Editor’s Choice Articles

Editor’s Choice articles are based on recommendations by the scientific editors of MDPI journals from around the world. Editors select a small number of articles recently published in the journal that they believe will be particularly interesting to readers, or important in the respective research area. The aim is to provide a snapshot of some of the most exciting work published in the various research areas of the journal.

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13 pages, 1514 KB  
Article
Establishment of a Sandwich ELISA for Detection of Pan-Merbecoviruses
by Kaixin Li, Misa Katayama, Ayano Ichikawa, Hiromichi Matsugo, Yuta Wakabayashi, Akiko Takenaka-Uema, Wataru Sekine, Taisuke Horimoto and Shin Murakami
Pathogens 2025, 14(6), 605; https://doi.org/10.3390/pathogens14060605 - 19 Jun 2025
Viewed by 1285
Abstract
Merbecovirus, a subgenus of Betacoronavirus, includes MERS-CoV and multiple bat-derived viruses with zoonotic potential. Given the unpredictable emergence of these viruses and their genetic diversity, development of broad-spectrum diagnostic tools is expected. In this study, we established a sandwich ELISA targeting [...] Read more.
Merbecovirus, a subgenus of Betacoronavirus, includes MERS-CoV and multiple bat-derived viruses with zoonotic potential. Given the unpredictable emergence of these viruses and their genetic diversity, development of broad-spectrum diagnostic tools is expected. In this study, we established a sandwich ELISA targeting the nucleocapsid (N) protein of merbecoviruses. We generated monoclonal antibodies (mAbs) using recombinant N protein of a bat merbecovirus, VsCoV-1, and selected cross-reactive clones for other merbecoviruses. Three mAbs showed strong reactivities with multiple merbecoviruses but not with SARS-CoV-2 or endemic human coronaviruses. Pairwise ELISA screening identified 1A8/10H6 mAbs as the optimal combination for detection of N protein from six merbecoviruses—VsCoV-1, EjCoV-3, MERS-CoV, NeoCoV, HKU4, and HKU5—with limits of detection (LODs) below 7.81 ng/mL, including 1.25 ng/mL for VsCoV-1. Infectious bat merbecovirus EjCoV-3 was detected at 1.3 × 103 PFU/mL. No cross-reactivity was observed with non-merbecoviruses, indicating its high specificity. This sandwich ELISA offers a rapid, reproducible, and cost-effective diagnostic platform with potential for high-throughput screening and automation. Moreover, its design is amenable to adaptation into point-of-care formats such as lateral flow assays, highlighting its value for field-based surveillance and pandemic preparedness. Full article
(This article belongs to the Special Issue The Epidemiology and Diagnosis of Acute Respiratory Infections)
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20 pages, 1074 KB  
Article
The Epidemiology of Coccidioidomycosis (Valley fever) and the Disease Ecology of Coccidioides spp. in New Mexico (2006–2023)
by Paris S. Salazar-Hamm, Sarah Shrum Davis, Jovani Catalán-Dibene, Adriana L. Romero-Olivares, Karen Edge, Andrew W. Bartlow, Donald O. Natvig and Morgan E. Gorris
Pathogens 2025, 14(6), 607; https://doi.org/10.3390/pathogens14060607 - 19 Jun 2025
Cited by 1 | Viewed by 1935
Abstract
Coccidioidomycosis (Valley fever), caused by Coccidioides spp., is a fungal infection endemic to semi-arid regions of the Americas. Despite 80 years of disease recognition in New Mexico, there is limited disease awareness. We incorporated clinical, epidemiological, and ecological datasets to summarize the knowledge [...] Read more.
Coccidioidomycosis (Valley fever), caused by Coccidioides spp., is a fungal infection endemic to semi-arid regions of the Americas. Despite 80 years of disease recognition in New Mexico, there is limited disease awareness. We incorporated clinical, epidemiological, and ecological datasets to summarize the knowledge of Valley fever in New Mexico. We analyzed 1541 human cases from 2006 to 2023. On average, 86 cases were reported each year (4.1 cases per 100,000 population per year). The highest levels of incidence were in southwestern New Mexico. American Indian or Alaska Natives in New Mexico had a 1.9 times higher incidence rate of coccidioidomycosis than White people, and among age groups, older populations in New Mexico had the highest incidence rates. We analyzed 300 soil samples near Las Cruces, New Mexico, for the presence of Coccidioides and reported the first known positive soil samples collected from the state, the majority of which were from grassland-dominated sites and from animal burrows. Sequence analyses in clinical specimens, wild animals, and soil samples confirmed that Coccidioides posadasii is the main causative species of coccidioidomycosis in New Mexico. Environmental surveillance validated that locally acquired infections could occur in, but are not limited to, Catron, Doña Ana, Sierra, and Socorro Counties. Full article
(This article belongs to the Special Issue An Update on Fungal Infections)
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29 pages, 2576 KB  
Review
A Comprehensive Review of the Mechanisms of Human Q Fever: Pathogenesis and Pathophysiology
by José-Luis Pérez-Arellano, Jose Curbelo and Cristina Carranza-Rodriguez
Pathogens 2025, 14(6), 589; https://doi.org/10.3390/pathogens14060589 - 14 Jun 2025
Cited by 2 | Viewed by 2252
Abstract
Coxiella burnetii infection has a worldwide distribution, although the incidence and clinical manifestations vary between and within countries. There are the following four basic forms: asymptomatic infection, acute Q fever, chronic Q fever, and post-Q fever fatigue syndrome. The aim of this review [...] Read more.
Coxiella burnetii infection has a worldwide distribution, although the incidence and clinical manifestations vary between and within countries. There are the following four basic forms: asymptomatic infection, acute Q fever, chronic Q fever, and post-Q fever fatigue syndrome. The aim of this review is to provide a comprehensive overview of the important aspects of its pathogenesis and pathophysiology. First, we provide a brief update of the taxonomic aspects, basic structures, and genotypes of C. burnetii necessary for the proper interpretation of the following sections. Routes of infection, different stages of pathogenesis (respiratory entry of C. burnetii; penetration into alveolar macrophages, life cycle, and effects; systemic dissemination), and innate, acquired humoral and cell-mediated immune responses in different forms of infection are described in detail. The pathophysiology and clinical manifestations of Q fever, such as the main mechanisms of injury, in isolation and in combination, are reviewed. The clinical and biological manifestations of the two main forms of Q fever (acute and chronic) are outlined, with a brief definition and mention of the mechanisms of post-Q fever fatigue syndrome. Full article
(This article belongs to the Section Bacterial Pathogens)
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60 pages, 6483 KB  
Review
The Challenge of Lyssavirus Infections in Domestic and Other Animals: A Mix of Virological Confusion, Consternation, Chagrin, and Curiosity
by Charles E. Rupprecht, Aniruddha V. Belsare, Florence Cliquet, Philip P. Mshelbwala, Janine F. R. Seetahal and Vaughn V. Wicker
Pathogens 2025, 14(6), 586; https://doi.org/10.3390/pathogens14060586 - 13 Jun 2025
Viewed by 4517
Abstract
Lyssaviruses are RNA viruses in the Family Rhabdoviridae, Genus Lyssavirus. They represent the causative agents of acute, progressive encephalitis, known historically as rabies. Regardless of specific etiology, their collective viral morphology, biochemistry, pathobiology, associated clinical signs, diagnosis, epizootiology, and management are essentially [...] Read more.
Lyssaviruses are RNA viruses in the Family Rhabdoviridae, Genus Lyssavirus. They represent the causative agents of acute, progressive encephalitis, known historically as rabies. Regardless of specific etiology, their collective viral morphology, biochemistry, pathobiology, associated clinical signs, diagnosis, epizootiology, and management are essentially the same. Despite centuries of clinical recognition, these quintessential neurotropic agents remain significant pathogens today, with substantive consequences to agriculture, public health, and conservation biology. Notably, the singular morbidity caused by lyssaviruses is incurable and constitutes the highest case fatality of any viral disease. All warm-blooded vertebrates are believed to be susceptible. The dog is the only domestic animal that serves as a reservoir, vector, and victim. In contrast, felids are effective vectors, but not reservoirs. All other rabid domestic species, such as livestock, constitute spillover infections, as a bellwether to local lyssavirus activity. Frequently, professional confusion abounds among the veterinary community, because although the viral species Lyssavirus rabies is inarguably the best-known representative in the Genus, at least 20 other recognized or putative members of this monophyletic group are known. Frequently, this is simply overlooked. Moreover, often the ‘taxonomic etiology’ (i.e., ‘Lyssavirus x’) is mistakenly referenced in a biopolitcal context, instead of the obvious clinical illness (i.e., ‘rabies’). Global consternation persists, if localities believe they are ‘disease-free’, when documented lyssaviruses circulate or laboratory-based surveillance is inadequate to support such claims. Understandably, professional chagrin develops when individuals mistake the epidemiological terminology of control, prevention, elimination, etc. Management is not simple, given that the only licensed veterinary and human vaccines are against rabies virus, sensu lato. There are no adequate antiviral drugs for any lyssaviruses or cross-reactive biologics developed against more distantly related viral members. While representative taxa among the mammalian Orders Chiroptera, Carnivora, and Primates exemplify the major global reservoirs, which mammalian species are responsible for the perpetuation of other lyssaviruses remains a seemingly academic curiosity. This zoonosis is neglected. Clearly, with such underlying characteristics as a fundamental ‘disease of nature’, rabies, unlike smallpox and rinderpest, is not a candidate for eradication. With the worldwide zeal to drive human fatalities from canine rabies viruses to zero by the rapidly approaching year 2030, enhanced surveillance and greater introspection of the poorly appreciated burden posed by rabies virus and diverse other lyssaviruses may manifest as an epidemiological luxury to the overall global program of the future. Full article
(This article belongs to the Special Issue Current Challenges in Veterinary Virology)
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16 pages, 1357 KB  
Article
EmsB Microsatellite Analysis of Echinococcus multilocularis Specimens Isolated from Belgian Patients with Alveolar Echinococcosis and from Animal Hosts
by Sabrina Egrek, Jenny Knapp, Rosalie Sacheli, Khalid El Moussaoui, Philippe Léonard, Eva Larranaga Lapique, Laurence Millon, Sara Engelskirchen, Olivier Detry, Annick Linden and Marie-Pierre Hayette
Pathogens 2025, 14(6), 584; https://doi.org/10.3390/pathogens14060584 - 12 Jun 2025
Viewed by 760
Abstract
Alveolar echinococcosis (AE), caused by Echinococcus multilocularis (E. multilocularis), is a severe parasitic zoonosis that is potentially fatal for humans. The parasite is primarily transmitted by wildlife, with red foxes acting as definitive hosts and rodents as intermediate hosts, while humans [...] Read more.
