Paramyxoviruses Research Articles

Paramyxovirus matrix proteins modulate host cell translation via exon-junction complex interactions in the cytoplasm.

Viruses have evolved myriad strategies to exploit the translation machinery of host cells to potentiate their replication. However, how paramyxovirus (PMVs) modulate cellular translation for their own benefit has not been systematically examined. Utilizing puromycylation labeling, overexpression of individual viral genes, and infection with wild-type virus versus its gene-deleted counterpart, we found that PMVs significantly inhibit host cells' nascent peptide synthesis during infection, with the viral matrix being the primary contributor to this effect. Using the rNiV-NPL replicon system, we discovered that the viral matrix enhances viral protein translation without affecting viral mRNA transcription and suppresses host protein expression at the translational level. Polysome profile analysis revealed that the HPIV3 matrix promotes the association of viral mRNAs with ribosomes, thereby enhancing their translation efficiency during infection. Intriguingly, our NiV-Matrix interactome identified the core exon-junction complex (cEJC), critical for mRNA biogenesis, as a significant component that interacts with the paramyxoviral matrix predominantly in the cytoplasm. siRNA knockdown of eIF4AIII simulated the restriction of cellular functions by the viral matrix, leading to enhanced viral gene translation and a reduction in host protein synthesis. Moreover, siRNA depletion of cEJC resulted in a 2-3 log enhancement in infectious virus titer for various PMVs but not SARS-CoV-2, enterovirus D68, or influenza virus. Our findings characterize a host translational interference mechanism mediated by viral matrix and host cEJC interactions. We propose that the PMV matrix redirects ribosomes to translate viral mRNAs at the expense of host cell transcripts, enhancing viral replication, and thereby enhancing viral replication. These insights provide a deeper understanding of the molecular interactions between paramyxoviruses and host cells, highlighting potential targets for antiviral strategies.

Corona- and Paramyxoviruses in Bats from Brazil: A Matter of Concern?

Chiroptera are one of the most diverse mammal orders. They are considered reservoirs of main human pathogens, where coronaviruses (CoVs) and paramyxoviruses (PMVs) may be highlighted. Moreover, the growing number of publications on CoVs and PMVs in wildlife reinforces the scientific community's interest in eco-vigilance, especially because of the emergence of important human pathogens such as the SARS-CoV-2 and Nipha viruses. Considering that Brazil presents continental dimensions, is biologically rich containing one of the most diverse continental biotas and presents a rich biodiversity of animals classified in the order Chiroptera, the mapping of CoV and PMV genetics related to human pathogens is important and the aim of the present work. CoVs can be classified into four genera: Alphacoronavirus, Betacoronavirus, Deltacoronavirus and Gammacoronavirus. Delta- and gammacoronaviruses infect mainly birds, while alpha- and betacoronaviruses contain important animal and human pathogens. Almost 60% of alpha- and betacoronaviruses are related to bats, which are considered natural hosts of these viral genera members. The studies on CoV presence in bats from Brazil have mainly assayed phyllostomid, molossid and vespertilionid bats in the South, Southeast and North territories. Despite Brazil not hosting rhinophilid or pteropodid bats, which are natural reservoirs of SARS-related CoVs and henipaviruses, respectively, CoVs and PMVs reported in Brazilian bats are genetically closely related to some human pathogens. Most works performed with Brazilian bats reported alpha-CoVs that were closely related to other bat-CoVs, despite a few reports of beta-CoVs grouped in the Merbecovirus and Embecovirus subgenera. The family Paramyxoviridae includes four subfamilies (Avulavirinae, Metaparamyxovirinae, Orthoparamyxovirinae and Rubulavirinae), and bats are significant drivers of PMV cross-species viral transmission. Additionally, the studies that have evaluated PMV presence in Brazilian bats have mainly found sequences classified in the Jeilongvirus and Morbillivirus genera that belong to the Orthoparamyxovirinae subfamily. Despite the increasing amount of research on Brazilian bats, studies analyzing these samples are still scarce. When surveying the representativeness of the CoVs and PMVs found and the available genomic sequences, it can be perceived that there may be gaps in the knowledge. The continuous monitoring of viral sequences that are closely related to human pathogens may be helpful in mapping and predicting future hotspots in the emergence of zoonotic agents.

