Search Results (325)

Porcine epidemic diarrhea virus (PEDV) imposes substantial economic losses on the global swine industry owing to its high pathogenicity and transmissibility. Although arginine (Arg) is known to support the integrity of intestinal barrier, it is not clear whether Arg can alleviate intestinal injury induced by PEDV. A total of 32 healthy 7-day-old piglets were randomly assigned to four groups (Control, Arg, PEDV, PEDV + Arg; eight replicates per group). From day 5, piglets in the Arg and PEDV + Arg groups were orally administered Arg at 400 mg/kg body weight until day 11; then, PEDV (1 × 10^5.5 TCID₅₀) was given orally for two PEDV-infected groups. On day 14, all piglets were slaughtered to obtain blood and intestine samples for further analysis. The results showed that PEDV infection significantly reduced T-SOD and CAT activities in plasma and intestine while elevating MPO levels. Arg supplementation restored T-SOD (plasma, duodenum, ileum), CAT (plasma, ileum), and GSH-Px (jejunum, ileum) activities and reduced MDA (jejunum) content in PEDV-infected piglets. Hematological analysis showed Arg alleviated PEDV-induced increases in MCV and RDW-SD, and significantly elevated MCHC. The real-time quantitative PCR analysis demonstrated that Arg further enhanced PEDV structural genes (M, N, S) expression in the duodenum, ileum, and colon. Concurrently, Arg significantly up-regulated interferon-stimulated genes (MX1, OASL, ISG15, IFITM3) in the ileum, IRF7 in the duodenum and colon, and IFN-β in the ileum. Arg also down-regulated the pro-inflammatory cytokines IL-6 and CXCL2 and the antimicrobial peptide REG3G in the colon, while up-regulating the tissue repair gene MMP13 in the ileum. In conclusion, oral Arg exhibits a unique dual role: it promotes PEDV replication to a certain extent while significantly enhancing antioxidant capacity, strengthening intestinal antiviral immunity, and attenuating intestinal inflammation. These findings highlight Arg’s role in promoting disease tolerance and offer a novel perspective for nutritional intervention strategies against PEDV infection. Full article

Background: Oncolytic viruses (OVs), a promising anti-cancer therapeutic, replicate more efficiently in cancer cells rather than in healthy cells due to the alterations in antiviral response mechanisms and dysregulation of signaling pathways. Vesicular stomatitis virus (VSV) is known for low pathogenicity, tropism to various cancer cells, and the ability to lyse cells in the hypoxic tumor microenvironment (TME). Targeted delivery of immune checkpoint and co-stimulatory molecules can enhance the anti-tumor immune response and remodel the immunosuppressive TME. The aim of this study was to compare the activity of rVSV-GFP with rVSV, encoding the programmed cell death protein 1 (PD-1) and tumor necrosis factor ligand superfamily member 9 (4-1BBL). Methods: The oncolytic efficacy of these rVSV variants used at 10⁵, 10⁶, and 10⁷ TCID50 was evaluated at 24 and 48 h post-infection by flow cytometry in a panel of mouse and human cancer cell lines. Quantitative real-time polymerase chain reaction (qPCR) was used to evaluate mRNA expression levels of certain genes at 12 and 48 h post-infection. Results: Murine hepatocellular carcinoma (H22) and human melanoma (A375) or human lung carcinoma (A549) were the most sensitive to rVSV therapy cell lines. The higher relative expression of the antiviral response genes RIG-I and IFIT1 within each biological species (mouse or human) correlated with lower sensitivity to rVSV. No such effect was observed for the type I interferons (IFNs), despite their proposed key role in resistance to OV therapy. Conclusions: H22, A375, and A549 are more susceptible to the oncolytic activity of the novel rVSV-PD-1-4-1BBL. Full article