Alveolar echinococcosis (AE), caused by Echinococcus multilocularis (E. multilocularis), is a severe parasitic zoonosis that is potentially fatal for humans. The parasite is primarily transmitted by wildlife, with red foxes acting as definitive hosts and rodents as intermediate hosts, while humans can become accidental but dead-end hosts. The aim of this study is to use EmsB typing on E. multilocularis isolates from human AE cases and local animals such as foxes and rodents. In this study, retrospective EmsB typing was performed on 39 samples, including 11 tissue samples from 10 patients, 18 fecal swabs from foxes, and 10 tissue samples from rodents. A dendrogram was created to determine the EmsB profiles present. The results showed that all the rodent samples were associated with the EmsB P1 profile (10/10), while the human and fox samples shared the EmsB profile P1 (5/11 humans and 8/18 foxes), a profile near P4 (2/11 humans and 3 foxes), and a profile near P8 (1/11 humans and 1/18 foxes). The study demonstrates that the same EmsB profiles circulate among humans and animals, confirming that wildlife reservoirs play a key role in transmission. Full article
(This article belongs to the Section Parasitic Pathogens)
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11 pages, 346 KB  
Article
The Effects of High-Dose Probiotic Supplementation on Immune Activation and Neurocognitive Disorders in People Living with HIV Undergoing Successful Antiretroviral Treatment: The Procog Study
by Matteo Vassallo, Margaux Zerlini, Roxane Fabre, Heloise Joly, Jacques Durant, Alain Makinson, Amandine Mauries, Jacqueline Capeau, Soraya Fellahi, Jean-Philippe Bastard, Christian Pradier and Christine Lebrun-Frenay
Pathogens 2025, 14(6), 568; https://doi.org/10.3390/pathogens14060568 - 6 Jun 2025
Viewed by 1401
Abstract
Background: The prevalence of HIV-associated neurocognitive disorders (HAND) remains high despite antiretroviral treatment (ART). Changes in gut microbiota and persistent immune activation have been suggested as possible causes, while the role of probiotic supplementation remains controversial. Methods: We included subjects with mild HAND [...] Read more.
Background: The prevalence of HIV-associated neurocognitive disorders (HAND) remains high despite antiretroviral treatment (ART). Changes in gut microbiota and persistent immune activation have been suggested as possible causes, while the role of probiotic supplementation remains controversial. Methods: We included subjects with mild HAND and successful ART. They were randomized to receive either 6 months of high-dose probiotic supplementation or to continue with only ART. Immune activation markers and neuropsychological testing were performed at baseline and the end of follow-up. Neuropsychological testing assessed learning, episodic memory, attention/concentration, executive functions, language, information processing speed, and motor skills. Z- and T-scores were calculated for all domains but motor skills, allowing the measurement of the global deficit score (GDS). The trajectories of neuropsychological performances and immune activation markers were compared between groups. Results: From September 2020 to July 2021, 31 PWHs were included (median age 62, 73% men, CD4 744 cc/mm3), and 28 completed the 6-month follow-up. The characteristics of the subjects and their neuropsychological performance at baseline in the two groups were similar. At the end of follow-up, probiotics did not have any impact on immune activation markers, while they were associated with better improvement in GDS (T-score 0.0 in controls vs. −0.3 in probiotics, p = 0.048) and the attention/concentration test (Z-score 0.4 in controls vs. 1.2 in probiotics, p = 0.035). Conclusions: Oral supplementation with high-dose probiotics for 6 months did not affect systemic immune activation but was associated with improved neurocognitive performance, suggesting benefits from probiotic supplementation for mild HAND. Full article
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11 pages, 1115 KB  
Article
Monitoring Multiple Sexually Transmitted Pathogens Through Wastewater Surveillance
by Balghsim Alshehri, Olivia N. Birch and Justin C. Greaves
Pathogens 2025, 14(6), 562; https://doi.org/10.3390/pathogens14060562 - 5 Jun 2025
Cited by 1 | Viewed by 1557
Abstract
Wastewater-based epidemiology (WBE) offers a promising tool for sexually transmitted infection (STI) surveillance, especially in settings where underdiagnosis or social stigma complicates conventional reporting. To assess its utility, we conducted a year-long study examining six STIs, Chlamydia trachomatis, Treponema pallidum, Neisseria [...] Read more.
Wastewater-based epidemiology (WBE) offers a promising tool for sexually transmitted infection (STI) surveillance, especially in settings where underdiagnosis or social stigma complicates conventional reporting. To assess its utility, we conducted a year-long study examining six STIs, Chlamydia trachomatis, Treponema pallidum, Neisseria gonorrhoeae, human immunodeficiency virus (HIV), hepatitis C virus (HCV), and herpes simplex virus (HSV), in weekly composite samples from the primary influent of a small-sized Midwestern wastewater treatment plant. Pathogen detection and quantification were performed via digital PCR. Among the tested targets, Gonorrhea, HIV, HCV, and HSV were detected at the highest frequencies, often in 40–50% of the samples, while Chlamydia and Syphilis appeared less frequently. Despite the variability in detection patterns, this study demonstrates that even infrequent signals can reveal community-level shedding of poorly reported or asymptomatic infections. Although month-to-month wastewater data were not strongly correlated with corresponding clinical records, which could potentially reflect delayed healthcare seeking and pathogen-specific shedding dynamics, the overall findings underscore WBE’s ability to complement existing surveillance by capturing infections outside traditional healthcare channels. These results not only advance our understanding of STI prevalence and population shedding but also highlight the practical benefits of WBE as an early warning and targeted intervention tool. Full article
(This article belongs to the Special Issue Wastewater Surveillance and Public Health Strategies)
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33 pages, 783 KB  
Systematic Review
Global Epidemiology and Antimicrobial Resistance of Metallo-β-Lactamase (MBL)-Producing Acinetobacter Clinical Isolates: A Systematic Review
by Matthew E. Falagas, Dimitrios S. Kontogiannis, Maria Zidrou, Charalampos Filippou and Giannoula S. Tansarli
Pathogens 2025, 14(6), 557; https://doi.org/10.3390/pathogens14060557 - 3 Jun 2025
Cited by 3 | Viewed by 1870
Abstract
This systematic review assessed the global epidemiology of metallo-β-lactamase (MBL)-producing Acinetobacter clinical isolates and the associated antimicrobial resistance. A total of 475 relevant articles from the Cochrane Library, Google Scholar, PubMed, Scopus, and Web of Science were identified and screened as potentially eligible [...] Read more.
This systematic review assessed the global epidemiology of metallo-β-lactamase (MBL)-producing Acinetobacter clinical isolates and the associated antimicrobial resistance. A total of 475 relevant articles from the Cochrane Library, Google Scholar, PubMed, Scopus, and Web of Science were identified and screened as potentially eligible articles. Data from 85 articles were extracted for the analysis. Most reports on MBL-producing Acinetobacter clinical isolates originated from Asia [68/85 (80%) studies] and Africa [14/85 (16.5%) studies]. There were also scarce reports from Europe and America. The blaVIM (in 31 studies), blaIMP (in 29 studies), and blaNDM (in 21 studies) genes were the most commonly identified genes. In 22 out of 28 (78.6%) studies with comparable data, the proportions of MBL-producing pathogens detected using phenotypic methods were numerically higher than those using genotypic methods. MBL-producing Acinetobacter isolates showed high resistance (up to 100%) to several antibiotic classes, including carbapenems, cephalosporins, fluoroquinolones, and monobactams. However, they showed low resistance to colistin [ranging from 0% (in six studies) to 14.3% (in one study)] and to tigecycline [0% (in three studies)]. No risk of bias assessment was conducted. The findings emphasize the global spread of MBL-producing Acinetobacter and the need for enhanced antimicrobial stewardship, infection control measures, and surveillance. Full article
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12 pages, 5734 KB  
Article
The Requirement of Turkey Herpesvirus (HVT) Glycoprotein C During Natural Infection in Chickens and Turkeys
by Huai Xu, Widaliz Vega-Rodriguez, Kathrine Van Etten and Keith Jarosinski
Pathogens 2025, 14(6), 538; https://doi.org/10.3390/pathogens14060538 - 28 May 2025
Viewed by 3239
Abstract
The glycoprotein C (gC) of gallid alphaherpesvirus 2—better known as Marek’s disease (MD) virus (MDV)—and gallid alphaherpesvirus 3 is required for horizontal transmission in chickens. Since gC is conserved within the Alphaherpesvirinae subfamily, we hypothesized that gC was also essential for the horizontal [...] Read more.
