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Partial characterization of a new adenovirus lineage discovered in testudinoid turtles
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... HVs have been associated with necrotizing and necrotic stomatitis, glossitis, rhinitis in tortoises (Hunt 2006;Okoh et al. 2021); proliferative and/or ulcerative lesions of the skin and shell in freshwater turtles (Okoh et al. 2021;Široký et al. 2018); and fibropapillomatosis and grey-patch disease in sea turtles (Cárdenas et al. 2019;Jacobson et al. 1986;Robben et al. 2023;Stacy et al. 2008). AdVs that have been found to infect chelonians include members of the genera Siadenovirus, Atadenovirus and Testadenovirus (Rivera et al. 2009;Doszpoly et al. 2013;Garcia-Morante et al. 2016;Salzmann et al. 2021). While siadenovirus (Sulawesi tortoise adenovirus 1) and atadenovirus (spur-thighed tortoise adenovirus 1) have been associated with anorexia, lethargy, ulcerations and erosions of the oral mucosa, stomatitis, esophagitis and mortality in tortoise (Rivera et al. 2009;Garcia-Morante et al. 2016), testadenoviruses have been frequently detected in clinically healthy chelonians (Doszpoly et al. 2013;Salzmann et al. 2021;Nieto-Claudin et al. 2022). ...
... AdVs that have been found to infect chelonians include members of the genera Siadenovirus, Atadenovirus and Testadenovirus (Rivera et al. 2009;Doszpoly et al. 2013;Garcia-Morante et al. 2016;Salzmann et al. 2021). While siadenovirus (Sulawesi tortoise adenovirus 1) and atadenovirus (spur-thighed tortoise adenovirus 1) have been associated with anorexia, lethargy, ulcerations and erosions of the oral mucosa, stomatitis, esophagitis and mortality in tortoise (Rivera et al. 2009;Garcia-Morante et al. 2016), testadenoviruses have been frequently detected in clinically healthy chelonians (Doszpoly et al. 2013;Salzmann et al. 2021;Nieto-Claudin et al. 2022). ...
... The data obtained in this study does not support SsTAdVs as the causative agent of the cutaneous lesions. This finding is typical of members in the genus Testadenovirus, which have so far not been associated with clinical diseases (Doszpoly et al. 2013;Salzmann et al. 2021). However, the occurrence of novel AdVs in this population of freshwater turtles should not be ignored since AdVs can potentially cause diseases of conservation concerns (Marschang 2011;Marschang et al. 2020). ...
Cutaneous lesions were observed in a wild population of freshwater turtles during routine disease surveillance at Alligator Creek, Townsville, Australia. Previous attempts to identify the causative agent of these lesions were unsuccessful; however, existing evidence suggests viral etiology. To further investigate these events, blood samples and lesion, oral and cloacal swabs collected from 128 freshwater turtles at two locations (Alligator Creek and Ross River) were screened for herpesvirus, adenovirus, poxvirus and papillomavirus by polymerase chain reaction. We detected three novel herpesviruses (chelid herpesvirus 1–3) as well as three adenoviruses (saw-shelled turtle adenovirus 1–3). Phylogenetic analyses showed that the herpesviruses formed a distinct clade with the tumour-associated chelonid alphaherpesvirus 5 (Scutavirus chelonidalpha5) within the subfamily Alphaherpesvirinae. The adenoviruses clustered with members of the genus Testadenovirus. Although the novel herpesviruses and adenoviruses could not be linked to the occurrence of cutaneous lesions, further characterisation will help provide better insights into their clinical, epidemiological and conservation significance.
... AdVs have been identified in more than 30 squamate species to date, and have been shown to cause morbidity and mortality in numerous captive-bred pet trade species (Frye et al., 1994;Kim et al., 2002;Marschang et al., 2003;Pasmans et al., 2008;Moormann et al., 2009;Ball et al., 2014). However, not all reptile species that harbor adenovirus show adverse health effects (Jacobson & Kollias, 1986;Ogawa, Ahne & Essbauer, 1992;Parkin et al., 2009;Doszpoly et al., 2013;Kubiak, 2013); the pathogenicity of adenovirus infection across reptiles is unclear. ...
... By contrast, across shallower host divergences AdV infection appears to be relatively species-specific. AdV sequences isolated from conspecific individuals tend to form monophyletic clades, such that members of the same host species are generally infected by the same (or very similar) AdV strain (Papp et al., 2009;Doszpoly et al., 2013). This pattern holds for both wild and captive squamates (Szirovicza et al., 2016), but it is important to note that the vast majority of this evidence comes from captive pet-trade animals, where pathogen exposure, transmission, and disease dynamics are likely to differ from wild populations. ...
... Our resulting phylogeny overall is consistent with recently constructed adenovirus trees (Fig. 2). It recovers monophyletic viral genera Aviadenovirus, Mastadenovirus, Atadenovirus, Siadenovirus (Harrach et al., 2011), and the recently proposed "Testudineinfecting AdV" genus (Doszpoly et al., 2013). Within the primarily reptile-infecting Atadenovirus clade, our tree shows support for previously established AdV subclades (e.g., Snakes + Eublepharids + Helodermatids, Anolis + chameleon, Agamids), and similar overall structure (i.e., Gekkonid AdV as sister to the remaining Atadenovirus) (Wellehan et al., 2004;Ascher et al., 2013;Ball et al., 2014;Szirovicza et al., 2016). ...
Adenoviruses (AdVs) infect a wide range of hosts, and they have undergone recent and ancient host transfers multiple times. In reptiles, AdVs have been found in many captive individuals, and have been implicated in morbidity and mortality in several species. Yet the pathogenicity, transmission, phylogenetic distribution, and source of AdVs in the environment are still unknown. We therefore chose to opportunistically sample deceased captive Anolis sagrei individuals that were collected from different populations in the Bahamas and the Cayman Islands, as well as fecal samples from one island population, to explore the disease dynamics and diversity of adenovirus infecting A. sagrei populations. We found that adenovirus infection was present in our captive colony at low prevalence (26%), and was likely not the primary cause of observed morbidity and mortality. Among the 10 individuals (out of 38 sampled) which tested positive for adenovirus, we identified four adenovirus clades, several of which are distantly related, despite the close relationships of the A. sagrei host populations. These results suggest that while adenovirus may not be highly prevalent in the wild, it is present at low levels across much of the range of A. sagrei . It may undergo frequent host switching across both deep and shallow host divergences.
... They are nonenveloped DNA viruses that replicate within host nuclei, with genomes of 26-45 kilobase pairs (Davidson et al. 2003). The family Adenoviridae may be divided into six genera that often show predilection for host clades (Doszpoly et al. 2013). Mastadenovirus is currently reported only in mammals and Aviadenovirus only in birds (Davidson et al. 2003). ...
... Atadenovirus likely originated in squamates but has been found in other host species including birds, ruminants, and marsupials (Wellehan et al. 2004). Siadenovirus has an unknown origin (Doszpoly et al. 2013). A single member of the genus Ichtadenovirus was characterized in the white sturgeon (Acipenser transmontanus) (Doszpoly et al. 2013). ...
... Siadenovirus has an unknown origin (Doszpoly et al. 2013). A single member of the genus Ichtadenovirus was characterized in the white sturgeon (Acipenser transmontanus) (Doszpoly et al. 2013). After the identification of a clade of novel viruses from testudinoid turtles, the provisional genus Testadenovirus has been proposed (Doszpoly et al. 2013). ...
Adenoviruses are nonenveloped, double-stranded DNA viruses, known to infect members of all tetrapod classes, with a similarity between phylogenies of hosts and viruses observed. We characterized bottlenose dolphin adenovirus 2 (BdAdV-2) found in a bottlenose dolphin (Tursiops truncatus) with enteritis. Virions were seen by negative staining electron microscopy of feces. Initial sequences obtained using conserved PCR primers were expanded using primer walking techniques, and the complete coding sequence was obtained. Phylogenetic analyses were consistent with coevolution of this virus and its bottlenose dolphin host, placing BdAdV-2 into a monophyletic group with other mastadenoviruses of Cetartiodactyla. When considering the low guanine/cytosine (G/C) content of BdAdV-2 with the phylogenetic data, this virus may represent a host-jumping event from another member of Cetartiodactyla. Analysis of partial polymerase indicated that bottlenose dolphin adenovirus 1, previously identified in Spain, and BdAdV-2 are sister taxa with harbor porpoise adenovirus 1, forming a cetacean clade. Bottlenose dolphin adenovirus 2 includes a highly divergent fiber gene. Two genes homologous to the dUTPase superfamily are also present which could play a role in enabling viral replication in nondividing cells. We used sequence data to develop a probe hybridization quantitative PCR assay specific to BdAdV-2 with a limit of detection of 10 copies.
... The positive samples were further tested by different PCRs depending on the virus type deduced from the partial pol sequences. Most of the additional (simple or nested) PCRs have been described for the amplification of fragments from the gene of the hexon, the major capsid protein (Meulemans et al., 2001; Raue et al., 2005a and b; Freick et al., 2008; Mase et al., 2009; Stenzel et al., 2012; Doszpoly et al., 2013). As shown in Table 1, we altered slightly the sequence of some primers in order to increase their sensitivity. ...
... It was originally designed for the recognition of novel atadenovirues in lizards, but its sensitivity and robustness allowed the discovery of many new aviadenoviruses (Wellehan et al., 2005; Bodewes et al., 2013), siadenoviruses (Katoh et al., 2009; Wellehan et al., 2009; Kovács et al., 2010; Park et al., 2012; Ballmann and Vidovszky, 2013; Joseph et al., 2014; Lee et al., 2014), atadenoviruses (Szirovicza et al., 2016 ) and mastadenoviruses (Vidovszky et al., 2015) in a wide range of hosts. Even the members of a hitherto unknown genus (with the proposed name Testadenovirus) could be detected (Doszpoly et al., 2013). We revealed the existence of four novel AdV types in pigeons. ...
... Even the nested primer set, originally designed on the basis of the most conserved aa stretches of the known siadenovirus hexons (Kovács, unpublished results) failed to recognise PiAdV-5. This was surprising, as this PCR had been successfully used for the demonstration of AdVs belonging to a different genus (Doszpoly et al., 2013). The relationship between the newly found PiAdVs and YPDS or other disease conditions remains to be elucidated. ...
Up to now, only a single adenovirus (AdV) isolate seemingly specific for pigeons, hence named pigeon AdV-1 (PiAdV-1), has been characterised at DNA sequence level. In the present work, the prevalence and diversity of AdVs occurring in domestic pigeon were examined by a survey performed on randomly collected samples using a very efficient, consensus nested PCR targeting the viral DNA polymerase gene. The newly detected viruses were characterised by sequencing and phylogeny analysis. Amplification of additional genome fragments was attempted by the use of several other PCR methods aiming at the hexon gene. During a 4-year survey, samples from dead or live, healthy pigeons originating from 27 lofts were examined in Hungary. Almost 50% of the samples (48 out of 97) proved to be positive for AdV. Sequence analysis revealed the presence of four hitherto unknown pigeon AdV types. PiAdV-1 was also identified in one sample. Two novel viruses named PiAdV-2 and -3 were found to belong to the genus Aviadenovirus, and two other novel types (PiAdV-4 and -5) to the genus Siadenovirus. This is the first report on the occurrence of siadenoviruses in birds belonging to the order Columbiformes. Approximately two-thirds of the PiAdV-2 genome was sequenced and analysed.
... Testadenovirus (Doszpoly et al. 2013). Adenoviruses infect a wide range of vertebrates, ...
... In general, this disease is self-limiting with debilitated or young animals affected (Nollens et al. 2006a, Nollens et al. 2006b). been proposed for the adenoviruses of testudinoid turtles (Doszpoly et al. 2013). ...
... Viral diseases have been reported to cause mortalities in wild populations of pinnipeds (Osterhaus 1989, Bostock et al. 1990, Osterhaus et al. 1990, Osterhaus et al. 1997, Anthony et al. 2012. Mortality events have occurred in Mediterranean monk seals (Monachus monachus), where morbillivirus was initially suspected, but later studies indicated that the most likely cause was a harmful algal bloom (Osterhaus et al. 1997, Hernández et al. 1998, Van de Bildt et al. 1999 , Doszpoly et al. 2013. Adenoviruses infect a wide range of vertebrates and show relatively high host specificity. ...
... plains box turtle, Terrapene ornata ornata) with hepatic degeneration. 5,8 In the present work, we describe a UATD in a spur-thighed tortoise (Testudo graeca) associated with an adenovirus infection. ...
... scripta scripta), a new adenovirus lineage was isolated and is being proposed as a candidate for a new genus. 5 This lineage did not show a clear pathogenic role and was therefore speculated to have coevolved with testudinoid turtles. 5 In the present case, we reported an adenoviral-like infection restricted to the upper digestive tract. ...
... 5 This lineage did not show a clear pathogenic role and was therefore speculated to have coevolved with testudinoid turtles. 5 In the present case, we reported an adenoviral-like infection restricted to the upper digestive tract. Regrettably, it was not possible to establish whether this animal suffered from a primary pathogenic adenoviral infection, or if it was a consequence of a stressful state produced by the intestinal parasitosis or other unrelated factors. ...
