scholarly journals Genetic characterization of a recombinant myxoma virus leap into the Iberian hare (Lepus granatensis)

2019 ◽  
Author(s):  
Ana Águeda-Pinto ◽  
Ana Lemos de Matos ◽  
Mário Abrantes ◽  
Simona Kraberger ◽  
Maria A. Risalde ◽  
...  
Keyword(s):  
Myxoma Virus ◽  
Open Reading Frames ◽  
Lepus Europaeus ◽  
European Rabbit ◽  
New Host ◽  
Tissue Samples ◽  
Lepus Granatensis ◽  
Domestic Rabbits ◽  
Iberian Hare

AbstractMyxomatosis is a lethal disease of wild European and domestic rabbits (Oryctolagus cuniculus) caused by a Myxoma virus (MYXV) infection, a leporipoxvirus that is found naturally in someSylvilagusrabbit species in South America and California. The introduction of MYXV in the early 1950s into feral European rabbit populations in Australia and Europe demonstrate the best documented field example of host-virus coevolution following a cross-species transmission. Recently, a new cross-species jump of MYXV has been suggested in both Great Britain and Spain, where European brown hares (Lepus europaeus) and Iberian hares (Lepus granatensis) were found dead with lesions consistent with those observed in myxomatosis. To investigate the possibility of a new cross-species transmission event by MYXV, tissue samples collected from a wild Iberian hare found dead in Spain (Toledo region) were analyzed and deep sequenced. Our results report a new MYXV strain (MYXV Toledo) in the tissues of this species. The genome of this new strain encodes three disrupted genes (M009L,M036LandM152R) and a novel 2.8 KB recombinant region that resulted from an insertion of four novel poxviral genes towards the 5’ end of its genome. From the open reading frames inserted into the MYXV Toledo strain, a new orthologue of a poxvirus host range gene family member was identified which is related to the MYXV geneM064R. Overall, we confirmed the identity of a new MYXV strain in Iberian hares that we hypothesize was able to more effectively counteract the host defenses in hares and start an infectious process in this new host.

scholarly journals Genetic Characterization of a Recombinant Myxoma Virus in the Iberian Hare (Lepus granatensis)

Viruses ◽  
2019 ◽  
Vol 11 (6) ◽  
pp. 530 ◽  
Author(s):  
Ana Águeda-Pinto ◽  
Ana Lemos de Matos ◽  
Mário Abrantes ◽  
Simona Kraberger ◽  
Maria A. Risalde ◽  
...  
Keyword(s):  
Myxoma Virus ◽  
Open Reading Frames ◽  
Lepus Europaeus ◽  
European Rabbit ◽  
New Host ◽  
Tissue Samples ◽  
Lepus Granatensis ◽  
Domestic Rabbits ◽  
Iberian Hare

Myxomatosis is a lethal disease in wild European and domestic rabbits (Oryctolagus cuniculus), which is caused by a Myxoma virus (MYXV) infection—a leporipoxvirus that is found naturally in some Sylvilagus rabbit species in South America and California. The introduction of MYXV into feral European rabbit populations of Australia and Europe, in the early 1950s, demonstrated the best-documented field example of host–virus coevolution, following a cross-species transmission. Recently, a new cross-species jump of MYXV has been suggested in both Great Britain and Spain, where European brown hares (Lepus europaeus) and Iberian hares (Lepus granatensis) were found dead with lesions consistent with those observed in myxomatosis. To investigate the possibility of a new cross-species transmission event by MYXV, tissue samples collected from a wild Iberian hare found dead in Spain (Toledo region) were analyzed and deep sequenced. Our results reported a new MYXV isolate (MYXV Toledo) in the tissues of this species. The genome of this new virus was found to encode three disruptive genes (M009L, M036L, and M152R) and a novel ~2.8 kb recombinant region, which resulted from an insertion of four novel poxviral genes towards the 3’ end of the negative strand of its genome. From the open reading frames inserted into the MYXV Toledo virus, a new orthologue of a poxvirus host range gene family member was identified, which was related to the MYXV gene M064R. Overall, we confirmed the identity of a new MYXV isolate in Iberian hares, which, we hypothesized, was able to more effectively counteract the host defenses in hares and start an infectious process in this new host.

scholarly journals Functional Analysis of Genes in the rfbLocus of Leptospira borgpetersenii Serovar Hardjo Subtype Hardjobovis