The glycoprotein C (gC) of gallid alphaherpesvirus 2—better known as Marek’s disease (MD) virus (MDV)—and gallid alphaherpesvirus 3 is required for horizontal transmission in chickens. Since gC is conserved within the Alphaherpesvirinae subfamily, we hypothesized that gC was also essential for the horizontal transmission of meleagrid alphaherpesvirus 1 (MeAHV1) or turkey herpesvirus (HVT). To test this hypothesis, we generated a fluorescent protein-tagged clone of recombinant (r)HVT (vHVT47G), removed the open reading frame of HVT gC from the genome (vHΔgC), and rescued the deletion by inserting an HA-epitope tagged HVT gC (vHΔgC-R) to test their ability to transmit in chickens and turkeys. We also tested whether MDV gC could compensate for HVT gC during transmission, where HVT gC was replaced with MDV gC (vH-MDVgC). Although all viruses replicated in chickens, none spread from chicken to chicken. However, when tested in turkeys, all viruses except vHΔgC transmitted from turkey to turkey. Importantly, the rescuent virus (vHΔgC-R) and HVT expressing MDV gC (vH-MDVgC) rescued transmission, showing that HVT gC is required and MDV gC can compensate for HVT gC for turkey-to-turkey transmission. These data confirm the host-specific transmission of HVT in turkeys and suggest that the essential function of alphaherpesvirus gC proteins is conserved. This information can be exploited while generating future vaccines against MD that will affect the poultry industry worldwide. Full article
(This article belongs to the Special Issue Current Challenges in Veterinary Virology)
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17 pages, 3320 KB  
Article
Comparative IP-MS Reveals HSPA5 and HSPA8 Interacting with Hemagglutinin Protein to Promote the Replication of Influenza A Virus
by Xingwei Feng, Mengfei Ning, Bin Chen, Xuan Li, Honglei Sun, Juan Pu, Jinhua Liu, Na Wang and Yinhua Huang
Pathogens 2025, 14(6), 535; https://doi.org/10.3390/pathogens14060535 - 27 May 2025
Viewed by 1067
Abstract
The influenza A viruses (IAV) are the principal pathogens for annual (seasonal) influenza, which cause world-wide outbreaks in poultry and pose a persistent threat to public health. The Hemagglutinin protein (HA) of IAV promotes virus infection by binding the host membrane receptor and [...] Read more.
The influenza A viruses (IAV) are the principal pathogens for annual (seasonal) influenza, which cause world-wide outbreaks in poultry and pose a persistent threat to public health. The Hemagglutinin protein (HA) of IAV promotes virus infection by binding the host membrane receptor and mediating virus–host membrane fusion. Immunoprecipitation–mass spectrometry (IP-MS) provides global insights into IAV HA–host protein interactions. However, various experimental conditions might affect the identification of interactions. Here, we performed a serial IP-MS to compare interactors of IAV HA in accidental host human, chicken and reservoir host duck cells. We found that the positive ratio of interactors identified by the IP-MS was improved when the transfected HA plasmid had a similar expression level to HA proteins found in IAV virus infection. Comparing interactors in human, chicken and duck cells, we found that HA–interacting host factors might play a role in the susceptibility of accidental hosts (human and chicken) to IAV infection compared to reservoir hosts (duck). We then focused on the function of two heat shock proteins (HSPA5 and HSPA8), which interacted with IAV HA proteins in all three species (human, chicken and duck). We found that both HSPA5 and HSPA8 promoted the IAV replication by enhancing the viral attachment and internalization. These findings extend our knowledge about the mechanisms of IAV entry to host cells and provide target genes to create chickens resistant to avian influenza. Full article
(This article belongs to the Section Epidemiology of Infectious Diseases)
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20 pages, 2323 KB  
Article
Temperature Influences Antimicrobial Resistance and Virulence of Vibrio parahaemolyticus Clinical Isolates from Quebec, Canada
by Feriel C. Mahieddine, Annabelle Mathieu-Denoncourt and Marylise Duperthuy
Pathogens 2025, 14(6), 521; https://doi.org/10.3390/pathogens14060521 - 23 May 2025
Cited by 1 | Viewed by 1460
Abstract
Climate change and ocean warming have a huge impact on microbial communities, leading to an increased prevalence of Vibrio parahaemolyticus infections in northern America. V. parahaemolyticus is an inhabitant of aquatic environments and is associated with fishes and shellfishes, particularly with oysters. It [...] Read more.
Climate change and ocean warming have a huge impact on microbial communities, leading to an increased prevalence of Vibrio parahaemolyticus infections in northern America. V. parahaemolyticus is an inhabitant of aquatic environments and is associated with fishes and shellfishes, particularly with oysters. It causes gastrointestinal infection through consumption of contaminated seafood, as well as wound infections or septicemia. Temperature is known to affect virulence and persistence factors in V. parahaemolyticus. In this study, twenty clinical strains isolated form sick patients in Quebec, Canada, were characterized for persistence and virulence factor production at different temperatures to assess the impact of a switch from sea water to the human body on them. Their capacity to produce biofilm, hemolysins, and membrane vesicles as well as their motility and antibiotic resistance at 20 °C and 37 °C were assessed. Our results revealed that while temperature had little effect on vesicle production, it significantly influenced their growth, antimicrobial resistance, biofilm formation, and motility. Additionally, all V. parahaemolyticus strains produced hemolysins at 37 °C but not under environmental conditions. Full article
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29 pages, 730 KB  
Review
Antimicrobial Resistant Staphylococcus spp., Escherichia coli, and Salmonella spp. in Food Handlers: A Global Review of Persistence, Transmission, and Mitigation Challenges
by Gustavo Guimarães Fernandes Viana, Marita Vedovelli Cardozo, Juliano Gonçalves Pereira and Gabriel Augusto Marques Rossi
Pathogens 2025, 14(5), 496; https://doi.org/10.3390/pathogens14050496 - 18 May 2025
Cited by 4 | Viewed by 2780
Abstract
Antimicrobial resistance in foodborne pathogens represents a critical global health challenge, with food handlers serving as key contributors in their transmission. This comprehensive review synthesizes evidence on the prevalence, transmission dynamics, and antimicrobial resistance patterns of three major pathogens, Staphylococcus spp., Escherichia coli [...] Read more.
Antimicrobial resistance in foodborne pathogens represents a critical global health challenge, with food handlers serving as key contributors in their transmission. This comprehensive review synthesizes evidence on the prevalence, transmission dynamics, and antimicrobial resistance patterns of three major pathogens, Staphylococcus spp., Escherichia coli, and Salmonella spp., among food handlers worldwide. Analysis of studies across diverse geographical regions reveals considerable variation in colonization rates, with Staphylococcus spp. prevalence ranging from 19.5% to 95.0%, Escherichia coli from 2.8% to 89.3%, and Salmonella spp. from 0.07% to 9.1%. Resistance profiles demonstrate alarming trends, including widespread β-lactam resistance and emerging resistance to last-resort antibiotics like carbapenems. Particularly concerning is the high occurrence of multidrug resistant (MDR) strains and extended spectrum β-lactamase (ESBL) producers in low- and middle-income countries. This review identified inadequate handwashing, poor hygiene infrastructure, and asymptomatic carriage as critical factors facilitating the transmission of antimicrobial resistant strains. These findings underscore the urgent need for enhanced surveillance systems, targeted decolonization strategies, improved hygiene protocols, and food handler education to mitigate the spread of resistant pathogens through the food chain. Full article
(This article belongs to the Special Issue Foodborne Pathogens: The Antimicrobial Resistance from Farm to Fork)
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16 pages, 295 KB  
Review
Methods of Controlling Microbial Contamination of Food
by Renata Urban-Chmiel, Jacek Osek and Kinga Wieczorek
Pathogens 2025, 14(5), 492; https://doi.org/10.3390/pathogens14050492 - 16 May 2025
Cited by 4 | Viewed by 3149
Abstract
The rapid growth of world population and increase in living standards have led to an increase in the demand for high-quality, safe food. The Food and Agriculture Organization of the United Nations (FAO) estimates that by 2050 the demand for food will increase [...] Read more.
The rapid growth of world population and increase in living standards have led to an increase in the demand for high-quality, safe food. The Food and Agriculture Organization of the United Nations (FAO) estimates that by 2050 the demand for food will increase by 60%, and production of animal protein will increase by 1.7% a year, with meat production to increase by nearly 70%, dairy products by 55%, and aquaculture by as much as 90%. Microbial contamination of food is a significant problem for the accessibility of safe food which does not pose a threat to the life and health of consumers. Campylobacter, Salmonella, and Yersinia are responsible for thousands of food-borne infections in humans. Currently, numerous programs are being developed to combat pathogenic bacteria in the food supply chain, especially at the primary production stage. These approaches include physical, chemical, biological, and other strategies and methods used to inhibit the bacterial growth of bacteria or completely eliminate the pathogens from the food chain. Therefore, an extremely important goal is to provide safe food and control its quality by eliminating pathogenic and spoilage microorganisms. However, the use of chemicals in food preservation has negative effects for both the consumption values of food and the natural environment. Therefore, it seems absolutely necessary to implement measures utilizing the most environmentally friendly and effective techniques for controlling microbial contamination in food. There is a great need to develop ecological methods in food production which guarantee adequate safety. One of these methods is the use of bacteriophages (bacterial viruses) naturally occurring in the environment. Given the above, the aim of this study was to present the most natural, ecological, and alternative methods of food preservation with regard to the most common foodborne zoonotic bacteria. We also present methods for reducing the occurrence of microbial contamination in food, thus to produce maximally safe food for consumers. Full article
25 pages, 12805 KB  
Article
From the Gut to the Brain: Transcriptomic Insights into Neonatal Meningitis Escherichia coli Across Diverse Host Niches
by Lekshmi K. Edison and Subhashinie Kariyawasam
Pathogens 2025, 14(5), 485; https://doi.org/10.3390/pathogens14050485 - 15 May 2025
Viewed by 1336
Abstract
Neonatal Meningitis-causing Escherichia coli (NMEC) is the leading cause of neonatal meningitis and exhibits remarkable adaptability to diverse host environments. Understanding its transcriptional responses across different host niches is crucial for deciphering pathogenesis and identifying potential therapeutic targets. We performed a comparative transcriptomic [...] Read more.
