Virological Studies Research Articles

Viral susceptibility and innate immune competency of Carollia perspicillata bat cells produced for virological studies.

Multiple viruses that are highly pathogenic in humans are known to have evolved in bats. How bats tolerate infection with these viruses, however, is poorly understood. As viruses engage in a wide range of interactions with their hosts, it is essential to study bat viruses in a system that resembles their natural environment like bat-derived in vitro cellular models. However, stable and accessible bat cell lines are not widely available for the broader scientific community. Here, we generated in vitro reagents for the Seba's short-tailed bat (Carollia perspicillata), tested multiple methods of immortalization, and characterized their susceptibility to virus infection and response to immune stimulation. Using a pseudotyped virus library and authentic virus infections, we show that these C. perspicillata cell lines derived from a diverse array of tissues are susceptible to viruses bearing the glycoprotein of numerous orthohantaviruses, including Andes and Hantaan virus and are also susceptible to live hantavirus infection. Furthermore, stimulation with synthetic double-stranded RNA prior to infection with VSV and MERS-CoV induced a protective antiviral response, demonstrating the suitability of our cell lines to study the bat antiviral immune response. Taken together, the approaches outlined here will inform future efforts to develop in vitro tools for virology from non-model organisms and these C. perspicillata cell lines will enable studies on virus-host interactions in bats.

Explicit description of viral capsid subunit shapes by unfolding dihedrons

Viral capsid assembly and the design of capsid-based nanocontainers critically depend on understanding the shapes and interfaces of constituent protein subunits. However, a comprehensive framework for characterizing these features is still lacking. Here, we introduce a novel approach based on spherical tiling theory that explicitly describes the 2D shapes and interfaces of subunits in icosahedral capsids. Our method unfolds spherical dihedrons defined by icosahedral symmetry axes, enabling systematic characterization of all possible subunit geometries. Applying this framework to real T = 1 capsid structures reveals distinct interface groups within this single classification, with variations in interaction patterns around 3-fold and 5-fold symmetry axes. We validate our classification through molecular docking simulations, demonstrating its consistency with physical subunit interactions. This analysis suggests different assembly pathways for capsid nucleation. Our general framework is applicable to other triangular numbers, paving the way for broader studies in structural virology and nanomaterial design.

IMPORTANCE OF VIROLOGICAL SURVEILLANCE AFTER MIGRANTS FROM POLIO HIGH-RISK TERRITORIES FOR POLIOMYELITIS MONITORING IN THE RUSSIAN FEDERATION

Abstract The results of virologically examined biological material collected from children and poliovirus-specific immunity arrived in the Russian Federation from polio high risk territories were analyzed. Over a ten-year period (2014-2023), more than 3,300 samples were examined at the Subnational WHO Laboratory in St. Petersburg. Most of the examined children arrived in different regions of the Russian Federation from the Republics of Tajikistan (56.5%) or Uzbekistan (6.2%) and from Ukraine (5.8%). Children who arrived from the North Caucasus were also examined comprising 22.2% examined cases. Polioviruses (65 PV) were isolated from 55 children during virological study. Most isolated strains were classified as types 1 and 3 polioviruses, with six strains classified as type 2. Only vaccine polioviruses were found, including type 2 polioviruses according to the ITD results. One PV2 strain was isolated from a child from Kyrgyzstan in 2014, and five PV2 strains from the new nOPV2 vaccine were isolated from Tajik children in 2021 after using nOPV2 vaccine to suppress cVDPV2 circulation in Tajikistan. Analysis of vaccination status in children arrived from high polio risk territories revealed problems poor immunization coverage in relevant place of residence. More than a third of children had no vaccination records, about 9% children had no polio vaccination for various reasons including medical exemptions and refusals of parents to vaccinate, another 12% were vaccinated incompletely. A comparatively analyzed effectiveness of polio vaccination for children resident in the Russian Federation and those who arrived from Tajikistan also evidenced poor polio immunization coverage in the latter. Many children from Tajikistan had no antibodies specific to polioviruses of different types, and 11-12% of children did not have antibodies to all three or two vaccine poliovirus types (according to the 2006-2010 or 2014-2020 vaccination schedules). The data obtained confirm the importance and need for epidemiological and virological surveillance for residents arrived from polio high risk territories in Russia within poliomyelitis control programme in the Russian Federation.