2000 ◽  
Vol 68 (7) ◽  
pp. 3793-3798 ◽  
Author(s):  
Dieter M. Bulach ◽  
Thareerat Kalambaheti ◽  
Alejandro de la Peña-Moctezuma ◽  
Ben Adler
Keyword(s):  
Sequence Similarity ◽  
Bacterial Species ◽  
Open Reading Frames ◽  
New Host ◽  
E Coli ◽  
Serovar Typhimurium ◽  
Genes Encoding ◽  
Leptospira Borgpetersenii ◽  
Nucleotide Sugars

ABSTRACT Lipopolysaccharide (LPS) is a key antigen in immunity to leptospirosis. Its biosynthesis requires enzymes for the biosynthesis and polymerization of nucleotide sugars and the transport through and attachment to the bacterial membrane. The genes encoding these functions are commonly clustered into loci; for Leptospira borgpetersenii serovar Hardjo subtype Hardjobovis, this locus, named rfb, spans 36.7 kb and contains 31 open reading frames, of which 28 have been assigned putative functions on the basis of sequence similarity. Characterization of the function of these genes is hindered by the fact that it is not possible to construct isogenic mutant strains in Leptospira. We used two approaches to circumvent this problem. The first was to clone the entire locus into a heterologous host system and determine if a “recombinant” LPS or polysaccharide was synthesized in the new host. The second approach used putative functions to identify mutants in other bacterial species whose mutations might be complemented by genes on the leptospiralrfb locus. This approach was used to investigate the function of three genes in the leptospiral rfb locus and demonstrated function for orfH10, which complemented awbpM strain of Pseudomonas aeruginosa, andorfH13, which complemented an rfbW strain ofVibrio cholerae. However, despite the similarity of OrfH11 to WecC, a wecC strain of E. coli was not complemented by orfH11. The predicted protein encoded byorfH8 is similar to GalE from a number of organisms. ASalmonella enterica serovar Typhimurium strain producing no GalE was used as a background in which orfH8 produced detectable GalE enzyme activity.

scholarly journals A Quadruplex qPCR for Detection and Differentiation of Classic and Natural Recombinant Myxoma Virus Strains of Leporids

2021 ◽  
Vol 22 (21) ◽  
pp. 12052
Author(s):  
Fábio A. Abade dos Santos ◽  
Carina L. Carvalho ◽  
Francisco Parra ◽  
Kevin P. Dalton ◽  
Maria C. Peleteiro ◽  
...  
Keyword(s):  
Real Time ◽  
Housekeeping Gene ◽  
Myxoma Virus ◽  
European Rabbit ◽  
Fast Method ◽  
Wild Rabbit ◽  
Viral Dna ◽  
Eukaryotic Organisms ◽  
Virus Strains ◽  
Iberian Hare

A natural recombinant myxoma virus (referred to as ha-MYXV or MYXV-Tol08/18) emerged in the Iberian hare (Lepus granatensis) and the European rabbit (Oryctolagus cuniculus) in late 2018 and mid-2020, respectively. This new virus is genetically distinct from classic myxoma virus (MYXV) strains that caused myxomatosis in rabbits until then, by acquiring an additional 2.8 Kbp insert within the m009L gene that disrupted it into ORFs m009L-a and m009L-b. To distinguish ha-MYXV from classic MYXV strains, we developed a robust qPCR multiplex technique that combines the amplification of the m000.5L/R duplicated gene, conserved in all myxoma virus strains including ha-MYXV, with the amplification of two other genes targeted by the real-time PCR systems designed during this study, specific either for classic MYXV or ha-MYXV strains. The first system targets the boundaries between ORFs m009L-a and m009L-b, only contiguous in classic strains, while the second amplifies a fragment within gene m060L, only present in recombinant MYXV strains. All amplification reactions were validated and normalized by a fourth PCR system directed to a housekeeping gene (18S rRNA) conserved in eukaryotic organisms, including hares and rabbits. The multiplex PCR (mPCR) technique described here was optimized for Taqman® and Evagreen® systems allowing the detection of as few as nine copies of viral DNA in the sample with an efficiency > 93%. This real-time multiplex is the first fast method available for the differential diagnosis between classic and recombinant MYXV strains, also allowing the detection of co-infections. The system proves to be an essential and effective tool for monitoring the geographical spread of ha-MYXV in the hare and wild rabbit populations, supporting the management of both species in the field.