Neonatal Meningitis-causing Escherichia coli (NMEC) is the leading cause of neonatal meningitis and exhibits remarkable adaptability to diverse host environments. Understanding its transcriptional responses across different host niches is crucial for deciphering pathogenesis and identifying potential therapeutic targets. We performed a comparative transcriptomic analysis of NMEC RS218, the prototype strain of NMEC, under four distinct host-mimicking conditions: colonic fluid (CF), serum (S), human brain endothelial cells (HBECs), and cerebrospinal fluid (CSF). Differential gene expression analysis was conducted to assess metabolic shifts, virulence factor regulation, and niche-specific adaptation strategies, in which RS218 demonstrated niche-specific transcriptional reprogramming. In CF, genes associated with biofilm formation, motility, efflux pumps, and cell division regulation were upregulated, aiding gut colonization. The serum environment triggered the expression of siderophore-mediated iron acquisition, enterobactin biosynthesis, and heme utilization genes, facilitating immune evasion and bacterial persistence. In HBECs, NMEC upregulated genes linked to nucleoside metabolism, membrane remodeling, pilus organization, and blood–brain barrier (BBB) traversal. In CSF, genes related to oxidative stress resistance, chemotaxis, DNA repair, biofilm formation, and amino acid biosynthesis were enriched, reflecting NMEC’s adaptive mechanisms for survival under nutrient-depleted conditions. Energy-intensive pathways were consistently downregulated across all niches, highlighting the need for an energy conservation strategy. This study provides novel insights into NMEC’s adaptive strategies across different host environments, emphasizing its metabolic flexibility, virulence regulation, and immune evasion mechanisms, offering potential targets for therapeutic intervention. Full article
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17 pages, 3975 KB  
Article
Orientia tsutsugamushi Modulates RIPK3 Cellular Levels but Does Not Inhibit Necroptosis
by Thomas E. Siff, Paige E. Allen, David L. Armistead, Jason R. Hunt, Steven J. Rolland, Hervé Agaisse and Jason A. Carlyon
Pathogens 2025, 14(5), 478; https://doi.org/10.3390/pathogens14050478 - 14 May 2025
Viewed by 1215
Abstract
Scrub typhus is an emerging chigger-borne disease caused by the obligate intracellular bacterium Orientia tsutsugamushi. Necroptosis is a form of programmed cell death (PCD) mediated by RIPK3 (serine/threonine kinase receptor interacting protein 3) and its downstream effector MLKL (mixed-lineage kinase domain-like). While [...] Read more.
Scrub typhus is an emerging chigger-borne disease caused by the obligate intracellular bacterium Orientia tsutsugamushi. Necroptosis is a form of programmed cell death (PCD) mediated by RIPK3 (serine/threonine kinase receptor interacting protein 3) and its downstream effector MLKL (mixed-lineage kinase domain-like). While O. tsutsugamushi modulates apoptosis, another form of PCD, its interplay with necroptosis is unknown. Much of Orientia pathobiology is linked to its ankyrin repeat (AR)-containing effectors (Anks). Two of these, Ank1 and Ank6, share similarities with the cowpox AR protein, vIRD (viral inducer of RIPK3 degradation) that prevents necroptosis. Here, we show that Ank1 and Ank6 reduce RIPK3 cellular levels although not as robustly as and mechanistically distinct from vIRD. Orientia infection lowers RIPK3 amounts and does not elicit necroptosis in endothelial cells. In HeLa cells ectopically expressing RIPK3, Orientia fails to inhibit RIPK3 and MLKL phosphorylation as well as cell death. MLKL colocalization with Orientia or Listeria monocytogenes, another intracytoplasmic pathogen, was not observed. Thus, O. tsutsugamushi reduces cellular levels of RIPK3 and does not elicit necroptosis but cannot inhibit this PCD pathway once it is induced. This study is a first step toward understanding how the relationship between Orientia and necroptosis contributes to scrub typhus pathogenesis. Full article
(This article belongs to the Special Issue Emerging Vector-Borne and Zoonotic Diseases—2nd Edition)
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20 pages, 3859 KB  
Article
Cryo-Electron Microscopy of BfpB Reveals a Type IVb Secretin Multimer Adapted to Accommodate the Exceptionally Wide Bundle-Forming Pilus
by Janay I. Little, Pradip Kumar Singh, Montserrat Samsó and Michael S. Donnenberg
Pathogens 2025, 14(5), 471; https://doi.org/10.3390/pathogens14050471 - 13 May 2025
Viewed by 1031
Abstract
Type IV pili (T4Ps) are multifunctional surface fibers essential for bacterial motility, adhesion, and virulence, found across Gram-negative and Gram-positive bacteria and archaea. Detailed descriptions of T4P structural biology are allowing progress in understanding T4P biogenesis. Secretins, large outer membrane channels, are crucial [...] Read more.
Type IV pili (T4Ps) are multifunctional surface fibers essential for bacterial motility, adhesion, and virulence, found across Gram-negative and Gram-positive bacteria and archaea. Detailed descriptions of T4P structural biology are allowing progress in understanding T4P biogenesis. Secretins, large outer membrane channels, are crucial for T4P extrusion in Gram-negative bacteria. Using cryo-EM and AlphaFold, we modeled the structure of BfpB, the secretin of the Bundle-Forming Pilus (BFP) of enteropathogenic Escherichia coli. BfpB exhibits a unique 17-fold symmetry, correlating with the thicker BFP filaments, and diverging from the 12–15 subunits typical of T4P, type 2 secretion (T2S), and type 3 secretion (T3S) systems. Additionally, we identified an extended β-hairpin loop in the N3 domain, resembling features of distantly related T3SS secretins, and an N-terminal helix where a C-terminal S-domain is seen in some T2S and T3S secretins. These findings reveal evolutionary parallels and structural adaptations in secretins, highlighting the link between oligomerization and pilus structure. This work advances our understanding of T4P biogenesis, secretin evolution, and bacterial secretion systems, offering insights into pathogenic diversity and future research directions. Full article
(This article belongs to the Special Issue Structural Biology Applied in the Study of Pathogenic Bacteria)
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36 pages, 885 KB  
Review
The Epithelial Immune Response to Human Papillomavirus Infection
by Shyantani Roy-Biswas and Merilyn Hibma
Pathogens 2025, 14(5), 464; https://doi.org/10.3390/pathogens14050464 - 9 May 2025
Cited by 1 | Viewed by 3913
Abstract
The skin is a complex organ, containing an intricate network of immune cells that are crucial for host barrier function and defence against pathogens. Human papillomavirus (HPV) exclusively infects the skin, and its lifecycle is intimately associated with epithelial cell division and differentiation. [...] Read more.
The skin is a complex organ, containing an intricate network of immune cells that are crucial for host barrier function and defence against pathogens. Human papillomavirus (HPV) exclusively infects the skin, and its lifecycle is intimately associated with epithelial cell division and differentiation. There are over 450 HPV types, 12 of which are classified as carcinogenic. The primary focus of this review is the epithelial immune response to HPV infection of the cervix during the initial stages of infection, productive infection, and disease progression. During the early stages of infection, cells are HPV-positive; however, there are no attributable histological changes to the epithelium. The HPV-infected cells have the capacity for innate sensing and signalling through toll-like receptors in response to viral nucleic acids. However, HPV has evolved multiple mechanisms to evade the innate response. During productive infection, all viral antigens are expressed and there are visible histological changes to the epithelium, including koilocytosis. Disease regression is associated with Tbet positive cells in the infected epithelium and the presence of CD4 and CD8 T cells in the lamina propria. Disease progression is associated with the overexpression of the E6 and E7 oncoproteins after integration of viral genomes into the host chromosomal DNA. Histologically, the epithelium is less differentiated, and changes to cells include a higher nuclear-to-cytoplasmic ratio and an increased mitotic index. Immune changes associated with disease progression include increased numbers of cells expressing suppressor molecules, such as FoxP3, Blimp-1, and HMGB1, and myeloid cell infiltrates with an M2-like phenotype. This review highlights the gaps in the understanding of the immune response in HPV-positive cervical neoplasia, and in regression and progression of disease. This knowledge is critical for the development of effective immunotherapies that reliably cause HPV-positive cervical neoplasia to regress. Full article
(This article belongs to the Section Viral Pathogens)
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15 pages, 3320 KB  
Article
Neglected Avian Blood Parasites (Onchocercidae and Trypanosomatidae) in Migratory Passerines of the Temperate Zone, Eastern Baltic Region
by Rasa Bernotienė, Tatjana Iezhova, Vytautas Eigirdas, Vytautas Jusys, Margarita Kazak and Rasa Binkienė
Pathogens 2025, 14(5), 452; https://doi.org/10.3390/pathogens14050452 - 5 May 2025
Viewed by 1124
Abstract
Passerine birds (n = 3335) of 19 species were caught and investigated for the presence of Trypanosomatidae and Onchocercidae parasites using the buffy coat method, microscopy and PCR in Ventės Ragas, Lithuania. Data on the spread patterns of these parasites are still lacking. [...] Read more.