A two-year-old female, spur-thighed tortoise (Testudo graeca) presented with poor body condition (1/5) and weakness. Fecal analysis revealed the presence of a large number of Oxyurid-like eggs, and radiographs were compatible with gastrointestinal obstruction. Despite supportive medical treatment, the animal died. At gross examination, an intestinal obstruction was confirmed. Histopathology revealed severe hyperplastic esophagitis and stomatitis with marked epithelial cytomegaly and enormous basophilic intranuclear inclusion bodies. Electron microscopy examination revealed a large number of 60-80 nm, non-enveloped, icosahedral virions arranged in crystalline arrays within nuclear inclusions of esophageal epithelial cells, morphologically compatible with adenovirus-like particles. PCR for virus identification was performed with DNA extracted from formalin-fixed paraffin-embedded tissues. A nested, consensus pan-adenovirus PCR and sequencing analysis showed a previously unidentified, novel adenovirus. According to phylogenetic calculations, it clustered to genus Atadenovirus in contrast with all other chelonian adenoviruses described to date. The present report details the pathological findings associated with an adenovirus infection restricted to the upper digestive tract.
... In contrast, animal adenoviruses appear to be associated with clinically important diseases in animals and birds [9,10]. AdVs, about 65-90 nm in size [1,11] with complex structural organization [11,12] have been grouped into six genera in Adenoviridae family [11,12]. Viruses in the Adenoviridae family contain a 26 to 48 kb [13] non-segmented linear doublestranded DNA genome [1,11], which transcribes at different times post-infection generating transcripts classified into early (E), intermediate (I) and late (L) regions [1,11]. ...
... In contrast, animal adenoviruses appear to be associated with clinically important diseases in animals and birds [9,10]. AdVs, about 65-90 nm in size [1,11] with complex structural organization [11,12] have been grouped into six genera in Adenoviridae family [11,12]. Viruses in the Adenoviridae family contain a 26 to 48 kb [13] non-segmented linear doublestranded DNA genome [1,11], which transcribes at different times post-infection generating transcripts classified into early (E), intermediate (I) and late (L) regions [1,11]. ...
Adenoviruses have served as a model for investigating viral-cell interactions and discovering different cellular processes, such as RNA splicing and DNA replication. In addition, the development and evaluation of adenoviruses as the viral vectors for vaccination and gene therapy has led to detailed investigations about adenovirus biology, including the structure and function of the adenovirus encoded proteins. While the determination of the structure and function of the viral capsid proteins in adenovirus biology has been the subject of numerous reports, the last few years have seen increased interest in elucidating the structure and function of the adenovirus core proteins. Here, we provide a review of research about the structure and function of the adenovirus core proteins in adenovirus biology.
... Siadenoviruses are known to occur commonly in birds, but have also been detected in tortoises and in a frog (Rivera et al., 2009;Kovács and Benkő, 2011;Ballmann and Vidovszky, 2013;Ballmann and Harrach, 2016;Lee et al., 2016). Recently, a sixth genus, Testadenovirus has been proposed for the AdVs discovered in testudinoid turtles (Doszpoly et al., 2013). Adenoviruses usually have a narrow host range restricted to a single, or several closely related, vertebrate species. ...
... Thanks to the increased sensitivity of broad-range PCRs, numerous novel AdVs have been discovered and described in the past couple of decades Wellehan et al., 2004;Papp et al., 2009;Rivera et al., 2009;Ballmann and Vidovszky, 2013;Doszpoly et al., 2013;Ballmann and Harrach, 2016;Garcia-Morante et al., 2016;Lee et al., 2016;Szirovicza et al., 2016;Podgorski et al., 2018) (https:// sites.google.com/site/adenoseq). In the majority of these cases however, actual isolation of the detected viruses has been hampered by the lack of appropriate cell lines. ...
The presence of a novel adenovirus (AdV) was detected by PCR and sequencing, in the internal organs of a captive polar bear that had died in the Budapest zoo. The virus content of the samples proved to be high enough to allow for conventional Sanger sequencing on PCR-amplified genomic fragments. With this approach, the sequence of the entire genome of the putative polar bear adenovirus 1 (PBAdV-1) was obtained. Although the genome was found to be short, consisting of 27,952 base pairs merely, with a relatively balanced G + C content of 46.3%, its organisation corresponded largely to that of a typical mastadenovirus. Every genus-common gene could be identified except that of protein IX. The short E3 region of the PBAdV-1 consisted of two novel, supposedly type-specific ORFs only, whereas no homologue of any of the E3 genes, usually conserved in mastadenoviruses, such as for example that of the 12.5 K protein, were present. In the E4 region, only the highly conserved gene of the 34 K protein was found besides two novel ORFs showing no homology to any known E4 ORFs. In silico sequence analysis revealed putative splicing donor and acceptor sites in the genes of the E1A, IVa2, DNA-dependent DNA polymerase, pTP, 33 K proteins, and also of U exon protein, all being characteristic for mastadenoviruses. Phylogenetic calculations, based on various proteins, further supported that the newly-detected PBAdV is the representative of a new species within the genus Mastadenovirus, and may represent the evolutionary lineage of adenoviruses that coevolved with carnivorans.
... This article is protected by copyright. All rights reserved A sixth AdV lineage has been discovered more recently in a number of testudinoid turtles, and was proposed, albeit not approved yet, to become an additional genus Testadenovirus [4]. The genera are further subdivided into AdV species, each of which may contain several virus types, designated by the host name with ascending numbers. ...
... Although originally, the adenoviral lineage co-evolving with reptiles was hypothesized to be Atadenovirus, it turned out soon that atadenoviruses, specifically those with genomic DNA of non-biased base composition, are confined to lizards, worm lizards and snakes, classified into the largest reptilian order Squamata (scaled reptiles). More recently, new virus sequences, representing multiple virus species of a putative novel AdV lineage have been discovered in red-eared and yellow-bellied sliders, box turtles, and pancake tortoises [4]. A similar virus was also detected in red-footed tortoises [9]. ...
Adenoviruses (AdVs) infect representatives of numerous species from almost every major vertebrate class, albeit their incidence shows great variability. AdVs infecting birds, reptiles and bats are the most common and diverse, whereas only one AdV has been so far isolated both from fish and amphibians. The family Adenoviridae is divided into five genera, each corresponding to an independent evolutionary lineage that supposedly co‐evolved with its respective vertebrate hosts. Members of genera Mastadenovirus and Aviadenovirus seem to infect exclusively mammals and birds, respectively. The genus Ichtadenovirus includes the single known AdV from fish. The majority of AdVs in the genus Atadenovirus originated from squamate reptiles (lizards and snakes), but also certain mammalian and avian AdVs are classified within this genus. The genus Siadenovirus contains the only AdV isolated from frog, along with numerous avian AdVs. In turtles, members of a sixth AdV lineage have been discovered, pending official recognition as an independent genus. The most likely scenario for AdV evolution includes long‐term co‐speciation with the hosts, as well as occasional switches between closely or, rarely, more distantly related hosts. (178).
... Members of the Adenoviridae family are middle-sized non-enveloped linear double stranded DNA viruses with an icosahedral nucleocapsid measuring between 80 and 110 nm in diameter (10). The family Adenoviridae contains five accepted genera: Mastadenovirus in mammals, Aviadenovirus in birds, Atadenovirus in squamates, birds and mammals (e. g. agamid adenovirus 1, which is the most commonly described in bearded dragons, snake adenovirus 1, 2, 3, Eublepharid adenovirus-1), Siadenovirus in chelonians, amphibians and birds, Ichtadenovirus in fishes (10) and recently, in 2013, a sixth genus, "Testadenovirus", which has been proposed to encompass adenoviruses found in chelonians (8). In central bearded dragons, clinical signs are non-specific (lethargy, anorexia, cachexia, diarrhea and encephalopathy) with the highest morbidity and mortality in neonates and juveniles (2,7,11,16). ...
... In central bearded dragons, clinical signs are non-specific (lethargy, anorexia, cachexia, diarrhea and encephalopathy) with the highest morbidity and mortality in neonates and juveniles (2,7,11,16). Atadenovirus infections have also been documented in several other species of squamates (1,3,4,8,11,16,18,21). ...
Zusammenfassung
Eine 2 Monate alte Bartagame wurde wegen Anorexie und Abmagerung vorgestellt. Ein anderes Tier aus dem gleichen Bestand war 2 Wochen zuvor plötzlich verstorben. Die Kotuntersuchung zeigte eine große Anzahl an Oozysten von Isospora amphiboluri und einige wenige Oxyureneier. Es wurden keine weiteren Untersuchungen durchgeführt und das Tier verstarb trotz unterstützender Behandlung wenige Tage später. Eine dritte Bartagame aus dem gleichen Terrarium, vorgestellt wegen Anorexie und vergrößertem Zölom, wurde euthanasiert. Die pathologische Untersuchung dieses Tieres ergab eine intestinale Kokzidiose, basophile intranukleäre Einschlüsse, die mit denen einer Adenovirusinfektion übereinstimmten, akute Lebernekrosen mit intra - hepatischen, für Mikrosporidien charakteristischen Organismen sowie eine Nierengicht. Die Diagnose Adenovirose wurde mittels PCR bestätigt. Bei der Sequenzierung war das PCR-Produkt zu 100% mit dem korrespondierenden Teil des Genoms von Agamid-Adenovirus 1 identisch. Eine PCR für den Nachweis von Encephalitozoon (E.) cuniculi war positiv. Die Sequenzierung eines Genabschnitts zeigte eine 100%ige Übereinstimmung mit einem E.-cuniculi-ähnlichen Organismus, der bereits bei Bartagamen nachgewiesen wurde. In Fällen, in denen sich Umweltfaktoren wie schlechte Hygiene oder Stress ausschließen lassen, kann eine systemische (virale) Infektion durch eine Immunsuppression zu einer verstärkten Besiedelung mit opportunistischen Krankheitserregern führen.
... The family Adenoviridae currently consists of five established genera: while the genera Mastadenovirus and Aviadenovirus have been recognized long ago [16], the genera Atadenovirus, Siadenovirus [17], and Ichtadenovirus [1] represent more recent additions. A sixth genus is being proposed: Testadenovirus [18]. ...
... There is one PCR [90] which is regarded the pan-adenovirus PCR, possibly detecting all adenoviruses, as it was tested with multiple samples belonging to all known and proposed genera in the family Adenoviridae [10,18,47,74,77,91,[100][101][102][103][104]. Thus, it simplifies the diagnostics of avian adenoviruses as it detects at-and siadenoviruses as well, not only aviadenoviruses. ...
... The evolutionary provenance of this genus is considered unclear at the moment (Kovács and Benkő, 2011). Recently, in different species of testudinoid turtles, candidate members of a sixth AdV lineage have been discovered, for which the establishment of a sixth genus (with the putative name Testadenovirus), has been proposed (Farkas and Gál, 2009; Doszpoly et al., 2013). ...
... The evolutionary provenance of this genus is considered unclear at the moment (Kovács and Benkő, 2011). Recently, in different species of testudinoid turtles, candidate members of a sixth AdV lineage have been discovered, for which the establishment of a sixth genus (with the putative name Testadenovirus), has been proposed (Farkas and Gál, 2009;Doszpoly et al., 2013). To further explore the biodiversity represented by AdVs, we conduct PCR screening of different vertebrate animals. ...
From over 1250 extant species of the order Chiroptera, 25 and 28 are known to occur in Germany and Hungary, respectively. Close to 350 samples originating from 28 bat species (17 from Germany, 27 from Hungary) were screened for the presence of adenoviruses (AdVs) using a nested PCR that targets the DNA polymerase gene of AdVs. An additional PCR was designed and applied to amplify a fragment from the gene encoding the IVa2 protein of mastadenoviruses. All German samples originated from organs of bats found moribund or dead. The Hungarian samples were excrements collected from colonies of known bat species, throat or rectal swab samples, taken from live individuals that had been captured for faunistic surveys and migration studies, as well as internal organs of dead specimens. Overall, 51 samples (14.73%) were found positive. We detected 28 seemingly novel and six previously described bat AdVs by sequencing the PCR products. The positivity rate was the highest among the guano samples of bat colonies. In phylogeny reconstructions, the AdVs detected in bats clustered roughly, but not perfectly, according to the hosts' families (Vespertilionidae, Rhinolophidae, Hipposideridae, Phyllostomidae and Pteropodidae). In a few cases, identical sequences were derived from animals of closely related species. On the other hand, some bat species proved to harbour more than one type of AdV. The high prevalence of infection and the large number of chiropteran species worldwide make us hypothesise that hundreds of different yet unknown AdV types might circulate in bats.
... AdVs have since been isolated from a large variety of hosts, including representatives of every major vertebrate class from fish to mammals (Harrach et al., 2011). Using polymerase chain reaction (PCR) technology, a large variety of novel AdVs have been detected, but isolation of such viruses and in vitro propagation is hampered in most cases by the lack of appropriate permissive cell cultures (Doszpoly et al., 2013). Some human and animal AdVs can cause diseases or even death, but most are not pathogenic in non-immuno-compromised, healthy individuals. ...
... Recently, very distinct AdVs were discovered in testudinoid turtles (Table 1) (Doszpoly et al., 2013). Partial DNA polymerase sequences proved their great distinctness from all other AdVs and also from each other suggesting a long independent evolution of this lineage. ...
Recombinant vectors based on human adenovirus serotype 5 (HAdV-5) have been extensively studied in pre-clinical models and clinical trials over the last two decades. However, the thorough understanding of the HAdV-5 interaction with human subjects has uncovered major concerns about its product applicability. High vector-associated toxicity and widespread pre-existing immunity have been shown to significantly impede the effectiveness of HAdV-5 mediated gene transfer. It is therefore that the in depth knowledge attained working on HAdV-5 is currently being used to develop alternative vectors. Here, we provide a comprehensive overview of data obtained in recent years disqualifying the HAdV-5 vector for systemic gene delivery as well as novel strategies being pursued to overcome the limitations observed with particular emphasis on the ongoing vectorization efforts to obtain vectors based on alternative serotypes.Molecular Therapy (2015); doi:10.1038/mt.2015.194.