Synchronicity of viral shedding in molossid bat maternity colonies.

Infection dynamics in vertebrates are driven by biological and ecological processes. For bats, population structure and reproductive cycles have major effects on RNA virus transmission. On Reunion Island, previous studies have shown that parturition of pregnant females and aggregation of juvenile Reunion free-tailed bats (Mormopterus francoismoutoui) are associated with major increase in the prevalence of bats shedding RNA viruses. The synchronicity of such shedding pulses, however, is yet to be assessed between viruses but also maternity colonies. Based on 3422 fresh faeces collected every 2-5 weeks during four consecutive birthing seasons, we report the prevalence of bats shedding astroviruses (AstVs), coronaviruses (CoVs) and paramyxoviruses (PMVs) in two maternity colonies on Reunion Island. We found that the proportion of bats shedding viruses is highly influenced by sampling collection periods, and therefore by the evolution of the population age structure. We highlight that virus shedding patterns are consistent among years and colonies for CoVs and to a lesser extent for PMVs, but not for AstVs. We also report that 1% of bats harbour co-infections, with two but not three of the viruses, and most co-infections were due to CoVs and PMVs.

Metagenomic analysis of endemic viruses in oral secretions from Chinese pigs.

BackgroundPigs are unique reservoirs for virus ecology. Despite the increased use of improved biosecurity measures, pig viruses readily circulate in Chinese swine farms.ObjectivesThe main objective of this study was to examine archived swine oral secretion samples with a panel of pan‐species viral assays such that we might better describe the viral ecology of swine endemic viruses in Chinese farms.MethodologyTwo hundred (n = 200) swine oral secretion samples, collected during 2015 and 2016 from healthy pigs on six swine farms in two provinces in China, were screened with molecular pan‐species assays for coronaviruses (CoVs), adenoviruses (AdVs), enteroviruses (EVs), and paramyxoviruses (PMV). Samples were also screened for porcine circovirus (PCV) 3, porcine reproductive and respiratory syndrome virus (PRRSV) and influenza A virus (IAV).ResultsAmong 200 swine oral secretion samples, 152 (76.0%) were found to have at least one viral detection. Thirty‐four samples (17%) were positive for more than one virus, including 24 (70.5%) with dual detection and 10 (29.5%) with triple detection. Seventy‐eight (39.0%) samples were positive for porcine AdVs, 22 (11.0%) were positive for porcine CoVs, 21 (10.5%) were positive for IAVs, 13 (6.5%) were positive for PCV, 7 (3.5%) were positive for PMV, six (3.0%) were positive for PRRSV and five (2.5%) were positive for porcine EV.ConclusionOur findings underscore the high prevalence of numerous viruses among production pigs in China and highlight the need for routine, periodic surveillance for novel virus emergence with the goal of protecting pigs.

Paramyxovirus Diversity within One Population of Miniopterus fuliginosus Bats in Sri Lanka.

Bats are known as typical reservoirs for a number of viruses, including viruses of the family Paramyxoviridae. Representatives of the subfamily Orthoparamyxovirinae are distributed worldwide and can cause mild to fatal diseases when infecting humans. The research on Paramyxoviruses (PMVs) from different bat hosts all over the world aims to understand the diversity, evolution and distribution of these viruses and to assess their zoonotic potential. A high number of yet unclassified PMVs from bats are recorded. In our study, we investigated bat species from the families Rhinolophidae, Hipposiderae, Pteropodidae and Miniopteridae that are roosting sympatrically in the Wavul Galge cave (Koslanda, Sri Lanka). The sampling at three time points (March and July 2018; January 2019) and screening for PMVs with a generic PCR show the presence of different novel PMVs in 10 urine samples collected from Miniopterus fuliginosus. Sequence analysis revealed a high similarity of the novel strains among each other and to other unclassified PMVs collected from Miniopterus bats. In this study, we present the first detection of PMVs in Sri Lanka and the presence of PMVs in the bat species M. fuliginosus for the first time.

Detection of novel paramyxoviruses in Chaerephon bat species in Nigeria and phylogenetics of paramyxoviruses co-evolution with bats in Africa.

