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15 pages, 1886 KB  
Article
Identification and Genetic Characterization of a Strain of African Horse Sickness Virus Serotype 1 and Its Safety Evaluation in a Mouse Model
by Min Zhang, Xue-Feng Wang, Si-Fan Guo, Lei Wang, Bo-Fan Fu, Jing-Wen Wang, Ya-Fen Song, Xiao-Yue Yang, Si-Yuan Hao, Qian-Yi Zhang, Bing Zhang and Cheng-Huai Yang
Microorganisms 2025, 13(10), 2314; https://doi.org/10.3390/microorganisms13102314 - 6 Oct 2025
Abstract
African horse sickness (AHS) is an arthropod-borne, severe equid disease caused by African horse sickness virus (AHSV). AHSV has high mortality and is endemic to sub-Saharan Africa. It has been classified into nine distinct serotypes (AHSV-1 to AHSV-9) based on VP2 immunogenicity. The [...] Read more.
African horse sickness (AHS) is an arthropod-borne, severe equid disease caused by African horse sickness virus (AHSV). AHSV has high mortality and is endemic to sub-Saharan Africa. It has been classified into nine distinct serotypes (AHSV-1 to AHSV-9) based on VP2 immunogenicity. The AHS outbreak in Thailand in 2020, caused by AHSV-1, marked the first occurrence of this disease in Southeast Asia. It posed a substantial threat to the security of the equine industry in the nations across the region. To ensure the emergency reserve for AHS prevention and control, the AHSV strain imported to China from abroad over 60 years ago was characterized in this study. The strain was passaged in mice and then blind-passaged in Vero cells. The plaque purification method was then used to purify the strain and obtain its cell-adapted version, named AHSV/C. Neutralization tests confirmed that the virus belongs to AHSV-1. Whole-genome sequencing revealed that AHSV/C was highly homologous to AHSV-1 isolate 1180, with over 95% homology of major antigenic protein VP2, as compared to other AHSV-1 strains, including the prevalent strain in Thailand. In the mouse models, AHSV/C exhibited no clinical signs or histopathological lesions, suggesting low virulence and safety. This research for the first time characterized the in vitro growth characteristics and viral subtypes of the AHSV in China, determined its complete whole-genome sequence, and evaluated its safety using a mouse model. It provides crucial experimental materials and scientific foundations for the development of diagnostic methods and vaccines against AHSV-1. Full article
(This article belongs to the Section Veterinary Microbiology)
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15 pages, 823 KB  
Article
Infectious Keratitis: A Tertiary Center’s Approach to Diagnosis, Management, and Enhanced Outcomes Through Microbiological Analysis
by Antonio Moramarco, Federico Cassini, Natalie di Geronimo, Giovanni Zanini, Michele Potenza, Matteo Farnè, Viviana Schisa, Erica De Carolis, Margherita Ortalli, Piera Versura, Tiziana Lazzarotto and Luigi Fontana
Microorganisms 2025, 13(10), 2308; https://doi.org/10.3390/microorganisms13102308 - 5 Oct 2025
Abstract
Background: The study aimed to assess the diagnostic and therapeutic management of infectious keratitis at a tertiary referral center, focusing on how microbiological analysis influences clinical outcomes. Methods: A retrospective review was conducted on 220 patients (221 eyes) with infectious keratitis treated between [...] Read more.
Background: The study aimed to assess the diagnostic and therapeutic management of infectious keratitis at a tertiary referral center, focusing on how microbiological analysis influences clinical outcomes. Methods: A retrospective review was conducted on 220 patients (221 eyes) with infectious keratitis treated between November 2021 and January 2025. Data collected included clinical presentation, microbiological findings, treatment approaches, and outcomes. Statistical analyses examined the relationships between microbiological results, improvements in visual acuity, and the need for rescue surgery. Results: Bacterial keratitis accounted for 64% of cases, followed by viral (20%), fungal (13%), and Acanthamoeba (3%). Microbiological testing was performed in 107 cases, with a culture positivity rate of 75.7%. Positive microbiological findings were significantly associated with better visual acuity (p = 0.019) and a reduced, though not statistically significant, need for rescue surgery. Use of contact lenses and ocular trauma were independent risk factors for culture positivity. Delayed referral (more than 15 days) was linked to longer treatment durations and a higher likelihood of surgical intervention (p < 0.001). Microbiological diagnosis correlates with improved visual outcomes and a decreased need for surgical procedures. Conclusion: Early referral and targeted therapy are essential for optimizing prognosis. The use of contact lenses and cases of ocular trauma should prompt early diagnostic sampling. Full article
(This article belongs to the Special Issue Mycosis and Antifungal Agents)
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19 pages, 1560 KB  
Article
Elimination of Airborne Microorganisms Using Compressive Heating Air Sterilization Technology (CHAST): Laboratory and Nursing Home Setting
by Pritha Sharma, Supriya Mahajan, Gene D. Morse, Rolanda L. Ward, Satish Sharma, Stanley A. Schwartz and Ravikumar Aalinkeel
Microorganisms 2025, 13(10), 2299; https://doi.org/10.3390/microorganisms13102299 - 3 Oct 2025
Abstract
Background: Airborne transmission of bacteria, viruses, and fungal spores poses a major threat in enclosed settings, particularly nursing homes where residents are highly vulnerable. Compressive Heating Air Sterilization Technology (CHAST) applies compressive heating to inactivate microorganisms without reliance on filtration or chemicals. Methods: [...] Read more.