... The identification of this virus in wild turtles in Indiana illustrates the importance of multiple pathogen surveillance in wildlife conservation studies and the benefit of studying previously unsampled populations. Although Sulawesi tortoise adenovirus has not been previously identified in freshwater turtles, other adenoviruses have been detected in multiple other chelonian species in Europe and North America, associated with varying clinical signs (Schumacher et al. 2012;Doszpoly et al. 2013;Salzmann et al. 2021). In the US, Terrapene adenovirus has been found in asymptomatic free-ranging eastern box turtles, which might indicate that the virus is hostadapted to this species (Archer et al. 2017;Franzen-Klein et al. 2019;Klein et al. 2021). ...
Freshwater turtles are sensitive to environmental changes and can serve as sentinel species for ecosystem health assessments. The Efroymson Restoration at Kankakee Sands in northwestern Indiana, USA has been restored in the past 25 yr from primarily agricultural land to a mosaic of prairie and wetland habitats. Health assessments of 40 free-ranging painted turtles (Chrysemys picta) at Kankakee Sands were performed in May 2021 to evaluate overall health status, screen for infectious diseases, and obtain baseline clinical pathology values in this population. Assessment of each turtle included a physical examination, complete blood count, plasma biochemistry panel, blood lactate level, venous blood gas analysis, serum trace mineral panel, serum vitamin D3 level, and plasma protein electrophoresis. Oral and cloacal swabs were tested for adenoviruses, herpesviruses, frog virus 3, and Mycoplasmopsis species by PCR in 39 painted turtles. Four turtles were positive for adenovirus, which shared 100% homology to Sulawesi tortoise adenovirus. Two turtles were herpesvirus-positive with 100% homology to emydid herpesvirus 1. No Mycoplasmopsis spp. or frog virus 3 was detected. Female turtles had significantly higher manganese, prealbumin, uric acid, triglycerides, and ionized calcium levels, while male turtles had significantly higher cholesterol, glutamate dehydrogenase, and CO2 levels. These baseline data can be used for future research into freshwater turtle health in restored wetland habitats.
... however, disease and pathogens are not usually listed as potential or current threats to reptile populations, as they remain poorly understood (Daszak et al., 2000;Gibbons & Steffes, 2013). Recent studies highlight the importance of expanding EID studies in turtles and tortoises to prevent mortality events of already compromised and declining populations of high conservation concern (Doszpoly et al., 2013;Kane et al., 2017;Kolesnik et al., 2017). In the Galapagos Archipelago, giant tortoises play key roles for maintaining healthy ecosystems (e.g., seed dispersal, habitat modifications) while also significantly contributing to the local economy based on eco-tourism activities, as these giants are considered one of the main attractions for visitors and researchers (Benitez-Capistros et al., 2019;Blake et al., 2012). ...
Emerging infectious diseases (EIDs) have been reported as causes of morbidity and mortality in free-living animal populations, including turtles and tortoises, and they have even resulted in species extinctions, with human activities contributing to the spread of many of these diseases. In the Galapagos, giant tortoises are endangered due to habitat change, invasive species, and other human impacts; however, the impact of EIDs on Galapagos tortoise conservation remains understudied. To fulfil this gap, we conducted health assessments of five tortoise species from the islands of Santa Cruz, Isabela, and Española. We performed health evaluations of 454 animals and PCR testing for pathogens known to be relevant in other tortoise species. We identified two novel sequences of adenoviruses and four of herpesviruses. Based on alignments of the DNA polymerase gene and maximum likelihood phylogenetic analyses, we found both novel adenoviruses to be most closely related to red footed tortoise adenovirus 2 by nucleotide sequence and red footed tortoise adenovirus 1 based on amino acid sequence. Three of the herpesvirus sequences translated into the same deduced amino acid sequence; therefore, they may be considered the same viral species, closely related to terrapene herpesvirus 2. The fourth herpesvirus sequence was highly divergent from any sequence previously detected and is related to an eagle owl herpesvirus based on nucleotide sequence and to loggerhead oro-cutaneous herpesvirus based on amino acids. These novel viruses seem to be pathogenic for giant tortoises under specific conditions (e.g., stress). Continued screening is crucial to determine if these viruses play a role in tortoise fitness, morbidity, and survival. This information allows us to provide recommendations to the Galapagos National Park Directorate and other institutions to improve the management of these unique species both in Galapagos and globally, and for tortoise reintroduction plans throughout the archipelago.
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... While the importance of these mechanistic differences for overall immunocompetence is unknown, anecdotal evidence suggests that they may play a role in disease susceptibility. A number of bacterial, viral, fungal, and parasitic infections have been documented in eastern box turtles including ranavirus (Adamovicz et al., 2018;Johnson et al., 2008), adenovirus (Doszpoly et al., 2013;Franzen-Klein et al., 2020), herpesviruses (Terrapene herpesvirus 1, Terrapene herpesvirus 2; Sim et al., 2015;Yonkers et al., 2015), Mycoplasma sp. (Feldman et al., 2006), phaeohyphomycosis (Perpiñán et al., 2016), and intranuclear coccidiosis of Testudines (Alvarez et al., 2013). ...
Eastern (Terrapene carolina carolina) and ornate (Terrapene ornata ornata) box turtles have robust plasma antibacterial activity, however, the mechanism behind this activity is unknown. We used sheep red blood cell (SRBC) hemolysis assays, mannan‐affinity chromatography, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS‐PAGE), and matrix‐assisted laser desorption ionization time‐of‐flight (MALDI‐TOF) to explore the mechanisms of complement activity in box turtles. Plasma from both species demonstrated volume, time, and temperature‐dependent SRBC hemolysis, with significantly greater hemolytic activity in ornate box turtle plasma. Hemolytic activity was highly attenuated following treatment with heat, EDTA, and salicylaldoxime in both species, but was unchanged after treatment with methylamine and ammonium hydroxide. Two abundant mannan‐binding proteins (presumed C‐type lectins) were identified in eastern box turtle plasma using SDS‐PAGE and MALDI‐TOF, but ornate box turtles did not express either protein. Eastern box turtles appear to rely on the lectin pathway of complement activation while ornate box turtles utilize the alternative pathway. This study provides further evidence that mechanisms underlying immune function are not always conserved between closely related species. This finding may have important implications for explaining species differences in susceptibility to emerging threats such as disease, toxicants, and climate change.
Research Highlights
• Box turtle plasma lyses sheep erythrocytes in a volume, time, and temperature‐dependent manner using the complement cascade.
• Eastern box turtles use the lectin pathway of complement activation while ornate box turtles use the alternative pathway.
... Five genera are presently recognized by the International Committee on Taxonomy of Viruses (ICTV) including Mastadenovirus (primarily occurring in mammals), Aviadenovirus (with birds as main hosts), Atadenovirus (found so far in birds, reptiles, ruminants and marsupials), Siadenovirus (detected in reptiles, birds, and a frog), and Ichtadenovirus (found exclusively in fish, so far) [1][2][3]. A sixth genus, Testadenovirus, has been proposed [4], but has not yet been accepted by the ICTV. Birds are generally regarded as common hosts for adenoviruses [2]. ...
Knowledge about adenoviruses in birds of the order Passeriformes is very scarce. Based on molecular characterizations, only two siadenoviruses, great tit adenovirus 1 and Gouldian finch adenovirus, have been described so far occurring in great tits and Gouldian finches, respectively. Assuming a broader occurrence of adenoviruses, various passeriform birds including pet, zoo, and wild birds were examined using a broad-range PCR targeting a fragment of the adenovirus DNA polymerase gene. Adenoviruses were detected in 25 individual birds belonging to 13 species and seven zoological families (Ploceidae, Fringillidae, Estrildidae, Paridae, Sylviidae, Turdidae, Muscicapidae). The putative viruses were further characterized by sequencing the PCR products and phylogenetic analyses. DNA of adenoviruses affiliating to 3 genera including aviadenovirus, siadenovirus, and atadenovirus was found. Viruses with sequences identical or closely related to great tit adenovirus 1 and Gouldian finch adenovirus 1 were detected in a great tit and in two zebra finches, respectively. Based on polymerase amino acid sequence comparisons, the viruses found in the remaining 22 birds revealed phylogenetic distances larger than 15% to adenoviruses known so far suggesting that they may belong to at least 14 different virus species. In some bird species (great tit, zebra finch, vitelline masked weaver) varying adenovirus genera were detected. These results suggest a broad variety of adenoviruses circulating in passeriform birds.
... There is limited information about other pathogens infecting the tortoise family Testudinidae. So far, viruses from several families have been identified as infecting members of Testudinidae, including picornaviruses [14,15], iridoviruses [16], herpesviruses [17], adenoviruses [18][19][20][21], paramyxoviruses [22,23], and the retrovirus Rous sarcoma virus [24]. There is no information on single-stranded DNA (ssDNA) viruses associated with members of the Testudinidae family and limited viral disease information on the Sonoran Desert tortoise (G. ...
The Sonoran Desert tortoise Gopherus morafkai is adapted to the desert, and plays an important ecological role in this environment. There is limited information on the viral diversity associated with tortoises (family Testudinidae), and to date no DNA virus has been identified associated with these animals. This study aimed to assess the diversity of DNA viruses associated with the Sonoran Desert tortoise by sampling their fecal matter. A viral metagenomics approach was used to identify the DNA viruses in fecal samples from wild Sonoran Desert tortoises in Arizona, USA. In total, 156 novel single-stranded DNA viruses were identified from 40 fecal samples. Those belonged to two known viral families, the Genomoviridae (n = 27) and Microviridae (n = 119). In addition, 10 genomes were recovered that belong to the unclassified group of circular-replication associated protein encoding single-stranded (CRESS) DNA virus and five circular molecules encoding viral-like proteins.
... The family Adenoviridae consists of non-enveloped, icosahedral, double-stranded Deoxyribonucleic acid (DNA) viruses which replicate in the nuclei of cells (Davison, Benkȍ & Balázs 2003;Harrach et al. 2011;Marchang 2011). At least five genera are known to exist (Doszpoly et al. 2013;Harrach et al. 2011). Adenoviruses detected in squamate reptiles fall into the genus Atadenovirus, which has been detected in birds, ruminants and marsupials (Harrach et al. 2011;Wellehan et al. 2004). ...
Adenoviral infections may cause mild to severe morbidity or fatality in a large array of animal species. In crocodilians, hatchlings under 5 months of age are usually affected. However, there is a paucity of information on actual incidences in hatchlings originating from South Africa. Two cases of adenoviral hepatitis in crocodile hatchlings about 2 weeks old, bred on a commercial farm in South Africa, are described. Both hatchlings showed typical clinical signs of hepatitis. The identification of intranuclear inclusion bodies in the liver was used to differentiate between adenoviral hepatitis and chlamydial hepatitis. Although vertical transmission has never been proven in crocodiles, the young age of the affected hatchlings raises the possibility of vertical transmission. The lack of epidemiological information on adenoviral hepatitis in crocodiles highlights the need for further characterisation of the virus and targeted surveillance.
... The family Adenoviridae consists of non-enveloped, icosahedral, double-stranded Deoxyribonucleic acid (DNA) viruses which replicate in the nuclei of cells (Davison, Benkȍ & Balázs 2003;Harrach et al. 2011;Marchang 2011). At least five genera are known to exist (Doszpoly et al. 2013;Harrach et al. 2011). Adenoviruses detected in squamate reptiles fall into the genus Atadenovirus, which has been detected in birds, ruminants and marsupials (Harrach et al. 2011;Wellehan et al. 2004). ...
... The single member of the genus Ichtadenovirus is the white sturgeon adenovirus. The sixth genus was proposed recently: testadenoviruses were detected in testudinoid turtles only until now ( Doszpoly et al. 2013). ...
Viruses have been infecting their host cells since the dawn of life, and this extremely long-term coevolution gave rise to some surprising consequences for the entire tree of life. It is hypothesised that viruses might have contributed to the formation of the first cellular life form, or that even the eukaryotic cell nucleus originates from an infection by a coated virus. The continuous struggle between viruses and their hosts to maintain at least a constant fitness level led to the development of an unceasing arms race, where weapons are often shuttled between the participants. In this literature review we try to give a short insight into some general consequences or traits of virus–host coevolution, and after this we zoom in to the viral clades of adenoviruses, herpesviruses, nucleo-cytoplasmic large DNA viruses, polyomaviruses and, finally, circoviruses.
... A sixth genus, Testadenovirus, is proposed to include members from the order Testudines. 6 Notably, all known adenoviruses in squamate hosts have been in the genus Atadenovirus. 17 While atadenovirus was initially discovered in birds and mammals, the nucleotide composition and phylogeny argue that squamates are the likely the hosts of origin. ...
