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Search Results (286)

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27 pages, 5867 KiB  
Article
Distinct Virologic Properties of African and Epidemic Zika Virus Strains: The Role of the Envelope Protein in Viral Entry, Immune Activation, and Neuropathogenesis
by Ashkan Roozitalab, Chenyu Zhang, Jiantao Zhang, Ge Li, Chengyu Yang, Wangheng Hou, Qiyi Tang and Richard Y. Zhao
Pathogens 2025, 14(7), 716; https://doi.org/10.3390/pathogens14070716 - 19 Jul 2025
Viewed by 325
Abstract
The 2016 Zika virus (ZIKV) epidemic has largely subsided, but a key question remains. How did ZIKV evolve to become a virulent human pathogen compared to the virus of its original discovery? What specific virologic and pathologic changes contributed to increased pathogenicity in [...] Read more.
The 2016 Zika virus (ZIKV) epidemic has largely subsided, but a key question remains. How did ZIKV evolve to become a virulent human pathogen compared to the virus of its original discovery? What specific virologic and pathologic changes contributed to increased pathogenicity in humans? Phylogenetic studies have identified two genetically distinct ZIKV, the African and Asian lineages, which differ in their pathogenicity. Previous studies including ours suggest that the envelope (E) protein plays a key role in viral entry, immune activation, and neuropathogenesis. This study aimed to further elucidate virologic and pathogenic differences between these lineages by assessing their ability to bind and replicate in host cells, induce apoptotic cell death, trigger inflammatory responses, and influence human neural progenitor cell (hNPC)-derived neurosphere formation. We compared a historic African ZIKV strain (MR766) with an epidemic Brazilian strain (BR15) and evaluated the effects of the E protein inhibitor quercetin-3-β-O-D-glucoside (Q3G) and an E protein-neutralizing antibody (AbII). Our results revealed distinct virologic properties and that MR766 exhibited stronger inhibition of neurosphere formation due to enhanced viral binding to neuronal SH-SY5Y cells, while BR15 infection triggered a heightened pro-inflammatory cytokine response with reduced viral binding. Chimeric virus studies suggested that the E protein likely influences viral binding, replication efficiency, immune activation, and neuropathogenesis. Notably, Q3G exhibited antiviral activities against both MR766 and BR15, whereas AbII preferentially inhibited MR766. These findings highlight the virological differences between ancestral and epidemic viral strains, as well as the critical role of E protein in viral permissiveness, immune response, and neuropathogenesis, providing insights for developing targeted antiviral strategies. Full article
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26 pages, 3044 KiB  
Article
Optimization of YF17D-Vectored Zika Vaccine Production by Employing Small-Molecule Viral Sensitizers to Enhance Yields
by Sven Göbel, Tilia Zinnecker, Ingo Jordan, Volker Sandig, Andrea Vervoort, Jondavid de Jong, Jean-Simon Diallo, Peter Satzer, Manfred Satzer, Kai Dallmeier, Udo Reichl and Yvonne Genzel
Vaccines 2025, 13(7), 757; https://doi.org/10.3390/vaccines13070757 - 16 Jul 2025
Viewed by 836
Abstract
Background: Modern viral vector production needs to consider process intensification for higher yields from smaller production volumes. However, innate antiviral immunity triggered in the producer cell may limit virus replication. While commonly used cell lines (e.g., Vero or E1A-immortalised cells) are already compromised [...] Read more.