Background: Airborne transmission of bacteria, viruses, and fungal spores poses a major threat in enclosed settings, particularly nursing homes where residents are highly vulnerable. Compressive Heating Air Sterilization Technology (CHAST) applies compressive heating to inactivate microorganisms without reliance on filtration or chemicals. Methods: CHAST efficacy was evaluated in laboratory and deployed for a feasibility and performance validation study of air sterilization in a nursing home environment. Laboratory studies tested prototypes (300–5000 CFM; 220–247 °C) against aerosolized surrogates including Bacillus globigii (Bg), B. stearothermophilus (Bst), B. thuringiensis (Bt), Escherichia coli, and MS2 bacteriophage. Viral inactivation thresholds were further assessed by exposing MS2 to progressively lower treatment temperatures (64.5–143 °C). Feasibility and performance validation evaluation involved continuous operation of two CHAST units in a nursing home, with pre- and post-treatment air samples analyzed for bacterial and fungal burden. Results: Laboratory testing demonstrated consistent microbial inactivation, with most prototypes achieving > 6-log (99.9999%) reductions across bacterial spores, vegetative bacteria, and viruses. A 5000 CFM prototype achieved > 7-log (99.99999%) elimination of B. globigii. MS2 was completely inactivated at 240 °C, with modeling suggesting a threshold for total viral elimination near 170 °C. In the feasibility study, baseline sampling revealed bacterial (35 CFU/m3) and fungal (17 CFU/m3) contamination, dominated by Bacillus, Staphylococcus, Cladosporium, and Penicillium. After 72 h of CHAST operation, discharge air contained no detectable viable organisms, and fungal spore counts showed a 93% reduction relative to baseline return air. Units maintained stable operation (464 °F ± 2 °F; 329–335 CFM) throughout deployment. Conclusion: CHAST reproducibly and scalably inactivated airborne bacteria, viruses, and fungi under laboratory and feasibility field studies, supporting its potential as a chemical-free strategy to improve infection control and indoor air quality in healthcare facilities. Full article
(This article belongs to the Section Public Health Microbiology)
23 pages, 3188 KB  
Article
Antiviral Activity of Origanum vulgare ssp. hirtum Essential Oil-Loaded Polymeric Micelles
by Neli Vilhelmova-Ilieva, Ivan Iliev, Katya Kamenova, Georgy Grancharov, Krasimir Rusanov, Ivan Atanassov and Petar D. Petrov
Biomedicines 2025, 13(10), 2417; https://doi.org/10.3390/biomedicines13102417 - 2 Oct 2025
Abstract
Background: Encapsulating essential oils in polymer-based nanocarriers can improve their stability, solubility, and bioavailability, while maintaining the biological activity of the oil’s active ingredients. In this contribution, we investigated the antiviral activity of Oregano Essential Oil (OEO) in its pure form and [...] Read more.