Adenoviruses are medium-sized DNA viruses with very high host fidelity. The phylogenetic relationships of the adenoviruses strongly resemble that of their hosts, consistent with evolutionary codivergence. The genus Atadenovirus appears to have evolved in squamate hosts. Perhaps the best known of the squamate adenoviruses is Agamid adenovirus 1 (AgAdV1), found most commonly in central bearded dragons (Pogona vitticeps), where it is a prevalent cause of hepatitis/enteritis, especially in young animals. All previous reports of adenoviruses in bearded dragons were AgAdV1. Helodermatid adenovirus 2 (HeAdV2) was first seen in Mexican beaded lizards (Heloderma horridus). Subsequently, partial adenoviral polymerase gene sequence from a western bearded dragon (Pogona minor) in Australia was found to share 99% nucleotide homology with HeAdV2. This article reports the discovery of a virus identical to HeAdV2 in a captive central bearded dragon in Florida and wild Gila monsters (Heloderma suspectum) in Arizona. Additionally, a partial adenoviral polymerase gene sharing 98% homology with this HeAdV2 was discovered in a death adder (Acanthophis antarcticus) in Australia. These findings call into question the provenance of HeAdV2. Further studies of atadenoviral host range, diversity of adenoviruses in captive animals, and characterization of adenoviruses from wild squamates are indicated.
... While FV3 represents a known cause of box turtle mortality, several novel pathogens including Ter- rapene herpesvirus 1 (TerHV1), box turtle adenovirus (BTADV), and an un-named Myco- plasma sp. (BTMyco) have been recently identified [28][29][30][31][32][33]. The impact of these pathogens on free-living populations and their relationship to mortality events is unclear. ...
Wildlife mortality investigations are important for conservation, food safety, and public health; but they are infrequently reported for cryptic chelonian species. Eastern box turtles (Terrapene carolina carolina) are declining due to anthropogenic factors and disease, and while mortality investigations have been reported for captive and translocated individuals, few descriptions exist for free-living populations. We report the results of four natural mortality event investigations conducted during routine health surveillance of three Illinois box turtle populations in 2011, 2013, 2014, and 2015. In April 2011, over 50 box turtles were found dead and a polymicrobial necrotizing bacterial infection was diagnosed in five survivors using histopathology and aerobic/anaerobic culture. This represents the first reported occurrence of necrotizing bacterial infection in box turtles. In August 2013, paired histopathology and qPCR ranavirus detection in nine turtles was significantly associated with occupation of moist microhabitats, identification of oral plaques and nasal discharge on physical exam, and increases in the heterophil count and heterophil to lymphocyte ratio (p < 0.05). In July 2014 and 2015, ranavirus outbreaks reoccurred within a 0.2km radius of highly-disturbed habitat containing ephemeral ponds used by amphibians for breeding. qPCR ranavirus detection in five individuals each year was significantly associated with use of moist microhabitats (p < 0.05). Detection of single and co-pathogens (Terrapene herpesvirus 1, adenovirus, and Mycoplasma sp.) was common before, during, and after mortality events, but improved sample size would be necessary to determine the impacts of these pathogens on the occurrence and outcome of mortality events. This study provides novel information about the causes and predictors of natural box turtle mortality events. Continued investigation of health, disease, and death in free-living box turtles will improve baseline knowledge of morbidity and mortality, identify threats to survival, and promote the formation of effective conservation strategies.
... Niewiele jest informacji dotyczących infekcji wirusowych u żółwi ozdobnych. W przypadku zwierząt odłowionych ze środowiska naturalnego stwierdzano obecność adenowirusów z prewalencją sięgającą 27,3%[17]. Nie wykazano natomiast zakażeń powodowanych przez rana-i herpeswirusy[69]pomimo, że infekcje ranawirusami są często notowane u płazów przebywających w tym samym środowisku, a żółwie czerwonolice są wrażliwe na zakażenie[2]. Inwazyjne gatunki żółwi mogą być nosicielami wirusów patogennych dla ryb, np. ...
Invasiveness of alien turtles results from their impact on the functioning of the local ecosystem. It is due to predation on or competing with resident species, but also transfer of new and unknown pathogenic bacteria, viruses, parasites, or fungi. Salmonella is the most often reported microorganism, both in free-living and captive turtles. Zoonotic aspect of Salmonella spp. carriage has led to the definition of RAS (Reptile Associated Salmonellosis) acquired from domestic pet reptiles. Mycobacterium spp., Leptospira spp. and aquatic bacteria are also found in turtles. Additionally, nematode transmissions from invasive turtles to the autochthonic ones have been described. Alien turtles were less affected by parasitic invasion than animals living in a native location, but the infestation of alien parasites in native turtle species was usually more severe. Reports on viral or fungal infections in turtles are scarce. The identified knowledge gaps justify the need for research which will provide basic and systematic data on microbial threats related to alien and invasive turtles present in the natural environment of Poland. It will also give more insight in the scope and the impact of the problem on epidemiology and public health.
1. Introduction. 2. Invasive turtle species. 3. Bacteria. 3.1. Salmonella spp. 3.1.1. Reptile Associated Salmonellosis (RAS). 3.2. Mycobacteria. 3.3. Other bacteria. 4. Parasitofauna of turtles. 4.1. Invasive turtles as a source of helminth invasion of European pond turtle. 4.2. Occurrence and invasiveness of alien parasites in native and alien turtles. 5. Viral infections. 6. Mycotic infection. 7. Conclusions
1. Wstęp. 2. Inwazyjne gatunki żółwi. 3. Bakterie. 3.1. Salmonella spp. 3.1.1. Reptile Associated Salmonellosis (RAS). 3.2. Prątki. 3.3. Inne bakterie. 4. Parazytofauna żółwi. 4.1. Żółwie inwazyjne źródłem zarażenia helmintami żółwia błotnego. 4.2. Występowanie i patogenność obcych pasożytów u natywnych i obcych gatunków żółwi. 5. Infekcje wirusowe. 6. Infekcje grzybicze. 7. Podsumowanie
... The family Adenoviridae is currently divided into five genera: Mastadeno-, Aviadeno-, Atadeno-, Siadeno-, and Ichtadenovirus (International Committee on Taxonomy of Viruses; http://www.ictvonline.org/). A sixth AdV lineage was described recently in testudinoid turtles in Hungary and USA [3,4]. Although several AdVs have been identified in different mammalian species belonging to the infraclass Placentalia, there is only a single known marsupial origin AdV, the Possum AdV 1, which was described in brushtail possums (Trichosurus vulpecula, order Diprotodontia) from New Zealand [5]. ...
A male kowari (Dasyuroides byrnei) originating from a zoo facility was delivered for post mortem evaluation in Hungary. Acute lobar pneumonia with histopathologic changes resembling an adenovirus (AdV) infection was detected by light microscopic examination. The presence of an AdV was confirmed by obtaining partial sequence data from the adenoviral DNA-dependent DNA-polymerase. Although the exact taxonomic position of this novel marsupial origin virus could not be determined, pairwise identity analyses and phylogenetic calculations revealed that it is distantly related to other members in the family Adenoviridae.
... Initially, the lineage of atadenoviruses had been thought to belong to reptiles in general [6]. However, targeted examinations revealed that atadenoviruses are prevalent among squamate reptiles only, whereas non-squamate reptiles seem likely to have their separate AdV lineages [50]. Interestingly however, the G+C content of the partial or full genomic DNA sequences obtained from snake and lizard AdVs, has been found usually in a non-biased range of 45−55% [8,25,51]. ...
Here, we report the results of a large-scale PCR survey on the prevalence and diversity of adenoviruses (AdVs) in samples, collected randomly from free-living reptiles. On the territories of the Guadarrama Mountains National Park in Central Spain and of the Chafarinas Islands in North Africa, cloacal swabs were taken from 318 specimens of eight native species representing five squamate reptilian families. The healthy-looking animals had been captured temporarily for physiological and ethological examinations, after which they were released. We found 22 AdV-positive samples in representatives of three species, all from Central Spain. Sequence analysis of the PCR products revealed the existence of three, hitherto unknown AdVs in 11 Carpetane rock lizards (Iberolacerta cyreni), nine Iberian worm lizards (Blanus cinereus), and two Iberian green lizards (Lacerta schreiberi), respectively. Phylogeny inference showed every novel putative virus to be member of the genus Atadenovirus. This is the very first description of the occurrence of AdVs in amphisbaenian and lacertid hosts. Unlike all squamate atadenoviruses examined previously, two of the novel putative AdVs had A+T rich DNA, a feature generally deemed to mirror previous host switch events. Our results shed new light on the diversity and evolution of atadenoviruses.
... Dream Taq DNA polymerase (Fermentas) was found to be optimal for the PCR amplification of the shorter fragments. The PCRs were performed as described previously (Doszpoly et al., 2013). For the amplification of longer (>1000 bp) fragments, the TaKaRa PrimeS-TAR Max DNA polymerase was used according to the manufacturer`s recommendations. ...
Within the family Adenoviridae presently, Simian mastadenovirus A is the single species approved officially for monkey AdVs, whilst the establishment of six further species (Simian mastadenovirus B to Simian mastadenovirus G) has been proposed in the last few years. We examined the genetic content and phylogenetic relationships of four Old World monkey (OWM) AdV types (namely SAdV 8, 11, 16 and 19) which had been proposed to be classified into different AdV species: SAdV 11 to Human mastadenovirus G, and the other three viruses into three novel species. By full genome sequencing, we identified gene contents characteristic for the genus Mastadenovirus. Among the 36 ORFs, two genes of different lengths, predicted to code for the adenoviral cellular attachment protein (the fibre), were found. The E3 regions contained six genes, present in every OWM AdV, but lacked the E3 19K gene which has seemingly appeared only in the ape (hominid) AdV lineages during evolution. For the first time in SAdVs, the two other exons belonging to the gene of the so called U exon protein were also predicted. Phylogenetic calculations, based on the fibre-1 and the major capsid protein, the hexon, implied that recombination events might have happened between different AdV species. Phylogeny inference, based on the viral DNA dependent DNA polymerase and the penton base protein, further supported the species classification proposed earlier.
... For example, although squamate-like ferlaviruses (Paramyxoviridae) have been detected in tortoises (Papp et al., 2010), an independent clade of ferlaviruses has also been described in a Hermann's tortoise. In the family Adenoviridae the genetic distance is more explicit; while all known adenoviruses of squamate reptiles are members of the genus Atadenovirus, the Sulawesi tortoise adenovirus (Rivera et al., 2009) belongs to the genus Siadenovirus whereas other known testudine adenoviruses form a distinct lineage and most probably are members of a separate newly proposed genus (Testadenovirus; Doszpoly et al., 2013). These findings are consistent with a very early diversification of homologous viruses within their recognized reptile hosts and co-evolution/co-speciation of these viruses. ...
In this study we report the sequence and phylogenetic characterization of an orthoreovirus strain, CH1197/96, isolated from a spur-thighed tortoise (Testudo graeca) on chicken embryo fibroblast cells. The 23,957 bp long genome sequence was obtained by combined use of semiconductor and capillary sequencing. Although the genomic characterization showed that the virus was most similar to the bush viper reovirus strain, 47/02, and in phylogenies performed with all segments the two strains formed a monophyletic group, the nucleotide (48.4 to 70.3%) and amino acid (39.2 to 80.7%) sequence identity values were moderate between the two reptile origin reoviruses. Based on our results and existing classification criteria for the genus Orthoreovirus, the tortoise reovirus strain CH1197/96 might be the first representative of a novel reptilian origin Orthoreovirus species.
... The evolutionary provenance of this genus is considered unclear at the moment (Kovács and Benkő, 2011). Recently, in different species of testudinoid turtles, candidate members of a sixth AdV lineage have been discovered, for which the establishment of a sixth genus (with the putative name Testadenovirus), has been proposed (Farkas and Gál, 2009; Doszpoly et al., 2013). To further explore the biodiversity represented by AdVs, we conduct PCR screening of different vertebrate animals. ...
From over 1250 extant species of the order Chiroptera, 25 and 28 are known to occur in Germany and Hungary, respectively. Close to 350 samples originating from 28 bat species (17 from Germany, 27 from Hungary) were screened for the presence of adenoviruses (AdVs) using a nested PCR that targets the DNA polymerase gene of AdVs. An additional PCR was designed and applied to amplify a fragment from the gene encoding the IVa2 protein of mastadenovi-ruses. All German samples originated from organs of bats found moribund or dead. The Hungarian samples were excrements collected from colonies of known bat species, throat or rectal swab samples, taken from live individuals that had been captured for faunistic surveys and migration studies, as well as internal organs of dead specimens. Overall, 51 samples (14.73%) were found positive. We detected 28 seemingly novel and six previously described bat AdVs by sequenc-ing the PCR products. The positivity rate was the highest among the guano samples of bat colonies. In phylogeny reconstructions, the AdVs detected in bats clustered roughly, but not perfectly, according to the hosts' families (Vespertilionidae, Rhinolophidae, Hipposideridae, Phyllostomidae and Pteropodidae). In a few cases, identical sequences were derived from animals of closely related species. On the other hand, some bat species proved to harbour more than one type of AdV. The high prevalence of infection and the large number of chiropteran species worldwide make us hypothesise that hundreds of different yet unknown AdV types might circulate in bats.
... Additionally, a sixth genus has been proposed (Testadenovirus), whose members were isolated from hosts in the order Testudines. 2 Many reptilian adenoviruses appear to have coevolved with the species they infect, leading to a fair amount of epidemiological species specificity. [17][18][19] Bearded dragons (Pogona sp.) are agamid lizards native to Australia that have become an increasingly popular pet and are frequently found in zoological collections. ...
