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

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

ICTV Virus Taxonomy Profile: Iridoviridae

The Iridoviridae is a family of large, icosahedral viruses with double-stranded DNA genomes ranging in size from 103 to 220 kbp. Members of the subfamily Alphairidovirinae infect ectothermic vertebrates (bony fish, amphibians and reptiles), whereas members of the subfamily Betairidovirinae mainly infect insects and crustaceans. Infections can be either covert or patent, and in vertebrates they can lead to high levels of mortality among commercially and ecologically important fish and amphibians. This is a summary of the current International Committee on Taxonomy of Viruses (ICTV) Report on the taxonomy of the Iridoviridae, which is available at www.ictv.global/report/iridoviridae.</p

The genome of a tortoise herpesvirus (testudinid herpesvirus 3) has a novel structure and contains a large region that is not required for replication in vitro or virulence in vivo

Testudinid herpesvirus 3 (TeHV-3) is the causative agent of a lethal disease affecting several tortoise species. The threat that this virus poses to endangered animals is focusing efforts on characterizing its properties, in order to enable the development of prophylactic methods. We have sequenced the genomes of the two most studied TeHV-3 strains (1976 and 4295). TeHV-3 strain 1976 has a novel genome structure and is most closely related to a turtle herpesvirus, thus supporting its classification into genus Scutavirus, subfamily Alphaherpesvirinae, family Herpesviridae. The sequence of strain 1976 also revealed viral counterparts of cellular interleukin-10 and semaphorin, which have not been described previously in members of subfamily Alphaherpesvirinae. TeHV-3 strain 4295 is a mixture of three forms (m1, m2, and M), in which, in comparison to strain 1976, the genomes exhibit large, partially overlapping deletions of 12.5 to 22.4 kb. Viral subclones representing these forms were isolated by limiting dilution, and each replicated in cell culture comparably to strain 1976. With the goal of testing the potential of the three forms as attenuated vaccine candidates, strain 4295 was inoculated intranasally into Hermann's tortoises (Testudo hermanni). All inoculated subjects died, and PCR analyses demonstrated the ability of the m2 and M forms to spread and invade the brain. In contrast, the m1 form was detected in none of the organs tested, suggesting its potential as the basis of an attenuated vaccine candidate. Our findings represent a major step towards characterizing TeHV-3 and developing prophylactic methods against it. IMPORTANCE: Testudinid herpesvirus 3 (TeHV-3) causes a lethal disease in tortoises, several species of which are endangered. We have characterized the viral genome, and used this information to take steps towards developing an attenuated vaccine. We have sequenced the genomes of two strains (1976 and 4295), compared their growth in vitro, and investigated the pathogenesis of strain 4295, which consists of three deletion mutants. The major findings are: (i) TeHV-3 has a novel genome structure; (ii) its closest relative is a turtle herpesvirus; (iii) it contains interleukin-10 and semaphorin genes, the first time these have been reported in an alphaherpesvirus; (iv) a sizeable region of the genome is not required for viral replication in vitro or virulence in vivo; and (v) one of the components of strain 4295, which has a deletion of 22.4 kb, exhibits properties indicating that it may serve as the starting point for an attenuated vaccine

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%

Complement protein C1q interacts with DC-SIGN via its globular domain, and thus may interfere with HIV-1 transmission

Dendritic Cells (DCs) are the most potent antigen presenting cells capable of priming naïve T cells. Its C-type lectin receptor, DC-SIGN, regulates a wide range of immune functions. Along with its role in HIV-1 pathogenesis through complement opsonization of the virus, DC-SIGN has recently emerged as an adaptor for complement protein C1q on the surface of immature DCs via a trimeric complex involving gC1qR, a receptor for the globular domain of C1q. Here, we have examined the nature of interaction between C1q and DC-SIGN in terms of domain localization, and implications of C1q-DC-SIGN-gC1qR complex formation on HIV-1 transmission. We first expressed and purified recombinant extracellular domains of DC-SIGN and its homologue SIGN-R as tetramers comprising of the entire extra cellular domain including the α-helical neck region, and monomers comprising of the carbohydrate recognition domain only. Direct binding studies revealed that both DC-SIGN and SIGN-R were able to bind independently to the recombinant globular head modules ghA, ghB and ghC, with ghB being the preferential binder. C1q appeared to interact with DC-SIGN or SIGN-R in a manner similar to IgG. Mutational analysis using single amino acid substitutions within the globular head modules showed that TyrB175 and LysB136 38 were critical for the C1q-DC-SIGN/SIGN-R interaction. Competitive studies revealed that gC1qR and ghB shared overlapping binding sites on DC-SIGN, implying that HIV- 1 transmission by DCs could be modulated due to the interplay of gC1qR-C1q with DC-SIGN. Since C1q, gC1qR and DC-SIGN can individually bind HIV-1, we examined how C1q and gC1qR modulated HIV-1-DC-SIGN interaction in an infection assay. Here, we report, for the first time, that C1q suppressed DC-SIGN-mediated transfer of HIV-1 to activated PBMCs, although the globular head modules did not. The protective effect of C1q was negated by the addition of gC1qR. In fact, gC1qR enhanced DC-SIGN-mediated HIV-1 transfer, suggesting its role in HIV-1 pathogenesis. Our results highlight the consequences of multiple innate immune pattern recognition molecules forming a complex that can modify their functions in a way which may be advantageous for the pathogen