Background: Encapsulating essential oils in polymer-based nanocarriers can improve their stability, solubility, and bioavailability, while maintaining the biological activity of the oil’s active ingredients. In this contribution, we investigated the antiviral activity of Oregano Essential Oil (OEO) in its pure form and encapsulated into nanosized polymeric micelles, based on a poly(ethylene oxide)-block-poly(ε-caprolactone) diblock copolymer. Methods: The effect of encapsulation was evaluated using three structurally different viruses: herpes simplex virus type 1 (HSV-1) (DNA—enveloped virus), human coronavirus (HCoV OC-43) (RNA—enveloped virus), and feline calicivirus (FCV) (RNA—naked virus). The effect on the viral replicative cycle was determined using the cytopathic effect inhibition (CPE) test. Inhibition of the viral adsorption step, virucidal activity, and protective effect on healthy cells were assessed using the final dilution method and were determined as Δlg compared to the untreated viral control. Results: In both studied forms (pure and nanoformulated), OEO had no significant effect on viral replication. In the remaining antiviral experiments, the oil embedded into nanocarriers showed a slightly stronger effect than the pure oil. When the oil was directly applied to extracellular virions, viral titers were significantly reduced for all three viruses, with the effect being strongest for HSV-1 and FCV (Δlg = 3.5). A distinct effect was also observed on the viral adsorption stage, with the effect being most significant for HSV-1 (Δlg = 3.0). Conclusions: Pretreatment of healthy cells with the nanoformulated OEO significantly protected them from viral infection, with the greatest reduction in viral titer for HCoV OC-43. Full article
(This article belongs to the Special Issue Recent Advances in Targeted Drug Delivery Systems)
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14 pages, 4597 KB  
Article
Exogenous Application of IR-Specific dsRNA Inhibits Infection of Cucumber Green Mottle Mosaic Virus in Watermelon
by Yanhui Wang, Liming Liu, Yongqiang Fan, Yanli Han, Zhiling Liang, Yanfei Geng, Fengnan Liu, Qinsheng Gu, Baoshan Kang and Chaoxi Luo
Agronomy 2025, 15(10), 2332; https://doi.org/10.3390/agronomy15102332 - 2 Oct 2025
Abstract
Cucumber green mottle mosaic virus (CGMMV) represents a serious threat in the production of watermelon. Small RNAs facilitate a mechanism known as RNA interference (RNAi), which regulates gene expression. RNAi technology employs foreign double-stranded RNAs (dsRNAs) to target and reduce the expression levels [...] Read more.
Cucumber green mottle mosaic virus (CGMMV) represents a serious threat in the production of watermelon. Small RNAs facilitate a mechanism known as RNA interference (RNAi), which regulates gene expression. RNAi technology employs foreign double-stranded RNAs (dsRNAs) to target and reduce the expression levels of specific genes in plants by interfering with their mRNAs. In this study, watermelon plants were treated with dsRNAs of CGMMV MET, IR, and HEL fragments that had been generated in E. coli HT115. We investigated variations in several factors, including viral accumulation, virus-derived small interfering RNAs (vsiRNAs), and symptom severity. MET-dsRNA, IR-dsRNA and HEL-dsRNA dramatically decreased the symptoms of CGMMV in plants in the growth chamber test. Plants treated with viral-derived dsRNA showed a considerable decrease in both virus titers and vsiRNA levels. We also explored the mobility of spray-on dsRNA-derived long dsRNA and discovered that it could be identified in both inoculated leaves and the systemic leaves. IR-dsRNA outperformed MET-dsRNA and HEL-dsRNA in dsRNA therapy. Illumina sequencing of small RNAs from watermelon plants treated with IR-dsRNA and those that were not treated showed that the decreased accumulation of vsiRNAs was consistent with interference with CGMMV infection in systemic leaves. dsRNA-treated plants showed a higher level of 24-nt viral siRNA and lower level of 22-nt viral siRNA accumulation, while 22-nt viral siRNA predominated in untreated plants, indicating that dsRNA treatment improved DCL3 activity. In conclusion, our research provides deeper insights into the mechanism of antiviral RNA interference and confirms the effectiveness of applying dsRNA locally to enhance plant antiviral activity. Full article
(This article belongs to the Section Pest and Disease Management)
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17 pages, 1654 KB  
Article
Post-COVID-19 Epidemiology of Viral Infections in Adults Hospitalized with Acute Respiratory Syndromes in Palermo, South of Italy
by Mariangela Pizzo, Floriana Bonura, Federica Cacioppo, Emilia Palazzotto, Chiara Filizzolo, Sharon Russo, Daniela Pistoia, Giuseppina Capra, Donatella Ferraro, Giovanni M. Giammanco and Simona De Grazia
Pathogens 2025, 14(10), 997; https://doi.org/10.3390/pathogens14100997 - 2 Oct 2025
Abstract
This study evaluated the epidemiology and seasonal patterns of respiratory viruses in adults hospitalized with acute respiratory tract infections during two consecutive post-COVID-19 pandemic seasons. A retrospective study was conducted at the University Hospital “P. Giaccone”, Palermo, from September 2022 to September 2024. [...] Read more.
