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

2020 ◽  
Author(s):  
Fábio A. Abade dos Santos ◽  
Carina L. Carvalho ◽  
Andreia Pinto ◽  
Ranjit Rai ◽  
Madalena Monteiro ◽  
...  
Keyword(s):  
Myxoma Virus ◽  
Virus Infectivity ◽  
Wild Rabbit ◽  
R Gene ◽  
Viral Particles ◽  
Domestic Rabbits ◽  
South Portugal ◽  
Iberian Hare ◽  
Continue Monitoring ◽  
Species Segregation

In late 2018, an epidemic myxomatosis outbreak emerged on the Iberian Peninsula leading to high mortality in Iberian hare populations. Soon, a recombinant virus (MYXV-Tol or ha-MYXV) was identified, harboring a 2.8 kb insertion containing evolved duplicates of M060L, M061L, M064L, and M065L from MYXV. Since 2017, 1616 rabbits and 82 hares were tested by a qPCR directed to M000.5L/R gene, conserved in MYXV and MYXV-Tol/ ha-MYXV strains. A subset (20%) of the positive samples was tested for the insert with MYXV being detected in rabbits and recombinant MYXV in hares. Recently, two wild rabbits found dead in South Portugal, showing skin oedema and pulmonary lesions tested positive for the 2.8 Kb insert. Sequencing showed 100% similarity with the insert sequences described in Iberian hares from Spain. Viral particles were observed in the lungs of both rabbits by electron microscopy, and isolation in RK13 cells showed virus infectivity. Despite the analysis of recombinant MYXV genomes may predict its 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, which raises serious concerns for the future of the Iberian wild leporids and emphasizes the need to continue monitoring MYXV and recombinant MYXV in both species.

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.

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 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 Phosphatidylserine within the Viral Membrane Enhances Chikungunya Virus Infectivity in a Cell-type Dependent Manner

2022 ◽  
Author(s):  
Kerri L Miazgowicz ◽  
Judith Mary Reyes Ballista ◽  
Marissa D Acciani ◽  
Ariana R Jimenez ◽  
Ryan S Belloli ◽  
...  
Keyword(s):  
Chikungunya Virus ◽  
Intervention Strategy ◽  
Host Cells ◽  
Virus Infectivity ◽  
Dependent Manner ◽  
Cell Type ◽  
Enveloped Viruses ◽  
Outer Leaflet ◽  
Viral Particles ◽  
In Cells

Chikungunya virus (CHIKV), an alphavirus of the Togaviridae family, is the causative agent of the human disease chikungunya fever (CHIKF), which is characterized by debilitating acute and chronic arthralgia. No licensed vaccines or antivirals exist for CHIKV. Preventing the attachment of viral particles to host cells is an attractive intervention strategy. Viral entry of enveloped viruses from diverse families including Filoviridae and Flaviviridae is mediated or enhanced by phosphatidylserine receptors (PSRs). PSRs facilitate the attachment of enveloped viruses to cells by binding to exposed phosphatidylserine (PS) in the viral lipid membrane - a process termed viral apoptotic mimicry. To investigate the role of viral apoptotic mimicry during CHIKV infection, we produced viral particles with discrete amounts of exposed PS on the virion envelope by exploiting the cellular distribution of phospholipids at the plasma membrane. We found that CHIKV particles containing high outer leaflet PS (produced in cells lacking flippase activity) were more infectious in Vero cells than particles containing low levels of outer leaflet PS (produced in cells lacking scramblase activity). However, the same viral particles were similarly infectious in NIH3T3 and HAP1 cells, suggesting PS levels can influence infectivity only in cells with high levels of PSRs. Interestingly, PS-dependent CHIKV entry was observed in mosquito Aag2 cells, but not C6/36 cells. These data demonstrate that CHIKV entry via viral apoptotic mimicry is cell-type dependent. Furthermore, viral apoptotic mimicry has a mechanistic basis to influence viral dynamics in vivo in both the human and mosquito host.

scholarly journals Myxomatosis in the Mallee region of Victoria, Australia

1978 ◽  
Vol 81 (2) ◽  
pp. 239-243 ◽  
Author(s):  
Rosamond C. H. Shepherd ◽  
J. W. Edmonds ◽  
I. F. Nolan ◽  
A. Gocs
Keyword(s):  
Late Summer ◽  
Nutritional Stress ◽  
Early Summer ◽  
Myxoma Virus ◽  
Direct Result ◽  
Healthy Population ◽  
Wild Rabbit ◽  
Population Reduction ◽  
In The Wild ◽  
Limited Food

SummarySharp reductions in the wild rabbit (Oryctolagus cuniculus (L.)) population in the Mallee are associated with annual myxomatosis epizootics. The extent to which the population reductionsare the direct result of the epizootics varies with time of epizootic occurrence. All grazing animals in the Mallee are under nutritional stress each summer and autumn. When the epizootic occurs during the early summer heavy losses occur in a previously healthy population. Similar losses which occur in the late summer and autumn are the result of a nutritional stress – epizootic complex. The end result in each case is a population reduction of about 80%.This reduction occurs in a population which is the most resistant to myxomatosis known in Victoria and in association with epizooties caused by field strains of myxoma virus of moderate virulence only.The earlier summer epizootics are of considerable economic importance because they sharply reduce the pressure on the limited food available for other grazing animals.