Background: Modern viral vector production needs to consider process intensification for higher yields from smaller production volumes. However, innate antiviral immunity triggered in the producer cell may limit virus replication. While commonly used cell lines (e.g., Vero or E1A-immortalised cells) are already compromised in antiviral pathways, the redundancy of innate signaling complicates host cell optimization by genetic engineering. Small molecules that are hypothesized to target antiviral pathways (Viral Sensitizers, VSEs) added to the culture media offer a versatile alternative to genetic modifications to increase permissiveness and, thus, viral yields across multiple cell lines. Methods: To explore how the yield for a chimeric Zika vaccine candidate (YF-ZIK) could be further be increased in an intensified bioprocess, we used spin tubes or an Ambr15 high-throughput microbioreactor system as scale-down models to optimize the dosing for eight VSEs in three host cell lines (AGE1.CR.pIX, BHK-21, and HEK293-F) based on their tolerability. Results: Addition of VSEs to an already optimized infection process significantly increased infectious titers by up to sevenfold for all three cell lines tested. The development of multi-component VSE formulations using a design of experiments approach allowed further synergistic titer increases in AGE1.CR.pIX cells. Scale-up to 1 L stirred-tank bioreactors and 3D-printed mimics of 200 or 2000 L reactors resulted in up to threefold and eightfold increases, respectively. Conclusions: Addition of single VSEs or combinations thereof allowed a further increase in YF-ZIK titers beyond the yield of an already optimized, highly intensified process. The described approach validates the use of VSEs and can be instructive for optimizing other virus production processes. Full article
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16 pages, 13905 KiB  
Article
Replication of Vectored Herpesvirus of Turkey (HVT) in a Continuous, Microcarrier-Independent Suspension Cell Line from Muscovy Duck
by Karoline Mähl, Deborah Horn, Sirine Abidi, Benedikt B. Kaufer, Volker Sandig, Alexander Karlas and Ingo Jordan
Vaccines 2025, 13(7), 714; https://doi.org/10.3390/vaccines13070714 - 30 Jun 2025
Viewed by 463
Abstract
Background/Objectives: More than 33 billion chickens are industrially raised for meat and egg production globally and vaccinated against Marek’s disease virus (MDV). The antigenically related herpesvirus of turkey (HVT) is used as a live-attenuated vaccine, commonly provided as a recombinant vector to protect [...] Read more.
Background/Objectives: More than 33 billion chickens are industrially raised for meat and egg production globally and vaccinated against Marek’s disease virus (MDV). The antigenically related herpesvirus of turkey (HVT) is used as a live-attenuated vaccine, commonly provided as a recombinant vector to protect chickens against additional unrelated pathogens. Because HVT replicates in a strictly cell-associated fashion to low levels of infectious units, adherent primary chicken or duck embryo fibroblasts are infected, dislodged from the cultivation surface and distributed as cryocultures in liquid nitrogen to the site of application. Although viable cells are complex products, application of infected cells in ovo confers protection even in presence of maternal antibodies. Methods/Results: The aim of our study was to determine whether a continuous cell line in a scalable cultivation format can be used for production of HVT-based vaccines. The AGE1.CR cell line (from Muscovy duck) was found to be highly permissive in adherent cultures. Propagation in suspension, however, initially gave very low yields. The induction of cell-to-cell contacts in carrier-independent suspensions and a metabolic shock improved titers to levels suitable for vaccine production (>105 infectious units/mL after infection with multiplicity of 0.001). Conclusions: Production of HVT is challenging to scale to large volumes and the reliance on embryonated eggs from biosecure facilities is complex. We demonstrate that a cell-associated HVT vector can be propagated in a carrier-independent suspension culture of AGE1.CR cells in chemically defined medium. The fed-batch production is independent of primary cells and animal-derived material and can be scaled to large volumes. Full article
(This article belongs to the Special Issue Animal Herpesviruses: 2nd Edition)
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25 pages, 11397 KiB  
Article
Impact of Airflow Disturbance from Human Motion on Contaminant Control in Cleanroom Environments: A CFD-Based Analysis
by Abiyeva Guldana, Sayat Niyetbay, Arman Zhanguzhinov, Gulbanu Kassabekova, Dilyara Jartayeva, Kulyash Alimova, Gulnaz Zhakapbayeva and Khalkhabay Bostandyk
Buildings 2025, 15(13), 2264; https://doi.org/10.3390/buildings15132264 - 27 Jun 2025
Viewed by 401
Abstract
The growing demands for sanitary regulations in medical facilities, particularly operating rooms, highlight the importance of ensuring high air quality and minimizing airborne hospital-acquired infections. Improperly designed ventilation systems may lead to contamination of up to 90–95% of patients, especially in light of [...] Read more.