This study evaluated the epidemiology and seasonal patterns of respiratory viruses in adults hospitalized with acute respiratory tract infections during two consecutive post-COVID-19 pandemic seasons. A retrospective study was conducted at the University Hospital “P. Giaccone”, Palermo, from September 2022 to September 2024. Multiplex molecular assays were used to detect the ten respiratory viruses most relevant from an epidemiological perspective in respiratory samples (n = 1110) of 1081 patients. A respiratory viral infection was identified in 29.6% of patients. The highest viral infection rate was observed in the 31–50 age group. Human rhinovirus/enterovirus (HRV/EV) was the most frequently detected (40%), followed by influenza A virus (IAV; 18.4%) and human coronaviruses (HuCoVs; 12.8%). Viral co-infections were identified in 10.9% of positive cases, with HRV/EV, adenovirus (ADV), and parainfluenza virus (PIV) being most frequently involved. Influenza and respiratory syncytial viruses (RSVs) showed a winter seasonality, while diverse circulation patterns were revealed for the other viruses. This study demonstrated a sustained circulation of respiratory viruses in adults hospitalized with severe respiratory symptoms, with HRV/EV accounting for most of them. Syndromic multiplex molecular testing, although limited to the detection of a small fraction of epidemiologically relevant known viruses, has proven to be a valuable tool, not only for diagnostic purposes but also for acquiring genotyping data and implementing epidemiological information from sentinel surveillance systems. Full article
(This article belongs to the Section Viral Pathogens)
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13 pages, 639 KB  
Article
Hepatitis E Virus Infection in a Hospital from Southern Romania—New Data About a Threat to Public Health
by Cristina Popescu, Alexandra Cireșă, Gabriel Adrian Popescu, Carmen Cristina Vasile, Leontina Mirela Bănică and Dragoș Florea
Microorganisms 2025, 13(10), 2290; https://doi.org/10.3390/microorganisms13102290 - 1 Oct 2025
Abstract
This study analyses all cases of acute hepatitis E diagnosed in a southern Romanian hospital from 2019 to 2023. Patients with positive anti-HEV IgM antibodies were included in three groups: group A1—96 patients with probable HEV infection and ALT levels over 2.5-fold the [...] Read more.
This study analyses all cases of acute hepatitis E diagnosed in a southern Romanian hospital from 2019 to 2023. Patients with positive anti-HEV IgM antibodies were included in three groups: group A1—96 patients with probable HEV infection and ALT levels over 2.5-fold the upper limit of normal (ULN); group A2—44 patients with probable HEV infection and ALT levels under 2.5-fold ULN; group B—43 patients with probable HEV coinfection with another hepatotropic virus. Between 2019 and 2023, 642 patients were diagnosed with acute viral hepatitis. Positive anti-HEV IgM antibodies were detected in 183 (28.5%) cases, HEV being the second most common cause of acute viral hepatitis. Patients from group A were older than those from group B (47.26 ± 15.13 years vs. 35.95 ± 14.83 years, p < 0.01). Patients from group A were less likely to present clinical features compared to those from group B: digestive symptoms (73.8% vs. 97.2%, p < 0.01), jaundice (38.9% vs. 88.4%, p < 0.01), hepatomegaly (64.1% vs. 88.6%, p = 0.02). Patients from group A, compared to patients from group B, had lower levels of ALT (18.2 ± 29.9 ULN vs. 83.7 ± 56.2 ULN, p < 0.01) and total bilirubin (3.08 ± 5.2 mg/dL vs. 7.82 ± 5.25 mg/dL, p < 0.01) at admission. Patients from group A had higher levels of anti-HEV IgM antibodies (4.3 ± 3.22 vs. 2.55 ± 1.34, p < 0.01) than those from group B. Nearly all patients had no history of travel; therefore, autochthonous origin of HEV is involved in a large majority of cases. Currently, hepatitis E virus is not an uncommon aetiology of acute hepatitis in Romania, more often in adults and elderly patients. The epidemiological and clinical features of HEV infections plead for a zoonotic transmission in most cases. The significant number of cases of hepatitis E diagnosed in a single centre in Bucharest justifies the need to include early testing for HEV in patients with acute hepatitis. Full article
(This article belongs to the Special Issue Infectious Disease Surveillance in Romania: Second Edition)
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22 pages, 2609 KB  
Review
A Review of Coronaviruses in Wild Birds and Opportunities for Future Research on Migratory Waterfowl
by Allison A. Vestal-Laborde, Rebecca C. Christofferson, Kevin M. Ringelman and Ashley M. Long
Birds 2025, 6(4), 52; https://doi.org/10.3390/birds6040052 - 1 Oct 2025
Abstract
Coronaviruses (CoVs) were first described in poultry in the early 1930s and formally recognized as pathogens of both animal and human populations in the late 1960s. They are now considered among the most abundant viral families in the world. Though their distribution and [...] Read more.
