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Article ; Online: Phenotypic and Genetic Evolutions of a Porcine Reproductive and Respiratory Syndrome Modified Live Vaccine after Limited Passages in Pigs

Julie Eclercy / Patricia Renson / Edouard Hirchaud / Mathieu Andraud / Véronique Beven / Frédéric Paboeuf / Nicolas Rose / Yannick Blanchard / Olivier Bourry

Vaccines, Vol 9, Iss 392, p

2021 Volume 392

Abstract: Modified live vaccines (MLVs) against the porcine reproductive and respiratory syndrome virus (PRRSV) have been regularly associated with safety issues, such as reversion to virulence. In order to characterize the phenotypic and genetic evolution of the ... ...

Abstract	Modified live vaccines (MLVs) against the porcine reproductive and respiratory syndrome virus (PRRSV) have been regularly associated with safety issues, such as reversion to virulence. In order to characterize the phenotypic and genetic evolution of the PRRSV-1 DV strain from the Porcilis ® PRRS MLV after limited passages in pigs, three in vivo experiments were performed. Trial#1 aimed (i) at studying transmission of the vaccine strain from vaccinated to unvaccinated contact pigs. Trial#2 and Trial#3 were designed (ii) to assess the reproducibility of Trial#1, using another vaccine batch, and (iii) to compare the virulence levels of two DV strains isolated from vaccinated (passage one) and diseased contact pigs (passage two) from Trial#1. DV strain isolates from vaccinated and contact pigs from Trial#1 and Trial#2 were submitted to Next-Generation Sequencing (NGS) full-genome sequencing. All contact animals from Trial#1 were infected and showed significantly increased viremia compared to vaccinated pigs, whereas no such change was observed during Trial#2. In Trial#3, viremia and transmission were higher for inoculated pigs with passage two of the DV strain, compared with passage one. In this study, we showed that the re-adaptation of the DV strain to pigs is associated with faster replication and increased transmission of the vaccine strain. Punctually, a decrease of attenuation of the DV vaccine strain associated with clinical signs and increased viremia may occur after limited passages in pigs. Furthermore, we identified three mutations linked to pig re-adaptation and five other mutations as potential virulence determinants.
Keywords	PRRS virus ; modified live vaccine ; attenuation ; variants ; mutation ; Medicine ; R
Subject code	630
Language	English
Publishing date	2021-04-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Rabbit haemorrhagic disease virus Lagovirus europaeus/GI.1d strain

Clément Droillard / Evelyne Lemaitre / Michel Amelot / Yannick Blanchard / Alassane Keita / Nicolas Eterradossi / Ghislaine Le Gall-Reculé

BMC Veterinary Research, Vol 17, Iss 1, Pp 1-

genome sequencing, in vivo virus replication kinetics, and viral dose effect

2021 Volume 13

Abstract: Abstract Background Rabbit haemorrhagic disease virus Lagovirus europaeus/GI.1d variant (GI.1d/RHDV) was identified in 1990 in France, and until the emergence of the new genotype GI.2, it was the main variant circulating in the country. The early stages ... ...

Abstract	Abstract Background Rabbit haemorrhagic disease virus Lagovirus europaeus/GI.1d variant (GI.1d/RHDV) was identified in 1990 in France, and until the emergence of the new genotype GI.2, it was the main variant circulating in the country. The early stages of RHDV infection have been described in a few studies of rabbits experimentally infected with earlier strains, but no information was given on the minimum infective dose. We report the genomic and phenotypic characterisation of a GI.1d/RHDV strain collected in 2000 in France (GI.1d/00–21). Results We performed in vivo assays in rabbits to study virus replication kinetics in several tissues at the early stage of infection, and to estimate the minimum infective dose. Four tested doses, negligible (10− 1 viral genome copies), low (104), high (107) and very high (1011) were quantified using a method combining density gradient centrifugation of the viral particles and an RT-qPCR technique developed to quantify genomic RNA (gRNA). The GI.1d/00–21 genome showed the same genomic organisation as other lagoviruses; however, a substitution in the 5′ untranslated region and a change in the potential p23/2C-like helicase cleavage site were observed. We showed that the liver of one of the two rabbits inoculated via the oral route was infected at 16 h post-infection and all tissues at 39 h post-infection. GI.1d/00–21 induced classical RHD signs (depression) and lesions (haemorrhage and splenomegaly). Although infective dose estimation should be interpreted with caution, the minimum infective dose that infected an inoculated rabbit was lower or equal to 104 gRNA copies, whereas between 104 and 107 gRNA copies were required to also induce mortality. Conclusions These results provide a better understanding of GI.1d/RHDV infection in rabbits. The genome analysis showed a newly observed mutation in the 5′ untranslated region of a lagovirus, whose role remains unknown. The phenotypic analysis showed that the pathogenicity of GI.1d/00–21 and the replication kinetics in infected ...
Keywords	Lagovirus ; RHDV ; GI.1d ; Replication kinetics ; Minimum infective dose ; RT-qPCR ; Veterinary medicine ; SF600-1100
Subject code	570 ; 630
Language	English
Publishing date	2021-07-01T00:00:00Z
Publisher	BMC
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Targeted Next Generation Sequencing to study insert stability in genetically modified plants