Bat paramyxoviruses (PmV) are a diverse group of viruses and include zoonotic viruses such as henipaviruses. Members of this group in other continents have been associated with severe respiratory and neurological infections in animals and humans. Furthermore, despite the richness of diverse bat species that can transmit this virus in African countries like Nigeria, there is very scanty information as to the presence and co-evolution of paramyxoviruses in bats. There is a need for continuous surveillance of zoonotic viruses and their biological reservoirs as this will help in the prevention and management of pathogens' spillovers. This study detected novel paramyxoviruses in Chaerephon nigeriae bat species found in Badagry, Lagos. Phylogenetic analyses of paramyxovirus sequences' co-evolution with frugivorous and insectivorous bats circulating in African countries were also performed using sequences of African origin available in the Database of Bat-Associated Viruses (DBatVir: http://www.mgc.ac.cn/DBatVir/). Oral swabs (n=18) and blood samples (n=32) were collected from C.nigeriae bats in Badagry, Lagos. The L gene of bat paramyxovirus was detected in all oral swabs using PCR techniques. Six of the amplicons were successfully sequenced. Estimated phylogenies placed the sequences in close relationship with those isolated from insectivorous bats. Phylogenetic analyses of previously sequenced isolates in the African region showed the likelihood of different co-evolution mechanisms of paramyxoviruses with frugivorous bats compared with insectivorous bats. This may be due to codon usage bias of the L gene. Spatial distribution of paramyxoviruses in African countries showed limited ongoing surveillance of this virus in the continent, especially in southern and northern countries. Extensive surveillance of paramyxoviruses with possible zoonotic potentials among bat species in the continent is recommended. This will provide further insights into co-evolution as well as prevent possible spillover into the human population.

Identification of Coronaviruses, Paramyxoviruses, Reoviruses, and Rotaviruses among Bats in Nigeria.

Bats are often consumed by some ethnic groups in Nigeria despite association of bats with many important emerging viruses. More than 300 bats representing eight species were captured during 2010-2011 in eight locations of northern Nigeria. Available fecal swabs (n = 95) were screened for the presence of arenaviruses, CoVs, paramyxoviruses (PMVs), reoviruses, rhabdoviruses, and influenza viruses using generic reverse transcription-polymerase chain reaction assays. Here, we document the detection of CoVs, PMVs, reoviruses, and rotaviruses (RVs) in Nigerian bats. The Nigerian bat CoVs are grouped within other bat SARS-CoV-like viruses identified from Ghana in a sister clade next to the human SARS-CoV clade. The phylogenetic analysis indicated a broad range of RVs present in Nigerian bats, some cluster with human RVs and some represent novel species. Our study adds that continuing global surveillance for viruses in bats to understand their origin, adaptation, and evolution is important to prevent and control future zoonotic disease outbreaks.

The Clinical Characteristics and Outcomes of Adult Patients With Pneumonia Related to Three Paramyxoviruses