Coronaviruses (CoVs) were first described in poultry in the early 1930s and formally recognized as pathogens of both animal and human populations in the late 1960s. They are now considered among the most abundant viral families in the world. Though their distribution and diversity remain understudied in wild animals, representatives from 13 orders of wild birds worldwide have tested positive for CoVs of the gamma and delta genera over the last 25 years. Many of these wild bird species are in the orders Charadriiformes (shorebirds and their relatives) and Anseriformes (waterfowl including ducks, geese, and swans). Waterfowl are particularly concerning as potential reservoirs for CoVs because they are globally distributed; often congregate in large, mixed-species flocks; and may exist in close proximity to humans and domesticated animals. This review describes the history and current knowledge of CoVs in birds, provides an updated list of global detections of CoVs in 124 species of wild birds as reported in the peer-reviewed literature since 2000, and highlights topics for future research that would help elucidate the role of waterfowl in CoV transmission. Our review reiterates the need for continuous surveillance to detect and monitor CoVs across all bird species and for standardization in data reporting and analysis of both negative and positive results. Such information is critical to understand the potential role of free-ranging birds in the maintenance, evolution, and transmission of the virus. Further, we believe that research on the potential impacts of coronavirus infections and coinfections on avian demographics, especially reproduction in waterfowl, is warranted given known consequences in domestic poultry. Full article
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13 pages, 887 KB  
Communication
Characterization of the Hemagglutinin Gene of Morbillivirus Canis in Domestic Dogs from the Mid-Western Area of Brazil
by Mayara Lima Kavasaki, Aneliza de Oliveira Souza, Amanda Noeli da Silva Campos, Isis Indaiara Gonçalves Granjeiro Taques, Rachel Vieira Paes de Barros, Sofia de Souza Pereira Gomes, Nathalia Assis Pereira, Tayane Bruna Soares Magalhães, Edson Viana Massoli Junior, Lucas Avelino D. Pavelegini, Luiz Donizete Campeiro Junior, Bruno Gomes de Castro, Michele Lunardi and Daniel Moura de Aguiar
Vet. Sci. 2025, 12(10), 948; https://doi.org/10.3390/vetsci12100948 - 30 Sep 2025
Abstract
Canine distemper virus (CDV) is a serious and often fatal disease caused by Morbillivirus canis, which affects domestic dogs and wild carnivores, with case-fatality rates reaching up to 47%. The hemagglutinin (H) protein mediates viral adsorption and shows high genetic variability, making [...] Read more.
Canine distemper virus (CDV) is a serious and often fatal disease caused by Morbillivirus canis, which affects domestic dogs and wild carnivores, with case-fatality rates reaching up to 47%. The hemagglutinin (H) protein mediates viral adsorption and shows high genetic variability, making it a valuable molecular marker. This study aimed to detect and characterize the H gene of CDV strains from 14 dogs with fatal neurological disease in the Brazilian states of Mato Grosso and Rondônia. Brain tissue was tested via RT-PCR for the nucleocapsid gene, and positive samples were amplified for the H gene. Ten complete H-gene sequences were obtained. Phylogenetic analysis revealed two distinct clusters within the South America I/Europe lineage: one related to strains from Uruguay and Argentina (with residues 530G/549Y) and another related to Brazilian strains (530S/549Y). One sequence (MT8) showed an intermediate position in the haplotype network but clustered phylogenetically with Uruguay/Argentina-related strains. Most sequences carried 530S/549Y, a pattern linked to altered SLAM receptor usage in wildlife. These findings demonstrate the co-circulation of two CDV clusters in Central–Western Brazil, their regional and international genetic connectivity, and amino acid substitutions potentially influencing host adaptation and antigenicity. Full article
(This article belongs to the Section Veterinary Microbiology, Parasitology and Immunology)
12 pages, 871 KB  
Article
First Serological Evidence of Crimean-Congo Hemorrhagic Fever Virus Infections in Croatia: A Multispecies Surveillance Approach Emphasising the Role of Sentinel Hosts
by Gorana Miletic, Ivona Coric, Snjezana Kovac, Alenka Skrinjaric, Magda Kamber Taslaman, Margarita Bozikovic, Ljubo Barbic, Viktor Masovic, Jelena Prpic, Lorena Jemersic and Vladimir Stevanovic
Viruses 2025, 17(10), 1335; https://doi.org/10.3390/v17101335 - 30 Sep 2025
Abstract
Crimean-Congo hemorrhagic fever virus (CCHFV) is a tick-borne zoonotic pathogen of growing public health concern in southeastern Europe. This study provides the first serological evidence of CCHFV circulation in Croatia, based on testing 1473 serum samples from farm and companion animals, including sheep, [...] Read more.