The growing demands for sanitary regulations in medical facilities, particularly operating rooms, highlight the importance of ensuring high air quality and minimizing airborne hospital-acquired infections. Improperly designed ventilation systems may lead to contamination of up to 90–95% of patients, especially in light of evolving threats, such as COVID-19. This study focuses on enhancing the energy efficiency and performance of air conditioning and ventilation systems for cleanrooms, where air recirculation is not permissible. A novel energy-efficient direct-flow air treatment scheme is proposed, integrating a heat pump system with adjustable thermal output. A computational fluid dynamics CFD model of a clean operating room was developed to assess the impact of inlet air velocity on aerosol particle removal and airflow stabilization time. The model also considers the effect of personnel movement. The results supported optimized air distribution, reducing microbial contamination risks, with less than 10 CFU/m3, and improved thermal performance. The proposed system was evaluated for energy and cost efficiency compared to conventional setups. Findings can inform the design and operation of cleanroom ventilation in surgical environments and other high-tech applications. This research contributes to improving indoor air quality and reducing infection risks while enhancing sustainability in healthcare infrastructure. Full article
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18 pages, 6973 KiB  
Article
TRIM5α/Cyclophilin A-Modified MDBK Cells for Lentiviral-Based Gene Editing
by Lijing Wo, Shuhui Qi, Yongqi Guo, Chao Sun and Xin Yin
Viruses 2025, 17(7), 876; https://doi.org/10.3390/v17070876 - 21 Jun 2025
Viewed by 450
Abstract
The human immunodeficiency virus 1 (HIV-1)-based lentivirus has been widely used for genetic modification. However, the efficiency of lentiviral-based gene modification in Madin–Darby bovine kidney (MDBK) cells is considerably limited. In this study, we have shown that siRNA-mediated depletion of TRIM5α, a [...] Read more.
The human immunodeficiency virus 1 (HIV-1)-based lentivirus has been widely used for genetic modification. However, the efficiency of lentiviral-based gene modification in Madin–Darby bovine kidney (MDBK) cells is considerably limited. In this study, we have shown that siRNA-mediated depletion of TRIM5α, a restriction factor in HIV-1 infection, can dramatically enhance HIV-1 infection in MDBK cells. Furthermore, we generated a doxycycline-inducible Cas9-overexpressing MDBK cell line (MDBK-iCas9) suitable for CRISPR/Cas9-mediated editing. On this basis, we created a TRIM5α knock-out MDBK-iCas9 cell line MDBK-iCas9TRIM5α−/− without additional genome insertions by combining sgRNA transfection and single-cell cloning. We found that MDBK-iCas9TRIM5α−/− displayed greater permissiveness to lentivirus infection compared with MDBK-WT cells. Notably, we found that treatment with the chemical compound cyclosporine A, which directly interacts with cell factor cyclophilin A (CypA), could markedly increase the infectivity of lentivirus in both MDBK-iCas9TRIM5α−/− and MDBK-WT cell lines, suggesting that CypA functions independently with TRIM5α as an inhibitor of the lentivirus in bovine cells. Therefore, combining bovine TRIM5α and CypA targeting could remarkably enhance lentivirus infection. In conclusion, our findings highlight a promising gene engineering strategy for bovine cells that can surmount the significant barriers to investigating the interplay between bovine viruses and their host cells. Full article
(This article belongs to the Special Issue Pestivirus 2025)
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12 pages, 4737 KiB  
Communication
Myricetin Restricts the Syncytial Development Triggered by Nipah Virus Envelope Glycoproteins In Vitro
by Ananda Murali Rayapati, Chanda Chandrasekhar, Sudarsana Poojari and Bhadra Murthy Vemulapati
Viruses 2025, 17(6), 827; https://doi.org/10.3390/v17060827 - 7 Jun 2025
Viewed by 506
Abstract
Background and Objectives: Myricetin, a flavonoid compound, was demonstrated to effectively arrest the cell-to-cell fusion and syncytial development triggered by Nipah virus (NiV) fusion (F) and attachment (G) envelope glycoproteins in vitro involving two permissive mammalian cell lines. Methods: Time-of-addition assays were carried [...] Read more.