Background: Respiratory syncytial virus (RSV), human metapneumovirus (hMPV), and human parainfluenza virus (hPIV) are paramyxoviruses (PMVs) that are important etiologies of community-acquired pneumonia. However, current knowledge about the clinical features and outcomes of PMV-related pneumonia (PMV-p) is limited. We aimed to investigate the clinical characteristics and disease severity in immunocompetent adults hospitalized with hMPV-related pneumonia (hMPV-p), hPIV-related pneumonia (hPIV-p), or RSV-related pneumonia (RSV-p).Methods: We retrospectively recruited 488 patients with PMV-p (153 with RSV-p, 137 with hMPV-p, and 198 with hPIV-p) from five teaching hospitals in China during 2011–2019. Univariate and multivariate analyses were performed to identify predictors to distinguish hMPV-p/hPIV-p from RSV-p and evaluate the effects of virus types on the clinical outcomes.Results: Compared with RSV-p, sputum production [odds ratio (OR) 5.029, 95% confidence interval (CI) 2.452–10.312, P < 0.001] was positively associated with hMPV-p, while solid malignant tumor (OR 0.346, 95% CI 0.126–0.945, P = 0.038), nasal congestion (OR 0.102, 95% CI 0.041–0.251, P < 0.001), and respiratory rate ≥ 30 breaths/min (OR 0.296, 95% CI 0.136–0.640, P = 0.002) were negatively related to hMPV-p. Sputum production (OR 13.418, 95% CI 6.769–26.598, P < 0.001) was positively associated with hPIV-p, while nasal congestion (OR 0.194, 95% CI 0.098–0.387, P < 0.001), dyspnea (OR 0.469, 95% CI 0.272–0.809, P < 0.001), and respiratory rate ≥30 breaths/min (OR 0.090, 95% CI 0.032–0.257, P < 0.001) on admission were negatively related to hPIV-p. After adjustment for confounders, multivariate logistic regression analysis suggested that hMPV-p (OR 0.355, 95% CI 0.135–0.932, P = 0.035) and hPIV-p (OR 0.311, 95% CI 0.121–0.784, P = 0.013) were associated with decreased 30-day mortality compared with RSV-p. RSV infection (OR 4.183, 95% CI 1.709–10.236, P = 0.002) was identified as an independent predictor of 30-day mortality in patients with PMV-p.Conclusion: RSV-p caused more severe disease than hMPV-p and hPIV-p. Although some clinical features are helpful for distinguishing the diseases, etiologic diagnosis is critical in the management of the PMV-p.

Data on known anti-virals in combating CoVid-19.

Design and development of effective anti-virals in combating CoVid-19 is a great challenge worldwide. Known drugs such as chloroquine, lopinavir, favipiravir and remdesivir are used in the management of CoVid - 19. It is known that Ivermectin and remdesivir both are effective against filoviruses, paramyxo viruses. Available data also shows that ivermectin and remedesivir repress the replication of SARS-CoV-2. Thus, we document the potential use of ivermectin and remdesivir in the management of CoVid -19.

Evaluation of 10 years of parainfluenza virus, human metapneumovirus, and respiratory syncytial virus infections in lung transplant recipients.

Respiratory tract infection with pneumoviruses (PVs) and paramyxoviruses (PMVs) are increasingly associated with chronic lung allograft dysfunction (CLAD) in lung transplant recipients (LTRs). Ribavirin may be a treatment option but its effectiveness is unclear, especially with respect to infection severity. We retrospectively analyzed 10 years of PV/PMV infections in LTRs. The main end points were forced expiratory volume in 1 second (FEV1) at 3 and 6 months postinfection, expressed as a percentage of pre‐infection FEV1 and incidence of new or progressed CLAD 6 months postinfection. A total of 139 infections were included: 88 severe infections (63%) (defined as >10% FEV1 loss at infection) and 51 mild infections (37%) (≤10% FEV1 loss). Overall postinfection CLAD incidence was 20%. Associations were estimated on postinfection FEV1 for ribavirin vs no ribavirin (+13.2% [95% CI: 7.79; 18.67]) and severe vs mild infection (−11.1% [95% CI: −14.76; −7.37]). Factors associated with CLAD incidence at 6 months were ribavirin treatment (odds ratio (OR [95% CI]) 0.24 [0.10; 0.59]), severe infection (OR [95% CI] 4.63 [1.66; 12.88]), and mycophenolate mofetil use (OR [95% CI] 0.38 [0.14; 0.97]). These data provide valuable information about the outcomes of lung transplant recipients with these infections and suggests possible associations of ribavirin use and infection severity with long‐term outcomes. Well‐designed prospective trials are needed to confirm these findings.

Differential Features of Fusion Activation within the Paramyxoviridae.

Paramyxovirus (PMV) entry requires the coordinated action of two envelope glycoproteins, the receptor binding protein (RBP) and fusion protein (F). The sequence of events that occurs during the PMV entry process is tightly regulated. This regulation ensures entry will only initiate when the virion is in the vicinity of a target cell membrane. Here, we review recent structural and mechanistic studies to delineate the entry features that are shared and distinct amongst the Paramyxoviridae. In general, we observe overarching distinctions between the protein-using RBPs and the sialic acid- (SA-) using RBPs, including how their stalk domains differentially trigger F. Moreover, through sequence comparisons, we identify greater structural and functional conservation amongst the PMV fusion proteins, as compared to the RBPs. When examining the relative contributions to sequence conservation of the globular head versus stalk domains of the RBP, we observe that, for the protein-using PMVs, the stalk domains exhibit higher conservation and find the opposite trend is true for SA-using PMVs. A better understanding of conserved and distinct features that govern the entry of protein-using versus SA-using PMVs will inform the rational design of broader spectrum therapeutics that impede this process.

