries as Algeria, Egypt, Libya,Mauritania, Morocco, Tunisia with Southern European ones as France, Italy, Portugal and Spain aims at improving the animal health in the Western Mediterranean Region since 2009. In this context, a first assessment of the diagnostic capacities of the laboratories involved in the RVF surveillance was performed. The first proficiency testing (external quality assessment— EQA) for the detection of the viral genome and antibodies of RVF virus (RVFV) was carried out from October 2013 to February 2014. Ten laboratories participated from 6 different countries (4 from North Africa and 2 from Europe). Six laboratories participated in the ring trial for both viral RNA and antibodies detection methods, while four laboratories participated exclusively in the antibodies detection ring trial. For the EQA targeting the viral RNA detection methods 5 out of 6 laboratories reported 100% of correct results. One laboratory misidentified 2 positive samples as negative and 3 positive samples as doubtful indicating a need for corrective actions. For the EQA targeting IgG and IgM antibodies methods 9 out of the 10 laboratories reported 100% of correct results, whilst one laboratory reported all correct results except one false-positive. These two ring trials provide evidence that most of the participating laboratories are capable to detect RVF antibodies and viral RNA thus recognizing RVF infection in affected ruminants with the diagnostic methods currently available

Ultra-Efficient PrPSc Amplification Highlights Potentialities and Pitfalls of PMCA Technology

In order to investigate the potential of voles to reproduce in vitro the efficiency of prion replication previously observed in vivo, we seeded protein misfolding cyclic amplification (PMCA) reactions with either rodent-adapted Transmissible Spongiform Encephalopathy (TSE) strains or natural TSE isolates. Vole brain homogenates were shown to be a powerful substrate for both homologous or heterologous PMCA, sustaining the efficient amplification of prions from all the prion sources tested. However, after a few serial automated PMCA (saPMCA) rounds, we also observed the appearance of PK-resistant PrPSc in samples containing exclusively unseeded substrate (negative controls), suggesting the possible spontaneous generation of infectious prions during PMCA reactions. As we could not definitively rule out cross-contamination through a posteriori biochemical and biological analyses of de novo generated prions, we decided to replicate the experiments in a different laboratory. Under rigorous prion-free conditions, we did not observe de novo appearance of PrPSc in unseeded samples of M109M and I109I vole substrates, even after many consecutive rounds of saPMCA and working in different PMCA settings. Furthermore, when positive and negative samples were processed together, the appearance of spurious PrPSc in unseeded negative controls suggested that the most likely explanation for the appearance of de novo PrPSc was the occurrence of cross-contamination during saPMCA. Careful analysis of the PMCA process allowed us to identify critical points which are potentially responsible for contamination events. Appropriate technical improvements made it possible to overcome PMCA pitfalls, allowing PrPSc to be reliably amplified up to extremely low dilutions of infected brain homogenate without any false positive results even after many consecutive rounds. Our findings underline the potential drawback of ultrasensitive in vitro prion replication and warn on cautious interpretation when assessing the spontaneous appearance of prions in vitro

Correlation between infectivity and disease associated prion protein in the nervous system and selected edible tissues of naturally affected scrapie sheep

<div><p>The transmissible spongiform encephalopathies (TSEs) or prion diseases are a group of fatal neurodegenerative disorders characterised by the accumulation of a pathological form of a host protein known as prion protein (PrP). The validation of abnormal PrP detection techniques is fundamental to allow the use of high-throughput laboratory based tests, avoiding the limitations of bioassays. We used scrapie, a prototype TSE, to examine the relationship between infectivity and laboratory based diagnostic tools. The data may help to optimise strategies to prevent exposure of humans to small ruminant TSE material via the food chain. Abnormal PrP distribution/accumulation was assessed by immunohistochemistry (IHC), Western blot (WB) and ELISA in samples from four animals. In addition, infectivity was detected using a sensitive bank vole bioassay with selected samples from two of the four sheep and protein misfolding cyclic amplification using bank vole brain as substrate (vPMCA) was also carried out in selected samples from one animal. Lymph nodes, oculomotor muscles, sciatic nerve and kidney were positive by IHC, WB and ELISA, although at levels 100–1000 fold lower than the brain, and contained detectable infectivity by bioassay. Tissues not infectious by bioassay were also negative by all laboratory tests including PMCA. Although discrepancies were observed in tissues with very low levels of abnormal PrP, there was an overall good correlation between IHC, WB, ELISA and bioassay results. Most importantly, there was a good correlation between the detection of abnormal PrP in tissues using laboratory tests and the levels of infectivity even when the titre was low. These findings provide useful information for risk modellers and represent a first step toward the validation of laboratory tests used to quantify prion infectivity, which would greatly aid TSE risk assessment policies.</p></div

Bases génétiques des neuropathologies à prions (le modèle ovin)