Anne-Laure Boutigny / Audrey Barranger / Claire De Boisséson / Yannick Blanchard / Mathieu Rolland

Scientific Reports, Vol 9, Iss 1, Pp 1-

2019 Volume 9

Abstract: Abstract The EU directive 2001/18/EC requires any genetically modified (GM) event to be stable. In the present work, a targeted Next-Generation Sequencing (NGS) approach using barcodes to specifically tag each individual DNA molecules during library ... ...

Abstract	Abstract The EU directive 2001/18/EC requires any genetically modified (GM) event to be stable. In the present work, a targeted Next-Generation Sequencing (NGS) approach using barcodes to specifically tag each individual DNA molecules during library preparation was implemented to detect mutations taking into account the background noise due to amplification and sequencing errors. The method was first showed to be efficient in detecting the mutations in synthetic samples prepared with custom-synthesized mutated or non-mutated P35S sequences mixed in different proportions. The genetic stability of a portion of the P35S promoter targeted for GM detection was then analyzed in GM flour samples. Several low frequency mutations were detected in the P35S sequences. Some mutated nucleotides were located within the primers and probes used in the P35S diagnostic test. If present not as somatic mutations but as the consensus sequence of some individuals, these mutations could influence the efficiency of the P35S real time PCR diagnostic test. This methodology could be implemented in genetic stability studies of GM inserts but also to detect single nucleotide mutant GM plants produced using “new breeding techniques”.
Keywords	Medicine ; R ; Science ; Q
Subject code	500
Language	English
Publishing date	2019-02-01T00:00:00Z
Publisher	Nature Publishing Group
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Whole genome sequencing and phylogenetic characterisation of rabies virus strains from Moldova and north-eastern Romania.

Mihaela Anca Dascalu / Evelyne Picard-Meyer / Emmanuelle Robardet / Alexandre Servat / Serghei Arseniev / Oxana Groza / Nicolae Starciuc / Vlad Vuta / Florica Barbuceanu / Oana Irina Tanase / Florentina Daraban Bocaneti / Helene Quenault / Edouard Hirchaud / Yannick Blanchard / Elena Velescu / Florence Cliquet

PLoS Neglected Tropical Diseases, Vol 17, Iss 7, p e

2023 Volume 0011446

Abstract: Background Rabies is the oldest fatal zoonotic disease recognised as a neglected tropical disease and is caused by an RNA virus belonging to the genus Lyssavirus, family Rhabdoviridae. Methodology/principal findings A deep molecular analysis was ... ...

Abstract	Background Rabies is the oldest fatal zoonotic disease recognised as a neglected tropical disease and is caused by an RNA virus belonging to the genus Lyssavirus, family Rhabdoviridae. Methodology/principal findings A deep molecular analysis was conducted on full-length nucleoprotein (N) gene and whole genome sequences of rabies virus from 37 animal brain samples collected between 2012 and 2017 to study the circulation of rabies virus (RABV) variants. The overall aim was to better understand their distribution in Moldova and north-eastern Romania. Both Sanger and high throughput sequencing on Ion Torrent and Illumina platforms were performed. Phylogenetic analysis of the RABV sequences from both Moldova and Romania revealed that all the samples (irrespective of the year of isolation and the species) belonged to a single phylogenetic group: north-eastern Europe (NEE), clustering into three assigned lineages: RO#5, RO#6 and RO#7. Conclusions/significance High throughput sequencing of RABV samples from domestic and wild animals was performed for the first time for both countries, providing new insights into virus evolution and epidemiology in this less studied region, expanding our understanding of the disease.
Keywords	Arctic medicine. Tropical medicine ; RC955-962 ; Public aspects of medicine ; RA1-1270
Subject code	590
Language	English
Publishing date	2023-07-01T00:00:00Z
Publisher	Public Library of Science (PLoS)
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Characterization of plasmids harboring blaCTX-M and blaCMY genes in E. coli from French broilers.