Background: Pseudorabies vaccines based on the live attenuated PRV Bartha-K61 strain remain essential for controlling pseudorabies in swine, but poor thermal stability during storage and transport limits field use. Objectives: This study aimed to develop a thermostable freeze-dried live pseudorabies vaccine through the integrated optimization of formulation and lyophilization and to preliminarily assess its humoral immunogenicity in piglets. Methods: Trehalose, mannitol, and glycine were screened as candidate lyoprotectants by single-factor experiments, followed by Box–Behnken response surface optimization using viral titer retention as the response. Critical thermal parameters were determined to establish a formulation-specific lyophilization cycle. Product quality was evaluated by post-lyophilization titer, cake appearance, residual moisture, and electron microscopy, and stability was compared with a commercial freeze-dried vaccine at 2–8 °C, 25 °C, 37 °C, and 45 °C. Conclusions: The optimized formulation, ST005, contained 9.5% trehalose, 2.0% mannitol, and 1.5% glycine. It showed a collapse-related critical temperature of approximately −34.0 °C and a dried-product glass transition temperature of 69.1 °C, produced an intact cake with residual moisture below 3.0%, and preserved viral morphology after lyophilization. Titer loss remained below 1.0 log10 TCID₅₀/mL for 24 months at 2–8 °C, 9 months at 25 °C, 21 days at 37 °C, and 9 days at 45 °C, outperforming the commercial comparator. In PRV-seronegative piglets, stored ST005 induced robust gB-specific and neutralizing antibody responses after storage under refrigerated, ambient, and accelerated conditions. Full article

Achieving long-term, continuous, and accurate localization for Unmanned Aerial Vehicles (UAVs) in outdoor GNSS-denied environments where pre-existing reference maps are available is challenging. To this end, this paper proposes a Cross-view Particle Filter Localization (CPFL) framework. Unlike existing particle filter approaches that rely on inertial sensors for state propagation or sparse semantic labels for observation updates, CPFL is a vision-driven solution. This framework introduces specific adaptations into the two core stages of particle filtering: In the motion propagation stage, it achieves visual state transition by calculating a feature-based inter-frame homography mapping to estimate the 2D global relative motion components, eliminating the dependency on inertial priors; in the observation correction stage, a Dual-Granularity Adaptive Gating (DGAG) cross-view network is designed to mitigate perceptual aliasing and generate discriminative absolute position weights for the particles. By fusing these two stages through a filter mechanism, the framework transforms unbounded cumulative drift into bounded absolute localization errors. Furthermore, addressing the measurement deficiencies of traditional single-frame metrics, this paper also proposes a Trajectory Continuity Index (TCI@d) tailored for continuous localization tasks. Experiments on the real-world MAFS dataset confirm that this framework achieves a mean localization error of 5.28 m and a localization success rate of 89.7% under a 10-m threshold. Compared with mainstream vision-only algorithms and IMU-fusion baselines, this framework demonstrates lower mean errors and improved trajectory continuity, validating its effectiveness for long-term robustness. Full article

Porcine circovirus type 3 (PCV3) capsid protein (Cap) is a key antigen for immunological studies and vaccine development. Different optimized PCV3 ORF2 sequences were used to construct six baculovirus transfer plasmids, with the pOET1.1-based design yielding the highest Cap level. Cap expression was confirmed by Western blot, IPMA and IFA. Recombinant baculovirus amplification was optimized, achieving the highest titer at an MOI of 0.1 with a 72 h harvest to 10^7.5TCID₅₀/0.1 mL, while maximal Cap production was obtained at an MOI of 0.1 with a 96 h harvest. PCV3 Cap virus-like particles (VLPs) were purified by sucrose density-gradient ultracentrifugation and cation-exchange chromatography, and TEM revealed spherical particles of approximately 17–20 nm. In mice, VLP immunization induced increasing antigen-specific IgG from day 14. Immunization increased both IgG1 and IgG2a without a significant difference, and post-immunization serum specifically recognized PCV3-positive passaged PK-15 cells in an indirect immunofluorescence assay. In splenic lymphocytes, IFN-γ, TNF-α, IL-4, and IL-10 mRNA levels were significantly upregulated (p < 0.01). Moreover, pig challenge data supported the protective potential of PCV3 Cap VLPs in the natural host. In our study, Cap assembled into VLPs and induced immune responses, providing a basis for PCV3 subunit vaccine development. Full article

Differentiation and detection of viruses causing vesicular/mucocutaneous lesions are essential for patient management. This study evaluated the analytical and clinical performance of the Novaplex™ HSV-1&2/VZV/MPXV multiplex real-time PCR assay (Novaplex), designed for the simultaneous detection of monkeypox virus (MPXV), herpes virus types 1 and 2 (HSV), and varicella-zoster virus (VZV). It was compared with Singleplex PCRs as reference methods. Analytical sensitivity was assessed only for MPXV using viral stocks of clades Ia, Ib, and IIb, while clinical performances for HSV, VZV and MPXV were evaluated on 93 residual clinical samples. Novaplex discriminated clade II from clade I viruses and demonstrated low limits of detection for MPXV (clade Ia: 2.9 Log TCID₅₀/mL; clade Ib: 1.9 Log TCID₅₀/mL; clade IIb: 1.4 Log TCID₅₀/mL). Clinically, the assay showed high overall sensitivity (97.1%) and specificity (100%), with almost perfect agreement with Singleplex PCR (κ = 0.947). Stratified results by viruses: HSV showed κ = 1.000 and 100% of sensitivity, MPXV showed a sensitivity of 97.8% with κ = 0.969, and VZV showed a κ = 0.914 with a sensitivity of 87.5%. Specificity was 100% for all three viruses. Novaplex offers a robust, efficient diagnostic approach for simultaneous detection of MPXV, HSV-1/2, and VZV, supporting timely clinical decision making and enhanced outbreak preparedness. Full article

