scholarly journals Experimental Reptarenavirus Infection of Boa constrictor and Python regius

2021 ◽  
Author(s):  
U. Hetzel ◽  
Y. Korzyukov ◽  
S. Keller ◽  
L. Szirovicza ◽  
T. Pesch ◽  
...  
Keyword(s):  
Inclusion Body ◽  
Intraperitoneal Injection ◽  
Cytoplasmic Inclusion ◽  
Antibody Responses ◽  
Python Regius ◽  
Boa Constrictor ◽  
Homologous Virus ◽  
Delivery Routes ◽  
Inclusion Body Disease ◽  
Virus Delivery

Boid inclusion body disease (BIBD) causes losses in captive snake populations globally. BIBD is associated with the formation of cytoplasmic inclusion bodies (IBs), which mainly comprise reptarenavirus nucleoprotein (NP). In 2017, BIBD was reproduced by cardiac injection of boas and pythons with reptarenaviruses, thus demonstrating a causative link between reptarenavirus infection and the disease. Here, we report experimental infections of Python regius (n = 16) and Boa constrictor (n = 16) with three reptarenavirus isolates. First, we used pythons (n = 8) to test two virus delivery routes: intraperitoneal injection and tracheal instillation. Viral RNAs but no IBs were detected in brains and lungs at 2 weeks postinoculation. Next, we inoculated pythons (n = 8) via the trachea. During the 4 months following infection, snakes showed transient central nervous system (CNS) signs but lacked detectable IBs at the time of euthanasia. One of the snakes developed severe CNS signs; we succeeded in reisolating the virus from the brain of this individual and could demonstrate viral antigen in neurons. In a third attempt, we tested cohousing, vaccination, and sequential infection with multiple reptarenavirus isolates on boas (n = 16). At 10 months postinoculation, all but one snake tested positive for viral RNA in lung, brain, and/or blood, but none exhibited the characteristic IBs. Three of the four vaccinated snakes seemed to sustain challenge with the same reptarenavirus; however, neither of the two snakes rechallenged with different reptarenaviruses remained uninfected. Comparison of the antibody responses in experimentally versus naturally reptarenavirus-infected animals indicated differences in the responses. IMPORTANCE In the present study, we experimentally infected pythons and boas with reptarenavirus via either intraperitoneal injection or tracheal instillation. The aims were to experimentally induce boid inclusion body disease (BIBD) and to develop an animal model for studying disease transmission and pathogenesis. Both virus delivery routes resulted in infection, and infection via the trachea could reflect the natural route of infection. In the experimentally infected snakes, we did not find evidence of inclusion body (IB) formation, characteristic of BIBD, in pythons or boas. Most of the boas (11/12) remained reptarenavirus infected after 10 months, which suggests that they developed a persistent infection that could eventually have led to BIBD. We demonstrated that vaccination using recombinant protein or an inactivated virus preparation prevented infection by a homologous virus in three of four snakes. Comparison of the antibody responses of experimentally and naturally reptarenavirus-infected snakes revealed differences that merit further studies.

scholarly journals Experimental Reptarenavirus Infection of Boa constrictor and Python regius

2020 ◽  
Author(s):  
U Hetzel ◽  
Y Korzyukov ◽  
S Keller ◽  
L Szirovicza ◽  
T Pesch ◽  
...  
Keyword(s):  
Inclusion Body ◽  
Intraperitoneal Injection ◽  
Cytoplasmic Inclusion ◽  
Antibody Responses ◽  
Viral Rna ◽  
Post Inoculation ◽  
Python Regius ◽  
Delivery Routes ◽  
Inclusion Body Disease ◽  
Virus Delivery