Fabrice Touzain / Laetitia Le Devendec / Claire de Boisséson / Sandrine Baron / Eric Jouy / Agnès Perrin-Guyomard / Yannick Blanchard / Isabelle Kempf

PLoS ONE, Vol 13, Iss 1, p e

2018 Volume 0188768

Abstract: Resistance to extended-spectrum cephalosporins (ESC) is a global health issue. The aim of this study was to analyze and compare plasmids coding for resistance to ESC isolated from 16 avian commensal and 17 avian pathogenic Escherichia coli (APEC) strains ...

Abstract	Resistance to extended-spectrum cephalosporins (ESC) is a global health issue. The aim of this study was to analyze and compare plasmids coding for resistance to ESC isolated from 16 avian commensal and 17 avian pathogenic Escherichia coli (APEC) strains obtained respectively at slaughterhouse or from diseased broilers in 2010-2012. Plasmid DNA was used to transform E. coli DH5alpha, and the resistances of the transformants were determined. The sequences of the ESC-resistance plasmids prepared from transformants were obtained by Illumina (33 plasmids) or PacBio (1 plasmid). Results showed that 29 of these plasmids contained the blaCTX-M-1 gene and belonged to the IncI1/ST3 type, with 27 and 20 of them carrying the sul2 or tet(A) genes respectively. Despite their diverse origins, several plasmids showed very high percentages of identity. None of the blaCTX-M-1-containing plasmid contained APEC virulence genes, although some of them were detected in the parental strains. Three plasmids had the blaCMY-2 gene, but no other resistance gene. They belonged to IncB/O/K/Z-like or IncFIA/FIB replicon types. The blaCMY-2 IncFIA/FIB plasmid was obtained from a strain isolated from a diseased broiler and also containing a blaCTX-M-1 IncI1/ST3 plasmid. Importantly APEC virulence genes (sitA-D, iucA-D, iutA, hlyF, ompT, etsA-C, iss, iroB-E, iroN, cvaA-C and cvi) were detected on the blaCMY-2 plasmid. In conclusion, our results show the dominance and high similarity of blaCTX-M-1 IncI1/ST3 plasmids, and the worrying presence of APEC virulence genes on a blaCMY-2 plasmid.
Keywords	Medicine ; R ; Science ; Q
Subject code	572
Language	English
Publishing date	2018-01-01T00:00:00Z
Publisher	Public Library of Science (PLoS)
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article: Lessons learnt from a porcine epidemic diarrhea (PED) case in France in 2014: Descriptive epidemiology and control measures implemented

Gallien, Sarah / Christelle Fablet / Lionel Bigault / Cécilia Bernard / Olivier Toulouse / Mustapha Berri / Yannick Blanchard / Nicolas Rose / Béatrice Grasland

Veterinary microbiology. 2018 Nov., v. 226

2018

Abstract: An acute epidemic of porcine epidemic diarrhea (PED) has affected the USA since 2013 and spread all around the world. In France, the immune status of the pig population against PED virus (PEDV) was expected to be low due to the absence of circulation of ... ...