Passerine birds (n = 3335) of 19 species were caught and investigated for the presence of Trypanosomatidae and Onchocercidae parasites using the buffy coat method, microscopy and PCR in Ventės Ragas, Lithuania. Data on the spread patterns of these parasites are still lacking. The prevalences of Trypanosoma parasites in birds of different species varied from 2.2% to 36.1%, while the prevalences of Onchocercidae parasites varied from 0% to 17.3%. Statistically significant differences between spring and autumn in the prevalences of Trypanosomatidae were determined for Acrocephalus schoenobaenus, Hirundo rustica and Turdus philomelos. No significant differences between the prevalences of Onchocercidae in spring and autumn were determined. The prevalence of Trypanosoma was significantly higher for long-distance migrant birds compared with short-distance migrants, for omnivorous birds compared with insectivorous birds, and for open-nesting birds compared with birds nesting in nest boxes. The prevalences of Onchocercidae parasites did not differ for the same bird groups except for the prevalence in omnivorous birds, which was higher compared with insectivorous birds. Both groups of parasites were detected in juveniles, showing the presence of transmission in the study area. The diet, breeding behaviour and migration features of avian hosts can influence the prevalence of avian blood parasites. Full article
(This article belongs to the Special Issue Updates on Zoonotic Parasites)
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15 pages, 10062 KB  
Article
A Practical Framework for ASFV Disinfectant Evaluation: Differentiating Cytopathic Effects from Cytotoxicity via Integrated Analytical Methods
by Sok Song, Kyu-Sik Shin, Su-Jeong Kim, Yong Yi Joo, Bokhee Han, So-Hee Park, Hyun-Ok Ku, Wooseog Jeong and Choi-Kyu Park
Pathogens 2025, 14(5), 451; https://doi.org/10.3390/pathogens14050451 - 4 May 2025
Viewed by 1288
Abstract
African swine fever virus (ASFV) is a highly virulent DNA virus that has spread globally since its introduction into Georgia in 2007, causing substantial economic losses in the swine industry. In the absence of an effective vaccine, chemical disinfection remains a key strategy [...] Read more.
African swine fever virus (ASFV) is a highly virulent DNA virus that has spread globally since its introduction into Georgia in 2007, causing substantial economic losses in the swine industry. In the absence of an effective vaccine, chemical disinfection remains a key strategy for disease control. However, in cell-based disinfectant efficacy testing, distinguishing between disinfectant-induced cytotoxicity and virus-induced cytopathic effects (CPEs) remains a major challenge, leading to the potential misinterpretation of results. To address this, we developed a multi-step analytical framework to differentiate CPEs from cytotoxicity using a Vero cell-adapted ASFV strain. Virkon® S was tested at three dilutions—375×, 275× (manufacturer-recommended), and 175×—and evaluated through CPE observation, lactate dehydrogenase (LDH) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays, and antigen detection via lateral flow immunoassay (p30) and immunofluorescence (p54). Notably, the 375× dilution achieved effective viral inactivation with significantly lower cytotoxicity, demonstrating that this framework can facilitate a more refined determination of disinfectant working dilutions. Furthermore, increased p30 signals after disinfection and the observation of lower cytotoxicity in virus-plus-disinfectant groups compared to disinfectant-only groups highlight the complexity of virus-disinfectant interactions and the potential for misinterpretation. This study provides a standardized and interpretable strategy for assessing ASFV disinfectant efficacy and offers a practical basis for evaluating other enveloped viruses in future disinfection studies. Full article
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18 pages, 2474 KB  
Article
Kalmusia variispora (Didymosphaeriaceae, Dothideomycetes) Associated with the Grapevine Trunk Disease Complex in Cyprus
by Georgios Makris, Marcelo Sandoval-Denis, Pedro W. Crous and Loukas I. Kanetis
Pathogens 2025, 14(5), 428; https://doi.org/10.3390/pathogens14050428 - 28 Apr 2025
Cited by 1 | Viewed by 1091
Abstract
Grapevine trunk diseases (GTDs) are widespread worldwide, causing serious economic losses to the vitiviniculture industry. The etiology of the complex pathogenic mycobiome associated with this group of diseases is critical to implementing appropriate management strategies. Diseased grapevines exhibiting typical GTD symptoms were collected [...] Read more.
Grapevine trunk diseases (GTDs) are widespread worldwide, causing serious economic losses to the vitiviniculture industry. The etiology of the complex pathogenic mycobiome associated with this group of diseases is critical to implementing appropriate management strategies. Diseased grapevines exhibiting typical GTD symptoms were collected from vineyards in different provinces of Cyprus, resulting in 19 pycnidial isolates. A subsequent multilocus sequence analysis of six genetic loci (ITS, LSU, SSU, b-tub, tef1-a, and rpb2) identified them as Kalmusia variispora, and twelve representative isolates are included in the phylogenetic analyses. According to pathogenicity trials on two-year-old potted vines (cv. Mavro), all tested isolates were pathogenic, exhibiting light to dark brown discoloration and lesions of varying levels, ranging from 4 to 12.3 cm long. The capacity of K. variispora isolates to produce cell-wall-degrading exoenzymes was qualitatively estimated on solid media. Cellulase, pectinase, and laccase production were evident for all the tested isolates, except isolate CBS 151329, where the latter enzyme was undetected. The severity of the symptoms was consistent with the laccase-producing capacity. The present study confirmed the association of K. variispora with grapevines as a pathogen and represents the first description of this ascomycete as a GTD causal agent in Cyprus. This highly virulent species may play a significant role in the GTD complex, and its biological cycle and epidemiology should be further investigated. Full article
(This article belongs to the Special Issue Filamentous Fungal Pathogens: 2nd Edition)
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23 pages, 5321 KB  
Article
Experimental Infection of Ferrets with Bartonella henselae: In Search of a Novel Animal Model for Zoonotic Bartonellosis
by Erin Lashnits, Cynthia Robveille, Pradeep Neupane, Toni Richardson, Keith Linder, Gabriel McKeon, Ricardo Maggi and Edward B. Breitschwerdt
Pathogens 2025, 14(5), 421; https://doi.org/10.3390/pathogens14050421 - 26 Apr 2025
Viewed by 2198
Abstract
Bartonella henselae is an important zoonotic pathogen with a wide range of clinical manifestations in humans. Despite advances in understanding its pathogenesis, there is no broadly applicable laboratory animal model for bartonellosis. This study aimed to assess the potential utility of an experimental [...] Read more.
Bartonella henselae is an important zoonotic pathogen with a wide range of clinical manifestations in humans. Despite advances in understanding its pathogenesis, there is no broadly applicable laboratory animal model for bartonellosis. This study aimed to assess the potential utility of an experimental model of chronic B. henselae infection using ferrets, a species previously utilized in various human pathogen studies. Six ferrets (n = 6) were divided into three groups: a control group (n = 2), a low-dose infection group (n = 2), and a high-dose infection group (n = 2). The two infection groups were inoculated intradermally with 105 (low dose) and 109 (high dose) CFU/mL B. henselae, respectively. Clinical signs, serological responses, and bacteriological findings were monitored over seven weeks; ferrets were then euthanized and tissues were examined histologically. Only minimal or transient systemic clinical signs and laboratory abnormalities developed in B. henselae inoculated ferrets. The high-dose group seroconverted to B. henselae antigen within two weeks, maintaining elevated titers throughout the study. Histopathological examination revealed that four B. henselae-infected ferrets had notable microscopic inflammatory lesions in the liver parenchyma (3/4), heart (1/4), and brain (1/4); similar lesions were not observed in the tissues of the two control ferrets. Despite the presence of microscopic lesions and seroconversion in the high-dose group, bacteremia was not documented and B. henselae DNA was not successfully amplified by quantitative PCR from lesional organs. This pilot study demonstrated that ferrets may serve as a promising model for investigating B. henselae pathogenesis, diagnosis, treatment, and prevention. Full article
(This article belongs to the Topic Zoonotic Vector-Borne Diseases of Companion Animals)
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18 pages, 1053 KB  
Review
The Main Arboviruses and Virus Detection Methods in Vectors: Current Approaches and Future Perspectives
by Amanda Montezano Cintra, Nathália Mayumi Noda-Nicolau, Milena Leite de Oliveira Soman, Pedro Henrique de Andrade Affonso, Guilherme Targino Valente and Rejane Maria Tommasini Grotto
Pathogens 2025, 14(5), 416; https://doi.org/10.3390/pathogens14050416 - 25 Apr 2025
Viewed by 3172
Abstract
Arthropod-borne viruses (arboviruses) represent a growing concern for global public and veterinary health, with cases reported across all continents. This review presents a broad overview of the geographic distribution of arboviruses transmitted by insect vectors, emphasizing the importance of early viral detection as [...] Read more.
Arthropod-borne viruses (arboviruses) represent a growing concern for global public and veterinary health, with cases reported across all continents. This review presents a broad overview of the geographic distribution of arboviruses transmitted by insect vectors, emphasizing the importance of early viral detection as a cornerstone of surveillance and outbreak preparedness. Special attention is given to the phenomenon of zoonotic spillover, where viruses maintained in natural transmission cycles often involving wildlife reservoirs and arthropod vectors cross into human populations, triggering emergent or re-emergent outbreaks. This article discusses key arboviral families of medical and veterinary significance, including Togaviridae, Flaviviridae, Nairoviridae, Phenuiviridae, Peribunyaviridae, and Orthomyxoviridae, highlighting their molecular and structural characteristics. These features are essential for guiding the development and implementation of specific and sensitive detection strategies. In addition, this work provides a comparative analysis of diverse laboratory methodologies for viral detection in vectors. From serological assays and viral isolation to advanced molecular tools and next-generation sequencing, we explore their principles, practical applications, and context-dependent advantages and limitations. By compiling this information, we aim to support researchers and public health professionals in selecting the most appropriate tools for vector surveillance, ultimately contributing to improved response strategies in the face of arboviral threats. Full article
(This article belongs to the Section Viral Pathogens)
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13 pages, 1648 KB  
Article
Does the Hfq Protein Contribute to RNA Cargo Translocation into Bacterial Outer Membrane Vesicles?
by Marisela Velez and Véronique Arluison
Pathogens 2025, 14(4), 399; https://doi.org/10.3390/pathogens14040399 - 21 Apr 2025
Viewed by 911
Abstract
Gram-negative bacteria release outer membrane vesicles (OMVs) that deliver various molecules, including virulence factors, to interact with their host. Recent studies have suggested that OMVs may also serve as carriers for RNAs, particularly small regulatory noncoding RNAs (sRNAs). For these RNAs to function [...] Read more.
