Clinical pattern characterisation of cattle naturally infected by BTV-8 - Clinical characterisation of BTV-8 infected cattle; Martinelle, Ludovic ; Guyot, Hugues et alin Transboundary and Emerging Diseases (in press) Detailed reference viewed: 16 (7 ULg) Molecular epidemiology of norovirus in symptomatic and asymptomatic population in Burkina FasoHUYNEN, Pascale ; Mauroy, Axel ; et alPoster (2012, September) Background Noroviruses (NoV), belonging to the family Caliciviridae, are now recognized as the leading cause of gastroenteritis outbreaks worldwide, and represent an important cause of sporadic ... [more ▼] Background Noroviruses (NoV), belonging to the family Caliciviridae, are now recognized as the leading cause of gastroenteritis outbreaks worldwide, and represent an important cause of sporadic gastroenteritis in both children and adults. Many studies describe NoV epidemiology. However, few data are available about the NoV strains circulating in most of African countries, in particular in Burkina Faso. The population of Burkina Faso is characterized by the young age of its habitants, and most are living in rural environment. Objectives The purpose of this epidemiological study was to determine the prevalence of NoV in Bobo Dioulasso (Southern part of Burkina Faso) by molecular diagnosis methods in patients presenting or not gastroenteritis symptoms, to quantify the excreted viral load, and to genotype the circulating strains. Methods Patients with and without gastro-intestinal disorders were selected in several Health Care Centres of Bobo Dioulasso. Clinical and epidemiological data, as well as stool samples, were collected during 8 weeks through March to April 2011. Viral genomic RNA was automatically extracted with a Maxwell® (Promega) instrument. Molecular detection of genogroups (G) I, II and IV NoV in stool samples was performed by a home-made real-time RT-PCR targeting the ORF1-ORF2 polymerase junction region. For each positive sample, viral load was estimated by using standard curves (successive dilutions of recombinant GI and GII plasmids). Molecular characterization was performed on the detected strains, using both polymerase and capsid regions. Results NoV were detected in 21.6% of the 453 collected stool samples, with a distribution of 21.0% and 23.1% in the samples from the 319 symptomatic (SP) and the 134 asymptomatic patients (AP) respectively. Genogroup distribution was 7.2% for GI, 10.7% for GII and 3.1% for both GI and GII among SP’s samples, and was 11.2% for GI, 10.4% for GII and 1.5% for both GI and GII among AP’s samples. Average viral load values were higher for GI NoV in SP than in AP (p=0.02), when they were higher for GII NoV in AP than in SP (p=0.04). Phylogenic analysis showed a high degree of genotypical diversity in both groups of patients. One recombinant strain GII.7/GII.6 was also detected, to our knowledge, for the first time. Conclusion Even if a true pathogenic role of NoV could not be showed from the study design, it allowed to precise the molecular epidemiology of NoV strains prevalent in a representative country of the East African region. It also showed that asymptomatic patients could play an important role as a NoV “reservoir”. Despite the fact that GII strains, and more precisely those belonging to GII.4 genotype, are nowadays highly reported worldwide, the surprising proportion of NoV GI detected in this study suggests that GI and GII strains should be excreted in equal proportion in the environment. The origin of this epidemiologic difference, even if partially explained by the difference in immunity and genetic sensitivity of the population, is still to be solved. [less ▲] Detailed reference viewed: 11 (1 ULg) Typage des souches de Norovirus circulant dans les populations symptomatiques et asymptomatiques au Burkina FasoHUYNEN, Pascale ; Mauroy, Axel ; et alPoster (2012) Appartenant à la famille des Caliciviridae, genre Norovirus, les norovirus (NoV) sont des virus non enveloppés dont le génome est composé d’un ARN monocaténaire de polarité positive ... [more ▼] Appartenant à la famille des Caliciviridae, genre Norovirus, les norovirus (NoV) sont des virus non enveloppés dont le génome est composé d’un ARN monocaténaire de polarité positive d’approximativement 7,5 kb. Les NoV infectent l’homme chez qui ils représentent au niveaumondial un agent majeur de