ABSTRACTBoid inclusion body disease (BIBD) causes losses in captive constrictor snake populations globally. BIBD associates with formation of cytoplasmic inclusion bodies (IB) which mainly comprise reptarenavirus nucleoprotein (NP). In 2017, BIBD was reproduced by cardiac injection of boas and pythons with reptarenaviruses, thus demonstrating a causative link between reptarenavirus infection and the disease. Herein, we report experimental infections of pythons (N=16) and boas (N=16) with three reptarenavirus isolates. First, we used pythons (N=8) to test two virus delivery routes: intraperitoneal injection and tracheal instillation. Independent of the delivery route, we detected viral RNA but no IBs in tissues two weeks post inoculation. Next, we inoculated pythons (N=8) via the trachea. During the four month following the infection snakes showed transient central nervous system (CNS) signs but lacked detectable IB at the time of euthanasia. One of the snakes developed severe CNS signs and we succeeded in re-isolating the virus from the brain of this individual, and could demonstrate viral antigen in neurons. In a third attempt, we tested co-housing, vaccination, and sequential infection with multiple reptarenavirus isolates on boas (N=16). At 10 months post inoculation all except one snake tested positive for viral RNA but none exhibited the characteristic IB. Analysis of the antibody responses demonstrated lower neutralizing but higher anti-reptarenavirus NP titers in experimentally versus naturally reptarenavirus infected boas. Our findings suggest that in addition to reptarenavirus infection, other factors, e.g. the antibody response, contribute to BIBD pathogenesis.IMPORTANCEA 2017 study demonstrated cardiac reptarenavirus injection to induce boid inclusion body disease (BIBD) in pythons and boas. In the present study, we experimentally infected pythons and boas with reptarenavirus via either intraperitoneal injection or tracheal instillation. We found both virus delivery routes to result in infection; though the latter could reflect the natural route of infection. In the experimentally infected snakes, we did not find evidence of inclusion body (IB) formation, characteristic to BIBD, in pythons or in boas. Most of the snakes (11/12) studied were reptarenavirus infected after ten-month follow up, which suggests that they could eventually have developed BIBD. We further found differences between the antibody responses of experimentally and naturally reptarenavirus infected snakes, which could indicate that the pathogenesis of BIBD involves factors additional to reptarenavirus infection. As snakes are poikilotherm, also the housing conditions could have an effect.

Boid Inclusion Body Disease in Captive Boas (Boa Constrictor) in Austria

2009 ◽  
Vol 141 (4) ◽  
pp. 305
Author(s):  
B. Richter ◽  
H. Eblinger ◽  
A. Kübber-Heiss
Keyword(s):  
Inclusion Body ◽  
Boa Constrictor ◽  
Inclusion Body Disease

scholarly journals Identification, Characterization, and In Vitro Culture of Highly Divergent Arenaviruses from Boa Constrictors and Annulated Tree Boas: Candidate Etiological Agents for Snake Inclusion Body Disease

mBio ◽  
2012 ◽  
Vol 3 (4) ◽  
Author(s):  
Mark D. Stenglein ◽  
Chris Sanders ◽  
Amy L. Kistler ◽  
J. Graham Ruby ◽  
Jessica Y. Franco ◽  
...  
Keyword(s):  
Inclusion Body ◽  
De Novo ◽  
Human Pathogens ◽  
Cytoplasmic Inclusions ◽  
Boa Constrictor ◽  
Behavioral Abnormalities ◽  
Etiologic Agents ◽  
Causative Agents ◽  
Inclusion Body Disease

ABSTRACTInclusion body disease (IBD) is an infectious fatal disease of snakes typified by behavioral abnormalities, wasting, and secondary infections. At a histopathological level, the disease is identified by the presence of large eosinophilic cytoplasmic inclusions in multiple tissues. To date, no virus or other pathogen has been definitively characterized or associated with the disease. Using a metagenomic approach to search for candidate etiologic agents in snakes with confirmed IBD, we identified andde novoassembled the complete genomic sequences of two viruses related to arenaviruses, and a third arenavirus-like sequence was discovered by screening an additional set of samples. A continuous boa constrictor cell line was established and used to propagate and isolate one of the viruses in culture. Viral nucleoprotein was localized and concentrated within large cytoplasmic inclusions in infected cells in culture and tissues from diseased snakes. In total, viral RNA was detected in 6/8 confirmed IBD cases and 0/18 controls. These viruses have a typical arenavirus genome organization but are highly divergent, belonging to a lineage separate from that of the Old and New World arenaviruses. Furthermore, these viruses encode envelope glycoproteins that are more similar to those of filoviruses than to those of other arenaviruses. These findings implicate these viruses as candidate etiologic agents of IBD. The presence of arenaviruses outside mammals reveals that these viruses infect an unexpectedly broad range of species and represent a new reservoir of potential human pathogens.IMPORTANCEInclusion body disease (IBD) is a common infectious disease of captive snakes. IBD is fatal and can cause the loss of entire animal collections. The cause of the disease has remained elusive, and no treatment exists. In addition to being important to pet owners, veterinarians, breeders, zoological parks, and aquariums, the study of animal disease is significant since animals are the source of virtually every emerging infectious human disease. We searched for candidate causative agents in snakes diagnosed with IBD and found a group of novel viruses distantly related mainly to arenaviruses but also to filoviruses, both of which can cause fatal hemorrhagic fevers when transmitted from animals to humans. In addition to providing evidence that strongly suggests that these viruses cause snake IBD, this discovery reveals a new and unanticipated domain of virus biology and evolution.

scholarly journals Identification and Characterization of Two Closely Related Unclassifiable Endogenous Retroviruses in Pythons (Python molurus and Python curtus)