Gram-negative bacteria release outer membrane vesicles (OMVs) that deliver various molecules, including virulence factors, to interact with their host. Recent studies have suggested that OMVs may also serve as carriers for RNAs, particularly small regulatory noncoding RNAs (sRNAs). For these RNAs to function effectively, they typically require a protein cofactor, Hfq, known as an RNA chaperone. In previous work, using molecular imaging, Circular Dichroism CD, and InfraRed FTIR spectroscopies, we demonstrated that Hfq interacts with the bacterial inner membrane and forms pores, suggesting a possible role in translocating RNA from the cytoplasm to periplasm and then to OMVs. In this study, we expand on our previous findings and provide evidence that RNA molecules bind to the Escherichia coli inner membrane in an Hfq-dependent manner. Moreover, we show that the lipid nature, in particular the presence of a cardiolipin-rich domain, is crucial for this interaction. These results reveal a new aspect of RNA translocation through the inner membrane, for further packaging in OMVs, and underscore the importance of Hfq in this mechanism. Full article
(This article belongs to the Section Bacterial Pathogens)
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13 pages, 442 KB  
Review
Change of Heart: Can Artificial Intelligence Transform Infective Endocarditis Management?
by Jack W. McHugh, Douglas W. Challener and Hussam Tabaja
Pathogens 2025, 14(4), 371; https://doi.org/10.3390/pathogens14040371 - 9 Apr 2025
Cited by 1 | Viewed by 1191
Abstract
Artificial intelligence (AI) has emerged as a promising adjunct in the diagnosis and management of infective endocarditis (IE), a disease characterized by diagnostic complexity and significant morbidity. Machine learning (ML) models such as SABIER and SYSUPMIE have demonstrated strong predictive accuracy for early [...] Read more.
Artificial intelligence (AI) has emerged as a promising adjunct in the diagnosis and management of infective endocarditis (IE), a disease characterized by diagnostic complexity and significant morbidity. Machine learning (ML) models such as SABIER and SYSUPMIE have demonstrated strong predictive accuracy for early IE diagnosis, embolic risk stratification, and postoperative mortality, surpassing traditional clinical scoring systems. In imaging, AI-enhanced echocardiography and advanced modalities like FDG-PET/CT offer improved sensitivity, specificity, and reduced inter-observer variability, potentially transforming clinical decision making. Additionally, AI-powered microbiological techniques, including MALDI-TOF mass spectrometry combined with ML and neural network-based metagenomic classifiers, show promise in rapidly identifying pathogens and predicting antimicrobial resistance. Despite encouraging early results, widespread adoption faces barriers, including data limitations, interpretability issues, ethical concerns, and the need for robust validation. Future directions include leveraging generative AI as clinical consultative tools, provided their capabilities and limitations are carefully managed. Ultimately, collaborative efforts addressing these challenges could transform IE care, enhancing diagnostic accuracy, clinical outcomes, and patient safety. Full article
(This article belongs to the Special Issue Updates in Infective Endocarditis—2nd Edition)
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19 pages, 3112 KB  
Article
Borrelia burgdorferi Strain-Specific Differences in Mouse Infectivity and Pathology
by Annabelle Pfeifle, Rose Anderson-Duvall, Levi A. Tamming, Wanyue Zhang, Sathya N. Thulasi Raman, Caroline Gravel, Jianguo Wu, Heather Coatsworth, Maarten J. Voordouw, Xu Zhang, Michael J. W. Johnston, Wangxue Chen, Simon Sauve, Lisheng Wang and Xuguang Li
Pathogens 2025, 14(4), 352; https://doi.org/10.3390/pathogens14040352 - 5 Apr 2025
Viewed by 1805
Abstract
Lyme disease (LD), caused by infection with the tick-borne bacteria, Borrelia burgdorferi, is associated with a wide array of symptoms in human patients. Variations in clinical manifestations are thought to be influenced by genetic differences among B. burgdorferi strains. In this study, [...] Read more.
Lyme disease (LD), caused by infection with the tick-borne bacteria, Borrelia burgdorferi, is associated with a wide array of symptoms in human patients. Variations in clinical manifestations are thought to be influenced by genetic differences among B. burgdorferi strains. In this study, we evaluated the infectivity, tissue bacterial load, pathology, and immunogenicity of five strains of B. burgdorferi sensu stricto (297 Ah130, Bb16-54, B31-A3, Bb16-126, JD1) in female C3H/HeN mice at three infectious doses (104, 105, 106 spirochetes). We found that strains Bb16-126 and JD1 were the most infectious, resulting in 100% infection across all the tested doses. Strain Bb16-126 caused the highest bacterial burden in the heart tissue and significant carditis, whereas JD1 exhibited the lowest spirochete load in the heart and minimal carditis. In comparison, strain B31-A3 demonstrated the highest abundance in the tibiotarsal joint. Infection with all the strains induced severe lymph node hyperplasia, with JD1 producing the greatest increase in cellularity. Using a diagnostic C6 peptide ELISA, all the strains induced significant anti-C6 IgM and IgG antibody titers at 14 days post-infection; however, strain B31-A3 elicited the highest anti-C6 IgM titers. Our findings demonstrate the importance of strain diversity in shaping B. burgdorferi pathogenesis in a mouse model and provide insights for developing strain-specific diagnostic, therapeutic, and vaccine strategies. Full article
(This article belongs to the Section Bacterial Pathogens)
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18 pages, 2025 KB  
Article
Immunogenicity of Trypanosoma cruzi Multi-Epitope Recombinant Protein as an Antigen Candidate for Chagas Disease Vaccine in Humans
by Christian F. Teh-Poot, Andrea Alfaro-Chacón, Landy M. Pech-Pisté, Miguel E. Rosado-Vallado, Oluwatoyin Ajibola Asojo, Liliana E. Villanueva-Lizama, Eric Dumonteil and Julio Vladimir Cruz-Chan
Pathogens 2025, 14(4), 342; https://doi.org/10.3390/pathogens14040342 - 3 Apr 2025
Cited by 1 | Viewed by 1490
Abstract
Chagas disease, caused by the protozoan Trypanosoma cruzi (T. cruzi), is the most significant neglected tropical disease affecting individuals in the Americas. Currently, available drugs, such as nifurtimox and benznidazole (BZN), are both toxic and ineffective in the chronic phase of [...] Read more.
Chagas disease, caused by the protozoan Trypanosoma cruzi (T. cruzi), is the most significant neglected tropical disease affecting individuals in the Americas. Currently, available drugs, such as nifurtimox and benznidazole (BZN), are both toxic and ineffective in the chronic phase of the disease. A promising alternative is the development of a Chagas disease vaccine, although this effort is hampered by the complexity of the parasite and HLA polymorphisms. In addition, the activation of epitope-specific CD8+ T cells is critical to conferring a robust cell-mediated immune response and protection by producing IFN-γ and perforin. Thus, the antigen (s) for the development of a Chagas vaccine or immunotherapy must include CD8+ T cell epitopes. In this study, we aimed to develop a multi-epitope recombinant protein as a novel human vaccine for Chagas disease. Sixteen database programs were used to predict de novo 40 potential epitopes for the HLA-A*02:01 allele. Nine out of the 40 predicted epitopes were able to elicit IFN-γ production in Peripheral Blood Mononuclear Cells (PBMCs) from Chagas patients. Molecular docking revealed a good binding affinity among the epitopes with diverse HLA molecules. Therefore, a recombinant multi-epitope protein including these nine T. cruzi CD8+ epitopes was expressed and demonstrated to recall an antigen-specific immune response in ex-vivo assays using PBMCs from Chagas patients with the HLA-A*02 allele. These findings support the development of this multi-epitope protein as a promising candidate human vaccine against Chagas disease. Full article
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11 pages, 2735 KB  
Article
Investigation of the Pathobiology of Edwardsiella piscicida—Septicemia in Largemouth Bass
by Grace Ramena, Suja Aarattuthodi, Gnanender Sriramoju and Yathish Ramena
Pathogens 2025, 14(4), 334; https://doi.org/10.3390/pathogens14040334 - 31 Mar 2025
Viewed by 1254
Abstract
Edwardsiella piscicida septicemia is a significant threat to aquaculture, causing substantial production and economic losses. The causative bacterium E. piscicida infects multiple fish species of aquaculture relevance. This study investigated the pathogenicity of catfish-derived E. piscicida in largemouth bass (Micropterus salmoides) [...] Read more.
Edwardsiella piscicida septicemia is a significant threat to aquaculture, causing substantial production and economic losses. The causative bacterium E. piscicida infects multiple fish species of aquaculture relevance. This study investigated the pathogenicity of catfish-derived E. piscicida in largemouth bass (Micropterus salmoides) fingerlings. The experimental infection of largemouth bass using genotypically distinct E. piscicida isolates resulted in significant fish mortality independent of the isolate genotypes. A specific correlation between discrete E. piscicida genotypes and fish mortalities was not identified. The histopathological assessment of tissues from infected fish revealed inflammatory lesions characteristic of bacterial septicemia. This study provides insights into the pathogenesis of heterologous E. piscicida isolates in largemouth bass fingerlings, which could be crucial in developing effective pathogen-targeted management strategies to combat a deadly disease. Full article
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24 pages, 1014 KB  
Review
The Dual-Edged Sword: Risks and Benefits of JAK Inhibitors in Infections
by Anders Jarneborn, Pradeep Kumar Kopparapu and Tao Jin
Pathogens 2025, 14(4), 324; https://doi.org/10.3390/pathogens14040324 - 27 Mar 2025
Cited by 3 | Viewed by 3732
Abstract
Janus kinase inhibitors (JAKis) represent a relatively new class of immunomodulatory drugs with potent effects on various cytokine signalling pathways. They have revolutionized the treatment landscape for autoimmune diseases such as rheumatoid arthritis, psoriatic arthritis, and ulcerative colitis. However, their ability to modulate [...] Read more.