gastroentérites épidémiques, d’origine souvent alimentaire mais aussi sporadique, et ce, toutes classes d'âges confondues. Les souches humaines sont classées génétiquement dans différents génotypes au sein de trois des cinq génogroupes, nommés (G) I, II et IV, composant le genre Norovirus. La voie de transmission des NoV est féco-orale. Les NoV sont très résistants dans l’environnement et la dose infectieuse est faible. Dans la population humaine, une grande diversité de souches appartenant principalement aux G I et II co-circulent. Parmi ces souches, le génotype Lordsdale (GII-4) est prédominant dans les épidémies actuelles, notamment lorsqu'une transmission de personne à personne est incriminée, alors que les souches du G I semblent plus fréquemment rapportées au cours des épidémies d’origine environnementale, comme celles liées à la consommation de fruits de mer. Si de nombreuses études d'épidémiologie moléculaire concernant ces virus ont été réalisées dans les pays industrialisés, les données sont par contre manquantes ou ténues pour bien des pays non industrialisés, et en particulier africains. Au cours d'une étude épidémiologique réalisée à Bobo Dioulasso au Burkina Faso et portant sur la prévalence des NoV dans les échantillons de selles de patients présentant ou non des symptômes de gastro-entérite, les souches détectées ont été quantifiées, leur génogroupe a été déterminé et pour certaines d'entre elles le génotype a été précisé. Quatre cent cinquante trois patients ont été prélevés, dont 319 présentant des symptômes diarrhéiques et 134 sujets témoins ne présentant pas de symptomatologie digestive. La détection des NoV et la quantification des charges virales excrétées ont été effectuées sur tous les échantillons par RT-PCR en temps réel permettant de discriminer les souches appartenant aux G I ou II. Une RT-PCR conventionnelle visant les régions de la polymérase (ORF1 du virus) ou de la capside (ORF2) a ensuite été réalisée sur une partie des échantillons détectés positifs en vue du séquençage de ces régions. Les relations phylogénétiques des souches circulant dans la population du Burkina Faso aux souches de référence ont aussi été inférées. Les résultats de RT-PCR en temps réel ont permis de mettre en évidence que les prévalences apparentes de l'infection par les NoV sont similaires dans les populations symptomatique et asymptomatique : une détection moléculaire de NoV chez 67 patients présentant de la diarrhée (21,0 %) et chez 31 des sujets témoins (23,1 %) a pu être observée. Les génotypes circulant détectés sont très variés dans les deux génogroupes, avec une proportion assez surprenante de NoV appartenant au G I. Université polytechnique de Bobo-Dioulasso, Institut supérieur des Sciences de la Santé (INSSA), Bobo-Dioulasso, Burkina Faso. Cette étude a permis de préciser l'épidémiologie moléculaire des souches de NoV circulant dans un pays représentatif de l'Afrique de l'Ouest. Elle a également montré que des individus asymptomatiques pourraient jouer un rôle assez important de réservoir du virus. Elle souligne enfin que, malgré le fait que les souches GII, et en particulier celles de génotype GII.4, soient à l'heure actuelle rapportées majoritairement au niveau mondial, les souches G I doivent être excrétées en égale proportion dans l'environnement. L'origine épidémiologique de la différence entre les prévalences apparentes des infections par les souches de GI et de GII, bien que partiellement expliquée par les différences de sensibilité génétique et d'immunité de population, reste donc à élucider. Remerciements: à la fondation A. Seghers, au Centre de Coopération au Développement de l'Université de Liège, à R. Boreux (assistance technique), aux membres du laboratoire du CMA de Dô et aux agents de santé de Bobo-Dioulasso (Burkina-Faso). [less ▲] Detailed reference viewed: 8 (0 ULg) A Review of Known and Hypothetical Transmission Routes for Noroviruses; ; et al in Food and Environmental Virology (2012), 4(4), 131-152 Human noroviruses (NoVs) are considered a worldwide leading cause of acute non-bacterial gastroenteritis. Due to a combination of prolonged shedding of high virus levels in feces, virus particle shedding ... [more ▼] Human noroviruses (NoVs) are considered a worldwide leading cause of acute non-bacterial gastroenteritis. Due to a combination of prolonged shedding of high virus levels in