Abstract	An acute epidemic of porcine epidemic diarrhea (PED) has affected the USA since 2013 and spread all around the world. In France, the immune status of the pig population against PED virus (PEDV) was expected to be low due to the absence of circulation of the virus since the 80′s and a compulsory notification of PED was set up in 2014. Here, we reported the first case of a PED outbreak in December 2014 in the North of France after a long absence of the disease, the monitoring of the excretion and the control measure implementation. The isolated strain in France in December 2014 was a PEDV “S-InDel” strain which was close to the “S-InDel” German PEDV strain isolated in May 2014. The individual shedding duration of PEDV in feces was estimated around 20 days for pigs of different ages. Biosecurity measures implemented allowed the limitation of PEDV spread to fattening and farrowing rooms without dissemination to the nursery block. Using strict biosecurity measures, direct shipment of infected fatteners to the slaughterhouse, strict decontamination protocols with a quarantine of 6 weeks for replacement gilts without voluntary contamination helped PEDV fade out within the herd and avoided the spread to other herds. PEDV presence in manure was investigated as well as the inactivation treatment of the virus present in the liquid manure. An increase to a pH 12 of liquid manure by liming led to the absence of PEDV detection by RT-PCR after seven days.
Keywords	biosecurity ; decontamination ; diarrhea ; epidemiology ; excretion ; farrowing ; feces ; finishing ; gilts ; herds ; liquid manure ; monitoring ; pH ; quarantine ; reverse transcriptase polymerase chain reaction ; slaughterhouses ; viruses ; France
Language	English
Dates of publication	2018-11
Size	p. 9-14.
Publishing place	Elsevier B.V.
Document type	Article
ZDB-ID	753154-0
ISSN	1873-2542 ; 0378-1135
ISSN (online)	1873-2542
ISSN	0378-1135
DOI	10.1016/j.vetmic.2018.09.023
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Recombination at the emergence of the pathogenic rabbit haemorrhagic disease virus Lagovirus europaeus/GI.2

Joana Abrantes / Clément Droillard / Ana M. Lopes / Evelyne Lemaitre / Pierrick Lucas / Yannick Blanchard / Stéphane Marchandeau / Pedro J. Esteves / Ghislaine Le Gall-Reculé

Scientific Reports, Vol 10, Iss 1, Pp 1-

2020 Volume 11

Abstract: Abstract Rabbit haemorrhagic disease is a viral disease that emerged in the 1980s and causes high mortality and morbidity in the European rabbit (Oryctolagus cuniculus). In 2010, a new genotype of the rabbit haemorrhagic disease virus emerged and ... ...

Abstract	Abstract Rabbit haemorrhagic disease is a viral disease that emerged in the 1980s and causes high mortality and morbidity in the European rabbit (Oryctolagus cuniculus). In 2010, a new genotype of the rabbit haemorrhagic disease virus emerged and replaced the former circulating Lagovirus europaeus/GI.1 strains. Several recombination events have been reported for the new genotype Lagovirus europaeus/GI.2, with pathogenic (variants GI.1a and GI.1b) and benign (genotype GI.4) strains that served as donors for the non-structural part while GI.2 composed the structural part; another recombination event has also been described at the p16/p23 junction involving GI.4 strains. In this study, we analysed new complete coding sequences of four benign GI.3 strains and four GI.2 strains. Phylogenetic and recombination detection analyses revealed that the first GI.2 strains, considered as non-recombinant, resulted from a recombination event between GI.3 and GI.2, with GI.3 as the major donor for the non-structural part and GI.2 for the structural part. Our results indicate that recombination contributed to the emergence, persistence and dissemination of GI.2 as a pathogenic form and that all described GI.2 strains so far are the product of recombination. This highlights the need to study full-genomic sequences of lagoviruses to understand their emergence and evolution.
Keywords	Medicine ; R ; Science ; Q
Subject code	570
Language	English
Publishing date	2020-09-01T00:00:00Z
Publisher	Nature Publishing Group
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article: Lleida Bat Lyssavirus isolation in Miniopterus schreibersii in France

Picard‐Meyer, Evelyne / Alexandre Servat / Cédric Guillaume / Edouard Hirchaud / Emmanuelle Robardet / Florence Cliquet / Gérald Larcher / Veronique Beven / Yannick Blanchard

Zoonoses and public health. 2019 Mar., v. 66, no. 2

2019

Abstract: Bat rabies cases are attributed in Europe to five different Lyssavirus species of 16 recognized Lyssavirus species causing rabies. One of the most genetically divergent Lyssavirus spp. has been detected in a dead Miniopterus schreibersii bat in France. ... ...