Sofosbuvir, Velpatasvir and Voxilaprevir for Treatment of Chronic Hepatitis C Virus Patients Non-Responder to Previous Sofosbuvir-Daclatasvir Therapy

Abstract Background Egypt is one of the countries with a high HCV burden; the percentage of HCV antibody positivity is 4.61 % of Egyptian general population. The advent of oral direct-acting antivirals (DAAs), as a treatment for chronic hepatitis C (CHC), has revolutionized the field with obvious safety and efficacy privileges. Despite the high rates of SVR achieved with DAAs, HCV is not eliminated from a considerable proportion of patients (1-15%). So, a new problem has emerged which is the need for re-treatment of patients who couldn’t be cured with DAAs in the first instance. Aim of the Work to evaluate the efficacy of sofosbuvir, velpatasvir, voxilaprevir for previously DAA-(Sofosbuvir–Daclatasvir)-treated Egyptian patients who failed to achieve SVR after their treatment. Patients and Methods This study was performed in Ahmed Maher Teaching Hospital, Cairo, Egypt, Internal Medicine Department. The study included 139 patients with chronic hepatitis C Non- responder to Previous Sofosbuvir-Daclatasvir Therapy. Patients enrolled in the study were prospectively evaluated as outpatients by the study staff at 4, 8, 12 weeks and at 12 weeks post-treatment. All the patients were subjected to history taking, clinical examination, complete biochemical and virological studies. Results The overall treatment outcome: there was a statistically highly significant difference in PCR before treatment (100%+ve) and after treatment (97.1%-ve) 3 month after treatment. Throughout the study, most of the patients were responders (97.1% achieved SVR 12). Conclusion Rescue therapy with SOF/VEL/VOX is safe and highly effective, for treatment of chronic hepatitis C virus patients non-responder to previous Sofosbuvir-Daclatasvir therapy achieving SVR12 rates 97.1%.

VirID: Beyond Virus Discovery-An Integrated Platform for Comprehensive RNA Virus Characterization.

RNA viruses exhibit vast phylogenetic diversity and can significantly impact public health and agriculture. However, current bioinformatics tools for viral discovery from metagenomic data frequently generate false positive virus results, overestimate viral diversity, and misclassify virus sequences. Additionally, current tools often fail to determine virus-host associations, which hampers investigation of the potential threat posed by a newly detected virus. To address these issues we developed VirID, a software tool specifically designed for the discovery and characterization of RNA viruses from metagenomic data. The basis of VirID is a comprehensive RNA-dependent RNA polymerase database to enhance a workflow that includes RNA virus discovery, phylogenetic analysis, and phylogeny-based virus characterization. Benchmark tests on a simulated data set demonstrated that VirID had high accuracy in profiling viruses and estimating viral richness. In evaluations with real-world samples, VirID was able to identify RNA viruses of all types, but also provided accurate estimations of viral genetic diversity and virus classification, as well as comprehensive insights into virus associations with humans, animals, and plants. VirID therefore offers a robust tool for virus discovery and serves as a valuable resource in basic virological studies, pathogen surveillance, and early warning systems for infectious disease outbreaks.

Characterizing Marine Medaka (Oryzias melastigma) Haploid Embryonic Stem Cells: A Valuable Tool for Marine Fish Genetic Research.

Haploid embryonic stem cells (ESCs), which combine the properties of haploidy and pluripotency, hold significant potential for advancing developmental biology and reproductive technology. However, while previous research has largely focused on haploid ESCs in freshwater species like Japanese medaka (Oryzias latipes), little is known about their counterparts in marine species. This study hypothesizes that haploid ESCs from marine fish could offer unique insights and tools for genetic and virological research. To address this, we successfully established and characterized a novel haploid ESC line, hMMES1, derived from marine medaka (Oryzias melastigma). The hMMES1 cells contain 24 chromosomes, exhibit core stem cell characteristics, and express key pluripotency markers. In vitro, hMMES1 cells form embryonic bodies (EBs) capable of differentiating into the three germ layers. In vivo, hMMES1 cells were successfully transplanted into marine medaka and zebrafish, resulting in the generation of interspecies and interordinal chimeras. Additionally, hMMES1 cells demonstrate high efficiency in transfection and transduction, and show susceptibility to major aquaculture viruses, nodavirus (NNV) and iridovirus (SGIV). These findings suggest that hMMES1 cells represent a valuable model for genetic manipulation and virological studies in marine fish species.