feces, virus particle shedding during asymptomatic infections, and a high environmental persistence, NoVs are easily transmitted pathogens. Norovirus (NoV) outbreaks have often been reported and tend to affect a lot of people. NoV is spread via feces and vomit, but this NoV spread can occur through several transmission routes. While person-to-person transmission is without a doubt the dominant transmission route, human infective NoV outbreaks are often initiated by contaminated food or water. Zoonotic transmission of NoV has been investigated, but has thus far not been demonstrated. The presented review aims to give an overview of these NoV transmission routes. Regarding NoV person-to-person transmission, the NoV GII. 4 genotype is discussed in the current review as it has been very successful for several decades but reasons for its success have only recently been suggested. Both pre-harvest and post-harvest contamination of food products can lead to NoV food borne illness. Pre-harvest contamination of food products mainly occurs via contact with polluted irrigation water in case of fresh produce or with contaminated harvesting water in case of bivalve molluscan shellfish. On the other hand, an infected food handler is considered as a major cause of post-harvest contamination of food products. Both transmission routes are reviewed by a summary of described NoV food borne outbreaks between 2000 and 2010. A third NoV transmission route occurs via water and the spread of NoV via river water, ground water, and surface water is reviewed. Finally, although zoonotic transmission remains hypothetical, a summary on the bovine and porcine NoV presence observed in animals is given and the presence of human infective NoV in animals is discussed. © 2012 Springer Science+Business Media New York. [less ▲] Detailed reference viewed: 13 (3 ULg) Molecular Detection and Genotyping of Noroviruses; ; et al in Food and Environmental Virology (2012), 4(4), 153-167 Noroviruses (NoVs) are a major cause of gastroenteritis worldwide in humans and animals and are known as very infectious viral agents. They are spread through feces and vomit via several transmission ... [more ▼] Noroviruses (NoVs) are a major cause of gastroenteritis worldwide in humans and animals and are known as very infectious viral agents. They are spread through feces and vomit via several transmission routes involving person-to-person contact, food, and water. Investigation of these transmission routes requires sensitive methods for detection of NoVs. As NoVs cannot be cultivated to date, detection of these viruses relies on the use of molecular methods such as (real-time) reverse transcriptase polymerase chain reaction (RT-PCR). Regardless of the matrix, detection of NoVs generally requires three subsequent steps: a virus extraction step, RNA purification, and molecular detection of the purified RNA, occasionally followed by molecular genotyping. The current review mainly focused on the molecular detection and genotyping of NoVs. The most conserved region in the genome of human infective NoVs is the ORF1/ORF2 junction and has been used as a preferred target region for molecular detection of NoVs by methods such as (real-time) RT-PCR, NASBA, and LAMP. In case of animal NoVs, broad range molecular assays have most frequently been applied for molecular detection. Regarding genotyping of NoVs, five regions situated in the polymerase and capsid genes have been used for conventional RT-PCR amplification and sequencing. As the expected levels of NoVs on food and in water are very low and inhibition of molecular methods can occur in these matrices, quality control including adequate positive and negative controls is an essential part of NoV detection. Although the development of molecular methods for NoV detection has certainly aided in the understanding of NoV transmission, it has also led to new problems such as the question whether low levels of human NoV detected on fresh produce and shellfish could pose a threat to public health. © 2012 Springer Science+Business Media New York. [less ▲] Detailed reference viewed: 17 (4 ULg) Complete genome sequence of a novel bovine norovirus: Evidence for slow genetic evolution in genogroup III genotype 2 norovirusesMauroy, Axel ; ; et alin Journal of Virology (2012), 86(22), 12449-12450 A new genogroup III genotype 2 bovine