Janus kinase inhibitors (JAKis) represent a relatively new class of immunomodulatory drugs with potent effects on various cytokine signalling pathways. They have revolutionized the treatment landscape for autoimmune diseases such as rheumatoid arthritis, psoriatic arthritis, and ulcerative colitis. However, their ability to modulate immune responses presents a dual-edged nature, influencing both protective immunity and pathological inflammation. This review explores the complex role of JAKis in infectious settings, highlighting both beneficial and detrimental effects. On the one hand, experimental models suggest that JAK inhibition can impair host defence mechanisms, increasing susceptibility to certain bacterial and viral infections. For example, tofacitinib-treated mice exhibited more severe joint erosions in Staphylococcus aureus (S. aureus) septic arthritis and showed impaired viral clearance in herpes simplex encephalitis. Additionally, clinical data confirm an increased risk of herpes zoster in patients receiving JAKis, underscoring the need for rigorous monitoring. On the other hand, JAK inhibition has demonstrated protective effects in certain infectious and hyperinflammatory conditions. In sepsis models, including cecal ligation and puncture (CLP) and S. aureus bacteraemia, tofacitinib improved survival by attenuating excessive inflammation. Furthermore, JAKis, particularly baricitinib, have shown substantial efficacy in mitigating cytokine storms during severe COVID-19 infections, leading to improved clinical outcomes and reduced mortality. These observations suggest that JAKis have a role in modulating hyperinflammatory responses in select infectious contexts. In conclusion, JAKis present a complex interplay between immunosuppression and immunomodulation. While they increase the risk of certain infections, they also show potential in managing hyperinflammatory conditions such as cytokine storms. The key challenge is determining which patients and situations benefit most from JAKis while minimizing risks, requiring a careful and personalized treatment approach. Full article
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20 pages, 1492 KB  
Review
The Role of Blastocystis spp. in the Etiology of Gastrointestinal and Autoimmune Diseases
by Oliwia Pawelec-Pęciak, Natalia Łanocha-Arendarczyk, Konrad Grzeszczak and Danuta Kosik-Bogacka
Pathogens 2025, 14(4), 313; https://doi.org/10.3390/pathogens14040313 - 25 Mar 2025
Cited by 3 | Viewed by 6017
Abstract
Blastocystis spp. has been linked to gastrointestinal symptoms, yet its pathogenicity remains uncertain. In addition, the roles of virulence factors, pathogenic potential, and host-specific traits associated with symptomatic infections are still not well understood. The growing number of immunocompromised patients has contributed to [...] Read more.
Blastocystis spp. has been linked to gastrointestinal symptoms, yet its pathogenicity remains uncertain. In addition, the roles of virulence factors, pathogenic potential, and host-specific traits associated with symptomatic infections are still not well understood. The growing number of immunocompromised patients has contributed to an increasing prevalence of Blastocystis spp. infections, which may be implicated in the development of various inflammatory diseases, including irritable bowel syndrome (IBS), colorectal cancer, and autoimmune disorders such as Hashimoto’s disease and ulcerative colitis. However, the presence of nonspecific symptoms often complicates diagnosis. This study aimed to present current data on the impact of Blastocystis spp. on the development and progression of gastrointestinal and autoimmune diseases, as well as to explore potential treatment options for Blastocystis spp. infections. A literature review was conducted to analyze the role of Blastocystis spp. in the pathogenesis of specific diseases and to investigate potential mechanisms of its interaction with the host organism. Advances in diagnostic techniques, particularly PCR, allow not only for the detection of Blastocystis spp. but also for the identification of specific subtypes, improving treatment precision. Beyond conventional therapies like metronidazole, there is a growing emphasis on alternative treatments, including the use of medicinal plants and probiotics. Full article
(This article belongs to the Section Parasitic Pathogens)
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14 pages, 2319 KB  
Article
Oral Vaccination with Attenuated Salmonella Expressing Viral M25 Protein Effectively Protects Mice Against Murine Cytomegalovirus Infection
by Yujun Liu, Hao Gong, Jiaming Zhu and Fenyong Liu
Pathogens 2025, 14(4), 314; https://doi.org/10.3390/pathogens14040314 - 25 Mar 2025
Cited by 1 | Viewed by 946
Abstract
Attenuated Salmonella strains are promising oral vectors for vaccination against human infectious diseases. Human cytomegalovirus (CMV) is among the most common causes of disability in children, including intellectual disability and sensorineural hearing loss. Developing an anti-CMV vaccine is a major public health priority. [...] Read more.
Attenuated Salmonella strains are promising oral vectors for vaccination against human infectious diseases. Human cytomegalovirus (CMV) is among the most common causes of disability in children, including intellectual disability and sensorineural hearing loss. Developing an anti-CMV vaccine is a major public health priority. We report in this study the construction of a new attenuated Salmonella strain to express murine cytomegalovirus (MCMV) M25 protein and its use for vaccination in mice against MCMV infection. In mice orally vaccinated with the constructed Salmonella vector carrying the M25 expression cassette, we revealed a substantial induction of anti-MCMV serum IgG and mucosal IgA humoral responses and a considerable elicitation of anti-MCMV T cell responses. When the vaccinated mice were challenged intraperitoneally and intranasally with MCMV, we observed a significant inhibition of virus infection and growth in various organs including spleens, livers, lungs, and salivary glands, compared to the non-vaccinated animals or those receiving a control vaccine without M25 protein expression. Moreover, we showed effective protection of these vaccinated mice from MCMV challenge. Our study provides the first direct evidence that an attenuated Salmonella-based vector with the MCMV M25 expression cassette can induce strong humoral and T cell responses and provide effective protection against MCMV infection. These results illustrate the feasibility of engineering Salmonella-based vectors expressing the M25 antigen for anti-CMV oral vaccine development. Full article
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13 pages, 231 KB  
Review
New β-Lactam/β-Lactamase Inhibitor Combination Antibiotics
by Maria Sargianou, Panagiotis Stathopoulos, Christos Vrysis, Iva D. Tzvetanova and Matthew E. Falagas
Pathogens 2025, 14(4), 307; https://doi.org/10.3390/pathogens14040307 - 24 Mar 2025
Cited by 8 | Viewed by 5449
Abstract
The growing problem of infections due to pathogens with antimicrobial resistance, especially Gram-negative bacteria, has led to the development of new β-lactam/β-lactamase inhibitor combination antibiotics. During the last 2 years from the writing of this article, cefepime/enmetazobactam, aztreonam/avibactam, and sulbactam/durlobactam were approved for [...] Read more.
The growing problem of infections due to pathogens with antimicrobial resistance, especially Gram-negative bacteria, has led to the development of new β-lactam/β-lactamase inhibitor combination antibiotics. During the last 2 years from the writing of this article, cefepime/enmetazobactam, aztreonam/avibactam, and sulbactam/durlobactam were approved for use in clinical practice. Cefepime/enmetazobactam targets extended-spectrum β-lactamase (ESBL)-producing Pseudomonas aeruginosa and Enterobacterales. It is indicated for the treatment of patients with complicated urinary tract infections, including pyelonephritis, in Europe and the USA, and also for hospital-acquired pneumonia, ventilator-associated pneumonia, and bacteremia associated with those infections (only in Europe). The antimicrobial spectrum of aztreonam/avibactam includes carbapenem-resistant Enterobacterales. Aztreonam/avibactam is indicated for the treatment of adult patients who suffer from complicated intra-abdominal infections, complicated urinary tract infections including pyelonephritis, hospital-acquired pneumonia, and ventilator-associated pneumonia due to aerobic Gram-negative infections with limited therapeutic options. Sulbactam/durlobactam, a combination of 2 β-lactamase inhibitors, is indicated for the treatment of adult patients with hospital-acquired bacterial pneumonia and ventilator-associated bacterial pneumonia due to the Acinetobacter baumannii–calcoaceticus complex [including carbapenem-resistant Acinetobacter baumannii (CRAB) infections]. Full article
(This article belongs to the Special Issue Beta Lactamases: A Weapon Against Antibiotics)
17 pages, 3053 KB  
Article
Virome of Terrestrial Mammals and Bats from Southern Brazil: Circulation of New Putative Members of the Togaviridae Family and Other Findings
by Julyana Sthéfanie Simões Matos, Meriane Demoliner, Juliana Schons Gularte, Micheli Filippi, Vyctoria Malayhka de Abreu Góes Pereira, Mariana Soares da Silva, Matheus Nunes Weber, Marcelo Pereira de Barros and Fernando Rosado Spilki
Pathogens 2025, 14(4), 310; https://doi.org/10.3390/pathogens14040310 - 24 Mar 2025
Cited by 1 | Viewed by 1549
Abstract
The surveillance of wildlife viromes is essential for identifying zoonotic threats within the One Health framework. This study analyzed rectal and oral swabs from 88 individuals representing 13 species as felids, wild rodents, marsupials and non-human primates in Southern Brazil using metagenomic sequencing. [...] Read more.