2002 ◽  
Vol 76 (15) ◽  
pp. 7607-7615 ◽  
Author(s):  
Jon B. Huder ◽  
Jürg Böni ◽  
Jean-Michel Hatt ◽  
Guido Soldati ◽  
Hans Lutz ◽  
...  
Keyword(s):  
Mononuclear Cells ◽  
Endogenous Retrovirus ◽  
Endogenous Retroviruses ◽  
Open Reading Frames ◽  
Peripheral Blood Mononuclear ◽  
Python Regius ◽  
Boa Constrictor ◽  
Inclusion Body Disease ◽  
Python Molurus

ABSTRACT Boid inclusion body disease (BIBD) is a fatal disorder of boid snakes that is suspected to be caused by a retrovirus. In order to identify this agent, leukocyte cultures (established from Python molurus specimens with symptoms of BIBD or kept together with such diseased animals) were assessed for reverse transcriptase (RT) activity. Virus from cultures exhibiting high RT activity was banded on sucrose density gradients, and the RT peak fraction was subjected to highly efficient procedures for the identification of unknown particle-associated retroviral RNA. A 7-kb full retroviral sequence was identified, cloned, and sequenced. This virus contained intact open reading frames (ORFs) for gag, pro, pol, and env, as well as another ORF of unknown function within pol. Phylogenetic analysis showed that the virus is distantly related to viruses from both the B and D types and the mammalian C type but cannot be classified. It is present as a highly expressed endogenous retrovirus in all P. molurus individuals; a closely related, but much less expressed virus was found in all tested Python curtus individuals. All other boid snakes tested, including Python regius, Python reticulatus, Boa constrictor, Eunectes notaeus, and Morelia spilota, were virus negative, independent of whether they had BIBD or not. Virus isolated from P. molurus could not be transmitted to the peripheral blood mononuclear cells of B. constrictor or P. regius. Thus, there is no indication that this novel virus, which we propose to name python endogenous retrovirus (PyERV), is causally linked with BIBD.

scholarly journals Replication of Boid Inclusion Body Disease-Associated Arenaviruses Is Temperature Sensitive in both Boid and Mammalian Cells

2014 ◽  
Vol 89 (2) ◽  
pp. 1119-1128 ◽  
Author(s):  
Jussi Hepojoki ◽  
Anja Kipar ◽  
Yegor Korzyukov ◽  
Lesley Bell-Sakyi ◽  
Olli Vapalahti ◽  
...  
Keyword(s):  
Body Temperature ◽  
Inclusion Body ◽  
Mammalian Cells ◽  
Cytoplasmic Inclusion ◽  
Reservoir Host ◽  
The Body ◽  
Species Barrier ◽  
Content Type ◽  
Inclusion Body Disease ◽  
Reservoir Hosts

ABSTRACTBoid inclusion body disease (BIDB) is a fatal disease of boid snakes, the etiology of which has only recently been revealed following the identification of several novel arenaviruses in diseased snakes. BIBD-associated arenaviruses (BIBDAV) are genetically divergent from the classical Old and New World arenaviruses and also differ substantially from each other. Even though there is convincing evidence that BIBDAV are indeed the etiological agent of BIBD, the BIBDAV reservoir hosts—if any exist besides boid snakes themselves—are not yet known. In this report, we use University of Helsinki virus (UHV; a virus that we isolated from aBoa constrictorwith BIBD) to show that BIBDAV can also replicate effectively in mammalian cells, including human cells, provided they are cultured at 30°C. The infection induces the formation of cytoplasmic inclusion bodies (IB), comprised mainly of viral nucleoprotein (NP), similar to those observed in BIBD and in boid cell cultures. Transferring infected cells from 30°C to 37°C ambient temperature resulted in progressive declines in IB formation and in the amounts of viral NP and RNA, suggesting that BIBDAV growth is limited at 37°C. These observations indirectly indicate that IB formation is linked to viral replication. In addition to mammalian and reptilian cells, UHV infected arthropod (tick) cells when grown at 30°C. Even though our findings suggest that BIBDAV have a high potential to cross the species barrier, their inefficient growth at mammalian body temperatures indicates that the reservoir hosts of BIBDAV are likely species with a lower body temperature, such as snakes.IMPORTANCEThe newly discovered boid inclusion body disease-associated arenaviruses (BIBDAV) of reptiles have drastically altered the phylogeny of the familyArenavirus. Prior to their discovery, known arenaviruses were considered mainly rodent-borne viruses, with each arenavirus species having its own reservoir host. BIBDAV have so far been demonstrated in captive boid snakes, but their possible reservoir host(s) have not yet been identified. Here we show, using University of Helsinki virus as a model, that these viruses are able to infect mammalian (including human) and arthropod cells. Our results providein vitroproof of the considerable ability of arenaviruses to cross species barriers. However, our data indicate that BIBDAV growth occurs at 30°C but is inhibited at 37°C, implying that crossing of the species barrier would be hindered by the body temperature of mammalian species.

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