Background and Objectives: Myricetin, a flavonoid compound, was demonstrated to effectively arrest the cell-to-cell fusion and syncytial development triggered by Nipah virus (NiV) fusion (F) and attachment (G) envelope glycoproteins in vitro involving two permissive mammalian cell lines. Methods: Time-of-addition assays were carried out using codon-optimized NiV wild type (WT) F and G plasmids followed by a challenge with the addition of myricetin 1 h and 6 h post-transfection in HEK 293T and Vero cells. Results: Upon evaluating different myricetin concentrations, it was determined that a 100 μM concentration of myricetin effectively inhibited 64–80% of syncytia in HEK and Vero cells. Interpretation & Conclusions: In this study, we concluded that myricetin mitigated the syncytial development in HEK and Vero cell lines. Given the flavonoid attributes of myricetin which is widely present in fruits, vegetables, tea, and wine, it may be regarded as a phytonutrient and a safer antiviral alternative against Nipah virus infections. Due to the BSL-4 nature of the virus, further research involving live virus culture is necessary to confirm myricetin as a potential antiviral compound for the mitigation of pathological effects of NiV infections. Full article
(This article belongs to the Section General Virology)
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13 pages, 2372 KiB  
Case Report
Reactivation of Latent Tuberculosis Following COVID-19 and Epstein-Barr Virus Coinfection: A Case Report
by Iryna Halabitska, Pavlo Petakh, Valentyn Oksenych and Oleksandr Kamyshnyi
Pathogens 2025, 14(5), 488; https://doi.org/10.3390/pathogens14050488 - 16 May 2025
Viewed by 942
Abstract
Background: This case is unique in demonstrating the reactivation of latent tuberculosis (TB) following co-infection with SARS-CoV-2 and Epstein–Barr virus (EBV) in an otherwise healthy young adult. It highlights a rare clinical scenario in which viral immune dysregulation likely facilitated TB progression. To [...] Read more.
Background: This case is unique in demonstrating the reactivation of latent tuberculosis (TB) following co-infection with SARS-CoV-2 and Epstein–Barr virus (EBV) in an otherwise healthy young adult. It highlights a rare clinical scenario in which viral immune dysregulation likely facilitated TB progression. To date, few reports have explored the complex interplay between COVID-19, EBV reactivation, and TB in a single patient, particularly with isolated extrapulmonary involvement. Case Presentation: A 24-year-old woman presented with persistent low-grade fever, fatigue, night sweats, unintentional weight loss, and progressive cervical and supraclavicular lymphadenopathy. These symptoms emerged shortly after a moderate COVID-19 infection. Laboratory studies revealed elevated inflammatory markers and pronounced lymphopenia. EBV reactivation was confirmed via serology and PCR. Despite antiviral therapy, symptoms persisted, and imaging revealed necrotic lymphadenopathy. Tuberculous lymphadenitis was diagnosed through fine-needle aspiration cytology and PCR detection of Mycobacterium tuberculosis. The patient was treated with a standard anti-tuberculosis regimen, resulting in clinical, radiological, and immunological improvement. Conclusions: This case underscores the importance of considering latent TB reactivation in patients with persistent lymphadenopathy and recent viral infections, particularly in regions with high TB prevalence. It also emphasizes the need for thorough immunological and microbiological assessment in complex post-viral syndromes. The main clinical takeaway is that COVID-19 and EBV co-infection may create a permissive environment for TB reactivation through immune system compromise. Full article
(This article belongs to the Special Issue Herpesvirus Latency and Reactivation)
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21 pages, 21042 KiB  
Article
Lassa Virus Infection of Primary Human Airway Epithelial Cells
by Helena Müller-Kräuter, Sarah Katharina Fehling, Lucie Sauerhering, Birthe Ehlert, Janine Koepke, Juliane Schilling, Mikhail Matrosovich, Andrea Maisner and Thomas Strecker
Viruses 2025, 17(5), 592; https://doi.org/10.3390/v17050592 - 22 Apr 2025
Viewed by 1024
Abstract
Lassa mammarenavirus (LASV), a member of the family Arenaviridae, is a highly pathogenic virus capable of causing severe systemic infections in humans. The primary host reservoir is the Natal multimammate mouse (Mastomys natalensis), with human infections typically occurring through mucosal exposure [...] Read more.
