Viruses Infecting Reptiles

A large number of viruses have been described in many different reptiles. These viruses include arboviruses that primarily infect mammals or birds as well as viruses that are specific for reptiles. Interest in arboviruses infecting reptiles has mainly focused on the role reptiles may play in the epidemiology of these viruses, especially over winter. Interest in reptile specific viruses has concentrated on both their importance for reptile medicine as well as virus taxonomy and evolution. The impact of many viral infections on reptile health is not known. Koch’s postulates have only been fulfilled for a limited number of reptilian viruses. As diagnostic testing becomes more sensitive, multiple infections with various viruses and other infectious agents are also being detected. In most cases the interactions between these different agents are not known. This review provides an update on viruses described in reptiles, the animal species in which they have been detected, and what is known about their taxonomic positions

Prevalence of inclusion body disease and associated comorbidity in captive collections of boid and pythonid snakes in Belgium

Inclusion body disease (IBD) is caused by reptarenaviruses and constitutes one of the most notorious viral diseases in snakes. Although central nervous system disease and various other clinical signs have been attributed to IBD in boid and pythonid snakes, studies that unambiguously reveal the clinical course of natural IBD and reptarenavirus infection are scarce. In the present study, the prevalence of IBD and reptarenaviruses in captive snake collections and the correlation of IBD and reptarenavirus infection with the clinical status of the sampled snakes were investigated. In three IBD positive collections, long-term follow-up during a three- to seven-year period was performed. A total of 292 snakes (178 boas and 114 pythons) from 40 collections in Belgium were sampled. In each snake, blood and buffy coat smears were evaluated for the presence of IBD inclusion bodies (IB) and whole blood was tested for reptarenavirus RNA by RT-PCR. Of all tested snakes, 16.5% (48/292) were positive for IBD of which all were boa constrictors (34.0%; 48/141) and 17.1% (50/292) were reptarenavirus RT-PCR positive. The presence of IB could not be demonstrated in any of the tested pythons, while 5.3% (6/114) were reptarenavirus positive. In contrast to pythons, the presence of IB in peripheral blood cells in boa constrictors is strongly correlated with reptarenavirus detection by RT-PCR (P<0.0001). Although boa constrictors often show persistent subclinical infection, long-term follow-up indicated that a considerable number (22.2%; 6/27) of IBD/reptarenavirus positive boas eventually develop IBD associated comorbidities

Export of Marcellus Shale Gas

The Marcellus Shale natural gas field that spans from West Virginia to New York is leading the recent surge in domestic energy production. Long an importer of natural gas, the United States will soon be able to export natural gas. Due to its low energy density however, natural gas must be converted to liquefied natural gas (LNG) before shipping to foreign markets. Liquefaction can occur at several different facilities: small-scale LNG plants, floating LNG operations, and retrofitted LNG import facilities. A design feasibility study is presented here to analyze the economics of retrofitting an existing LNG import facility into an LNG export plant. The existing import facility is the Dominion Cove Point LNG plant located near Lusby, Maryland. This study sizes the export facility at 5 to 6 million tons per annum (MMTPA), which corresponds to a feed of about 750 million standard cubic feet per day of natural gas (MMscfd). In this process, natural gas is first precooled by propane and then liquefied with a mixed refrigerant blend of methane, ethane, propane, and nitrogen. One challenge is to minimize the large amount of mixed refrigerant used in this process. This can be done by optimizing the composition of the mixed refrigerant to reduce the amount needed to liquefy the natural gas. After a comprehensive economic analysis, this proposed design is economically viable. This process has an estimated IRR of 23.5% and NPV of $219 million at a 20$650 per ton. This 23.5% IRR is possible due to the retrofit advantages of some existing equipment and reduced construction time. Without these advantages, the IRR would be much less favorable at about 9.1%

Detection and Characterization of Invertebrate Iridoviruses Found in Reptiles and Prey Insects in Europe over the Past Two Decades

Invertebrate iridoviruses (IIVs), while mostly described in a wide range of invertebrate hosts, have also been repeatedly detected in diagnostic samples from poikilothermic vertebrates including reptiles and amphibians. Since iridoviruses from invertebrate and vertebrate hosts differ strongly from one another based not only on host range but also on molecular characteristics, a series of molecular studies and bioassays were performed to characterize and compare IIVs from various hosts and evaluate their ability to infect a vertebrate host. Eight IIV isolates from reptilian and orthopteran hosts collected over a period of six years were partially sequenced. Comparison of eight genome portions (total over 14 kbp) showed that these were all very similar to one another and to an earlier described cricket IIV isolate, thus they were given the collective name lizard–cricket IV (Liz–CrIV). One isolate from a chameleon was also subjected to Illumina sequencing and almost the entire genomic sequence was obtained. Comparison of this longer genome sequence showed several differences to the most closely related IIV, Invertebrate iridovirus 6 (IIV6), the type species of the genus Iridovirus, including several deletions and possible recombination sites, as well as insertions of genes of non-iridoviral origin. Three isolates from vertebrate and invertebrate hosts were also used for comparative studies on pathogenicity in crickets (Gryllus bimaculatus) at 20 and 30 C. Finally, the chameleon isolate used for the genome sequencing studies was also used in a transmission study with bearded dragons. The transmission studies showed large variability in virus replication and pathogenicity of the three tested viruses in crickets at the two temperatures. In the infection study with bearded dragons, lizards inoculated with a Liz–CrIV did not become ill, but the virus was detected in numerous tissues by qPCR and was also isolated in cell culture from several tissues. Highest viral loads were measured in the gastro-intestinal organs and in the skin. These studies demonstrate that Liz–CrIV circulates in the pet trade in Europe. This virus is capable of infecting both invertebrates and poikilothermic vertebrates, although its involvement in disease in the latter has not been proven