scholarly journals Viral Disease in Lagomorphs: A Molecular Perspective

2021 ◽  
Author(s):  
Kevin P. Dalton ◽  
Ana Podadera ◽  
José Manuel Martin Alonso ◽  
Inés Calonge Sanz ◽  
Ángel Luis Álvarez Rodríguez ◽  
...  
Keyword(s):  
Viral Disease ◽  
Myxoma Virus ◽  
Lepus Europaeus ◽  
European Rabbit ◽  
Great Effort ◽  
Brown Hare ◽  
European Brown Hare ◽  
European Brown Hare Syndrome ◽  
Molecular Aspects ◽  
Species Specific

Our understanding of molecular biology of the viruses that infect lagomorphs is largely limited to the leporipoxvirus myxoma virus (MYXV) and the lagoviruses rabbit haemorrhagic disease virus (RHDV) and European brown hare syndrome virus (EBHSV) that infect the European rabbit (Oryctolagus cuniculus) and the European brown hare (Lepus europaeus) respectively. Thanks to the great effort of historic surveillance studies and careful sample archiving, the molecular evolution of these viruses is being resolved. Although historically considered viruses that cause species specific diseases recent reports show that several lagomorphs may now face the threat of these maladies. The driving factors behind these changes has not been determined and the effect of these species jumps on lagomorph populations has yet to be seen. Lagomorphs are also affected by several other lesser studied viral diseases. In addition, recent metagenomic studies have led to the identification of novel lagomorph viruses the importance of these to lagomorph health remains to be fully determined. In this chapter we summarize molecular aspects of viruses that infect lagomorphs, paying particular attention to recent interspecies infections.

scholarly journals Detection of Recombinant Hare Myxoma Virus in Wild Rabbits (Oryctolagus cuniculus algirus)

Viruses ◽  
2020 ◽  
Vol 12 (10) ◽  
pp. 1127
Author(s):  
Fábio A. Abade dos Santos ◽  
Carina L. Carvalho ◽  
Andreia Pinto ◽  
Ranjit Rai ◽  
Madalena Monteiro ◽  
...  
Keyword(s):  
Myxoma Virus ◽  
Virus Infectivity ◽  
Sequencing Analysis ◽  
Wild Rabbit ◽  
Mainland Portugal ◽  
Viral Particles ◽  
Domestic Rabbits ◽  
Virus Strains ◽  
Iberian Hare ◽  
Species Segregation

In late 2018, an epidemic myxomatosis outbreak emerged on the Iberian Peninsula leading to high mortality in Iberian hare populations. A recombinant Myxoma virus (strains MYXV-Tol and ha-MYXV) was rapidly identified, harbouring a 2.8 kbp insertion containing evolved duplicates of M060L, M061L, M064L, and M065L genes from myxoma virus (MYXV) or other Poxviruses. Since 2017, 1616 rabbits and 125 hares were tested by a qPCR directed to M000.5L/R gene, conserved in MYXV and MYXV-Tol/ha-MYXV strains. A subset of the positive samples (20%) from both species was tested for the insert with MYXV being detected in rabbits and the recombinant MYXV in hares. Recently, three wild rabbits were found dead South of mainland Portugal, showing skin oedema and pulmonary lesions that tested positive for the 2.8 kbp insert. Sequencing analysis showed 100% similarity with the insert sequences described in Iberian hares from Spain. Viral particles were observed in the lungs and eyelids of rabbits by electron microscopy, and isolation in RK13 cells attested virus infectivity. Despite that the analysis of complete genomes may predict the recombinant MYXV strains’ ability to infect rabbit, routine analyses showed species segregation for the circulation of MYXV and recombinant MYXV in wild rabbit and in Iberian hares, respectively. This study demonstrates, however, that recombinant MYXV can effectively infect and cause myxomatosis in wild rabbits and domestic rabbits, raising serious concerns for the future of the Iberian wild leporids while emphasises the need for the continuous monitoring of MYXV and recombinant MYXV in both species.