Lassa mammarenavirus (LASV), a member of the family Arenaviridae, is a highly pathogenic virus capable of causing severe systemic infections in humans. The primary host reservoir is the Natal multimammate mouse (Mastomys natalensis), with human infections typically occurring through mucosal exposure to virus-containing aerosols from rodent excretions. To better understand the molecular mechanisms underlying LASV replication in the respiratory tract, we utilized differentiated primary human airway epithelial cells (HAECs) grown under air–liquid interface conditions, closely mimicking the bronchial epithelium in vivo. Our findings demonstrate that HAECs are permissive to LASV infection and support productive virus replication. While LASV entry into polarized HAECs occurred through both apical and basolateral surfaces, progeny virus particles were predominantly released from the apical surface, consistent with an intrinsic apical localization of the envelope glycoprotein GP. This suggests that apical virus shedding from infected bronchial epithelia may facilitate LASV transmission via airway secretions. Notably, limited basolateral release at later stages of infection was associated with LASV-induced rearrangement of the actin cytoskeleton, resulting in compromised epithelial barrier integrity. Finally, we demonstrate that LASV-infected HAECs exhibited a pronounced type III interferon response. A detailed understanding of LASV replication and host epithelial responses in the respiratory tract could facilitate the development of targeted future therapeutics. Full article
(This article belongs to the Special Issue Viral Infection in Airway Epithelial Cells)
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21 pages, 11598 KiB  
Article
Organoids with a Type 1 Collagen Scaffold to Model Bacterial Cancer Therapy
by Lydia Farrell, Cleo Bonnet, Alethea Tang, Severina Peneva, Non G. Williams, Sunil Dolwani, Lee Parry and Paul Dyson
Cells 2025, 14(7), 524; https://doi.org/10.3390/cells14070524 - 1 Apr 2025
Cited by 1 | Viewed by 928
Abstract
Bacterial cancer therapy (BCT) is emerging as an important option for the treatment of solid tumours, with promising outcomes in preclinical trials. Further progress is hampered by an incomplete understanding of how oncotropic bacteria, such as attenuated strains of Salmonella enterica serovar Typhimurium, [...] Read more.
Bacterial cancer therapy (BCT) is emerging as an important option for the treatment of solid tumours, with promising outcomes in preclinical trials. Further progress is hampered by an incomplete understanding of how oncotropic bacteria, such as attenuated strains of Salmonella enterica serovar Typhimurium, colonise tumours and the responses of both the bacteria and tumour cells to this colonisation. To model this, we developed organoids that are permissive for bacterial colonisation, replacing the conventional commercially available extracellular matrix (e.g., Matrigel) with a type I collagen matrix scaffold. A comparison of the two extracellular matrices indicated that type 1 collagen permitted an initial infection efficiency more than 5-times greater than with Matrigel. In addition, subsequent growth within type 1 collagen expanded bacterial cell numbers by over 10-fold within 4 days of infection. These organoids allow for the visualisation of bacterial chemoattraction, cell invasion and subsequent population of the interior lumen, and will permit the future optimisation of BCT. In addition, by establishing patient-derived organoids, we demonstrate a platform for developing future personalised treatments exploiting BCT. Full article
(This article belongs to the Section Cell Methods)
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15 pages, 2074 KiB  
Article
The ACE2 Receptor from Common Vampire Bat (Desmodus rotundus) and Pallid Bat (Antrozous pallidus) Support Attachment and Limited Infection of SARS-CoV-2 Viruses in Cell Culture
by Abhijeet Bakre, Ryan Sweeney, Edna Espinoza, David L. Suarez and Darrell R. Kapczynski
Viruses 2025, 17(4), 507; https://doi.org/10.3390/v17040507 - 31 Mar 2025
Viewed by 579
Abstract
During the COVID-19 pandemic, severe acute respiratory syndrome coronavirus 2 (SC2) infection was confirmed in various animal species demonstrating a wide host range of the virus. Prior studies have shown that the ACE2 protein is the primary receptor used by the virus to [...] Read more.