The long isoform of ZAP widely restricts Paramyxoviruses

Paramyxoviruses (PVs) are negative-sense RNA viruses that are important in human and animal health, and can cause severe zoonotic diseases. We screened a wide range of animal and human PV matrix proteins for host interactions, and found that the long (L) isoform of the host protein ZAP (zinc-finger antiviral protein) interacted with all the tested viral matrixes. ZAP-L is constitutively expressed, has a prenlyation motif for membrane-localization, and primarily mediates antiviral activity by binding and degrading viral RNAs through the exosome complex. However, ZAP restriction of PVs has not been demonstrated in the literature. We found that knockdown of ZAP-L results in increased replication of a panel of five human and animal PVs. Overexpression of ZAP-L (but not ZAP-short) restricts replication of PVs in a reciprocal pattern – with the notable exception of Sendai virus (SeV) – and as with other viruses, mutating the prenylation motif of ZAP-L abolishes restriction. RT-qPCR of PV RNAs and pulled-down RNAs does not indicate specific targeting of a viral transcript by ZAP-L, although overall genome abundance is reduced. Finally, immunoprecipitation of ZAP shows an additional RNA-independent interaction between ZAP-L and SeV-nucleocapsid not found with HPIV3-nucleocapsid, a closely-related PV. Thus, we have observed that ZAP-L interacts with the matrix of, and restricts replication of, a wide range of paramyxoviruses. A PV that is not restricted (SeV), has an additional interaction with ZAP via its nucleocapsid protein that may ameliorate ZAP restriction. Investigating ZAP-related restriction differences between closely-related PVs may shed light on anti-viral mechanisms of ZAP.

Efficacy of Prepared Oil Inactivated Pigeon Paramyxo Vaccine

Control of pigeon paramyxo virus infection relays on the production of a vaccine that is able to protect the birds from the lethal virulent infection as well as reducing shedding level to prevent virus circulation in the environment. In the current study inactivated vaccine was prepared from the recent field isolate (PPMV-1 YA/14) which is extensively characterized and identified. The vaccine was prepared in two formulas. The aluminum hydroxide gel (given S/C and I/M) and the oil adjuvanted formula. Evaluation of the 2 formulas revealed that the gel-based formula gave a higher antibody titer than the oil-based preparation during the course of the vaccination period and after challenge. The cellular immune response was also evaluated by the QRT-PCR analysis of the IFN-γ transcripts and it was found that the oil-based formula gave a high fold change than the gel-based formula although it was not reflected in decreasing the shedding level. Studying the shedding level by the EID50 or QRT-PCR of the cloacal swaps after challenge revealed that the gel-based formula administrated S/C gave lesser degree of shedding than the oil-based formula. In conclusion the gel-based formula of the vaccine prepared from the recent characterized PPMV-1 YA/14 isolate could protect pigeon from lethal infection with the virulent virus and could reduce but not eliminate the shedding level.