Agamid adenovirus 1 (AgAdv-1) is a significant cause of disease in bearded dragons (Pogona sp.). Clinical manifestations of AgAdv-1 infection are variable and often nonspecific; the manifestations range from lethargy, weight loss, and inappetence, to severe enteritis, hepatitis, and sudden death. Currently, diagnosis of AgAdv-1 infection is achieved through a single published method: standard nested polymerase chain reaction (nPCR) and sequencing. Standard nPCR with sequencing provides reliable sensitivity, specificity, and validation of PCR products. However, this process is comparatively expensive, laborious, and slow. Probe hybridization, as used in a TaqMan assay, represents the best option for validating PCR products aside from the time-consuming process of sequencing. This study developed a real-time PCR (qPCR) assay using a TaqMan probe-based assay, targeting a highly conserved region of the AgAdv-1 genome. Standard curves were generated, detection results were compared with the gold standard conventional PCR and sequencing assay, and limits of detection were determined. Additionally, the qPCR assay was run on samples known to be positive for AgAdv-1 and samples known to be positive for other adenoviruses. Based on the results of these evaluations, this assay allows for a less expensive, rapid, quantitative detection of AgAdv-1 in bearded dragons.
© 2015 The Author(s).
... The family is divided into five genera: Mastadenovirus, Aviadenovirus, Atadenovirus, Siadenovirus and Ichtadenovirus . Recently, a sixth genus, Testadenovirus, has been proposed for the adenoviruses of testudinoid turtles (Doszpoly et al., 2013). Adenoviruses are often considered to be host specific viruses that co-evolved along with their hosts . ...
... The family Adenoviridae consists of five accepted genera: (1) Mastadenovirus, which contains mammalian AdVs; (2) Aviadenovirus, containing AdVs of birds; (3) Siadenovirus, with chelonian, amphibian and avian AdVs; (4) Ichtadenovirus, with a single AdV isolated from a white sturgeon (Acipenser tranmontanus); and (5) Atadenovirus, containing all squamate AdVs described so far, as well as AdVs of birds and several mammals (Harrach et al., 2012). Recently, a sixth genus, Testadenovirus, has been proposed for AdVs in testudinoid hosts (Doszpoly et al., 2013). Atadenoviruses appear to have coevolved with reptiles and, subsequently, have made several host switches (Harrach, 2000). ...
We describe gross pathology, histopathology, ultrastructure, and molecular characterization of skunk adenovirus 1 (SkAdV-1) in 3 ferrets (Mustela putorius) submitted to the Infectious Diseases Laboratory, Zoo and Exotic Animal Pathology Service at the University of Georgia. Case 1 came from a cohort with a 6-week history of multiple ferrets with respiratory disease and mortality. Cases 2 and 3 were submitted 19 days later; they died 3 days after arrival at the vendor's facility. Histopathological evaluation found evidence of suppurative bronchopneumonia, with intralesional gram-positive cocci in case 1. The others had evidence of subacute multifocal histiocytic and lymphoplasmacytic interstitial pneumonia. Viral isolation, polymerase chain reaction (PCR), and DNA sequencing identified SkAdV-1 in lung tissue. In situ hybridization confirmed adenovirus within the lung lesion, and adenovirus particles were visible under an electron microscope. This is the seventh species in which SkAdV-1 has been identified. Cross-species transmission events have important implications for wildlife management and multispecies facilities.
This chapter presents an overview of the clinical signs and major disease syndromes affecting captive and wild reptiles (marine and freshwater turtles, crocodilians, marine iguanas and sea snakes).
Crane‐associated adenovirus 1 (CrAdV‐1) is a proposed novel virus in the genus Aviadenovirus, first detected in fecal samples from hooded cranes (a vulnerable crane species) on the Izumi plain, a major overwintering site for migratory cranes in Japan. CrAdV‐1 was genetically characterized in that study; however, its virological characteristics remain largely unclear. To investigate the prevalence and organ tropism of CrAdV‐1, we collected swab and organ samples from dead or debilitated cranes on the Izumi plain. CrAdV‐1 gene was detected in 47% (45/95) of tested cranes, comprising mainly hooded cranes but also white‐naped and sandhill cranes. These results indicate that CrAdV‐1 shedding is widespread among cranes overwintering on the Izumi plain. Phylogenetic analyses revealed that the 68 nucleotide sequences determined from the positive swabs formed a single cluster, suggesting phylogenetic differences between CrAdV‐1 and other aviadenoviruses. CrAdV‐1 prevalence showed a significant linear increase with time through the overwintering period (November to February), especially among juveniles. These findings indicate that CrAdV‐1 spreads mainly by transmission between juveniles progressively through the overwintering period. The CrAdV‐1 gene‐positive rate was significantly higher in cloacal swabs than conjunctival or tracheal swabs. Copy numbers for the partial CrAdV‐1 gene sequence were markedly high in the colon samples from three of the four cranes investigated for organ tropism. We also detected relatively high copy numbers in the cerebrum, trachea, lung, and heart, suggesting that CrAdV‐1 mainly targets these four organs and transmitted via the fecal‐oral route and airborne transmission. These results contribute to further understanding of the virological characteristics of CrAdV‐1. This article is protected by copyright. All rights reserved
Adenoviruses have been identified in a wide variety of avian species, and in some species, they have been shown to cause disease and increase mortality. As part of an endeavor to investigate viruses associated with common terns (Sterna hirundo), a novel adenovirus was identified in fecal samples from two common terns on Gull Island, Lake Ontario, Canada. The coding-complete genome sequence of the new adenovirus is 31,094 bp, containing 28 putative genes, and this is the first adenovirus to be associated with terns. The virus was identified in two out of 13 fecal samples from tern chicks, and it was found to be most closely related to duck adenovirus 1, with the DNA polymerase sharing 58% amino acid sequence identity. Phylogenetic analysis based on DNA polymerase protein sequences showed that the new virus forms a distinct sub-branch within the atadenovirus clade and likely represents a new species in this genus.
Although reptiles have often been overlooked in research, information on viruses of reptiles has been growing steadily in recent decades as has our understanding of the importance of these animals in the ecosystem. As ectotherms, their immune systems are dependent on temperature, among other factors, and interactions between infection and disease are complex and dependent on host, pathogen, and environmental factors. This chapter provides an overview of the viruses described in reptiles so far, as well as insight into some of the diseases caused by viruses in this group of animals. It also discusses the reptile immune system and the host reaction to infection. Influences of the environment on development of disease are in many cases not well understood, and this chapter includes a discussion of some important progress in this field. Studies of the effects of viruses on wild, pet, and farmed reptiles are limited, but indicate that viral disease can strongly affect individual populations in the wild, and that human action and the animal trade likely play a role in disease epidemiology.
Adenoviruses are ubiquitous infectious agents associated with respiratory, gastrointestinal and conjunctival diseases that usually cause mild and self-limiting infection in immunocompetent individuals, but can result in severe pneumonia, hepatitis or encephalitis in neonates and immunocompromised patients. Soon after their initial identification adenoviruses were shown to induce malignant tumors in newborn rodents, and although they have not been associated with malignant disease in humans they represent one of the primary model systems of tumor virology. Adenoviruses are also common candidate vectors for gene therapies and anti-cancer therapies, and they are often used as platforms for vaccine development. The study of adenovirus-infected cells has contributed to our understanding of fundamental aspects of cellular processes, including regulation of gene expression, organization of the cell nucleus and regulation of cell proliferation. This chapter describes general aspects of the biology of adenoviruses, including their classification, virion structure, organization of the viral genome, and functions of the gene products that have been studied in more detail. An overview of the molecular events during the viral replication cycle is followed by a brief description of the virus interactions with the host cell.
Infectious diseases are the prime cause of morbidity and mortality in animals leading to heavy economic losses to the livestock/poultry industry. One of the most effective ways to control these losses is by use of recombinant vaccines. However, one of the requirements for the development of efficient recombinant veterinary vaccines is the availability of an efficient vaccine delivery system. The basic characteristic of the viruses to successfully deliver their genetic material into the host cells makes them a powerful tool for introducing foreign DNA into a variety of eukaryotic cells. While recombinant vectors from a number of viruses are available, vectors based on adenoviruses are being evaluated in the highest number of vaccination and gene therapy clinical trials globally. Interestingly, the last decade has witnessed a significant increase in the development and evaluation of species- or non-species-specific adenoviruses as vectors for veterinary vaccines. In fact, an adenovirus-vectored foot and mouth disease vaccine has been conditionally licensed for use in cattle in the USA. This chapter focuses on the current state of the research related to the development of adenovirus-vectored veterinary vaccines.
Since the first report on adenoviruses in the early 1950s, more than a hundred serotypes of this virus have been reported from reptiles, fish, mammalian, and avian species. These viruses exhibit different lineages based on the differences noticed in the genes located in the terminal regions. Adenoviruses are grouped into five genera including Mastadenovirus, Aviadenovirus, Atadenovirus, Siadenovirus, and Ichtadenovirus. Recently, a sixth genus, Testadenovirus, is proposed to include adenoviruses from turtles. Bats have been identified as prospective reservoir hosts of emerging and re-emerging diseases and playing an important role in the evolution of adenoviruses. This chapter details the information on epidemiology, clinical signs, pathology, diagnosis, prevention, and control aspects of various species-specific adenoviruses affecting bovine, ovine, porcine, canine, and equine that are reported from both healthy animals and those suffering from diarrhea and pneumoenteritis. There is no specific treatment or vaccine available for adenoviruses.
This chapter provides a brief summary of natural history of terrestrial and aquatic chelonians and practical information on medical management, including the most prevalent concerns for each species and the epidemiology of infectious and parasitic diseases. The most common medical issues seen in freshwater turtles presented for rehabilitation are injuries sustained from automobile strikes and fishhook and line ingestion. Both of these issues are depleting populations drastically. Sick and injured turtles are usually given broad‐spectrum antibiotics as a treatment for established bacterial infections or as a preventive measure. Wound management techniques used for the various chelonian species are similar to those for humans and domestic animals. Anesthetic and analgesic protocols should provide muscle relaxation and analgesia when performing surgery on chelonian patients. It is the goal of every rehabilitation setting to provide care to injured or ill wildlife that results in release of the patient to conserve populations.
The Chinese soft-shelled turtle ( Pelodiscus sinensis) has become one of the leading cultured organisms in China and South East Asia. The objectives of the present study were to use next generation sequencing to identify viral genomes present in liver tissues from Chinese soft-shelled turtle in China. BLAST analysis of viral sequences from liver samples showed high homology with the human adenovirus (HAdV) penton base and encapsidation proteins. This homology points to possible existence of HAdV in freshwater environments used for the culture of soft-shelled turtles. Therefore, our findings merit further investigations to determine possible contamination of HAdV in aquaculture environments and the possible role of the Chinese soft-shelled turtle in transmitting HAdV to humans.
The scarcity or complete lack of information on the adenoviruses (AdVs) occurring in the most ancient non-human primates resulted in the initiation of a study for exploring their abundance and diversity in prosimians and New World monkeys (NWMs). In order to assess the variability of these AdVs and the possible signs of the hypothesised virus-host co-evolution, samples from almost every family of NWMs and prosimians were screened for the presence of AdVs. A PCRscreening of 171 faecal or organ samples from live or dead, captive or wild-living prosimians and NWMs was performed. The PCR products from the gene of the IVa2 protein were sequenced and used in phylogeny calculations. The presence of 10 and 15 new AdVs in seven and ten different species of prosimians and NWMs was revealed, respectively. Phylogenetic analysis indicated that the tentative novel AdVs cluster into two separate groups, which form the most basal branches among the primate AdVs, and therefore support the theory on the co-evolution of primate AdVs with their hosts. This is the first report that provides a comprehensive overview of the AdVs occurring in prosimians and NWMs, and the first insight into the evolutionary relationships among AdVs from all major primate groups.
During the period between 2001 and 2010, the authors observed respiratory diseases relating to adenovirus infection in eight large flocks (6, 500-17, 000) of young turkeys. The disease occurred at 6-10 weeks of age of the birds and lasted for 2-3 weeks in the flocks. The clinical signs included malaise, anorexia, sneezing and hampered respiration (dyspnoe). The mortality attributable to the disease was around 2-5.5%. At necropsy, gross pathology findings consisted of acute mucous-fibrinous tracheitis, multifocal catarrhal pneumonia, as well as mucous-fibrinous air-sacculitis and pericardititis. By histopathology, mucous-fibrinous tracheo-bronchitis with deciliation of epithelium in the respiratory tract, accompanied by epithelial proliferation and metaplasia, and catarrhal pneumonia were revealed. In the epithelial cells of the respiratory tract, basophil and/or acidophil nuclear inclusion bodies could be observed. In samples from the affected organs, the presence of aviadenoviruses was demonstrated by PCR. The viruses were also isolated in cell culture. The detected viruses, determined as turkey adenovirus 5, differed from every previously known adenovirus but seemed to be identical with each other. The novel virus was found to be a close relative of fowl adenoviruses, but different from turkey adenovirus 1, sequenced recently. Most likely, turkey adenovirus 5 represents a novel adenovirus species. Bacteria, that can be considered as primary pathogen such as Bordetella, Riemerella, Ornithobacterium, Pasteurella, etc., or Mycoplasma were not found. By virological and serological tests (with paired serum samples), the etiological role of the Newcastle disease virus (NDV), avian influenza virus (AIV) and the turkey rhinotracheitis (TRT) virus could be excluded except one flock where also a low pathogenic H9N2 AIV strain was isolated.