During the COVID-19 pandemic, severe acute respiratory syndrome coronavirus 2 (SC2) infection was confirmed in various animal species demonstrating a wide host range of the virus. Prior studies have shown that the ACE2 protein is the primary receptor used by the virus to gain cellular entry and begin the replication cycle. In previous studies, we demonstrated that human and various bat ACE2 proteins can be utilized by SC2 viruses for entry. Bats are a suspected natural host of SC2 because of genetic homology with other bat coronaviruses. In this work, we demonstrate that expression of ACE2 genes from the common vampire bat (CVB) (Desmodus rotundus) and the pallid bat (PB) (Antrozous pallidus), supports infection and replication of some SC2 viruses in cell culture. Two cell lines were produced, CVB-ACE2 and PB-ACE2, expressing ACE2 from these bat species along with human TMPRSS2, in a model previously established using a non-permissive chicken DF-1 cell line. Results demonstrate that the original Wuhan lineage (WA1) virus and the Delta variant were able to infect and replicate in either of the bat ACE2 cell lines. In contrast, the Lambda and Omicron variant viruses infected both cell lines, but viral titers did not increase following infection. Viral detection using immunofluorescence demonstrated abundant spike (S) protein staining for the WA1 and Delta variants but little signal for the Lambda and Omicron variants. These studies demonstrate that while ACE2 from CVB and PB can be utilized by SC2 viruses to gain entry for infection, later variants (Lambda and Omicron) replicate poorly in these cell lines. These observations suggest more efficient human adaption in later SC2 variants that become less fit for replication in other animal species. Full article
(This article belongs to the Special Issue Multiple Hosts of SARS-CoV-2, 3rd Edition)
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17 pages, 21045 KiB  
Article
Experimental Infection of Rats with Influenza A Viruses: Implications for Murine Rodents in Influenza A Virus Ecology
by Long Li, Rirong Chen, Zhigang Yan, Qinglong Cai, Yi Guan and Huachen Zhu
Viruses 2025, 17(4), 495; https://doi.org/10.3390/v17040495 - 29 Mar 2025
Cited by 1 | Viewed by 1077
Abstract
Rattus norvegicus (brown rat), a widely distributed rodent and common biomedical model, is a known reservoir for many zoonotic pathogens but has not been traditionally recognized as a host for influenza A virus (IAV). To evaluate their susceptibility, we intranasally inoculated Sprague-Dawley rats [...] Read more.
Rattus norvegicus (brown rat), a widely distributed rodent and common biomedical model, is a known reservoir for many zoonotic pathogens but has not been traditionally recognized as a host for influenza A virus (IAV). To evaluate their susceptibility, we intranasally inoculated Sprague-Dawley rats with various IAV subtypes, including H5Nx, H7N9, H9N2, H10N8 and the 2009 pandemic H1N1. All strains productively infected the rats, inducing seroconversion without overt clinical signs. While replication efficiency varied, all viruses caused significant lung injury with a preferential tropism for the upper respiratory tract. Investigation of receptor distribution revealed a predominance of α2,3-linked sialic acid (SA) in the nasal turbinates and trachea, whereas α2,6-linked SA was more abundant in the lungs. Notably, both receptor types coexisted throughout the respiratory tract, aligning with the observed tissue-specific replication patterns and broad viral infectivity. These findings demonstrate that rats are permissive hosts for multiple IAV subtypes, challenging their exclusion from IAV ecology. The asymptomatic yet pathogenic nature of infection, combined with the global synanthropy of rats, underscores their potential role as cryptic reservoirs in viral maintenance and transmission. This study highlights the need for expanded surveillance of rodents in influenza ecology to mitigate zoonotic risks. Full article
(This article belongs to the Special Issue Advances in Animal Influenza Virus Research: Third Edition)
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14 pages, 5306 KiB  
Article
Environmental Monitoring and Thermal Data Analysis Related to Mortality Rates in a Commercial Pig House
by Hyo-Jae Seo, Byung-Wook Oh and Il-Hwan Seo
Agriculture 2025, 15(6), 635; https://doi.org/10.3390/agriculture15060635 - 17 Mar 2025
Cited by 1 | Viewed by 707
Abstract
Diseases in pig houses not only hinder the growth and productivity of pigs but also result in significant economic losses for farmers due to high mortality rates. Although viral infections, including PRRS and PCV-2, are the primary causes, the likelihood of disease onset [...] Read more.