Coronavirus and paramyxovirus in bats from Northwest Italy

BackgroundBat-borne virus surveillance is necessary for determining inter-species transmission risks and is important due to the wide-range of bat species which may harbour potential pathogens. This study aimed to monitor coronaviruses (CoVs) and paramyxoviruses (PMVs) in bats roosting in northwest Italian regions. Our investigation was focused on CoVs and PMVs due to their proven ability to switch host and their zoonotic potential. Here we provide the phylogenetic characterization of the highly conserved polymerase gene fragments.ResultsFamily-wide PCR screenings were used to test 302 bats belonging to 19 different bat species. Thirty-eight animals from 12 locations were confirmed as PCR positive, with an overall detection rate of 12.6% [95% CI: 9.3–16.8]. CoV RNA was found in 36 bats belonging to eight species, while PMV RNA in three Pipistrellus spp. Phylogenetic characterization have been obtained for 15 alpha- CoVs, 5 beta-CoVs and three PMVs; moreover one P. pipistrellus resulted co-infected with both CoV and PMV. A divergent alpha-CoV clade from Myotis nattereri SpA is also described. The compact cluster of beta-CoVs from R. ferrumequinum roosts expands the current viral sequence database, specifically for this species in Europe. To our knowledge this is the first report of CoVs in Plecotus auritus and M. oxygnathus, and of PMVs in P. kuhlii.ConclusionsThis study identified alpha and beta-CoVs in new bat species and in previously unsurveyed Italian regions. To our knowledge this represents the first and unique report of PMVs in Italy. The 23 new bat genetic sequences presented will expand the current molecular bat-borne virus databases. Considering the amount of novel bat-borne PMVs associated with the emergence of zoonotic infections in animals and humans in the last years, the definition of viral diversity within European bat species is needed. Performing surveillance studies within a specific geographic area can provide awareness of viral burden where bats roost in close proximity to spillover hosts, and form the basis for the appropriate control measures against potential threats for public health and optimal management of bats and their habitats.

Synthesis and antiviral study of 4-(7,7-dimethyl-4-(piperazin-1-yl)-5,6,7,8-tetrahydroquinazolin-2-yl) morpholine derivatives

A series of 4-(7,7-dimethyl-4-(piperazin-1-yl)-5,6,7,8-tetrahydroquinazolin-2-yl)morpholine substituted sulfonamide and urea derivatives has been synthesized and characterized using spectral techniques. The antiviral activity of these compounds against an avian paramyxo virus (AMPV-1) was screened using MTT assay and found that one of the sulfonamide derivatives (2d) show three-fold higher antiviral activity than Ribavirin, a commercial antiviral drug substance.

Molecular detection of viruses in Kenyan bats and discovery ofnovel astroviruses, caliciviruses and rotaviruses

This is the first country-wide surveillance of bat-borne viruses in Kenya spanningfrom 2012–2015 covering sites perceived to have medium to high level bat-humaninteraction. The objective of this surveillance study was to apply a non-invasiveapproach using fresh feces to detect viruses circulating within the diverse speciesof Kenyan bats. We screened for both DNA and RNA viruses; specifically, astroviruses(AstVs), adenoviruses (ADVs), caliciviruses (CalVs), coronaviruses (CoVs),flaviviruses, filoviruses, paramyxoviruses (PMVs), polyomaviruses (PYVs) androtaviruses. We used family-specific primers, amplicon sequencing and furthercharacterization by phylogenetic analysis. Except for filoviruses, eight virusfamilies were detected with varying distributions and positive rates across the fiveregions (former provinces) studied. AstVs (12.83%), CoVs (3.97%), PMV (2.4%), ADV(2.26%), PYV (1.65%), CalVs (0.29%), rotavirus (0.19%) and flavivirus (0.19%). NovelCalVs were detected in Rousettus aegyptiacus andMops condylurus while novelRotavirus-A-related viruses were detected in Taphozous bats and R.aegyptiacus. The two Rotavirus A (RVA)strains detected were highly related to human strains with VP6 genotypes I2 and I16.Genotype I16 has previously been assigned to human RVA-strain B10 from Kenya only,which raises public health concern, particularly considering increased human-batinteraction. Additionally, 229E-like bat CoVs were detected in samples originatingfrom Hipposideros bats roosting in sites withhigh human activity. Our findings confirm the presence of diverse viruses in Kenyanbats while providing extended knowledge on bat virus distribution. The detection ofviruses highly related to human strains and hence of public health concern,underscores the importance of continuous surveillance.