Herpesviruses are important pathogens of chelonians, and include Chelonid herpesvirus 5, which is associated with fibropapillomatosis in sea turtles. Spirorchid trematodes are blood flukes that reside within the cardiovascular system of marine turtles and may be associated with severe disease. An eastern box turtle (Terrapene carolina) at the South Florida Wildlife Care Center (Fort Lauderdale, Florida) was presented to the facility with papillomatous growths behind both rear legs. Surgical removal resulted in remission for 8 months; however, lesions recurred, prompting a second surgery and acyclovir therapy. Surgical biopsies revealed subacute superficial inflammation associated with the supporting stroma of the cutaneous papillomas and granulomas within the superficial dermis containing fragmented and collapsed brown trematode eggs surrounded by multinucleated giant cells and epithelioid macrophages. Pan-herpesviral and pan-trematode consensus polymerase chain reaction and sequencing were run on tissue samples. Comparative sequence analysis revealed a novel alphaherpesvirus and a novel trematode in the genus Spirorchis. The animal became anorexic and was euthanized due to poor quality of life. While we do not yet have a complete understanding of the effects of herpesvirus and trematode infections in eastern box turtles, the findings thus presented provide initial insights into the disease relationships among these chelonians.
© 2015 The Author(s).
The authors screened samples, collected randomly from house-dwelling bats, for the presence of adenoviruses by PCR. Two novel adenoviruses were found in guano samples originating from the Aggtelek National Park and its surrounding area. Considering the results of previous studies, four new types of adenoviruses have been found in the representatives of four different bat species in Hungary. To date, samples have been collected from eight house-dwelling bat species, out of which four were found to harbour new adenoviruses. These include the Grey Long-eared Bat (Plecotus austriacus) and the Common Noctule (Nyctalus noctula) belonging to the family of Common bats (Vespertilionidae), as well as the Lesser Horseshoe Bat (Rhinolophus hipposideros) and the Greater Horseshoe Bat (R. ferrumequinum) from the family of Horseshoe bats (Rhinolophidae). Based on preliminary phylogenic tree reconstructions the bat adenoviruses from different host species appear on distinct branches.
In 2009, 26 clinical samples (organs and oral/cloacal swabs) from a total of 24 corn snakes (Pantherophis guttatus) from a single owner were sent to our laboratory to be tested for the presence of viruses. Paramyxoviruses (PMV), adenoviruses (AdV) and reoviruses were detected by RT-PCR, PCR and virus isolation methods. Three snakes were infected with all three viruses at the same time, while two other snakes had a double infection (PMV and reo, AdV and reo) and nine other snakes had a single infection with any of the three viruses. No viruses were detected in 10 animals. All isolated reoviruses were identical to one another and to the reptilian orthoreovirus isolate 55-02 in the partial RNA dependent RNA polymerase (RDRP) gene sequence. AdV partial polymerase sequences represented four different types, one of which was first described here: most similar to SnAdV-1, while the other three were identical to known types: SnAV-1, -2 and -3. However, the detected single PMV differed distinctly from described reptile PMV and was a new type. According to partial L gene, HN gene and U gene sequences it may be the first described representative of a third squamatid PMV cluster: "group C" within the proposed reptilian PMV genus "Ferlavirus". Nucleotide identity values for the L gene of the new PMV compared to group A viruses range between 76.5 and 80.3%, and between 80.5 and 81.2% compared to group B viruses. For the HN gene, these values were similar: 78.2-80% (A) and 79.9-80.5% (B) and somewhat lower for the U gene: 72.7-75.4% (A) and 69.7-70% (B). No reports on the prevalence of concurrent viral infection in captive snake populations have been published so far. The possibility of concurrent infection with several different viruses and subsequent consequences for animal health should be kept in mind when testing reptile samples for viruses.
Samples from native Hungarian or captive bats were tested by PCR for the presence of adenoviruses and herpesviruses. Two novel adenoviruses from a common noctule (Nyctalus noctula) and a greater horseshoe (Rhinolophus ferrum-equinum) bat were detected. In captive Egyptian fruit bats (Rousettus aegyptiacus), DNA from two novel herpesviruses was demonstrated. Phylogenetic analysis facilitated provisional taxonomic placement of the newly detected viruses. Such analysis and the existence of unique, shared early proteins (E3 and E4) suggest that canine adenoviruses may have originated in vespertilinoid bats.
Adenoviruses have been identified in humans and a wide range of vertebrate animals, but not previously from the polar region. Here, we report the entire 26,340-bp genome of a novel adenovirus, detected by PCR, in tissues of six of nine South Polar skuas (Catharacta maccormicki), collected in Lake King Sejong, King George Island, Antarctica, from 2007 to 2009. The DNA polymerase, penton base, hexon and fiber genes of the South Polar skua adenovirus (SPSAdV) exhibited 68.3%, 75.4%, 74.9% and 48.0% nucleotide sequence similarity with their counterparts in turkey hemorrhagic enteritis virus. Phylogenetic analysis based on the entire genome revealed that SPSAdV belonged to the genus Siadenovirus, family Adenoviridae. This is the first evidence of a novel adenovirus, SPSAdV, from a large polar seabird (family Stercorariidae) in Antarctica.
One of the major threats to biodiversity involves biological invasions with direct consequences on the stability of ecosystems. In this context, the role of parasites is not negligible as it may enhance the success of invaders. The red-eared slider, Trachemys scripta elegans, has been globally considered among the worst invasive species. Since its introduction through the pet trade, T. s. elegans is now widespread and represents a threat for indigenous species. Because T. s. elegans coexists with Emys orbicularis and Mauremys leprosa in Europe, it has been suggested it may compete with the native turtle species and transmit pathogens. We examined parasite transfer from American captive to the two native species that co-exist in artificial pools of a Turtle Farm in France. As model parasite species we used platyhelminth worms of the family Polystomatidae (Monogenea) because polystomes have been described from American turtles in their native range. Phylogenetic relationships among polystomes parasitizing chelonian host species that are geographically widespread show patterns of diversification more complex than expected. Using DNA barcoding to identify species from adult and/or polystome eggs, several cases of host switching from exotic to indigenous individuals were illustrated, corroborating that parasite transmission is important when considering the pet trade and in reintroduction programmes to reinforce wild populations of indigenous species.
We give the first published description of the pathology and molecular findings associated with adenovirus infection in lizards in Australia. A central netted dragon (Ctenophorus nuchalis) exhibited severe necrotising hepatitis with abundant intranuclear inclusion bodies within hepatocytes and rarely within intestinal epithelial cells. Polymerase chain reaction (PCR) using pooled tissues yielded an amplicon that shared strong nucleotide identity with an agamid adenovirus (EU914203). PCR on the liver of a bearded dragon (Pogona minor minor) with illthrift, coccidiosis, nematodiasis and hepatic lipidosis yielded an amplicon with strong nucleotide identity to a helodermatid adenovirus (EU914207).
Partial genome sequences were determined and subjected to comparative analyses from two fish herpesviruses (HVs). Acipenserid (Aci) HV-2, originating from the white sturgeon (Acipenser transmontanus), and ictalurid (Ic) HV-2, isolated from the black bullhead (Ameiurus melas), are recently approved species of the genus Ictalurivirus of the family Alloherpesviridae.
An almost 8,000-base-pair fragment, spanning between the genes of the DNA polymerase and the ATPase subunit of the terminase, was sequenced from each virus.
The size, position and orientation of 2 partial and 3 full open reading frames, contained in the studied genome fragment, proved to be similar to their counterparts in IcHV-1, the type species of the genus Ictalurivirus. Thus, a well-conserved genus-specific gene block was identified. In the members of two other genera (Cyprinivirus and Batrachovirus) of the family Alloherpesviridae, no such gene block could be found; the location and orientation of the homologous genes showed significant divergence.
The results of phylogenetic calculations were in good agreement with the genome arrangements inasmuch as AciHV-2, IcHV-1 and -2 are monophyletic and separated from the lineages of the other two genera. The new sequence enabled the inclusion of a hitherto unassigned HV, that of the Australian pilchard, into a phylogenetic calculation.
Consensus polymerase chain reaction was used to identify a novel adenovirus from two psittacine birds: a plum-headed parakeet (Psittacula cyanocephala) with lethargy, weight loss, and marked leukocytosis; and an umbrella cockatoo (Cacatua alba) with lethargy, weight loss, and feather abnormalities. Phylogenetic and comparative sequence analysis suggested that this virus is a member of the genus Siadenovirus, and is here termed psittacine adenovirus 2. This extends the characterized adenoviruses of psittacine birds beyond Aviadenovirus to include the genus Siadenovirus. Identification and further study of adenoviral types and species will provide useful diagnostic, prognostic, and epidemiologic information for the clinician. Like other known members of the genus Siadenovirus, Psittacine adenovirus 2 is AT-rich over the region sequenced, and it is hypothesized that this may be associated with shorter host-virus evolutionary association.
A novel siadenovirus was identified in the Sulawesi tortoise (Indotestudo forsteni). A group of 105 Sulawesi tortoises was obtained by the Turtle Survival Alliance. Many of the tortoises were in poor health. Clinical signs included anorexia, lethargy, mucosal ulcerations and palatine erosions of the oral cavity, nasal and ocular discharge, and diarrhea. Initial diagnostic tests included fecal testing for parasites, complete blood count and plasma biochemical analysis, mycoplasma serology, and polymerase chain reaction (PCR) testing for intranuclear coccidia and chelonian herpesvirus. Treatment included administration of antibiotics, antiparasitic medications, parenteral fluids, and nutritional support. Tissue samples from animals that died were submitted for histopathologic evaluation. Histopathologic examination revealed systemic inflammation and necrosis associated with intranuclear inclusions consistent with a systemic viral infection in 35 tortoises out of 50 examined. Fecal testing results and histopathologic findings revealed intestinal and hepatic amoebiasis and nematodiasis in 31 animals. Two of 5 tortoises tested by PCR were positive for Chlamydophila sp. Aeromonas hydrophila and Escherichia coli were cultured from multiple organs of 2 animals. The mycoplasma serology and PCR results for intranuclear coccidia and chelonian herpesvirus were negative. Polymerase chain reaction testing of tissues, plasma, and choanal/cloacal samples from 41 out of 42 tortoises tested were positive for an adenovirus, which was characterized by sequence analysis and molecular phylogenetic inference as a novel adenovirus of the genus Siadenovirus. The present report details the clinical and anatomic pathologic findings associated with systemic infection of Sulawesi tortoises by this novel Siadenovirus, which extends the known reptilian adenoviruses to the chelonians and extends the known genera of reptilian Adenoviridae beyond Atadenovirus to include the genus Siadenovirus.
Turkey hemorrhagic enteritis virus (THEV) is a member of the genus Siadenovirus and causes disease in turkey poults characterized by splenomegaly, bloody diarrhoea and death. The mechanism responsible for intestinal lesion formation and mortality is not known, although there is strong evidence that it is immune-mediated. All strains of THEV are serologically indistinguishable, although there are naturally occurring avirulent strains of THEV that replicate efficiently in turkeys without the intestinal haemorrhage or mortality associated with more virulent strains. The purpose of this study was to determine which viral genes are involved in virulence. The full-length genome of an avirulent vaccine strain was sequenced and compared with the genome of a virulent field isolate from Israel that was sequenced in 1998. Comparison of the two 26.3 kb genomes revealed 49 nucleotide differences resulting in 14 putative amino acid changes within viral proteins. Sequencing of the regions surrounding the 14 missense mutations revealed variations in ORF1, E3 and the fiber (fib) knob domain in five additional strains with varying degrees of virulence. Complete sequences of these genes were determined in a total of 11 different strains of THEV. All strains had at least one missense mutation in ORF1, and all but two of the strains had one missense mutation in E3. At least one missense mutation was found in the fiber knob domain in six out of seven virulent strains. Sequence variation of ORF1, E3 and fib in strains of THEV with different phenotypes strongly indicates that these genes are the key factors affecting virulence.
Our understanding of the origins, the functions and/or the structures of biological sequences strongly depends on our ability to decipher the mechanisms of molecular evolution. These complex processes can be described through the comparison of homologous sequences in a phylogenetic framework. Moreover, phylogenetic inference provides sound statistical tools to exhibit the main features of molecular evolution from the analysis of actual sequences. This chapter focuses on phylogenetic tree estimation under the maximum likelihood (ML) principle. Phylogenies inferred under this probabilistic criterion are usually reliable and important biological hypotheses can be tested through the comparison of different models. Estimating ML phylogenies is computationally demanding, and careful examination of the results is warranted. This chapter focuses on PhyML, a software that implements recent ML phylogenetic methods and algorithms. We illustrate the strengths and pitfalls of this program through the analysis of a real data set. PhyML v3.0 is available from (http://atgc_montpellier.fr/phyml/).
TOPALi v2 simplifies and automates the use of several methods for the evolutionary analysis of multiple sequence alignments. Jobs are submitted from a Java graphical user interface as TOPALi web services to either run remotely on high-performance computing clusters or locally (with multiple cores supported). Methods available include model selection and phylogenetic tree estimation using the Bayesian inference and maximum likelihood (ML) approaches, in addition to recombination detection methods. The optimal substitution model can be selected for protein or nucleic acid (standard, or protein-coding using a codon position model) data using accurate statistical criteria derived from ML co-estimation of the tree and the substitution model. Phylogenetic software available includes PhyML, RAxML and MrBayes.
Availability: Freely downloadable from http://www.topali.org for Windows, Mac OS X, Linux and Solaris.
Contact: iain.milne@scri.ac.uk
No abstract available.