Development and characterization of segment-specific enteroids from the pig small intestine in Matrigel and transwell inserts: insights into susceptibility to porcine epidemic diarrhea Virus.

The critical early stages of infection and innate immune responses to porcine epidemic diarrhea virus (PEDV) at the intestinal epithelium remain underexplored due to the limitations of traditional cell culture and animal models. This study aims to establish a porcine enteroid culture model to investigate potential differences in susceptibility to infection across segments of the porcine small intestine (duodenum, jejunum, and ileum). Intestinal crypt cells from nursery pigs were cultured in Matrigel to differentiate into porcine enteroid monolayer cultures (PEMCs). Following characterization, PEMCs were enzymatically dissociated and subcultured on transwell inserts (PETCs) for apical surface exposure and infection studies. Characterization of region-specific PEMCs and PETCs included assessment of morphology, proliferation, viability, and cellular phenotyping via immunohistochemistry/immunocytochemistry and gene expression analysis. Subsequently, PETCs were inoculated with 105 TCID50 (50% tissue culture infectious dose)/mL of a high pathogenic PEDV non-S INDEL strain and incubated for 24h. Infection outcomes were assessed by cytopathic effect, PEDV N protein expression (immunofluorescence assay, IFA), and PEDV N-gene detection (quantitative reverse transcription polymerase chain reaction, RT-qPCR). No significant morphological and phenotypical differences were observed among PEMCs and PETCs across intestinal regions, resembling the porcine intestinal epithelium. Although PETCs established from different segments of the small intestine were susceptible to PEDV infection, jejunum-derived PETCs exhibited higher PEDV replication, confirmed by IFA and RT-qPCR. This segment-specific enteroid culture model provides a reliable platform for virological studies, offering a controlled environment that overcomes the limitations of in vivo and traditional cell culture methods. Standardizing culture conditions and characterizing the model are essential for advancing enteroid-based infection models.

Establishment of a humanized mouse model of HIV-1 infection and quantification of integrated HIV proviral DNA in vivo

In recent years, virological, pathological, and immunological studies need to be carried out for the emerging anti-human immunodeficiency virus (HIV) therapies such as gene therapy, broadly neutralizing antibodies, and the derived chimeric antigen receptor (CAR)-T immunotherapy, which necessitates suitable, simple, and inexpensive small-animal models and methods for accurate quantification of the viral genome in the HIV-1 infected. In our research, the HIV-∆ENV-Jurkat-EGFP-mCherry cell line was engineered through the infection with a dual-labelled HIV pseudovirus. A nested quantitative PCR (nested-qPCR) method with the cellular genome as the integrated standard was established for the quantification of HIV proviral copies. We administered intravenous injections of healthy human peripheral blood mononuclear cell (PBMC) into NOD/Prkdcscid/IL2rgnull (NPG) mice. To verify engraftment kinetics, we analyzed the percentages of hCD45+, hCD3+, hCD4+, and hCD8+ cells in the peripheral blood of hu-PBMC-NPG mice. To evaluate HIV-1 infection in hu-PBMC-NPG mice, we inoculated these mice with HIV NL4-3-NanoLuc by intraperitoneal (IP) injection. We then monitored the luciferase expression by the small animal imaging system and measured the viral load in the spleen by qPCR. The infiltration of human PBMCs in mice was detected 3-5 weeks after intravenous injection, and the percentage of hCD45 in humanized mouse PBMCs were more than 25% five weeks after IP inoculation. The expression of the virus-associated luciferase protein was detected by luciferase imaging 27 days post infection. Moreover, the viral total DNA, RNA, and proviral DNA copies reached 18 000 copies/106 cells, 15 000 copies/μg RNA, and 15 000 copies/106 cells, respectively, in the mouse spleen. Taken together, we reported a convenient method for building a simple humanized mouse model of HuPBMC-NPG/severe combined immunodeficiency (SCID) by intravenous injection with hu-PBMCs without advanced surgical skills and irradiation. Furthermore, we established a convenient method for the efficient determination of proviral DNA to assess HIV replication in vivo, viral reservoir sizes, and efficacy of novel anti-HIV therapies including CAR-T immunotherapy and gene therapy.