norovirus, B309/2003/BE, was entirely sequenced and genetically compared to the original Newbury2/1976/UK strain and to Dumfries/1994/UK, detected in 1976 and 1994 ... [more ▼] A new genogroup III genotype 2 bovine norovirus, B309/2003/BE, was entirely sequenced and genetically compared to the original Newbury2/1976/UK strain and to Dumfries/1994/UK, detected in 1976 and 1994, respectively. Interestingly, except in welldefined coding regions (N-terminal protein, 3A-like protease, hypervariable region of the capsid protein, and C-terminal part of the minor structural protein), very low genetic differences were noted between the entire genomes of these three strains along a 30-year-long period. It allowed some hypotheses of hotspots of genetic evolution through a low genetic evolution background in genotype 2 genogroup III bovine noroviruses. © 2012, American Society for Microbiology. [less ▲] Detailed reference viewed: 13 (3 ULg) Alternative attachment factors and internalization pathways for GIII.2 bovine noroviruses.Mauroy, Axel ; Gillet, Laurent ; Mathijs, Elisabeth et alin Journal of General Virology (The) (2011) Bovine noroviruses belong to the family Caliciviridae, genus Norovirus. Two genotypes are described and viruses genetically related to the Jena and Newbury-2 strains are classified into genotypes 1 and 2 ... [more ▼] Bovine noroviruses belong to the family Caliciviridae, genus Norovirus. Two genotypes are described and viruses genetically related to the Jena and Newbury-2 strains are classified into genotypes 1 and 2 respectively. In this study, virus-like particles (VLP) of the previously detected B309 Belgian strain, genetically related to genotype 2 bovine noroviruses, were used to investigate virus-host interactions in vitro. B309 VLP were shown to bind to several bovine cell lines. This binding was not affected by heparinase or chondroitinase treatment but was significantly inhibited by both sodium periodate, alpha-galactosidase, trypsin and phospholipase C treatment. Cell treatment by neuraminidase also moderately affected this binding. Taken together, these results show that, in addition to a galactosyl residue, sialic acid could also be involved in binding to susceptible cells. In addition, both the cholesterol-dependent pathway and macropinocytosis are used for B309 VLP internalisation by Madin-Darby Bovine Kidney cells. The data increase the knowledge on bovine norovirus cell interactions. [less ▲] Detailed reference viewed: 27 (8 ULg) Les Norovirus : grands coupables méconnus de gastro-entéritesMauroy, Axel ; HUYNEN, Pascale ; De Mol, Patrick et alin Revue de la Médecine Générale [=RMG] (2011), (285), 316-321 Recently, noroviruses emer- ged worldwide as a main and frequent cause of sporadic and epidemic gastroenteritis. The symptomatology they cause is usually benign. Their real impact lies on Public Health ... [more ▼] Recently, noroviruses emer- ged worldwide as a main and frequent cause of sporadic and epidemic gastroenteritis. The symptomatology they cause is usually benign. Their real impact lies on Public Health and Food Safety levels. [less ▲] Detailed reference viewed: 14 (0 ULg) Virus de la diahrée virale bovine : de la diversité à la singularitéMauroy, Axel ; Thiry, Etienne ![]() in Point Vétérinaire (2011) Detailed reference viewed: 13 (1 ULg) Experimental evidence of recombination in murine norovirusesMathijs, Elisabeth ; Muylkens, Benoît ; Mauroy, Axel et alin Journal of General Virology (The) (2010) Detailed reference viewed: 17 (7 ULg) Etude du risque zoonotique lié aux calicivirus entériques animaux par l'analyse des séquences détectées dans les espèces porcine et bovine et des interactions entre les norovirus bovins et les cellulesMauroy, Axel ![]() Doctoral thesis (2010) Detailed reference viewed: 26 (2 ULg) Molecular detection of kobuviruses and recombinant noroviruses in cattle in continental EuropeMauroy, Axel ; ; Mathijs, Elisabeth et alin Archives of Virology (2009), 154 Detailed reference viewed: 49 (4 ULg) Epidemiological study of bovine norovirus infection by RT-PCR and a VLP-based antibody ELISAMauroy, Axel ; Scipioni, Alexandra ; Mathijs, Elisabeth et alin Veterinary Microbiology (2009), 137 Detailed reference viewed: 64 (21 ULg) Spécificité et diversité virales : implications en médecine vétérinaireThiry, Etienne ; Dal Pozzo, Fabiana ; Muylkens, Benoît et alin Bulletin