Cospeciation generally increases the similarity between host and parasite phylogenies. Incongruence between host and parasite phylogenies has previously been explained in terms of host switching, sorting, and duplication events. Here, we describe an additional process, failure of the parasite to speciate in response to host speciation, that may be important in some host-parasite systems. Failure to speciate is likely to occur when gene flow among parasite populations is much higher than that of their hosts. We reconstructed trees from mitochondrial and nuclear DNA sequences for pigeons and doves (Aves: Columbiformes) and their feather lice in the genus Columbicola (Insecta: Phthiraptera). Although comparisons of the trees from each group revealed a significant amount of cospeciation, there was also a significant degree of incongruence. Cophylogenetic analyses generally indicated that host switching may be an important process in the history of this host-parasite association. Using terminal sister taxon comparisons, we also identified three apparent cases where the host has speciated but the associated parasite has not. In two of these cases of failure to speciate, these comparisons involve allopatric sister taxa of hosts whose lice also occur on hosts sympatric with both of the allopatric sisters. These additional hosts for generalist lice may promote gene flow with lice on the allopatric sister species. Relative rate comparisons for the mitochondrial cytochrome oxidase I gene indicate that molecular substitution occurs about 11 times faster in lice than in their avian hosts.
From adenovirus-like viruses originating from a fish and a snake species, a conserved part of the adenoviral DNA polymerase
gene was PCR amplified, cloned and sequenced. Phylogenetic analysis showed that the snake adenovirus is closely related to
the members of the proposed genus Atadenovirus, whereas the fish isolate seems to represent a separate cluster, likely a new genus.
This paper describes a hypothesis on the origin of the members of the recently established adenovirus genus, Atadenovirus, invading cattle, sheep, deer, duck and poultry. Comparison of the phylogenetic trees of adenoviruses and their hosts suggests a very ancient but common origin for the atadenoviruses. The surprisingly large difference between these virus types and other adenoviruses infecting the same host can be easily understood by assuming their separate evolution in different hosts (e.g., in reptiles versus a coevolution with mammals and birds, respectively) followed by a later host switch.
MrBayes 3 performs Bayesian phylogenetic analysis combining information from different data partitions or subsets evolving under different stochastic evolutionary models. This allows the user to analyze heterogeneous data sets consisting of different data types—e.g. morphological, nucleotide, and protein—and to explore a wide variety of structured models mixing partition-unique and shared parameters. The program employs MPI to parallelize Metropolis coupling on Macintosh or UNIX clusters.
Availability: http://morphbank.ebc.uu.se/mrbayes
Contact: fredrik.ronquist@ebc.uu.se
*
To whom correspondence should be addressed.
This review provides an update of the genetic content, phylogeny and evolution of the family Adenoviridae. An appraisal of the condition of adenovirus genomics highlights the need to ensure that public sequence information is interpreted accurately. To this end, all complete genome sequences available have been reannotated. Adenoviruses fall into four recognized genera, plus possibly a fifth, which have apparently evolved with their vertebrate hosts, but have also engaged in a number of interspecies transmission events. Genes inherited by all modern adenoviruses from their common ancestor are located centrally in the genome and are involved in replication and packaging of viral DNA and formation and structure of the virion. Additional niche-specific genes have accumulated in each lineage, mostly near the genome termini. Capture and duplication of genes in the setting of a 'leader-exon structure', which results from widespread use of splicing, appear to have been central to adenovirus evolution. The antiquity of the pre-vertebrate lineages that ultimately gave rise to the Adenoviridae is illustrated by morphological similarities between adenoviruses and bacteriophages, and by use of a protein-primed DNA replication strategy by adenoviruses, certain bacteria and bacteriophages, and linear plasmids of fungi and plants.
A consensus nested-PCR method was designed for investigation of the DNA polymerase gene of adenoviruses. Gene fragments were
amplified and sequenced from six novel adenoviruses from seven lizard species, including four species from which adenoviruses
had not previously been reported. Host species included Gila monster, leopard gecko, fat-tail gecko, blue-tongued skink, Tokay
gecko, bearded dragon, and mountain chameleon. This is the first sequence information from lizard adenoviruses. Phylogenetic
analysis indicated that these viruses belong to the genus Atadenovirus, supporting the reptilian origin of atadenoviruses. This PCR method may be useful for obtaining templates for initial sequencing
of novel adenoviruses.
Lice in the genus Pectinopygus parasitize a single order of birds (Pelecaniformes). To examine the degree of congruence between the phylogenies of 17 Pectinopygus species and their pelecaniform hosts, sequences from mitochondrial 12S rRNA, 16S rRNA, COI, and nuclear wingless and EF1-α
genes (2290 nucleotides) and from mitochondrial 12S rRNA, COI, and ATPases 8 and 6 genes (1755 nucleotides) were obtained
for the lice and the birds, respectively. Louse data partitions were analyzed for evidence of incongruence and evidence of
long-branch attraction prior to cophylogenetic analyses. Host-parasite coevolution was studied by different methods: TreeFitter,
TreeMap, ParaFit, likelihood-ratio test, data-based parsimony method, and correlation of coalescence times. All methods agree
that there has been extensive cospeciation in this host-parasite system, but the results are sensitive to the selection of
different phylogenetic hypotheses and analytical methods for evaluating cospeciation. Perfect congruence between phylogenies
is not found in this association, probably as a result of occasional host switching by the lice. Errors due to phylogenetic
reconstruction methods, incorrect or incomplete taxon sampling, or to different loci undergoing different evolutionary histories
cannot be rejected, thus emphasizing the need for improved cophylogenetic methodologies.
Bovine adenovirus (BAV) serotypes 4 through 8 have been found clearly distingushable from BAVs 1,2,3, and 9 and from all other mastadenoviruses, and were therefore classified as subgroup 2 BAVs (1) and were considered as candidate members of a new taxon (2). The distinction was originally based on biological properties, such as the requirement of primary cell culture for propagation, the special appearance of the caused inclusion bodies, and the lack of antigenic crossreaction with other mastadenoviruses (1). The separation was later strengthened by DNA studies revealing special restriction-enzyme pattern, smaller genome size (3) and lack of cross-DNA hybridization with subgroup 1 BAVs (4). Similarly, the egg-drop syndrome (EDS) virus (5) seemed to be an atypical aviadenovirus (6–9) and was described as a candidate member of a new genus (2). The official classification of subgroup 2 BAVs into a new genus and the EDS virus into a new Aviadenovirus genus was, however postponed, until further evidence is gathered (2,10). Recently, a new ovine adenovirus isolate (OAV287) emerged (11) that differed from the offcially accepted OAV serotypes (12,13). The genome of OAV287 has been completely sequenced and was found to have a genomtc organization different from that of HAV-2 and most of other mastadenoviruses (14,15). Because of the close genetic relationship found (based on comparative study of a single gene sequence), we have informally proposed at different adenovirus meetings that subgroup 2 BAVs, OAV287, and EDS virus should all be classified into a common taxon. This could be a new (third) genus, with a proposed name of A Tadenovirus describing the characteristic high AT content found in their genomes (16). The idea evoked considerable oppositton, perhaps mainly because all the 51 types of human adenovnuses (HAVs) (17,18) are very simtlar to each other compared to the differences between the two subgroups of BAVs containing only nine offictally accepted serotypes (10). Furthermore, the sequencing and genetic study of BAVs (and generally of all animal adenoviruses) were missing, but were required before a decision could have been made. The situatton changed significantly recently, since two animal adenovirus genomes have been completely sequenced (15,19), and our group also have sequenced and analyzed characteristic genome parts of different animal adenoviruses.
Cospeciation generally increases the similarity between host and parasite phylogenies. Incongruence between host and parasite phylogenies has previously been explained in terms of host switching, sorting, and duplication events. Here, we describe an additional process, failure of the parasite to speciate in response to host speciation, that may be important in some host-parasite systems. Failure to speciate is likely to occur when gene flow among parasite populations is much higher than that of their hosts. We reconstructed trees from mitochondrial and nuclear DNA sequences for pigeons and doves (Aves: Columbiformes) and their feather lice in the genus Columbicola (Insecta: Phthiraptera). Although comparisons of the trees from each group revealed a significant amount of cospeciation, there was also a significant degree of incongruence. Cophylogenetic analyses generally indicated that host switching may be an important process in the history of this host-parasite association. Using terminal sister taxon comparisons, we also identified three apparent cases where the host has speciated but the associated parasite has not. In two of these cases of failure to speciate, these comparisons involve allopatric sister taxa of hosts whose lice also occur on hosts sympatric with both of the allopatric sisters. These additional hosts for generalist lice may promote gene flow with lice on the allopatric sister species. Relative rate comparisons for the mitochondrial cytochrome oxidase I gene indicate that molecular substitution occurs about 11 times faster in lice than in their avian hosts.
Inclusion body hepatitis (IBH) is an acute disease in chickens caused by fowl adenoviruses (FAdVs). Among twelve known FAdV types (FAdV-1 to 8a, and FAdV-8b to 11), classified into five different species (A-E), all of them were already involved in naturally occurring cases. The disease is mainly distributed in areas with intensive poultry industry. In Slovenia the etiological agent of IBH has not been confirmed to date. An outbreak of acute mortality affected a broiler flock of 12,000 animals. In two waves of elevated mortality rate, a total of 264 (2.2%) chickens were found dead in one week time. Affected birds showed ruffled feathers, depression, watery droppings and some of them limping. The most common pathological lesions seen on necropsy were pale, swollen and friable livers with subcapsular hemorrhages. On histological examination, acute hepatitis characterized by necrosis and hepatocytes, with large basophilic intranuclear inclusion bodies, were observed. The histological results were characteristic for IBH caused by adenovirus infection. The causative agent was identified as fowl adenovirus (FAdV) type 8b, a member of the Fowl adenovirus E species, based on PCR results of partial sequence of adenoviral polymerase and hexon gene. The confirmed type in our case is one of the most common causative agents involved in IBH. In addition, infectious bursal disease virus and infectious bronchitis virus were detected in the same flock.
An outbreak of adenoviral disease occurred in 2 separate raptor collections in the United Kingdom during August and September of 2004, involving a Harris hawk (Parabuteo unicinctus), a Bengal eagle owl (Bubo bengalensis), and a Verreaux's eagle owl (Bubo lacteus). The cases were diagnosed by results of necropsy, histologic examination, and polymerase chain reaction (PCR). Although virus isolation and electron microscopy were unsuccessful in identifying adenovirus as the causative agent, PCR testing with consensus primers resulted in amplicons of specific size. DNA sequencing of the PCR products identified the detected virus as a new member of the genus Siadenovirus. To our knowledge, this is the first report of adenovirus infection in these avian species, and we propose that this virus be called “raptor adenovirus.”
Murine adenoviruses (MAdV) are supposedly the oldest members of the genus Mastadenovirus. Currently, there are three distinct MAdV types known with rather different tropism and pathology. Here we report and annotate the DNA sequence of the full genome of MAdV-2. It was found to consist of 35,203 bp thus being considerably larger than the genomes of the other two MAdV types. The increased size of the MAdV-2 genome is generally due to larger genes and ORFs, although some differences in the number of ORFs were observed for the early regions E1, E3 and E4. The homologue of the 19K gene of E1B from MAdV-2 codes for 330 amino acids (aa) and is almost twice as large as from other mastadenoviruses. Accordingly, only the N-terminal half (155aa) has homology to the 19K protein. A homologue of the gene of the 12.5K protein was identified in the E3 region of MAdV-2, but not in MAdV-1 or MAdV-3. The other gene of yet unknown function in the E3 region of MAdV-2 seems to be unique. The E4 region of MAdV-2 contains three ORFs. One has similarity to the 34K gene of other AdVs. Two unique ORFs in the E4 region of MAdV-2 have no homology to any of the five and six ORFs in the E4 region of MAdV-1 or MAdV-3, respectively. Phylogenetic analyses showed that the three murine AdVs have a close common ancestor. They likely formed the first branching of the lineage of mastadenoviruses, and seem to be the most ancient representatives of this genus.
Adenoviruses are frequent infectious agents in different poultry species. The traditional, serological typing of new isolates by virus neutralisation tests is now in transition to be replaced by PCR and sequencing. The first PCRs, recommended for the detection of adenoviruses, had been designed to target the gene of the major capsid protein, the hexon. In birds, members of three different genera of the family Adenoviridae may occur. Accordingly, three specific hexon PCRs had to be elaborated for the detection of adenoviruses in poultry. A significantly more sensitive PCR, targeting the viral DNA-dependent DNA polymerase gene, has been described recently. This method proved to be an efficient alternative for the general detection of adenoviruses irrespective of their genus affiliation. Fowl adenoviruses (FAdVs), isolated from chicken to date, comprise twelve serotypes classified into five virus species (FAdV-A to E). The polymerase gene sequence has been determined yet only from three FAdV types representing three species. In the present work, the panel of polymerase gene sequences was completed with those of the rest of FAdVs. The newly determined sequences will facilitate the identification of new FAdV isolates as an existing species or as a putative new FAdV. Once the polymerase sequence is known, more specific PCRs for the amplification of the hexon and other genes can be designed and performed according to the preliminary species classification.
Currently, the family Adenoviridae contains five genera, out of which Siadenovirus is one of the two least densely populated ones. A new member representing a new species in this genus has been detected in various birds of prey. The virus, named raptor adenovirus 1 (RAdV-1), could not be isolated, probably because no appropriate permissive cell-line was available. Partial genomic sequences, obtained by PCR and suggesting that the virus is a new siadenovirus species, have been published earlier. In the present paper, determination and analysis of the complete RAdV-1 genome are reported. This is the first complete genome sequence acquired from a non-isolated adenovirus (AdV). The sole source was a mixture of the internal organs of the diseased and dead birds. Until now, the genomic organization considered characteristic to siadenoviruses had been deduced from the detailed study of only two virus species, one of which originated from birds and the other from a frog. The present analysis of RAdV-1 confirmed the genus-specific genetic content and genomic features of siadenoviruses, and a putative novel gene was found as well. In general, AdVs and most of the AdV genera are thought to be strictly host specific. In the genus Siadenovirus, however, two virus species of rather divergent (avian and amphibian) host origin were present when the genus was found. Although by now the greatest number of known siadenoviruses infect birds, the original hosts of the genus remain unknown.