Real World Use of Tixagevimab/Cilgavimab Pre-Exposure Prophylaxis of COVID-19 in Immunocompromised Individuals: Data from the OCTOPUS Study.

Objective. We aimed to report the real-world use and outcomes over time in immunocompromised individuals receiving tixagevimab/cilgavimab (T/C) pre-exposure prophylaxis (PrEP). Methods. This observational study included participants who received T/C PrEP, categorized into three groups: (i) No COVID-19 (NoC), i.e., participants who never had COVID-19; (ii) Hybrids (H), i.e., participants who had COVID-19 before PrEP; and (iii) Break-through Infections (BTIs), i.e., participants who had COVID-19 after PrEP. The study measured several immune markers at the administration of T/C (T0) at 3 (T1), 6 (T2), and 9 (T3) months afterward. These markers included: anti-receptor-binding domain (RBD) IgG antibodies; BA.5-neutralizing antibodies (nAbs); mucosal IgG; and T cell immunity. The incidence rate ratios for BTIs were analyzed using a Poisson regression model. Results. A total of 231 participants with a median age of 63 years (IQR 54.0-73.0). were included. Among these, 84% had hematological diseases and received a median of three vaccine doses. N = 72 participants belonged to the NoC group, N = 103 to the H group, and n = 56 to the BTI group (24%), with most BTIs being mild/moderate. The incidence rate (IR) of BTIs was 4.2 per 100 patient-months (95% CI 3.2-5.4), with no associated risk factors identified. There was a significant increase in anti-RBD IgG levels 3 months after the T/C administration in all groups, followed by a decline at 6 months, whereas at the same time points, geometric mean titers (GMTs) of anti-BA.5 nAbs were low for all groups and were around or below the detection threshold. No significant changes were observed in IFN-γ levels. The mucosal immune response was observed only 3 months after the PrEP administration. Conclusion. We provided a real-world experience model on the clinical efficacy of T/C PrEP in preventing severe COVID-19 during the Omicron wave through a comprehensive virological and immunological study. While waiting for the arrival of new monoclonal antibodies that can effectively neutralize the most recent variants, T/C PrEP remains the only viable strategy in the available armamentarium today to prevent COVID-19 complications in an extremely fragile population with suboptimal immune responses to COVID-19 vaccines.

Future research avenues for the study of fibropapillomatosis in sea turtles

Fibropapillomatosis (FP) is a debilitating tumoral disease affecting sea turtles worldwide. While mainly afflicting immature individuals and potentially altering vital functions, the precise impact of this panzootic on turtle health and survival remains unclear. Moreover, the etiological factors implicated in the FP emergence, development and transmission are not yet definitively identified. Among them, an infection by a spreading herpesvirus and the contamination by pollutants (either organic pollutants and trace elements) are suspected. Here, we provide an overview of discoveries, knowledge and propose hypotheses related to FP within five key FP research areas, i.e., virology studies, transmission studies, contamination studies, host genomic studies, and veterinary treatment assays. Moreover, we recommend urgent research avenues to develop at the interface of virology, epidemiology, ecotoxicology, oncology, physiology, immunology, cellular and evolutionary biology, in order to characterize the dynamics of FP and to predict its consequences on sea turtle populations. Importantly, extending the implementation and development of strong collaborations between rehabilitation centers, field biologists and research laboratories at large geographical scale is required to rapidly increase our knowledge on FP and work towards its effective management.

Characterization and arbovirus susceptibility of cultured CERNI cells derived from sika deer (Cervus nippon).