Abstract	Bat rabies cases are attributed in Europe to five different Lyssavirus species of 16 recognized Lyssavirus species causing rabies. One of the most genetically divergent Lyssavirus spp. has been detected in a dead Miniopterus schreibersii bat in France. Brain samples were found positive for the presence of antigen, infectious virus and viral RNA by classical virological methods and molecular methods respectively. The complete genome sequence was determined by next‐generation sequencing. The analysis of the complete genome sequence confirmed the presence of Lleida bat lyssavirus (LLEBV) in bats in France with 99.7% of nucleotide identity with the Spanish LLEBV strain (KY006983).
Keywords	antigens ; brain ; Chiroptera ; high-throughput nucleotide sequencing ; Lyssavirus ; nucleotide sequences ; rabies ; RNA ; viruses ; France
Language	English
Dates of publication	2019-03
Size	p. 254-258.
Publishing place	John Wiley & Sons, Ltd
Document type	Article
Note	JOURNAL ARTICLE
ZDB-ID	2271118-1
ISSN	1863-2378 ; 1863-1959
ISSN (online)	1863-2378
ISSN	1863-1959
DOI	10.1111/zph.12535
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Modulation of chromatin remodelling induced by the freshwater cyanotoxin cylindrospermopsin in human intestinal caco-2 cells.

Antoine Huguet / Aurélie Hatton / Romain Villot / Hélène Quenault / Yannick Blanchard / Valérie Fessard

PLoS ONE, Vol 9, Iss 6, p e

2014 Volume 99121

Abstract: Cylindrospermopsin (CYN) is a cyanotoxin that has been recognised as an emerging potential public health risk. Although CYN toxicity has been demonstrated, the mechanisms involved have not been fully characterised. To identify some key pathways related ... ...

Abstract	Cylindrospermopsin (CYN) is a cyanotoxin that has been recognised as an emerging potential public health risk. Although CYN toxicity has been demonstrated, the mechanisms involved have not been fully characterised. To identify some key pathways related to this toxicity, we studied the transcriptomic profile of human intestinal Caco-2 cells exposed to a sub-toxic concentration of CYN (1.6 µM for 24hrs) using a non-targeted approach. CYN was shown to modulate different biological functions which were related to growth arrest (with down-regulation of cdkn1a and uhrf1 genes), and DNA recombination and repair (with up-regulation of aptx and pms2 genes). Our main results reported an increased expression of some histone-modifying enzymes (histone acetyl and methyltransferases MYST1, KAT5 and EHMT2) involved in chromatin remodelling, which is essential for initiating transcription. We also detected greater levels of acetylated histone H2A (Lys5) and dimethylated histone H3 (Lys4), two products of these enzymes. In conclusion, CYN overexpressed proteins involved in DNA damage repair and transcription, including modifications of nucleosomal histones. Our results highlighted some new cell processes induced by CYN.
Keywords	Medicine ; R ; Science ; Q
Subject code	570
Language	English
Publishing date	2014-01-01T00:00:00Z
Publisher	Public Library of Science (PLoS)
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Virus persistence in pig herds led to successive reassortment events between swine and human influenza A viruses, resulting in the emergence of a novel triple-reassortant swine influenza virus

Amélie Chastagner / Emilie Bonin / Christelle Fablet / Stéphane Quéguiner / Edouard Hirchaud / Pierrick Lucas / Stéphane Gorin / Nicolas Barbier / Véronique Béven / Emmanuel Garin / Yannick Blanchard / Nicolas Rose / Séverine Hervé / Gaëlle Simon

Veterinary Research, Vol 50, Iss 1, Pp 1-

2019 Volume 9

Abstract: Abstract This report describes the detection of a triple reassortant swine influenza A virus of H1avN2 subtype. It evolved from an avian-like swine H1avN1 that first acquired the N2 segment from a seasonal H3N2, then the M segment from a 2009 pandemic ... ...

Abstract	Abstract This report describes the detection of a triple reassortant swine influenza A virus of H1avN2 subtype. It evolved from an avian-like swine H1avN1 that first acquired the N2 segment from a seasonal H3N2, then the M segment from a 2009 pandemic H1N1, in two reassortments estimated to have occurred 10 years apart. This study illustrates how recurrent influenza infections increase the co-infection risk and facilitate evolutionary jumps by successive gene exchanges. It recalls the importance of appropriate biosecurity measures inside holdings to limit virus persistence and interspecies transmissions, which both contribute to the emergence of new potentially zoonotic viruses.
Keywords	Veterinary medicine ; SF600-1100
Language	English
Publishing date	2019-10-01T00:00:00Z
Publisher	BMC
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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