Crimean-Congo hemorrhagic fever virus (CCHFV) is a tick-borne zoonotic pathogen of growing public health concern in southeastern Europe. This study provides the first serological evidence of CCHFV circulation in Croatia, based on testing 1473 serum samples from farm and companion animals, including sheep, horses, cattle, goats, dogs, and cats. A total of 109 samples (7.4%) tested positive for CCHFV antibodies using a commercially available enzyme-linked immunosorbent assay (ELISA) kit. The highest seroprevalence was recorded in sheep (28.3%), followed by horses (4.3%) and a single cat (0.5%), with no antibodies detected in cattle, goats, or dogs. Almost all seropositive animals originated from coastal and subcoastal Croatia, where Hyalomma ticks are present. Only two seropositive cases were detected in continental areas. Sheep samples from several farms in Zadar County showed intra-farm seropositivity rates of up to 85.7%, suggesting localised virus circulation likely influenced by vector distribution and farm-level practices. No viral ribonucleic acid (RNA) was detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR), consistent with the transient nature of viremia in most animal hosts. These findings confirm the silent circulation of CCHFV in Croatia and reinforce the need for targeted, regionally adapted surveillance strategies that integrate multiple hosts and support early warning systems aligned with the One Health concept. Full article
(This article belongs to the Special Issue Emerging and Re-Emerging Viral Zoonoses)
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18 pages, 4107 KB  
Article
Intertypic Recombination Between Coxsackievirus A16 and Enterovirus A71 Structural and Non-Structural Genes Modulates Virulence and Protection Efficacy
by Hooi Yee Chang, Han Kang Tee, Kien Chai Ong, Kartini Jasni, Syahril Abdullah, I.-Ching Sam and Yoke Fun Chan
Vaccines 2025, 13(10), 1017; https://doi.org/10.3390/vaccines13101017 - 29 Sep 2025
Abstract
Background/Objectives: Enterovirus A71 (EV-A71) and coxsackievirus A16 (CVA16) are major causative agents of hand, foot and mouth disease (HFMD), often co-circulating and occasionally undergoing genetic recombination. While natural recombinants often involve genomic regions encoding non-structural proteins, their effects on replication and pathogenesis [...] Read more.
Background/Objectives: Enterovirus A71 (EV-A71) and coxsackievirus A16 (CVA16) are major causative agents of hand, foot and mouth disease (HFMD), often co-circulating and occasionally undergoing genetic recombination. While natural recombinants often involve genomic regions encoding non-structural proteins, their effects on replication and pathogenesis remain unclear. Methods: To address this, four chimera viruses (Chi-CCE, Chi-ECE, Chi-EEC, and Chi-CEC) were constructed with 5′UTR, capsid P1, and non-structural P2 and P3 genes, from CVA16 (denoted as C) or EV-A71 (denoted as E). These chimeras were tested for replication kinetics and cytopathic effects in rhabdomyosarcoma cells while in vivo virulence and protection efficacy were evaluated using a newborn BALB/c mouse model. Results: All chimeric viruses remained viable and exhibited higher replication than CVA16. In vivo, all chimeric viruses were avirulent except Chi-CCE and CVA16, which showed high virulence and viral titres in the brains and limbs of infected newborn mice. This suggests that 5′UTR and capsid P1 genes of CVA16 are critical genetic determinants of virulence. Notably, only the anti-inflammatory cytokine IL-10 was elevated, suggesting potential immune modulation during infection. Inactivated Chi-CCE immunisation conferred 100% protection against lethal CVA16 or mouse-adapted EV-A71 challenge revealing its potential as a bivalent vaccine candidate. Conclusions: Our study demonstrates that recombination between CVA16 and EV-A71 influences viral virulence and protection efficacy with implications for future development of multivalent vaccines. Full article
(This article belongs to the Section Vaccines against Tropical and other Infectious Diseases)
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16 pages, 654 KB  
Review
Effect of Microgravity and Space Radiation Exposure on Human Oral Health: A Systematic Review
by Shahnawaz Khijmatgar, Matteo Pellegrini, Martina Ghizzoni and Massimo Del Fabbro
Biophysica 2025, 5(4), 45; https://doi.org/10.3390/biophysica5040045 - 29 Sep 2025
Abstract
A systematic review was conducted to assess the effects of microgravity and space radiation on astronauts’ oral health. This review aimed to determine if these conditions increase the risk of dental and periodontal diseases, identify pre-mission dental care strategies, and specify relevant dental [...] Read more.