Cervus nippon (sika deer) are widely distributed throughout eastern Asia. Deer possess a variety of antibodies against several zoonotic pathogens, indicating that they act as reservoir of zoonoses. In this study, we reported the characterization of cultured cells derived from sika deer and evaluated their susceptibility to arthropod-borne viruses to clarify their usefulness in virological studies. Cells derived from testicular tissue in Dulbecco's modified eagle medium with 16% fetal bovine serum started growing as primary cultured cells. The diploid cells consisted of 68 chromosomes, consistent with those of Japanese sika deer previously reported. The phylogenetic analysis showed the cells formed a robust clade with Japanese population of C. nippon, indicating that the cultured cells established in this study were originated from the Japanese sika deer. The cells immortalized by the simian virus 40 T-antigen were predominantly spindle-shaped cells exhibiting adhesive properties, and cultivated at 37°C and 5% CO2, which are common culture conditions for many mammalian cell lines. Western blotting analysis indicated that the cultured cells were multiple types of cells that coexist, including at least epithelial, fibroblast, and also Leydig cells. We confirmed that the cells have susceptibility to several arboviruses distributed in Japan: Getah virus, Japanese encephalitis virus, Oz virus, and severe fever with thrombocytopenia syndrome virus, but not to Tarumiz tick virus. From these results, the cells contribute to clarify the role of sika deer as a reservoir of zoonoses in nature and deer-associated experimental research at the cellular and molecular levels.

Structure-Guided Design of Potent Coronavirus Inhibitors with a 2-Pyrrolidone Scaffold: Biochemical, Crystallographic, and Virological Studies.

Zoonotic coronaviruses are known to produce severe infections in humans and have been the cause of significant morbidity and mortality worldwide. SARS-CoV-2 was the largest and latest contributor of fatal cases, even though MERS-CoV has the highest case-fatality ratio among zoonotic coronaviruses. These infections pose a high risk to public health worldwide warranting efforts for the expeditious discovery of antivirals. Hence, we hereby describe a novel series of inhibitors of coronavirus 3CLpro embodying an N-substituted 2-pyrrolidone scaffold envisaged to exploit favorable interactions with the S3-S4 subsites and connected to an invariant Leu-Gln P2-P1 recognition element. Several inhibitors showed nanomolar antiviral activity in enzyme and cell-based assays, with no significant cytotoxicity. High-resolution crystal structures of inhibitors bound to the 3CLpro were determined to probe and identify the molecular determinants associated with binding, to inform the structure-guided optimization of the inhibitors, and to confirm the mechanism of action of the inhibitors.

Application of advanced bioimaging technologies in viral infections

Viruses are major pathogens responsible for a wide range of infectious diseases that may lead to global pandemics. The understanding of structural underpinnings of virions, molecular mechanisms of infection and the pathogenesis of viral diseases is crucial for exploring prophylactic and therapeutic approaches for viral disorders. Though the latest developments in imaging techniques have equipped scientists with tools to investigate viral infections in unprecedented detail with multiple spatial and temporal resolutions, the unveiling of how viral particles dynamically interact with host cells requires integrations of strategies connecting the structural complexity with functional properties. Here, we aim to delineate (1) the diversity of imaging devices that have been applied in the investigation of viral morphology and structure, (2) the contribution of current imaging techniques in revealing features of viral life-cycle and the associated host reactions, and (3) the potential of imaging systems in the diagnosis and prognosis of viral diseases. This review provides an essential overview of bioimaging techniques in scenarios of viral infections and comes up with instructions of how to select appropriate bioimaging methods for virological studies based on specific objectives of investigations. Additionally, we discuss the challenges and opportunities of microscopy imaging in promoting more insightful comprehension of complexities between viruses and hosts in the nano-scale range.