Single-Centre Experience with Oral Ribavirin in Lung Transplant Recipients with Paramyxovirus Infections

Paramyxovirus (PV) infections are increasingly recognized in lung transplant recipients and have been linked to subsequent graft failure and bronchiolitis obliterans syndrome (BOS). Ribavirin represents a possible treatment option although the outcome on graft function and BOS incidence is unknown. We analysed outcomes of all PV infections in lung/heart-lung recipients between September 2006 and April 2009 in a single centre. PV-infected recipients treated with oral ribavirin were compared with those unable to receive ribavirin due to contraindications. Recovery of graft function, time to recovery and new development of BOS were compared. A total of 38 patients (ribavirin group) were treated with ribavirin for a median of 9 days (IQR 8-12), whilst 29 patients (non-ribavirin group) received best supportive care including corticosteroids. The median forced expiratory volume in 1 s dropped 20% (IQR 15-32) from baseline in the ribavirin group versus 18% (IQR 13-30) in the non-ribavirin group during infection. In 84% of patients treated with ribavirin and 59% of the non-ribavirin group, graft function recovered within 30 days (P=0.02). New onset of BOS developed within 6 months in 5% of the ribavirin group versus 24% of the non-ribavirin group (P=0.02). Treatment of PV after lung/heart-lung transplantation with oral ribavirin seems to be associated with earlier recovery of graft function and to prevent BOS.

A unique novel reptilian paramyxovirus, four atadenovirus types and a reovirus identified in a concurrent infection of a corn snake ( Pantherophis guttatus) collection in Germany

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.

Expression of paramyxovirus V proteins promotes replication and spread of hepatitis C virus in cultures of primary human fetal liver cells.

Here we demonstrate that primary cultures of human fetal liver cells (HFLC) reliably support infection with laboratory strains of hepatitis C virus (HCV), although levels of virus replication vary significantly between different donor cell preparations and frequently decline in a manner suggestive of active viral clearance. To investigate possible contributions of the interferon (IFN) system to control HCV infection in HFLC, we exploited the well-characterized ability of paramyxovirus (PMV) V proteins to counteract both IFN induction and antiviral signaling. The V proteins of measles virus (MV) and parainfluenza virus 5 (PIV5) were introduced into HFLC using lentiviral vectors encoding a fluorescent reporter for visualization of HCV-infected cells. V protein-transduced HFLC supported enhanced (10 to 100-fold) levels of HCV infection relative to untransduced or control vector-transduced HFLC. Infection was assessed by measurement of virus-driven luciferase, by assays for infectious HCV and viral RNA, and by direct visualization of HCV-infected hepatocytes. Live cell imaging between 48 and 119 hours postinfection demonstrated little or no spread of infection in the absence of PMV V protein expression. In contrast, V protein-transduced HFLC showed numerous HCV infection events. V protein expression efficiently antagonized the HCV-inhibitory effects of added IFNs in HFLC. In addition, induction of the type III IFN, IL29, following acute HCV infection was inhibited in V protein-transduced cultures. Conclusion: These studies suggest that the cellular IFN response plays a significant role in limiting the spread of HCV infection in primary hepatocyte cultures. Strategies aimed at dampening this response may be key to further development of robust HCV culture systems, enabling studies of virus pathogenicity and the mechanisms by which HCV spreads in its natural host cell population. (Hepatology 2011;54:1901-1912)

Paramyxovirus Infection in a Leopard Tortoise (Geochelone pardalis babcocki) with Respiratory Disease

Paramyxoviruses (PMVs) are well documented as a cause of respiratory disease in snakes, but they have only been documented in tortoises in a very few cases. This study describes a leopard tortoise (Geochelone pardalis babcocki) with lethargy and respiratory disease. Examination of tracheal swabs for bacteria resulted in strong growth of Pseudomonas aeruginosa and Morganella morganii. Despite antibiotic therapy, the animal died. Gross pathology noted severe diffuse consolidation of the lungs. Fourteen different tissue samples were collected for virological examinations. Viruses commonly detected in tortoises (herpesviruses, picorna-like viruses, and ranaviruses) or recently reported in chelonians (adenoviruses) could not be detected in the samples by virus isolation or by molecular methods (polymerase chain reaction [PCR]). The samples were tested by reverse transcriptase-PCR for PMVs, and cloaca, heart, liver, and small intestine were found to be positive. Sequence analysis of the PMV amplicons revealed that the tortoise was coinfected with at least two different squamatid PMV species and not with a chelonian host-specific PMV. The contribution of these viruses to the observed clinical signs and pathology is unknown.