In 2023, a highly immunogenic live attenuated vaccine based on the Nigeria 75/1 strain was introduced in Kazakhstan to provide protection against PPR. This study presents the results of a three-year animal trial evaluating the vaccine’s efficacy, safety, and immunogenicity. The novelty of this study lies in the long-term (up to 36 months) evaluation of protective immunity in adult animals, as well as in the comparative analysis of immune responses across different age groups and the assessment of viral suppression following challenge infection. Sheep and goats of different age groups were included, including lambs and kids aged 1.5 and 3 months, as well as adult animals aged 2–3 years. The vaccine was well tolerated following a single immunization, and no clinically significant adverse effects were observed in vaccinated animals, apart from only mild transient local reactions. A strong humoral (IgG) response to PPRV antigens was detected in all groups, with the highest antibody titers observed in young animals. Seroconversion was detected in 100% of vaccinated animals by day 21 post-vaccination. Long-term protective immunity (at least 36 months) was demonstrated in adult animals, whereas in young animals early protection was confirmed at 21 days post-vaccination along with subsequent humoral immune dynamics following a single immunization with a 1.0 mL dose of the vaccine (Nigeria 75/1 strain, titer 10^3.0 TCID₅₀/mL). These findings indicate that the vaccine is well tolerated, highly immunogenic, and provides sustained protection in adult animals while inducing early immune responses in young animals. Full article

Despite increased interest in virus survival on surfaces, data on bovine herpesvirus type 1 (BoHV-1) interactions with metal surfaces remain limited. This study aimed to assess the effects of copper, zinc, and iron on BoHV-1 viability, viral titre, and DNA stability under different conditions. MDBK-adapted BoHV-1 was used to investigate the virucidal effect of copper, zinc and stainless steel surfaces. The virus was exposed for 1 and 24 h under both wet and dry conditions. Inactivation was assessed based on changes in TCID₅₀ log₁₀ values, qPCR Ct results, and calculating half-lives of the virus and its DNA. Virus stability varied depending on surface type, environmental conditions, and duration of exposure. Copper demonstrated the strongest virucidal effect, significantly reducing viral titres and DNA levels under all conditions. After 1 h in wet conditions, copper reduced viral titre to 4.7 log₁₀, while zinc and stainless steel showed minimal impact. Under dry conditions, copper reduced viral titres to the limit of detection after 24 h. Half-life analysis confirmed rapid inactivation on copper, with the shortest persistence observed across all conditions. Zinc showed moderate virucidal activity but required longer exposure times. These findings highlight copper’s superior antiviral properties and suggest its potential application in reducing viral transmission on surfaces. Full article

Getah virus (GETV), a mosquito-borne virus capable of infecting multiple economically important animal species, poses a potential epidemic risk. In May 2024, one pig farm from Heyuan, Guangdong Province, China, suffered reproductive disorders in sows and diarrhea in newborn piglets. Out of the six blood samples that were collected, three tested strongly positive for GETV, yielding a positivity rate of 50%. Moreover, a GETV strain (designated GDHYLC2024) was successfully isolated and identified. The viral titer of GDHYLC2024 was 10^7.687 TCID₅₀/mL in Vero cells. Its genome was composed of 11,688 bases in length. Interestingly, compared with GDHYLC23, it had no unique 32-nucleotide repeat insertion in 3′ non-coding region. However, phylogenetic analysis showed that GDHYLC2024 and GDHYLC23 clustered in genotype III. Animal infection experiments demonstrated that the GDHYLC2024 strain was highly pathogenic to 4-day-old piglets, which caused obvious clinical symptoms including fever, depression, anorexia, periorbital edema, ataxia, and three deaths out of a total of five individuals in the infection group. This study reported re-emergence of GETV in the same region of Guangdong Province, China. The above findings suggest that GETV continuously poses a threat to farm pig’s health and has genetic diversity. Full article