Multifunctional nanorobot system in precise evaluation and manipulation of virus capsid

It is imperative to have high adaptive techniques for sensing and manipulating biological targets at the nanoscale. This necessity becomes particularly crucial when dealing with fragile living bio-organisms like viruses, where the expression of capsids is closely linked to viral functions and genome constitution. Therefore, the development of a comprehensive system for dissecting and measuring viruses holds significant implications for the pharmaceutical industry and drug manufacturing. Leveraging the sub-nanometer spatial resolution and controllable tip-cantilever architecture of atomic force microscopy (AFM), a probe-laser system has been integrated as a self-sensing robotic end effector. To address intrinsic challenges in AFM-based robotic systems such as the lack of real-time monitoring, low scanning rates, and nonlinear motion caused by piezoelectric actuators, an augmented reality robotic system has been implemented. This system incorporates stereoscopic vision, a haptic feedback controller, a position recovery scheme, and a real-time force control algorithm. The integration of these components enhances the system’s capability to accurately dissect virus capsids. Operators can now perform highly efficient nanoscale tasks with multidimensional perception, utilizing the combination of stereoscopic vision and haptic force control. The position correction during manipulation can achieve a frame rate of over 30 frames per second, imperceptible to the operator, enabling closed-loop operation control. By adopting the proposed nanorobotic system in virology studies, it becomes possible to achieve accurate manipulation and dissection of SARS-CoV-2 pseudovirus capsids, and derive multi-parametric properties such as structural integrity, protein fragment thickness, and adhesive forces. The established nanobot system and experimental results serve as a guiding platform for high-accuracy evaluation in drug manufacturing development.

Establishment and characterization of a kidney cell line from hybrid snakehead (male Channa argus × female Channa maculata) and its susceptibility to hybrid snakehead rhabdovirus (HSHRV)

Hybrid snakehead (male Channa argus × female Channa maculata) is an emerging fish breed with increasing production levels. However, infection with hybrid snakehead rhabdovirus (HSHRV) critically affects hybrid snakehead farming. In this study, a fish cell line called CAMK, derived from the kidneys of hybrid snakehead, was established and characterized. CAMK cells exhibited the maximum growth rate at 28 °C in Leibovitz's-15 medium supplemented with 10% fetal bovine serum(FBS). Karyotyping revealed diploid chromosomes in 54% of the cells at the 50th passage (2n = 66), and 16S rRNA sequencing validated that CAMK cells originated fromhybrid snakehead, and the detection of kidney-specific antibodies suggested that it originated from kidney. .The culture was free from mycoplasma contamination, and the green fluorescent protein gene was effectively transfected into CAMK cells, indicating their potential use for in vitro gene expression investigations. Furthermore, qRT-PCR and immunofluorescence analysis revealed that HSHRV could replicate in CAMK cells, indicating that the cells were susceptible to the virus. Transmission electron microscopy revealed that the viral particles had bullet-like morphology. The replication efficiency of HSHRV was 107.33 TCID50/mL. Altogether, we successfully established and characterized a kidney cell line susceptible to the virus. These findings provide a valuable reference for further genetic and virological studies.

Association between hepatitis C virus genotype 4 and renal cell carcinoma: Molecular and virological studies.

Hepatitis C virus (HCV) is the most common infection worldwide. The correlation between HCV and renal cell carcinoma (RCC) is still mysterious. Therefore, the relationship between HCV and RCC was investigated. The study included 100 patients with RCC; 32 with HCV infection, and 68 without HCV infection. Expressions of viral proteins (NS3 and NS5A) were tested using an immune electron-microscope (IEM) and immunohistochemistry (IHC). IHC and quantitative real time-PCR investigated the presentation of human proteins TP53 and p21 genes. Transmission electron (TEM) detected viral-like particles in infected RCC tissues. The gene and protein expression of P53 was higher in HCV positive versus HCV negative patients and p21 was lower in HCV positive versus HCV negative in both tumor and normal tissue samples. Viral like particles were observed by TEM in the infected tumor and normal portion of the RCC tissues and the plasma samples. The IEM showed the depositions of NS3 and NS5A in infected renal tissues, while in noninfected samples, were not observed. The study hypothesizes that a correlation between HCV and RCC could exist through successfully detecting HCV-like particles, HCV proteins, and (p53 and p21) in RCC-infected patients.