First detection of the amphibian chytrid fungus (Batrachochytridium dendrobatidis) in free- living anuran populations in Greece

Ο χυτριδιομύκητας των αμφιβίων (Batrachochytrium dendrobatidis) αποτελεί έναν ευρείας εξάπλωσης, κοσμοπολίτικο μικροοργανισμό, που επηρεάζει τους άγριους πληθυσμούς των αμφιβίων. Μέχρι στιγμής δεν έχει δημοσιοποιηθεί κάποια έρευνα για την ύπαρξη του μύκητα στην Ελλάδα. Σε αυτή την προκαταρκτική έρευνα δειγματίσαμε 59 μεταμορφωμένα άνουρα από τέσσερις ελληνικούς υγροτόπους. Πέντε δείγματα διαγνώστηκαν θετικά με την μέθοδο της αλυσιδωτής αντίδρασης της πολυμεράσης σε πραγματικό χρόνο (real-time PCR). Το Βατραχοχυτρίδιο (Batrachochytrium dendrobatidis) εντοπίστηκε σε τρία είδη βατράχων (Bufo viridis, Pelophylax epeiroticus, Pelophylax ridibundus) όχι όμως και στο απειλούμενο ενδημικό είδος του βατράχου της Καρπάθου (Pelophylax cerigensis). Αυτή αποτελεί την πρώτη αναφορά ανίχνευσης του βατραχοχυτριδίου (Batrachochytrium dendrobatidis) από άγριους πληθυσμούς αμφιβίων από την Ελλάδα.The amphibian chytrid fungus (Batrachochytrium dendrobatidis) is a widespread, cosmopolitan pathogen largely affecting free-living amphibian populations. So far there are no published studies for the presence of B. dendrobatidis in Greece. In this preliminary study we sampled 59 metamorphosed anurans from four Greek wetlands. Five samples were positive for the fungus by real-time PCR. B. dendrobatidis was detected in three species (Bufo viridis, Pelophylax epeiroticus, Pelophylax ridibundus) but not in endangered endemic Karpathos water frogs (Pelophylax cerigensis). This is the first report of the amphibian chytrid fungus (Batrachochytrium dendrobatidis) in free-living anuran populations from Greece

Mass mortality of unknown etiology in alpine newts (Ichtyosaura alperstris veluchiensis) in an alpine lake in Greece

ΔΕΝ ΔΙΑΤΙΘΕΤΑΙ ΠΕΡΙΛΗΨΗA mass mortality in alpine newts (Ichthyosaura alpestris veluchiensis) was observed in May/June 2013, in Drakolimni lake on Smolikas Mountain, Northwest Greece. 1300 alpine newts were found dead in two events. In 1998 a similar incident was recorded in the nearby lake of Timfi Mt. Newts of every stage and sex were affected, presenting incoordination and inability to float evenly. Ten animals were submitted for complete pathological examination. Field environmental measurements (water temperature, oxygen saturation, pH, conductance, nitric/phosphate concentration) and samples (water, snow, benthos) were collected for ecotoxicological and quality analysis. Necropsy, microbiology (parasitology, bacteriology, mycology), histopathology, molecular investigations (Ranavirus spp, Batrachochytridium dendrobatidis, Batrachochytridium salamandrivorans), quality and ecotoxicological examinations did not indicate a causative source for the mortality. To the author´s knowledge this is the biggest mortality of unknown etiology reported in free-living alpine newts in Europe