Vaporized free chlorine, primarily present as hypochlorous acid (HOCl), is increasingly used for indoor microbial control; however, virus-dependent susceptibility and its molecular determinants remain unclear. We evaluated virucidal effects under controlled indoor conditions (0–9 ppb) against echovirus 30 (E30), influenza A/H1N1, and human adenovirus type 3 (HAdV3). Infectious titers were quantified by TCID₅₀ assays. Computational fluid dynamics (CFD) simulations and gas-sensor measurements assessed spatial dispersion, and structural analyses examined oxidation-sensitive amino acid residues. Significant reductions in infectivity were observed for E30 (99.0%, p = 0.00727) and influenza A/H1N1 (99.9%, p = 0.000597), whereas no significant reduction was detected for HAdV3 (p = 0.142). Analyses including all data points without outlier exclusion confirmed the robustness of these findings. CFD indicated uniform dispersion, although spatial heterogeneity within the indoor environment cannot be excluded. These findings suggest that viral susceptibility to vaporized HOCl is associated with residue-level composition and structural context; however, this relationship should be interpreted as correlative rather than causal. Moreover, integration of molecular and structural analyses provides a plausible mechanistic framework, although direct biochemical validation remains necessary. Structural analyses showed lower proportions of oxidation-sensitive residues in adenoviral proteins compared with influenza A hemagglutinin (OR = 0.34–0.40, adjusted p < 0.001) and the E30 VP1 intermediate. Residues were clustered in surface-exposed functional domains in susceptible viruses. Full article

Lassa virus (LASV), the causative agent of Lassa fever, must be handled under biosafety level 4 (BSL-4) conditions, requiring validated inactivation protocols to ensure laboratory and public safety. Although LASV is an enveloped virus theoretically susceptible to physical and chemical inactivation methods, quantitative data on its inactivation kinetics remain limited. This study systematically evaluated the efficacy of thermal treatment (56 °C, 70 °C, 95 °C), laboratory chemical inactivants (beta-propiolactone, formaldehyde, methanol, TRIzol), and five commercial disinfectants against infectious LASV. Viral infectivity was determined by titrating residual virus in Vero E6 cells, and complete inactivation was verified by three consecutive blind passages. Thermal inactivation was achieved at 56 °C for 40 min, 70 °C for 5 min, and 95 °C for 2 min. Both 0.1% and 0.05% beta-propiolactone completely inactivated LASV after 24 h at 4 °C, while 4% formaldehyde, 50% methanol, and 25% TRIzol achieved complete inactivation within 15 min, 10 min, and 2 min, respectively. For surface disinfection, 2% and 5% Micro-Chem Plus™ and 75% ethanol reduced viral titers by ≥4 log₁₀ TCID₅₀/mL within 30 s; 1% sodium hypochlorite and 0.25% Virkon required 1 min, whereas 3% hydrogen peroxide required 3 min to achieve the same reduction. These results provide quantitative, evidence-based parameters that can serve as a valuable reference for the safe handling of LASV under controlled BSL-4 laboratory conditions. Full article

Polysaccharides derived from cyanobacteria and microalgae have attracted increasing interest as natural virucidal agents. Among them, polysaccharides from the cyanobacterium Arthrospira platensis (A. platensis), and the green microalgae Dunaliella salina (D. salina) have shown virucidal activities, mainly against enveloped viruses, while evidence on non-enveloped viruses is still limited. In this study, the virucidal activity of purified polysaccharides extracted from A. platensis (APPs) and from D. salina (DSPs) was evaluated in vitro against human adenovirus type 5 (HAdV5), a non-enveloped pathogenic virus with high persistence in the environment and resistance to disinfection. The in vitro assays were carried out at concentrations previously verified as non-toxic by morphological evaluation of A549 cells after 24 and 48 h of incubation, testing two viral loads, namely, 10³ and 10⁴ tissue culture infectious dose 50% per milliliter (TCID₅₀/mL). For APPs, a possible time-dependent effect was also assessed at different contact times (15, 30 and 60 min). DSPs showed a limited virucidal effect related to the starting viral concentration, while APPs induced a consistent viral reduction (up to 98.8%) at both viral concentrations. The virucidal effect of APPs occurred rapidly and was not significantly influenced by contact time, thus suggesting that prolonged exposure is not a determining factor for polysaccharide virucidal activity. These findings demonstrate the virucidal activity of APPs against a highly resistant non-enveloped virus and provide preliminary in vitro evidence of their potential application as natural virucidal agents, particularly for environmental disinfection purposes. Further investigations are warranted to elucidate the underlying mechanisms of action and to optimize their practical use. Full article