First cases of myxomatosis in Iberian hares (Lepus granatensis) in Portugal

2020 ◽  
Vol 8 (2) ◽  
pp. e001044 ◽  
Author(s):  
Carina Luísa Carvalho ◽  
Fábio Alexandre Abade dos Santos ◽  
Madelena Monteiro ◽  
Paulo Carvalho ◽  
Paula Mendonça ◽  
...  
Keyword(s):  
Body Condition ◽  
Myxoma Virus ◽  
Common Origin ◽  
Lepus Granatensis ◽  
Short Course ◽  
Good Body Condition ◽  
Myxoid Matrix ◽  
Gross Lesions ◽  
Perineal Region ◽  
Iberian Hare

Myxomatosis was detected in Iberian hares (Lepus granatensis) in Portugal, October 2018, following its emergence in Spain 3 months earlier. Here, we describe the epidemiological, molecular and anatomo-histopathological data of the first two cases. Myxoma virus DNA was detected in the eyelids, nose and perineal region in both hares. It was also detected in the lungs of hare 1 and in the spleen and liver of hare 2. The genomic insertion identified in strains from Spain was confirmed in both strains suggesting a common origin for the Iberian viruses. Gross lesions in hare 1 included palpebral oedema and conjunctival mucopurulent discharge, common in both forms of the disease in rabbits. Hare 2 presented eyelid thickening with small diffuse nodules. Histopathology of the eyelids showed extracellular myxoid matrix in hare 1 and purulent dermatitis in hare 2. Both animals exhibited good body condition, suggesting a short course of the disease and higher virulence of the virus towards the Iberian hare.

scholarly journals Genome Scale Evolution of Myxoma Virus Reveals Host-Pathogen Adaptation and Rapid Geographic Spread

2013 ◽  
Vol 87 (23) ◽  
pp. 12900-12915 ◽  
Author(s):  
Peter J. Kerr ◽  
Matthew B. Rogers ◽  
Adam Fitch ◽  
Jay V. DePasse ◽  
Isabella M. Cattadori ◽  
...  
Keyword(s):  
Laboratory Strain ◽  
Myxoma Virus ◽  
Open Reading Frames ◽  
European Rabbit ◽  
Content Type ◽  
Multiple Introductions ◽  
Virulence Proteins ◽  
Host Pathogen ◽  
Attenuated Viruses ◽  
Early Radiation

The evolutionary interplay between myxoma virus (MYXV) and the European rabbit (Oryctolagus cuniculus) following release of the virus in Australia in 1950 as a biological control is a classic example of host-pathogen coevolution. We present a detailed genomic and phylogeographic analysis of 30 strains of MYXV, including the Australian progenitor strain Standard Laboratory Strain (SLS), 24 Australian viruses isolated from 1951 to 1999, and three isolates from the early radiation in Britain from 1954 and 1955. We show that in Australia MYXV has spread rapidly on a spatial scale, with multiple lineages cocirculating within individual localities, and that both highly virulent and attenuated viruses were still present in the field through the 1990s. In addition, the detection of closely related virus lineages at sites 1,000 km apart suggests that MYXV moves freely in geographic space, with mosquitoes, fleas, and rabbit migration all providing means of transport. Strikingly, despite multiple introductions, all modern viruses appear to be ultimately derived from the original introductions of SLS. The rapidity of MYXV evolution was also apparent at the genomic scale, with gene duplications documented in a number of viruses. Duplication of potential virulence genes may be important in increasing the expression of virulence proteins and provides the basis for the evolution of novel functions. Mutations leading to loss of open reading frames were surprisingly frequent and in some cases may explain attenuation, but no common mutations that correlated with virulence or attenuation were identified.

scholarly journals Lack of polymorphism of the agouti signaling protein (ASIP) gene among four different brown hare (Lepus europaeus Pallas 1778) populations

2016 ◽  
pp. 81-85
Author(s):  
Noémi Soós ◽  
Szilvia Kusza
Keyword(s):  
Mammalian Species ◽  
Lepus Europaeus ◽  
European Rabbit ◽  
Brown Hare ◽  
Base Pairs ◽  
Game Species ◽  
Genetic Studies ◽  
Tissue Samples ◽  
Signaling Protein ◽  
Minimal Information