Diseases in pig houses not only hinder the growth and productivity of pigs but also result in significant economic losses for farmers due to high mortality rates. Although viral infections, including PRRS and PCV-2, are the primary causes, the likelihood of disease onset is closely linked to the pigs’ immune status, which is often compromised by environmental stressors. This study aimed to investigate the relationship between environmental conditions and pig mortality through detailed field monitoring in a commercial pig house with 600 growing pigs. The facility, which experienced a surge in mortality after a ventilation system change, was analyzed for various environmental parameters, including ammonia concentration (range: 7.0–10.7 ppm), dust levels (PM10: 106 µg/m3, PM2.5: 45 µg/m3), ventilation rates (0.49 AER, 67% of design capacity), air temperature (mean: 22.3 °C, range: 18.1–28.7 °C), and relative humidity (mean: 67.4%, range: 55.3–83.2%). Pig mortality and its spatial distribution were recorded, while viral infections were identified using RT-PCR, detecting pathogens such as PRRS, PCV-2, Mycoplasma hyopneumoniae, and Salmonella. Our findings revealed that although dust and ammonia concentrations remained within permissible limits, mortality was significantly correlated with thermal instability. Chronic respiratory diseases were observed in regions where ventilation was concentrated, resulting in daily temperature variations as high as 6.64 °C. The combination of improper ventilation and frequent temperature fluctuations weakened the pigs’ immunity, facilitating the onset of disease. This research underscores the critical role of maintaining stable microclimatic conditions in reducing mortality and highlights the need for advanced automated environmental control systems in smart livestock barns. The insights gained from this study provide a foundational framework for developing precision ventilation and thermal management strategies to enhance productivity and animal welfare. Full article
(This article belongs to the Section Artificial Intelligence and Digital Agriculture)
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12 pages, 1082 KiB  
Article
Serological Assays Reveal No Evidence of Natural SARS-CoV-2 Infection in US Cattle
by Santhamani Ramasamy, Meysoon Quraishi, Swastidipa Mukherjee, Sonalika Mahajan, Lindsey C. LaBella, Shubhada K. Chothe, Padmaja Jakka, Abhinay Gontu, Sougat Misra, Meera Surendran-Nair, Ruth H. Nissly and Suresh V. Kuchipudi
Microorganisms 2025, 13(3), 600; https://doi.org/10.3390/microorganisms13030600 - 5 Mar 2025
Viewed by 777
Abstract
Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) continues to pose a significant threat to public health. Notably, SARS-CoV-2 demonstrates the capacity to infect various non-human animal species, including both captive and free-living animals. Earlier experimental studies revealed low susceptibility of domestic cattle (Bos [...] Read more.
Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) continues to pose a significant threat to public health. Notably, SARS-CoV-2 demonstrates the capacity to infect various non-human animal species, including both captive and free-living animals. Earlier experimental studies revealed low susceptibility of domestic cattle (Bos taurus) to ancestral B.1 lineage; however, recent experimental findings indicate greater permissiveness of cattle to SARS-CoV-2 Delta variant. While some studies detected evidence of SARS-CoV-2 infection in cattle in Italy, Germany, India, and Nigeria, currently, there is no evidence of SARS-CoV-2 infections in US cattle. We have investigated over 600 samples, including pre-pandemic and pandemic cattle sera collected from Pennsylvania for the presence of SARS-CoV-2 antibodies. Since serological tests have inherent problems of false positives and negatives, we conducted a comprehensive assessment of multiple serological assays. As there are no known SARS-CoV-2 positive cattle serum samples, we used hyperimmune serum raised in cattle with SARS-CoV-2-spike receptor binding domain (RBD) as positive control for the test validation. We found that pseudovirus neutralization assays with a luciferase reporter system can produce false positive results, and care must be taken to interpret serological diagnosis using these assays. We found no serological evidence of natural SARS-CoV-2 infection or transmission among cattle in the US. This study underscores the importance of robust evaluation when employing serological assays for SARS-CoV-2 detection in cattle populations. Full article
(This article belongs to the Section Virology)
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25 pages, 5366 KiB  
Article
Characterization of Human Cytomegalovirus (HCMV) Long Non-Coding RNA1.2 During Lytic Replication
by Salomé Manska, Andrew Hagemann, Janna Magana, Cyprian C. Rossetto and Subhash C. Verma
Viruses 2025, 17(2), 149; https://doi.org/10.3390/v17020149 - 23 Jan 2025
Viewed by 1033
Abstract
During lytic replication of human cytomegalovirus (HCMV), the most abundant viral transcripts are long non-coding RNAs (lncRNAs). Viral lncRNAs can have a variety of functions, some of which are necessary for viral production and the modulation of host processes during infection. HCMV produces [...] Read more.