A systematic review was conducted to assess the effects of microgravity and space radiation on astronauts’ oral health. This review aimed to determine if these conditions increase the risk of dental and periodontal diseases, identify pre-mission dental care strategies, and specify relevant dental emergencies for astronauts to manage during missions. Following PRISMA guidelines, the review was registered on PROSPERO (CRD42023472765). Databases including PubMed, Scopus, Web of Science, Cochrane Library, and OVID Medline were searched. Of the 13 studies identified, 7 were eligible for qualitative synthesis. The included studies revealed that space conditions compromise oral health. Findings indicate changes in saliva composition, with a significant decline in salivary lysozyme levels during missions lasting 28 to 84 days. Salivary IgA levels also increased before and peaked after flights (microgravity alters fluid shear and protein folding). Viral reactivation was a key finding, with latent viruses such as Epstein–Barr virus (EBV), cytomegalovirus (CMV), and varicella zoster virus (VZV) being reactivated during missions (immune suppression and gene expression shifts under spaceflight stress). Data from a study found that 50% of crew members shed viruses in their saliva or urine, and 38% tested positive for herpesviruses. The included studies also documented alterations in the oral microbiome, including increased gastrointestinal and decreased nasal microbial diversity. This suggests alterations in salivary biomarkers, viral shedding, and microbiome changes in astronauts during long-duration missions. These changes appear associated with immune dysregulation and stress, but causality remains uncertain due to observational designs, small heterogeneous samples, and confounding factors. Although current evidence is indicative rather than definitive, these findings highlight the need for preventive dental measures prior to missions and preparedness for managing oral emergencies in-flight. Future studies should address the mechanistic separation of microgravity and radiation effects, with implications for upcoming Moon and Mars missions. Full article
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18 pages, 2648 KB  
Article
Whole-Blood Cellular Responses: A Promising Indicator of SARS-CoV-2 Immunity Compared to Serology
by Lucas M. Zhou, Elizabeth H. Duncan, Rupsa C. Boelig, Margaret Costanzo, Jeffrey R. Currier and Elke S. Bergmann-Leitner
J. Clin. Med. 2025, 14(19), 6889; https://doi.org/10.3390/jcm14196889 - 29 Sep 2025
Abstract
Background: Currently available immunological tests for SARS-CoV-2 assess only antibody responses. Despite the growing evidence that T cells play a crucial role in protection, especially against emerging viral variants, no routine test is available to determine T cell immunity. The prognostic value of [...] Read more.
Background: Currently available immunological tests for SARS-CoV-2 assess only antibody responses. Despite the growing evidence that T cells play a crucial role in protection, especially against emerging viral variants, no routine test is available to determine T cell immunity. The prognostic value of SARS-CoV-2-specific antibodies for determining whether individuals are immune and protected against disease remains uncertain. This is in part due to the following: (a) specificity and limitations such as the sensitivity of antibody tests, and (b) the lack of correlation between antibody titers (quantity) and the antiviral function of antibodies (quality). Approximately a quarter of SARS-CoV-2-infected patients with symptoms fail to show seroconversion in serological assays. Methods: The current report describes the development and application of a whole-blood-based assay to detect previous exposure to SARS-CoV-2. Whole blood is stimulated with SARS-CoV-2-derived peptides identified during assay development and stimulation-induced cytokines quantified using a multiplex testing platform. The resulting cytokine profiles are generated using computational tools to identify previous exposure to the virus. Results: The application of the assay revealed a lack of self-awareness of individuals’ COVID-19 infection history and demonstrated the value of this new assay to assess the prevalence of SARS-CoV-2 exposure history and immunity in populations. Conclusions: Positive responses in this assay can facilitate the identification of underlying causes of unexplained symptoms and provide clinically actionable insights for healthcare applications, including in the continued conundrum of post-acute sequela of SARS-CoV-2 infection (PASC or “long COVID”), for which both diagnosis and management remain challenging. Full article
(This article belongs to the Section Immunology & Rheumatology)
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11 pages, 914 KB  
Article
Seroprevalence of Hepatitis A in Oman Prior to National Vaccination
by Halima Al Shuaili, Mohamed El-Kassas, Khalid M. Al-Naamani, Zakaryia Al Muharrmi and Muhannad Al-Kobaisi
J. Clin. Med. 2025, 14(19), 6857; https://doi.org/10.3390/jcm14196857 - 28 Sep 2025
Abstract
Background: The hepatitis A virus (HAV) is a major cause of acute viral hepatitis and a significant global health concern. This study provides a pre-vaccination baseline for Oman, enabling longitudinal comparison with post-hepatitis A vaccination cohorts. This study aimed to determine the pre-vaccination [...] Read more.