A seemingly novel siadenovirus species was detected by PCR and sequencing in the sample of a great tit (Parus major) found dead in Hungary. Since the genus Siadenovirus has very few known members so far, further study of the virus was intriguing not only from epizootiological but also from taxonomical aspects. The sample, which had been tested in another PCR survey previously, consisted of less than 50 microl of extracted nucleic acid. To ensure sufficient target DNA for an extended study, the viral genome had to be preserved. To this end, the sample was subjected to a novel method of non-specific DNA amplification. Using the amplified DNA as target, different PCR and sequencing strategies were applied with consensus or specific primers for the study of the central genome part of the putative tit adenovirus. The sequence of supposedly one half (13,628 bp) of the genome was determined including eight full genes between the genes of the IVa2 and hexon proteins. The gene content of the viral genome fragment as well as the results of the phylogenetic analyses with different proteins confirmed the discovery of a new species in the genus Siadenovirus. This is the first report on the detection of an adenovirus in great tits. The methods, described in this work, proved suitable for the recovery of nucleic acid samples that contain irreplaceable microbial genomic DNA but are only available in limited quantities.
Five budgerigars in the same breeding facility died or showed ruffled feathers. To determine the cause, five dead or euthanized budgerigars were examined. Splenomegaly was observed at necropsy in all birds examined. Histopathology of the spleen revealed a slight-to-moderate deletion of lymphocytes and increase of macrophages. Concurrent congestions in several tissues such as liver, lung, kidney, and/or brain and basophilic intranuclear inclusion bodies in the epithelial cells of renal tubules were found in all the birds examined. Psittacine adenoviral DNA was detected in the kidney of one of the five budgerigars by PCR. Sequencing and phylogenetic analysis of the hexon gene revealed that the adenovirus gene detected in the budgerigar was derived from an unknown adenovirus belonging to the genus Siadenovirus. Using a new pair of primers based on the obtained sequence, we confirmed the presence of the newly found adenovirus in all five birds. The newly found unknown adenovirus is designated as Budgerigar Adenovirus 1.
A female, adult ornate box turtle (Terrapene ornata ornata) with fatty liver was submitted for virologic examination in Hungary. Signs of an adenovirus infection including degeneration of the liver cells, enlarged nuclei and intranuclear inclusion bodies were detected by light microscopic examination. The presence of an adenovirus was later confirmed by obtaining partial sequence data from the adenoviral DNA-dependent DNA-polymerase. Phylogenetic analyses revealed that this novel chelonian adenovirus was distinct from previously described reptilian adenoviruses, not belonging to any of the recognized genera of the family Adenoviridae. As a part of the routine diagnostic procedure for chelonians the detection of herpes-, rana- and iridoviruses together with Mycoplasma spp. was attempted. Amplicons were generated by a general mycoplasma polymerase chain reaction (PCR) targeting the 16S/23S ribosomal RNA (rRNA) intergenic spacer region, as well as, a specific Mycoplasma agassizii PCR targeting the 16S rRNA gene. Based on the analyses of partial sequences of the 16S rRNA gene, the Mycoplasma sp. of the ornate box turtle seemed to be identical with the recently described eastern box turtle (Terrapene carolina carolina) Mycoplasma sp. This is the first report of a novel chelonian adenovirus and a mycoplasma infection in an ornate box turtle (T. ornata ornata) in Europe.
A consensus nested PCR was used to screen diagnostic samples from approximately 70 reptiles for the presence of adenoviruses (AdV) in the years 2006-2007. Classical virus isolation methods were also used with all samples. After adenoviruses were detected in a group of helodermatid lizards in a Danish zoo, a follow-up study was also carried out on lizards from this group (10 Mexican beaded lizards and 24 Gila monsters) over the period of a year. Adenoviruses were detected in a total of 26 lizards and snakes by PCR. The PCR amplicons from all positive animals were sequenced and the resulting polymerase gene sequences were used for phylogenetic analysis. Altogether six Agamid AdVs were amplified, with a minimal sequence variation between one another and between these and GenBank Agamid AdVs. The sequence obtained from one of the Gila monsters is identical with the GenBank Helodermatid AdV, while the sequences from the Mexican beaded lizards differ from this. In a snake collection we have detected a new AdV from an Asp viper. All of the above mentioned adenoviruses cluster in the Atadenovirus genus. However, the sequence from a new Varanid AdV detected in this study clusters outside this genus. On cell culture, viruses were isolated from three of the AdV positive helodermatid lizards (one Mexican beaded lizard and two Gila monsters) and identified as AdVs based on electron microscopy and PCR and sequencing using cell culture supernatant. This is the first report of the successful isolation of a lizard AdV.
An algorithm is presented for the multiple alignment of sequences, either proteins or nucleic acids, that is both accurate
and easy to use on microcomputers. The approach is based on the conventional dynamic-programming method of pairwise alignment.
Initially, a hierarchical clustering of the sequences is performed using the matrix of the pairwise alignment scores. The
closest sequences are aligned creating groups of aligned sequences. Then close groups are aligned until all sequences are
aligned in one group. The pairwise alignments included in the multiple alignment form a new matrix that is used to produce
a hierarchical clustering. If it is different from the first one, iteration of the process can be performed. The method is
illustrated by an example : a global alignment of 39 sequences of cytochrome c.
The three-dimensional crystal structure of the adenovirus major coat protein is presented. Adenovirus type 2 hexon, at 967 residues, is now the longest polypeptide whose structure has been determined crystallographically. Taken with our model for hexon packing, which positions the 240 trimeric hexons in the capsid, the structure defines 60% of the protein within the 150 X 10(6) dalton virion. The assembly provides the first details of a DNA-containing animal virus that is 20 times larger than the spherical RNA viruses previously described. Unexpectedly, the hexon subunit contains two similar beta-barrels whose topology is identical to those of the spherical RNA viruses, but whose architectural role in adenovirus is very different. The hexon structure reveals several distinctive features related to its function as a stable protective coat, and shows that the type-specific immunological determinants are restricted to the virion surface.
A serially propagated epithelial cell line (TH-1) was established from the heart of a box turtle (Terrapene carolina). TH-1 cells grew in mammalian media at well as in media of reduced toncity designed for amphibian cells. TH-1 cells routinely were grown and maintained at room temperature (23 °C). An incubation temperature of 30 °C was tolerated for only a few passages, while a temperature of 36 °C supported no cell growth.TH-1 cells could be maintained for brief periods at 36 °C. At this temperature replication of vaccinia, herpes simplex, pseudorabies and vesicular stomatitis viruses was obtained. TH-1 cells supported the replication of frog viruses at 23 °C. It is suggested that poikilothermic cell lines may offer unusual opportunities to study the growth of homothermic vertebrate viruses in host cells phylogenetically far removed from the normal host.
Hemorrhagic enteritis virus (HEV) belongs to the Adenoviridae family, a subgroup of adenoviruses (Ads) that infect avian species. In this article, the complete DNA sequence and the genome organization of the virus are described. The full-length of the genome was found to be 26,263 bp, shorter than the DNA of any other Ad described so far. The G + C content of the genome is 34.93%. There are short terminal repeats (39 bp), as described for other Ads. Genes were identified by comparison of the DNA and predicted amino acid sequences with published sequences of other Ads. The organization of the genome in respect to late genes (52K, IIIa, penton base, core protein, hexon, endopeptidase, 100K, pVIII, and fiber), early region 2 genes (polymerase, terminal protein, and DNA binding protein), and intermediate gene IVa2 was found to be similar to that of other human and avian Ad genomes. No sequences similar to E1 and E4 regions were found. Very low similarity to ovine E3 region was found. Open reading frames were identified with no similarity to any published Ad sequence.
This article presents a proposal for the establishment of a new adenovirus genus to accommodate certain bovine, ovine, and avian adenoviruses with special characteristics which differentiate them from members of the existing genera Mastadenovirus and Aviadenovirus. This proposal has been developed from earlier versions with advice from the Adenovirus Study Group of the International Committee on Taxonomy of Viruses (ICTV).
New advances in the field of genetic characterization of adenoviruses originating from different animal species are summarized. Variations seen in the host range and specificity, pathogenicity, genomic arrangement or gene complement are much wider than expected based on previous studies of human adenoviruses. Several exceptional adenoviruses from the two traditional conventional genera are now removed, and proposed to form at least two new genera. The eventual host origin of the new genera, however, is not clarified. Novel results from the genomic and phylogenetic analyses of adenoviruses originating from lower vertebrate species (including reptiles, amphibians and fish) seem to imply that probably five major clusters of adenoviruses exist corresponding to the five major classes of Vertebrata. Adenoviruses, which are now suspected to have common origin with enterobacterium phages from the family Tectiviridae, are perhaps very ancient indeed, and may have undergone a co-evolution with vertebrate hosts.
Partial genome characterisation of a non-cultivable marsupial adenovirus is described. Adenovirus-like particles were found by electron microscopy (EM) in the intestinal contents of brushtail possums (Trichosurus vulpecula) in New Zealand. Using degenerate PCR primers complementary to the most conserved genome regions of adenoviruses, the complete nucleotide sequence of the penton base gene, and partial nucleotide sequences of the DNA polymerase, hexon, and pVII genes were obtained. Phylogenetic analysis of the penton base gene strongly suggested that the brushtail possum adenovirus (candidate PoAdV-1) belongs to the recently proposed genus Atadenovirus. Sequence analysis of the PCR products amplified from the intestinal contents of brushtail possums originating from different geographical regions of New Zealand identified a single genotype. This is the first report of molecular confirmation of an adenovirus in a marsupial.
The genome of frog adenovirus (FrAdV-1) was sequenced and found to be the smallest of all known adenovirus genomes. The sequence obtained was 26163 bp in size and contains a substantial direct repeat near the right terminus, implying that it was derived by recombination from a parental genome of only 25517 bp. The closest relative of FrAdV-1 proved to be turkey adenovirus 3, an avian adenovirus with no previously known near relative. Sequence comparisons showed that the two viruses have equivalent gene complements, including one gene the product of which is related to sialidases. Phylogenetic analyses supported the establishment of a fourth adenovirus genus containing these two viruses, in addition to the established genera Mastadenovirus: and Aviadenovirus: and the proposed genus Atadenovirus: Sixteen genes were identified as being conserved between these four lineages and were presumably inherited from an ancestral adenovirus.
Genome sequencing and analysis of snake adenovirus type 1 (SnAdV-1), originating from corn snake, were completed. This is the first full genomic sequence of an adenovirus from reptilian hosts. The presence of characteristic genus-common genes and transcription units, showed that SnAdV-1 shares similar genome organisation with members of the recently established genus Atadenovirus. Three novel open reading frames of yet unknown functions were found. One of these seemed to be related to a putative gene, the so-called 105R that has recently been described from the genome of the tree shrew adenovirus. The other two putative genes were found to be unique for SnAdV-1. On phylogenetic trees, SnAdV-1 clustered within the atadenovirus clade. Thereby the hypothesis on the reptilian origin of atadenoviruses was further strengthened. Interestingly, however, one of the most striking features of atadenoviruses, namely the base content heavily biased towards A+T, is not characteristic for SnAdV-1 having a genome of balanced composition with a G+C value of 50.21%.
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No. 5,pp000.85–000.164. 112 A. Doszpoly et al./Infection, Genetics and Evolution 17 (2013) 106–112
Molecular evolution of adenoviruses Adenoviruses: Model and Vectors in Virus Host Interactions
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Benk} o, M., Harrach, B., 2003. Molecular evolution of adenoviruses. In: Doerfler, W., Bohm, P. (Eds.), Adenoviruses: Model and Vectors in Virus Host Interactions. Curr. Top. Microbiol. Springer, Berlin, pp. 3–35.
Turtles of the world, 2010 update: annotated checklist of taxonomy, synonymy, distribution, and conservation status Conservation Biology of Freshwater Turtles and Tortoises: A Compilation Project of the IUCN/SSC Tortoise and Freshwater Turtle Specialist Group
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- A G J Dijk
- P P Iverson
- J B Shaffer
- H B Rhodin
- A G J Pritchard
- P C H Dijk
- P P Saumure
- R A Buhlmann
- K A Iverson
- J B Mittermeier
Turtle Taxonomy Working Group Rhodin, A.G.J., van Dijk, P.P., Iverson, J.B., Shaffer, H.B., 2010. Turtles of the world, 2010 update: annotated checklist of taxonomy, synonymy, distribution, and conservation status. In: Rhodin, A.G.J., Pritchard, P.C.H., van Dijk, P.P., Saumure, R.A., Buhlmann, K.A., Iverson, J.B., Mittermeier, R.A. (Eds.), Conservation Biology of Freshwater Turtles and Tortoises: A Compilation Project of the IUCN/SSC Tortoise and Freshwater Turtle Specialist Group. Chelonian Research Monographs No. 5, pp 000.85–000.164. doi:10.3854/crm.5.000.checklist.v3.2010, http://www.iucn–tftsg.org/cbftt/.