Selection of antigens for use in a virus-vectored immunocontraceptive vaccine: PH-20 as a case study

1997 ◽  
Vol 9 (1) ◽  
pp. 117 ◽  
Author(s):  
Michael K. Holland ◽  
Jason Andrews ◽  
Hannah Clarke ◽  
Clayton Walton ◽  
Lyn A. Hinds
Keyword(s):  
Oryctolagus Cuniculus ◽  
Genetically Engineered ◽  
Myxoma Virus ◽  
Wild Rabbit ◽  
Pest Species ◽  
Genes Encoding ◽  
Overall Efficiency ◽  
Selection Of ◽  
Sperm Antigen

The European wild rabbit (Oryctolagus cuniculus) has become the major agricultural and environmental pest species in Australia. Current methods of rabbit control are lethal procedures which are increasingly questioned for their overall efficiency, applicability, specificity, cost and humaneness. New initiatives are required. One such initiative is virus-vectored immunocontraception. In this approach, the lagomorph-specific myxoma virus will be genetically engineered to include genes encoding components of rabbit gametes which can induce an immune response that causes infertility. Central to such a strategy is the ability to identify antigens capable of inducing an immunocontraceptive response. A strategy for identifying such antigens has been described previously. A case study of one sperm antigen, PH-20, is reported here. The issues involved in developing this antigen to the stage where it could be considered as a candidate for insertion into a recombinant myxoma virus with the ultimate goal of testing for immunocontraceptive efficacy are discussed. Techniques for inserting genes into myxoma virus have been described previously. The knowledge gained from research with this particular antigen are broadly applicable to other antigens used for both immunocontraceptive vaccines in general and, specifically, for virus-vectored immunocontraception.

Progress towards using recombinant myxoma virus as a vector for fertility control in rabbits

1997 ◽  
Vol 9 (1) ◽  
pp. 77 ◽  
Author(s):  
A. J. Robinson ◽  
R. Jackson ◽  
P. Kerr ◽  
J. Merchant ◽  
I. Parer ◽  
...  
Keyword(s):  
Recombinant Virus ◽  
Fertility Control ◽  
Myxoma Virus ◽  
Wild Rabbit ◽  
Recombinant Viruses ◽  
History Of ◽  
Insertion Sites ◽  
Influenza Virus Haemagglutinin ◽  
Double Recombinant ◽  
Virus Haemagglutinin

The history of myxoma virus, its use in Australia as a mortality agent and the development of the virus as a vector for controlling fertility in wild rabbit populations in Australia is reviewed. Myxoma virus recombinants have been constructed to express model antigens. Four potential insertion sites in the genome have been identified and two have been used to construct single and double recombinant viruses expressing Escherichia colienzymes β-galactosidase and β-glucuronidase. Another recombinant expressing an influenza virus haemagglutinin gene (A/PR8/34) induced high and sustained antibody responses following intradermal inoculation in rabbits. To demonstrate the potential of introducing a recombinant virus into wild rabbit populations, a virus containing a natural deletion was released at four field locations. Preliminary analysis of the data has shown that the introduced virus spread well on 3 of the 4 locations. The steps being taken to address the ethical and safety implications of the introduction of a recombinant virus into the field are discussed.

scholarly journals Myxoma virus jumps species to the Iberian hare

2019 ◽  
Vol 66 (6) ◽  
pp. 2218-2226 ◽  
Author(s):  
Kevin P. Dalton ◽  
José M. Martín ◽  
Inés Nicieza ◽  
Ana Podadera ◽  
Daniel Llano ◽  
...  
Keyword(s):  
Myxoma Virus ◽  
Iberian Hare

scholarly journals Zoonotic Microsporidia in Wild Lagomorphs in Southern Spain

Animals ◽  
2020 ◽  
Vol 10 (12) ◽  
pp. 2218
Author(s):  
Anabel Martínez-Padilla ◽  
Javier Caballero-Gómez ◽  
Ángela Magnet ◽  
Félix Gómez-Guillamón ◽  
Fernando Izquierdo ◽  
...  
Keyword(s):  
Fungal Pathogens ◽  
Enterocytozoon Bieneusi ◽  
Southern Spain ◽  
Health Concern ◽  
Wild Rabbit ◽  
Specific Pcr ◽  
Encephalitozoon Intestinalis ◽  
Wild Lagomorphs ◽  
Species Specific ◽  
Iberian Hare

Microsporidia are obligate intracellular protist-like fungal pathogens that infect a broad range of animal species, including humans. This study aimed to assess the presence of zoonotic microsporidia (Enterocytozoon bieneusi, Encephalitozoon intestinalis, Encephalitozoon hellem, and Encephalitozoon cuniculi) in organ meats of European wild rabbit (Oryctolagus cuniculus) and Iberian hare (Lepus granatensis) consumed by humans in Spain. Between July 2015 and December 2018, kidney samples from 383 wild rabbits and kidney and brain tissues from 79 Iberian hares in southern Spain were tested by species-specific PCR for the detection of microsporidia DNA. Enterocytozoon bieneusi infection was confirmed in three wild rabbits (0.8%; 95% CI: 0.0–1.7%) but not in hares (0.0%; 95% CI: 0.0–4.6%), whereas E. intestinalis DNA was found in one wild rabbit (0.3%; 95% CI: 0.0–0.8%) and three Iberian hares (3.8%; 95% CI: 0.0–8.0%). Neither E. hellem nor E. cuniculi infection were detected in the 462 (0.0%; 95% CI: 0.0–0.8%) lagomorphs analyzed. The absence of E. hellem and E. cuniculi infection suggests a low risk of zoonotic foodborne transmission from these wild lagomorph species in southern Spain. To the authors’ knowledge, this is the first report of E. intestinalis infection in wild rabbits and Iberian hares. The presence of E. bieneusi and E. intestinalis in organ meats from wild lagomorphs can be of public health concern. Additional studies are required to determine the real prevalence of these parasites in European wild rabbit and Iberian hare.

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