des Groupements Techniques Vétérinaires (2008), hors série Detailed reference viewed: 54 (7 ULg) Les mécanismes et les conséquences sur l'organisme des infections virales chez les équidésThiry, Etienne ; Dal Pozzo, Fabiana ; Mathijs, Elisabeth et alin Nouveau Praticien Vétérinaire Equine (2008), 5 Detailed reference viewed: 30 (6 ULg) Noroviruses and sapoviruses in pigs in Belgium.Mauroy, Axel ; Scipioni, Alexandra ; Mathijs, Elisabeth et alin Archives of Virology (2008), 153(10), 1927-31 Porcine noroviruses and sapoviruses belong to the family Caliciviridae and are rarely reported in European countries. In this study, swine stools from a region representative of northern Europe were ... [more ▼] Porcine noroviruses and sapoviruses belong to the family Caliciviridae and are rarely reported in European countries. In this study, swine stools from a region representative of northern Europe were screened for these viruses by RT-PCR. Both porcine noroviruses and sapoviruses were detected, showing their circulation in this region. The porcine norovirus strains were genetically related to genotype 19 strains in the genogroup II of the genus Norovirus. The porcine sapovirus strains were genetically related to the porcine enteric calicivirus Cowden reference strain and to newly described porcine strains in the genus Sapovirus. [less ▲] Detailed reference viewed: 42 (6 ULg) A SYBR Green RT-PCR assay in single tube to detect human and bovine noroviruses and control for inhibition.Scipioni, Alexandra ; Mauroy, Axel ; Ziant, Dominique et alin Virology journal (2008), 5 BACKGROUND: Noroviruses are single-stranded RNA viruses belonging to the family Caliciviridae. They are a major cause of epidemic and sporadic gastroenteritis in humans and clinical signs and lesions of ... [more ▼] BACKGROUND: Noroviruses are single-stranded RNA viruses belonging to the family Caliciviridae. They are a major cause of epidemic and sporadic gastroenteritis in humans and clinical signs and lesions of gastroenteritis were reported in bovines. Due to their genetic proximity, potential zoonotic transmission or animal reservoir can be hypothesized for noroviruses. RT-PCR has become the "gold standard" for the detection of noroviruses in faecal and environmental samples. With such samples, the control for inhibition of the reaction during amplification and detection is crucial to avoid false negative results, which might otherwise not be detected. The aim of the reported method is to detect, with a SYBR Green technology, a broad range of noroviruses with a control for inhibition. RESULTS: A SYBR Green real-time RT-PCR assay was developed making use of a foreign internal RNA control added in the same tube. This assay is able to detect human and bovine noroviruses belonging to genogroups I, II and III and to distinguish between norovirus and internal control amplicons using melting curve analysis. A 10-fold dilution of samples appears to be the method of choice to remove inhibition. This assay was validated with human and bovine stool samples previously tested for norovirus by conventional RT-PCR. CONCLUSION: This SYBR Green real-time RT-PCR assay allows the detection of the most important human and bovine noroviruses in the same assay, and avoids false negative results making use of an internal control. Melting curves allow the discrimination between the internal control and norovirus amplicons. It gives preliminary information about the species of origin. The sensitivity of the developed assay is higher than conventional RT-PCR and a 10-fold dilution of samples showed a better efficiency and reproducibility to remove RT-PCR inhibition than addition of bovine serum albumin. [less ▲] Detailed reference viewed: 60 (6 ULg) Detection and quantification of human and bovine novoviruses by a TaqMan RT-PCR assay with a control for inhibition.Scipioni, Alexandra ; ; Mauroy, Axel et alin Molecular and Cellular Probes (2008), 22 Noroviruses are single-stranded RNA viruses belonging to the family Caliciviridae. They are a major cause of epidemic and sporadic gastroenteritis in humans and calves. Reverse transcription-polymerase ... [more ▼] Noroviruses are single-stranded RNA viruses belonging to the family Caliciviridae. They are a major cause of epidemic and sporadic gastroenteritis in humans and calves. Reverse transcription-polymerase chain reaction (RT-PCR) has become the ‘‘gold standard’’ for detection of noroviruses in faecal and environmental samples. However, false negative results due to co-concentration of RT-PCR inhibitors are a continuous concern. A TaqMan real-time RT-PCR assay making use of a foreign internal RNA control and a RNA standard was developed. Very interestingly, this method is capable of detecting human noroviruses belonging to genogroups I and II, and bovine noroviruses belonging to genogroup III. Inhibitors were removed efficiently by 1/10 dilution of the sample or addition of bovine serum albumin to the RT-PCR mix. This assay was validated with human and bovine stool samples previously tested for norovirus by conventional RT-PCR. The ability to detect norovirus in stool samples that were negative by conventional RT-PCR assay demonstrate the higher sensitivity of the TaqMan assay compared to the conventional RT-PCR assay. This real-time RT-PCR assay allows the detection of both human and bovine noroviruses, avoids false negative results and is able to quantify the level of norovirus contamination. [less ▲] Detailed reference viewed: 66 (17 ULg) Bluetongue in captive yaks.Mauroy, Axel ; Guyot, Hugues ; et alin Emerging Infectious Diseases (2008), 14(4), 675-6 In August 2006, several Northern European countries including Belgium reported their first cases of bluetongue (BT). Surprisingly, it was the first time that BT was diagnosed so far in the northern ... [more ▼] In August 2006, several Northern European countries including Belgium reported their first cases of bluetongue (BT). Surprisingly, it was the first time that BT was diagnosed so far in the northern hemisphere (1). BT is a non contagious, arthropod borne animal disease. The causal virus belongs to the genus Orbivirus in the family Reoviridae. The genome of the bluetongue virus (BTV) consists of 10 segments of double-stranded RNA and 24 serotypes have been reported (2). Serotype 8 (BTV-8) was implied in the emergence in Belgium (3). All ruminant species are thought to be susceptible to BT (2) but lack of data remains for certain species. We report here laboratory confirmed clinical cases of BT in yaks. [less ▲] Detailed reference viewed: 31 (8 ULg) Animal noroviruses.Scipioni, Alexandra ; Mauroy, Axel ; et alin Veterinary Journal (2008), 178(1), 32-45 Among enteric caliciviruses, noroviruses belong to the genus Norovirus, one of the four accepted genera in the family Caliciviridae. These single-stranded, positive-sense RNA viruses are highly variable ... [more ▼] Among enteric caliciviruses, noroviruses belong to the genus Norovirus, one of the four accepted genera in the family Caliciviridae. These single-stranded, positive-sense RNA viruses are highly variable both genetically and antigenically. Several animal enteric caliciviruses that are morphologically indistinguishable and genetically closely related to human noroviruses have been identified. The first bovine enteric noroviruses were described in Great Britain and are known as Newbury Agent 2. At least three genetic clusters of porcine noroviruses join together within genogroup II noroviruses. Human noroviruses are the most important cause of acute gastroenteritis illness in people of all ages. In the USA, they are associated with approximately 30-50% of all food-borne outbreaks. Until now, noroviruses have not been associated with gastroenteritis outbreaks in immunocompetent animals. Neither bovine nor porcine noroviruses can replicate in cell culture, although human norovirus can grow in a complex 3D culture system. However, the recently discovered murine noroviruses can replicate in cell culture and are therefore used as model viruses to study human noroviruses. This review focusses on virus classification, virion structure, pathogenesis, epidemiology, immune response and diagnosis of animal noroviruses in comparison with human noroviruses. The classification of animal enteric caliciviruses within the Norovirus genus raises the question of whether transmission from an animal reservoir to humans could occur. Answering this question is important in determining the risk of cross-species infections affecting the epidemiology and evolution of these viruses and so complicating the control of human norovirus infections. [less ▲] Detailed reference viewed: 67 (16 ULg) |
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