Background: The hepatitis A virus (HAV) is a major cause of acute viral hepatitis and a significant global health concern. This study provides a pre-vaccination baseline for Oman, enabling longitudinal comparison with post-hepatitis A vaccination cohorts. This study aimed to determine the pre-vaccination seroprevalence of HAV antibodies (anti-HAV) in Oman and explore the associated demographic factors. Methods: A cross-sectional study was conducted from April 2014 to August 2015 among patients attending the medical outpatient clinic of the Medical City Hospital for Military and Security Services. Demographic data were collected via a structured questionnaire, and serum samples were tested for anti-HAV immunoglobulin IgG and IgM using enzyme-linked immunosorbent assays. Multivariate analysis was performed to identify the predictors of anti-HAV seroprevalence. Results: Among 1975 participants, 88.1% were positive for anti-HAV IgG. The mean age was 37.4 ± 16.1 years; however, those negative for anti-HAV IgG were considerably younger (mean age: 24.8 ± 15.7 years). Anti-HAV IgG seroprevalence was 37% in individuals aged ≤18 years and 91% in those >18 years (p < 0.001). The factors associated with seropositivity included older age (p < 0.001), consuming food prepared outside the home (p < 0.001), occupation (p < 0.001), and education level (p = 0.003). In the multivariable analysis, only age showed a strong independent association with serostatus: per 10-year increase, the aOR for anti-HAV IgG seropositivity was 2.87 (95% CI 2.25–3.63; p < 0.001). Conclusions: Our study estimates show high anti-HAV IgG seroprevalence and serve as a pre-vaccination baseline for evaluating the hepatitis A vaccination program in Oman over time. Given the lower natural exposure among younger cohorts, continued routine vaccination, scheduled serosurveys, and strengthened surveillance are required to identify emerging immunity gaps and prevent future HAV outbreaks. Full article
(This article belongs to the Section Epidemiology & Public Health)
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35 pages, 2417 KB  
Review
Insights into Persistent SARS-CoV-2 Reservoirs in Chronic Long COVID
by Swayam Prakash, Sweta Karan, Yassir Lekbach, Delia F. Tifrea, Cesar J. Figueroa, Jeffrey B. Ulmer, James F. Young, Greg Glenn, Daniel Gil, Trevor M. Jones, Robert R. Redfield and Lbachir BenMohamed
Viruses 2025, 17(10), 1310; https://doi.org/10.3390/v17101310 - 27 Sep 2025
Abstract
Long COVID (LC), also known as post-acute sequelae of COVID-19 infection (PASC), is a heterogeneous and debilitating chronic disease that currently affects 10 to 20 million people in the U.S. and over 420 million people globally. With no approved treatments, the long-term global [...] Read more.
Long COVID (LC), also known as post-acute sequelae of COVID-19 infection (PASC), is a heterogeneous and debilitating chronic disease that currently affects 10 to 20 million people in the U.S. and over 420 million people globally. With no approved treatments, the long-term global health and economic impact of chronic LC remains high and growing. LC affects children, adolescents, and healthy adults and is characterized by over 200 diverse symptoms that persist for months to years after the acute COVID-19 infection is resolved. These symptoms target twelve major organ systems, causing dyspnea, vascular damage, cognitive impairments (“brain fog”), physical and mental fatigue, anxiety, and depression. This heterogeneity of LC symptoms, along with the lack of specific biomarkers and diagnostic tests, presents a significant challenge to the development of LC treatments. While several biological abnormalities have emerged as potential drivers of LC, a causative factor in a large subset of patients with LC, involves reservoirs of virus and/or viral RNA (vRNA) that persist months to years in multiple organs driving chronic inflammation, respiratory, muscular, cognitive, and cardiovascular damages, and provide continuous viral antigenic stimuli that overstimulate and exhaust CD4+ and CD8+ T cells. In this review, we (i) shed light on persisting virus and vRNA reservoirs detected, either directly (from biopsy, blood, stool, and autopsy samples) or indirectly through virus-specific B and T cell responses, in patients with LC and their association with the chronic symptomatology of LC; (ii) explore potential mechanisms of inflammation, immune evasion, and immune overstimulation in LC; (iii) review animal models of virus reservoirs in LC; (iv) discuss potential T cell immunotherapeutic strategies to reduce or eliminate persistent virus reservoirs, which would mitigate chronic inflammation and alleviate symptom severity in patients with LC. Full article
(This article belongs to the Special Issue SARS-CoV-2, COVID-19 Pathologies, Long COVID, and Anti-COVID Vaccines)
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