Detection and Characterization of Invertebrate Iridoviruses Found in Reptiles and Prey Insects in Europe over the Past Two Decades

Invertebrate iridoviruses (IIVs), while mostly described in a wide range of invertebrate hosts, have also been repeatedly detected in diagnostic samples from poikilothermic vertebrates including reptiles and amphibians. Since iridoviruses from invertebrate and vertebrate hosts differ strongly from one another based not only on host range but also on molecular characteristics, a series of molecular studies and bioassays were performed to characterize and compare IIVs from various hosts and evaluate their ability to infect a vertebrate host. Eight IIV isolates from reptilian and orthopteran hosts collected over a period of six years were partially sequenced. Comparison of eight genome portions (total over 14 kbp) showed that these were all very similar to one another and to an earlier described cricket IIV isolate, thus they were given the collective name lizard–cricket IV (Liz–CrIV). One isolate from a chameleon was also subjected to Illumina sequencing and almost the entire genomic sequence was obtained. Comparison of this longer genome sequence showed several differences to the most closely related IIV, Invertebrate iridovirus 6 (IIV6), the type species of the genus Iridovirus, including several deletions and possible recombination sites, as well as insertions of genes of non-iridoviral origin. Three isolates from vertebrate and invertebrate hosts were also used for comparative studies on pathogenicity in crickets (Gryllus bimaculatus) at 20 and 30 C. Finally, the chameleon isolate used for the genome sequencing studies was also used in a transmission study with bearded dragons. The transmission studies showed large variability in virus replication and pathogenicity of the three tested viruses in crickets at the two temperatures. In the infection study with bearded dragons, lizards inoculated with a Liz–CrIV did not become ill, but the virus was detected in numerous tissues by qPCR and was also isolated in cell culture from several tissues. Highest viral loads were measured in the gastro-intestinal organs and in the skin. These studies demonstrate that Liz–CrIV circulates in the pet trade in Europe. This virus is capable of infecting both invertebrates and poikilothermic vertebrates, although its involvement in disease in the latter has not been proven

Three genetically distinct ferlaviruses have varying effects on infected corn snakes (Pantherophis guttatus)

Ferlaviruses are important pathogens in snakes and other reptiles. They cause respiratory and neurological disease in infected animals and can cause severe disease outbreaks. Isolates from this genus can be divided into four genogroups-A, B, and C, as well as a more distantly related sister group, "tortoise". Sequences from large portions (5.3 kb) of the genomes of a variety of ferlavirus isolates from genogroups A, B, and C, including the genes coding the surface glycoproteins F and HN as well as the L protein were determined and compared. In silico analyses of the glycoproteins of genogroup A, B, and C isolates were carried out. Three isolates representing these three genogroups were used in transmission studies with corn snakes (Pantherophis guttatus), and clinical signs, gross and histopathology, electronmicroscopic changes in the lungs, and isolation of bacteria from the lungs were evaluated. Analysis of the sequences supported the previous categorization of ferlaviruses into four genogroups, and criteria for definition of ferlavirus genogroups and species were established based on sequence identities (80% resp. 90%). Analysis of the ferlavirus glycoprotein models showed parallels to corresponding regions of other paramyxoviruses. The transmission studies showed clear differences in the pathogenicities of the three virus isolates used. The genogroup B isolate was the most and the group A virus the least pathogenic. Reasons for these differences were not clear based on the differences in the putative structures of their respective glycoproteins, although e.g. residue and consequential structure variation of an extended cleavage site or changes in electrostatic charges at enzyme binding sites could play a role. The presence of bacteria in the lungs of the infected animals also clearly corresponded to increased pathogenicity. This study contributes to knowledge about the structure and phylogeny of ferlaviruses and lucidly demonstrates differences in pathogenicity between strains of different genogroups

Repeated Detection of an Invertebrate Iridovirus (IIV) in Amphibians

Invertebrate iridoviruses (IIVs) (family: Iridoviridae) are known pathogens for invertebrates, causing high mortality and reduced fertility in affected insects. Over the past 2 decades, IIVs have also been increasingly found in lizards in association with nonspecific clinical signs. It has been hypothesized that IIVs from insects can also infect reptiles. From 2010-2011, IIVs were repeatedly detected via polymerase chain reaction testing and virus isolation methods in routine diagnostic samples from different amphibians: 3 blue poison dart frogs (Dendrobates tinctorius azureus), 4 edible frogs (Pelophylax kl. esculentus), a giant ditch frog (Leptodactylus fallax), an Amazon milk frog (Trachycephalus resinifictrix), mixed organs from agile frogs (Rana dalmatina), a black-spined toad (Bufo melanostictus), and one Lake Urmia newt (Neurergus crocatus). IIVs were found in skin swabs from apparently healthy animals, as well as in multiple organs of frogs that died of unknown causes. Prey insects (crickets) from one owner also tested positive for the presence of IIV. The obtained partial sequences from the major capsid protein (MCP) gene (222nt) from each of these were 100% identical to each other and 98% identical to IIV-6, the type species of the genus Iridovirus. Although the pathogenicity of IIV in amphibians remains unclear, these findings provide further evidence that IIVs may be able to infect vertebrates under some conditions and underline the importance of the genus Iridovirus in vertebrates