The brown hare (Lepus europaeus Pallas 1778) is a common palearctic and a popular game species therefore it has been an obvious subject for population genetic studies since the second part of the 20th century. Among the several mitochondrial DNA studies some have been carried out concerning nuclear genes as well. The agouti signaling protein gene (ASIP) is involved in regulating the synthesis of eumelanin and pheomelanin in melanocytes of mammals. Though many studies focused on it in relation with several mammalian species, minimal information is available on this topic concerning the brown hare. Here we present a short communication concerning the agouti signaling protein (ASIP) gene in four different country’s L. europaeus populations, namely Lithuania, Hungary, Serbia and Georgia. N=45 tissue samples have been investigated from overall 17 sampling sites of the different countries. There has not been found any polymorphism among the sequences. In an alignment with other Leporid species’ partial ASIP sequences downloaded from ENA we have found that based on a 178 base pairs long DNA sequence the haplotype of our samples contains three other Lepus species as well. This is concordant with the findings of a previous study focusing predominantly on the European rabbit (Orycto lagus cuniculus Linnaeus 1758) and the several mutations of its ASIP gene.

scholarly journals Colonization history of Mallorca Island by the European rabbit,Oryctolagus cuniculus, and the Iberian hare,Lepus granatensis(Lagomorpha: Leporidae)

2014 ◽  
Vol 111 (4) ◽  
pp. 748-760 ◽  
Author(s):  
Fernando A. Seixas ◽  
Javier Juste ◽  
Paula F. Campos ◽  
Miguel Carneiro ◽  
Nuno Ferrand ◽  
...  
Keyword(s):  
Oryctolagus Cuniculus ◽  
European Rabbit ◽  
Colonization History ◽  
Lepus Granatensis ◽  
History Of ◽  
Mallorca Island ◽  
Iberian Hare

scholarly journals A Potential Atypical Case of Rabbit Haemorrhagic Disease in a Dwarf Rabbit

Animals ◽  
2020 ◽  
Vol 11 (1) ◽  
pp. 40
Author(s):  
Fábio A. Abade dos Santos ◽  
Carolina Magro ◽  
Carina L. Carvalho ◽  
Pedro Ruivo ◽  
Margarida D. Duarte ◽  
...  
Keyword(s):  
Clinical Signs ◽  
Myxoma Virus ◽  
European Rabbit ◽  
Rabbit Haemorrhagic Disease Virus ◽  
Structural Genes ◽  
Bivalent Vaccine ◽  
Rabbit Haemorrhagic Disease ◽  
Cross Immunity ◽  
Domestic Rabbits ◽  
Haemorrhagic Disease

Rabbit haemorrhagic disease (RHD) is a highly contagious infectious disease of European wild and domestic rabbits. Rabbit haemorrhagic disease virus (RHDV, GI.1) emerged in 1986 in Europe, rapidly spreading all over the world. Several genotypes of RHDV have been recognised over time, but in 2010, a new virus (RHDV2/RHDVb, GI.2) emerged and progressively replaced the previous RHDV strains, due to the lack of cross-immunity conferred between RHDV and RHDV2. RHDV2 has a high mutation rate, similarly to the other calivirus and recombines with strains of RHDV and non-pathogenic calicivirus (GI.4), ensuring the continuous emergence of new field strains. Although this poses a threat to the already endangered European rabbit species, the available vaccines against RHDV2 and the compliance of biosafety measures seem to be controlling the infection in the rabbit industry Pet rabbits, especially when kept indoor, are considered at lower risk of infections, although RHDV2 and myxoma virus (MYXV) constitute a permanent threat due to transmission via insects. Vaccination against these viruses is therefore recommended every 6 months (myxomatosis) or annually (rabbit haemorrhagic disease). The combined immunization for myxomatosis and RHDV through a commercially available bivalent vaccine with RHDV antigen has been extensively used (Nobivac® Myxo-RHD, MSD, Kenilworth, NJ, USA). This vaccine however does not confer proper protection against the RHDV2, thus the need for a rabbit clinical vaccination protocol update. Here we report a clinical case of hepatitis and alteration of coagulation in a pet rabbit that had been vaccinated with the commercially available bivalent vaccine against RHDV and tested positive to RHDV2 after death. The animal developed a prolonged and atypical disease, compatible with RHD. The virus was identified to be an RHDV2 recombinant strain, with the structural backbone of RHDV2 (GI.2) and the non-structural genes of non-pathogenic-A1 strains (RCV-A1, GI.4). Although confirmation of the etiological agent was only made after death, the clinical signs and analytic data were very suggestive of RHD.

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