Ce projet de recherche avait pour but d'identifier des gènes capables d'influencer la sensibilité/résistance à la scrapie chez le mouton, autres que PRNP. Nous avons abordé cette problématique par 2 voies complémentaires et consécutives. Nous avons tout d'abord utilisé u protocole classique de détection de QTL (Quantitative Trait Loci). Nous avons identifié par cette approche un QTL sur le chromosome ovi 18 (OAR18). Nous avons ainsi pu identifier de nouveaux microsatellites de façon ciblée, qui, une fois génotypes, ont confirmé l'associatio génétique avec le QTL et ont significativement raffiné sa localisation dans un intervalle d'environ 7 Mb. En parallèle aux progrès de l'étude génétique, nous avons développé un programme de recherche pour mettre en évidence les modifications des profils d'expression des gènes induits par la maladie dans le cerveau, en utilisant la technique de l'hybridation soustractive et suppressive (SSH). De cette façon, nous avons identifié six gènes influencés par la progression de la maladie, dont 5 son induits transcriptionnellement (CHN1, LRFN5, PPP2AC, CaMK2 and COX1) et l'un diminué (Rab9p40). Chez la souris, CHN1 est localisé 74 Mb sur le chromosome 2 (MMU2), précisément au centre de l'intervalle QTL identifié par Manolakou et coll. (2001 ). LRFN5 est localisé 40 Mb sur le chromosome 14, position correspondant à l'intervalle que nous avons identifié sur le chromosome 18. En ce qui concerne CHN1, nous avons pu démontrer l'existence d'un épissage alternatif, responsable de l'existence de deux isoformes, dont l'une est spécifiquement présente dans les cerveaux des animaux atteints.The objective of this research project was to identify gènes affecting thé genetic sensibility/resistance to scrapie in sheep, outside thé PRNI gène. We dealt with this scientific problem following two complementary ways. First, we carried out a classical approach of positional clonin for Quantitative Trait Loci (QTL). We could thus identify a QTL on sheep chromosome 18 (OAR18), linked to thé TGLA122 and BM7243 markers. Subsequently, by thé targeted production of new microsatellites, we could confirm thé genetic association with thé QTL and significantly refine ils localisation into a ~7 Mb interval. In parallel with thé progresses of this genetic study, we developed a research program aiming at analysing thé modifications of gènes expression profiles caused by thé disease in thé brain, using thé Subtractive Suppressive Hybridization (SSH) technique. This way we identified six gènes influenced by thé development of thé disease, 5 of them are stimulated (CHN1, LRFN5, PPP2AC, CaMK2 and COX1 ) and another one is down-regulated (Rab9p40). In mice CHN1 is located at 74 M on chromosome 2 (MMU2), exactly in thé center of a QTL interval identified by Manolakou el al in 2001. LRFN5 is located at 40 Mb on HSA14, a position corresponding to thé interval previously identified on OAR18. Concerning CHN1, we could demonstrate thé existence ( an alternative splicing, responsible for two isoforms, one of which is specifically présent in affected brain.VERSAILLES-BU Sciences et IUT (786462101) / SudocSudocFranceF

Episodi di peste equina in Namibia dal 2006 al 2013: Rilievi clinici, patologici e molecolari

African horse sickness (AHS) is a vector-borne viral disease of equids, endemic in Sub-Saharan Africa. This article reports the clinic-pathological and laboratory findings observed in the framework of passive surveillance during the AHS outbreaks which occurred in Namibia between 2006 and 2013. This study was conducted in the framework of the collaboration among the Istituto Zooprofilattico Sperimentale dell’Abruzzo e del Molise (Teramo, Italy), the Namibian Ministry of Agriculture Water and Forestry, and the Namibian National Veterinary Association. A total of 92 horses were investigated, showing different clinical form of AHS: peracute/acute (n = 43), sub-acute (n = 21) and mild AHS fever (n = 19). Clinical data were not available for 9 horses, because they were found dead. Pathological findings have been recorded for 35 horses. At necropsy, pulmonary and subcutaneous oedema, haemorrhages and enlargement of lymph nodes were mainly observed. Diagnosis was confirmed by laboratory testing, AHS virus (AHSV) was isolated from 50 horses and the identified serotypes were: 1, 2, 4, 6, 7, 8, and 9. The phylogenetic analysis of the S10 genome sequences segregated the Namibian AHSV strains in the same clusters of those circulating in South Africa in recent years. The description of AHS clinical, pathological, and laboratory features of AHS provided in this article is of value for differential diagnosis and control of AHS, especially in areas currently free from this disease

Health Status of the Eastern Grey Squirrel (Sciurus carolinensis) Population in Umbria: Results of the LIFE Project &lsquo;U-SAVEREDS&rsquo;

The introduction of the Eastern grey squirrel (Sciurus carolinensis) in Europe is one of the best-known cases of invasive alien species (IAS) colonisation, that poses a severe risk to the conservation of biodiversity. In 2003, it was released in a private wildlife park near the city of Perugia (Italy), where it is replacing the native Eurasian red squirrel (Sciurus vulgaris). The LIFE13 BIO/IT/000204 Project (U-SAVEREDS) was set up for the Sciurus vulgaris conservation in Umbria through an eradication campaign of grey squirrels. One hundred and fifty-four animals were analysed for bacteriological, mycological, virological, and serological investigations (C4 action). Sanitary screening showed that Sciurus carolinensis is a dermatophyte carrier, and therefore, it could cause public health issues for humans, considering its confident behaviour. Moreover, it has been marginally responsible for the spreading of Candida albicans, Coxiella burnetii, and Borrelia lusitaniae. Health status evaluation conducted on the Sciurus carolinensis population indicated that it is necessary to raise awareness of its impacts on biodiversity and human health. Moreover, the health status and behaviours of the IAS must be considered when control or eradication campaigns are planned