Respiratory infections caused by influenza viruses are frequently associated with coinfection by other infectious agents. Torque teno viruses (TTVs) are small DNA viruses that can function as opportunistic pathogens and are epidemiologically linked to influenza viruses as well as a broad spectrum of infectious and immune-mediated diseases. Among TTVs, swine torque teno viruses (TTSuVs) are unique in that they have been shown to act as primary pathogens. With the long-term objective of developing experimental tools to better understand inter-viral interactions, this study aimed to optimize a murine model of TTV and influenza virus coinfection. Experimental mice were inoculated with TTSuV1 on day 1 post infection (DPI 1), while phosphate-buffered saline (PBS)-treated mice served as negative controls. A subset of TTSuV1-infected mice was subsequently coinfected with the influenza A virus H1N1 (IAV) at either 12 or 27 days following TTSuV1 infection. An additional group of mice was maintained as an IAV only control. Mice infected with IAV were euthanized 72–84 h post-IAV infection, corresponding to DPI 15 and 30, respectively. Unexpectedly, gross and histopathological examination of lung tissues revealed that prior TTSuV1 infection significantly attenuated IAV-induced pathology in coinfected mice. Coinfected animals also exhibited a tendency toward reduced IAV replication in the lungs as measured by qPCR, TCID₅₀ and HAs compared to mice infected with IAV alone, accompanied by lower levels of virus-specific antibodies to IAV at DPI 30 and TTSuV1 at DPI 15 respectively. At DPI 30, TTSuV1 genomic DNA levels in lung tissue and whole blood were higher in coinfected mice, suggestive of prolonged viremia in the coinfected group. Collectively, these findings establish baseline parameters for a murine TTV and influenza coinfection model and provide a foundation for future studies aimed at elucidating the molecular and immunological mechanisms underlying viral coinfections. Full article

Caprine herpesvirus 1 (CpHV-1) is responsible for significant economic losses in goat farming. The CpHV-1 genital infection in goats has been used as a homologous animal model for the study of human herpes simplex virus type 2 (HSV-2). This study aimed to investigate the in vitro virucidal and antiviral effect of lemon juice (LJ) and its main component, citric acid (CA), against CpHV-1 on Madin-Darby Bovine Kidney (MDBK) cells. Cytotoxicity was assessed using an XTT assay, while viral titers were determined by the Reed–Muench method and viral DNA was quantified via qPCR. Pure LJ (pH 2.3) and its corresponding CA solution demonstrated potent and rapid virucidal activity, reducing the viral titer by over 5.0 log10 TCID₅₀/50 µL within 1 min. When applied after viral entry, a non-cytotoxic dilution of LJ (pH 4.32) significantly inhibited viral replication, causing a 2.5 log10 TCID₅₀/50 µL reduction in viral titer and a corresponding decrease in viral DNA. The antiviral effects were minimal at a near-neutral pH of 6.67, probably interacting with envelope structures. These results suggest that LJ could be a potential low-cost topical agent or disinfectant for controlling CpHV-1 in goat populations and offer a basis for translational research on human herpesviruses. Full article

Background/Objectives: Recombinant adeno-associated virus (rAAV) has become a leading vector in gene therapy. However, manufacturing limitations may result in replication-competent AAV (rcAAV) contamination of clinical rAAV products, posing safety risks. Rigorous testing is therefore essential, and the use of accurately calibrated rcAAV reference standard materials is critical for ensuring assay stability and reliability. A disadvantage of the widely used Tissue Culture Infectious Dose 50 (TCID₅₀) assay is its high variability. This study introduces an optimized TCID₅₀ assay for the precise quantification of infectious rcAAV particles. Methods: We developed a TCID₅₀ assay tailored to rep2-based rcAAV, optimizing key aspects such as viral infection conditions, qPCR reaction systems, and standard curve preparation. We employed an innovative strategy to prepare the standard curve using serial dilutions of rcAAV in cell lysate, ensuring alignment with the test sample matrices. Results: The rcAAV-derived standard curve demonstrated exceptional linearity (R² > 0.99), sensitivity (LOQ ≈ 38 copies), and reproducibility, enabling robust endpoint qPCR analysis. The optimized assay significantly improved the precision of the TCID₅₀ assay, as an inter-assay coefficient of variation (CV) of 11.4% was achieved. Conclusions: This refined TCID₅₀ assay is a reliable method for calibrating infectious titers of rcAAV reference standard materials, thereby enabling the standardization of rcAAV testing. Full article