The cell cultures in virology: from the past to the future

The aim of the review is to give a brief characteristic of cell cultures obtained from mammalian tissues and to consider the current possibilities and prospects for their use in virology.
 The analysis of the literature data presented in the main databases, such as Web of Science, PubMed, Scopus, Elsevier, Google Scholar and RSCI (as of July 2023), indicates that various types of cell cultures are currently used in virological studies. The use of cell culture has a number of advantages over other in vitro and in vivo methods of virological research. The review provides numerous examples on the development of new methods of obtaining cell cultures for the cultivation of viruses. Among them are sensitive and reporter cell systems, the design of which can be a promising tool for diagnostics of existing and new unknown viral infections. Cell cultures are characterized as potential in vitro models in virology for developing new diagnostic test-systems and antiviral drugs. An important area of cell cultures application is their use as a substrate for the production of culture-derived vaccines. Another aspect of the cell cultures application is also highlighted, such as the study of the effect of the viruses on the host immune system or the mechanisms of immunopathogenesis of viral infections. It is concluded that the use of cell cultures remains currently and in the near future one of the most important methods in practical virology and in scientific research.

Virome characterization of diarrheic red-crowned crane (G. japonensis)

BackgroundThe red-crowned crane is one of the vulnerable bird species. Although the captive population has markedly increased over the last decade, infectious diseases can lead to the death of young red-crowned cranes while few virological studies have been conducted.MethodsUsing a viral metagenomics approach, we analyzed the virome of tissues of the dead captive red-crowned crane with diarrhea symptoms in Dongying Biosphere Reserve, Shandong Province, China and feces of individual birds breeding at the corresponding captive breeding center, which were pooled separately.ResultsThere is much more DNA and RNA viruses in the feces than that of the tissues. RNA virus belonging to the families Picornaviridae, and DNA viruses belonging to the families Parvoviridae, associated with enteric diseases were detected in the tissues and feces. Genomes of the picornavirus, genomovirus, and parvovirus identified in the study were fully characterized, which further suggested that infectious viruses of these families were possibly presented in the diseased red-crowned crane.ConclusionRNA virus belonging to the families Picornaviridae, and DNA viruses belonging to the families Genomoviridae and Parvoviridae were possibly the causative agent for diarrhea of red-crowned crane. This study has expanded our understanding of the virome of red-crowned crane and provides a baseline for elucidating the etiology for diarrhea of the birds.

Effective plant virus enrichment using carbon nanotubes and microfluidics

Plant virus detection and identification in crops is a pillar for disease management, import of crop material, production of clean stock plants and basic plant virology studies. In this report, we present a platform for the enrichment and isolation of known or unknown viruses. This platform is based on carbon nanotube arrays inside a microfluidic device that can be a solution for the identification of low titer viruses from plants. Using our microfluidic devices, we achieved enrichment of two economically important viruses, the orthotospovirus, tomato spotted wilt orthotospovirus (TSWV) and the potyvirus, zucchini yellow mosaic virus (ZYMV). The carbon nanotube arrays integrated in these microfluidic devices are capable of trapping viruses discriminated by their size; the virus rich arrays can be then analyzed by common downstream techniques including immunoassays, PCR, HTS and electron microscopy. This procedure offers a simple to operate and portable sample preparation device capable of trapping viruses from raw plant extracts while reducing the host contamination.

Indication of Phages and Their Role in the Regulation of the Purification Process at the Govsany Aeration Station in Baku

Due to the epidemic situation and the emergence of enteroviral diseases, we set ourselves the task of conducting comprehensive virological and bacteriological studies of the wastewater, its sludge and contaminated soil from the Govsany aeration station. The fact of constant detection of enteroviruses in samples of wastewater and soil in combination with bacteriophages is considered by us as a possibility of detection of bacteriophages at WWTP sites. The 26 wastewater samples contain enteroviruses at various stages of treatment, 18 of which were characterised by high levels of faecal contamination (thermoletarian coliforms), C. perfringens and Enterococcus faecalis. In samples with high contamination (TCB up to 106 per 100 ml of wastewater), which are potentially dangerous in terms of the content of infectious disease pathogens, both bacterial and viral, the number of coliphages was below the norm. Consequently, coliphages cannot be used as a sanitary indicator of microorganisms of viral contamination. Intestinal phages are common where humans and animals live and therefore intestinal bacteria are present. Coliphages (bacteriophages) can be formed from lysogenic enterobacteria during the process of stepwise wastewater treatment at the Govsany WWTP.