During lytic replication of human cytomegalovirus (HCMV), the most abundant viral transcripts are long non-coding RNAs (lncRNAs). Viral lncRNAs can have a variety of functions, some of which are necessary for viral production and the modulation of host processes during infection. HCMV produces four lncRNAs, Beta2.7 (RNA2.7), RNA4.9, RNA5.0 and RNA1.2. While there has been research on these viral lncRNAs, many of their functions remain uncharacterized. To determine the function of RNA1.2, we explored its requirement during lytic infection by generating viral mutants containing either a full or partial deletion of the RNA1.2 locus. Within permissive fibroblasts, the RNA1.2 deletion mutants showed no defects in viral DNA synthesis, transcript expression, protein production, or generation of viral progeny. Further investigation to identify potential cellular and viral protein binding partners of RNA1.2 was performed using liquid chromatography-mass spectrometry (LC-MS). A significant number of cellular proteins were identified and associated with RNA1.2. Specifically those associated with the innate immune response, mitochondrial processes, and cell cycle regulation. While RNA1.2 is dispensable for lytic replication, these findings suggest it may play a pivotal role in modulating the host response. Full article
(This article belongs to the Special Issue Epigenetic and Transcriptional Regulation of DNA Virus Infections)
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16 pages, 1709 KiB  
Article
Intra- and Interspecies Conjugal Transfer of Plasmids in Gram-Negative Bacteria
by Julia R. Savelieva, Daria A. Kondratieva and Maria V. Golikova
Biomedicines 2025, 13(1), 238; https://doi.org/10.3390/biomedicines13010238 - 20 Jan 2025
Viewed by 1105
Abstract
Background/Objectives: Plasmid-mediated resistance is a significant mechanism that contributes to the gradual decrease in the efficacy of antibiotics from various classes, including carbapenems. The aim of this study is to investigate the frequency of transfer of carbapenemase-encoding plasmids from K. pneumoniae to E. [...] Read more.
Background/Objectives: Plasmid-mediated resistance is a significant mechanism that contributes to the gradual decrease in the efficacy of antibiotics from various classes, including carbapenems. The aim of this study is to investigate the frequency of transfer of carbapenemase-encoding plasmids from K. pneumoniae to E. coli and P. aeruginosa. Methods: Matings were performed on agar with subsequent isolation of transconjugant, recipient, and donor colonies. The frequency of conjugation (CF) and minimum inhibitory concentrations (MICs) of meropenem were determined for the PCR-confirmed transconjugants. A pharmacodynamic study was conducted using a hollow-fiber infection model on E. coli transconjugant in order to evaluate its viability in the presence of therapeutic concentrations of meropenem. Results: CF for K. pneumoniae-K. pneumoniae was similar to that for K. pneumoniae-E. coli and was higher the higher was meropenem MIC of the K. pneumoniae donor. The meropenem MICs for K. pneumoniae and E. coli transconjugants were higher (0.25–4 μg/mL) compared to recipients (0.03–0.06 μg/mL). P. aeruginosa did not acquire plasmids from K. pneumoniae. In pharmacodynamic experiments, an E. coli transconjugant with MIC of 2 mg/L within the “susceptibility range”, failed to respond to meropenem treatment. Conclusions: The frequency of conjugation between K. pneumoniae and E. coli falls within a similar range. A higher permissiveness of K. pneumoniae for plasmids from K. pneumoniae, i.e., within the same species, was observed. Conjugation did not occur between K. pneumoniae and P. aeruginosa. The transconjugants with meropenem MICs with borderline susceptibility may pose a potential threat to the efficacy of meropenem. Full article
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