The surveillance of wildlife viromes is essential for identifying zoonotic threats within the One Health framework. This study analyzed rectal and oral swabs from 88 individuals representing 13 species as felids, wild rodents, marsupials and non-human primates in Southern Brazil using metagenomic sequencing. Akodon montensis (n = 15 individuals) and Coendou spinosus (n = 4) harbored Chikungunya virus (ChikV, Togaviridae), marking its first detection in these hosts. Molossus molossus (n = 17) presented Coronaviridae and Orthoherpesviridae, while Eptesicus furinalis (n = 1) also carried Coronaviridae. A broad virome diversity, including Togaviridae and Adenoviridae members, was identified in Didelphis albiventris (n = 43), with significant relevance to human health. Additional species, such as Callithrix jacchus (n = 1), Leopardus guttulus (n = 1), Myocastor coypus (n = 1), Monodelphis iheringi (n = 1), Thaptomys nigrita (n = 1), Sooretamys angouya (n = 1), Brucepattersonius iheringi (n = 1), and Lasiurus blossevillii (n = 1), contributed to insights into viral reservoirs. These results underscore the importance of virome studies in regions harboring high biodiversity, emphasizing genomic surveillance as a vital tool for monitoring zoonotic viruses and safeguarding global health. Full article
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15 pages, 538 KB  
Review
Comprehensive Insights into Highly Pathogenic Avian Influenza H5N1 in Dairy Cattle: Transmission Dynamics, Milk-Borne Risks, Public Health Implications, Biosecurity Recommendations, and One Health Strategies for Outbreak Control
by Henrietta Owusu and Yasser M. Sanad
Pathogens 2025, 14(3), 278; https://doi.org/10.3390/pathogens14030278 - 13 Mar 2025
Cited by 10 | Viewed by 4869
Abstract
Highly pathogenic avian influenza (HPAI) H5N1 has been traditionally linked to poultry and wild birds, which has recently become a serious concern for dairy cattle, causing outbreaks all over the United States. The need for improved surveillance, biosecurity protocols, and interagency collaboration is [...] Read more.
Highly pathogenic avian influenza (HPAI) H5N1 has been traditionally linked to poultry and wild birds, which has recently become a serious concern for dairy cattle, causing outbreaks all over the United States. The need for improved surveillance, biosecurity protocols, and interagency collaboration is highlighted by the discovery of H5N1 in dairy herds in several states and its human transmission. The epidemiology, transmission dynamics, and wide-ranging effects of H5N1 in cattle are reviewed in this paper, with particular attention paid to the disease’s effects on agricultural systems, public health, and animal health. Nonspecific clinical symptoms, such as decreased milk production and irregular milk consistency, are indicative of infection in dairy cows. Alarmingly, significant virus loads have been discovered in raw milk, raising worries about potential zoonotic transmission. The dangers of viral spillover between species are further highlighted by cases of domestic cats experiencing severe neurological symptoms after ingesting raw colostrum and milk from infected cows. Even though human cases remain rare, and they are mostly related to occupational exposure, constant attention is required due to the possibility of viral adaptability. The necessity of a One Health approach that integrates environmental, animal, and human health efforts is further supported by the broad occurrence of H5N1 across multiple species. For early detection, containment, and mitigation, cooperation between veterinary clinics, public health organizations, and agricultural stakeholders is crucial. Controlling the outbreak requires stringent movement restrictions, regular testing of dairy cows in reference labs, and adherence to biosecurity procedures. This review highlights the importance of thorough and coordinated efforts to manage H5N1 in dairy cattle by combining existing knowledge and pointing out gaps in surveillance and response strategies. Additionally, it sheds light on the potential risk of consumption of cow’s milk contaminated with H5N1 virus by humans and other companion animals like cats. In the face of this changing threat, proactive monitoring, strict biosecurity protocols, and cross-sector cooperation are crucial for reducing financial losses and protecting human and animal health. Full article
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19 pages, 6831 KB  
Article
Conformational and Stability Analysis of SARS-CoV-2 Spike Protein Variants by Molecular Simulation
by Gustavo E. Olivos-Ramirez, Luis F. Cofas-Vargas, Tobias Madl and Adolfo B. Poma
Pathogens 2025, 14(3), 274; https://doi.org/10.3390/pathogens14030274 - 12 Mar 2025
Cited by 1 | Viewed by 2670
Abstract
We performed a comprehensive structural analysis of the conformational space of several spike (S) protein variants using molecular dynamics (MD) simulations. Specifically, we examined four well-known variants (Delta, BA.1, XBB.1.5, and JN.1) alongside the wild-type (WT) form of SARS-CoV-2. The conformational states of [...] Read more.
We performed a comprehensive structural analysis of the conformational space of several spike (S) protein variants using molecular dynamics (MD) simulations. Specifically, we examined four well-known variants (Delta, BA.1, XBB.1.5, and JN.1) alongside the wild-type (WT) form of SARS-CoV-2. The conformational states of each variant were characterized by analyzing their distributions within a selected space of collective variables (CVs), such as inter-domain distances between the receptor-binding domain (RBD) and the N-terminal domain (NTD). Our primary focus was to identify conformational states relevant to potential structural transitions and to determine the set of native contacts (NCs) that stabilize these conformations. The results reveal that genetically more distant variants, such as XBB.1.5, BA.1, and JN.1, tend to adopt more compact conformational states compared to the WT. Additionally, these variants exhibit novel NC profiles, characterized by an increased number of specific contacts distributed among ionic, polar, and nonpolar residues. We further analyzed the impact of specific mutations, including T478K, N500Y, and Y504H. These mutations not only enhance interactions with the human host receptor but also alter inter-chain stability by introducing additional NCs compared to the WT. Consequently, these mutations may influence the accessibility of certain protein regions to neutralizing antibodies. Overall, these findings contribute to a deeper understanding of the structural and functional variations among S protein variants. Full article
(This article belongs to the Section Viral Pathogens)
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13 pages, 1850 KB  
Article
Acid Tolerance of Coxiella burnetii Is Strain-Specific and Might Depend on Stomach Content
by Katharina Sobotta, Jan Schulze-Luehrmann, Martha Ölke, Katharina Boden and Anja Lührmann
Pathogens 2025, 14(3), 272; https://doi.org/10.3390/pathogens14030272 - 12 Mar 2025
Viewed by 1707
Abstract
Q fever is a zoonotic disease caused by the obligate intracellular bacterium Coxiella (C.) burnetii. Human infections occur mainly via inhalation, but infections via the oral route have been observed. Gastric acidic conditions (pH 2–4) are the first defense mechanism [...] Read more.
Q fever is a zoonotic disease caused by the obligate intracellular bacterium Coxiella (C.) burnetii. Human infections occur mainly via inhalation, but infections via the oral route have been observed. Gastric acidic conditions (pH 2–4) are the first defense mechanism to limit food-associated infections. In this study, we tested the ability of C. burnetii to survive extremely acidic conditions (pH 2–3) to assess the risk of oral infection in humans. We treated different C. burnetii strains with different pH values and calculated the recovery rate by counting colony-forming units. The analysis of an additional eight C. burnetii strains showed that some strains are acid-resistant, while others are not. Importantly, the presence of pepsin, an endopeptidase and the main digestive enzyme in the gastrointestinal tract, increases the survival rate of C. burnetii. Similarly, the presence of milk might also increase the survival rate. These results suggest that oral infections by C. burnetii are possible and depend on the bacterial strain and the stomach microenvironment. Consequently, the digestive infection route of C. burnetii could play a role in the transmission of the pathogen. Full article
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13 pages, 6620 KB  
Review
Encystment and Excystment Processes in Acanthamoeba castellanii: An Emphasis on Cellulose Involvement
by Mathew Choaji, Ascel Samba-Louaka, Zineb Fechtali-Moute, Willy Aucher and Sébastien Pomel
Pathogens 2025, 14(3), 268; https://doi.org/10.3390/pathogens14030268 - 10 Mar 2025
Cited by 1 | Viewed by 2713
Abstract
The free-living amoeba Acanthamoeba castellanii is a unicellular eukaryote distributed in a wide range of soil or aquatic environments, either natural or human-made, such as rivers, lakes, drinking water, or swimming pools. Besides its capacity to transport potential pathogens, such as bacteria or [...] Read more.
The free-living amoeba Acanthamoeba castellanii is a unicellular eukaryote distributed in a wide range of soil or aquatic environments, either natural or human-made, such as rivers, lakes, drinking water, or swimming pools. Besides its capacity to transport potential pathogens, such as bacteria or viruses, Acanthamoeba spp. can have intrinsic pathogenic properties by causing severe infections at the ocular and cerebral level, named granulomatous amoebic encephalitis and amoebic keratitis, respectively. During its life cycle, A. castellanii alternates between a vegetative and mobile form, named the trophozoite, and a resistant, latent, and non-mobile form, named the cyst. The cyst wall of Acanthamoeba is double-layered, with an inner endocyst and an outer ectocyst, and is mainly composed of cellulose and proteins. The resistance of cysts to many environmental stresses and disinfection treatments has been assigned to the presence of cellulose. The current review aims to present the importance of this glycopolymer in Acanthamoeba cysts and to further report the pathways involved in encystment and excystment. Full article
(This article belongs to the Special Issue Acanthamoeba Infections)
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21 pages, 2438 KB  
Article
Development of Low-Cost In-House Assays for Quantitative Detection of HBsAg, HBeAg, and HBV DNA to Enhance Hepatitis B Virus Diagnostics and Antiviral Screening in Resource-Limited Settings
by Simmone D’souza, Layla Al-Yasiri, Annie Chen, Dan T. Boghici, Guido van Marle, Jennifer A. Corcoran, Trushar R. Patel and Carla S. Coffin
Pathogens 2025, 14(3), 258; https://doi.org/10.3390/pathogens14030258 - 5 Mar 2025
Viewed by 2161
Abstract
Globally, an estimated 254 million people are living with chronic hepatitis B virus (HBV) infection, yet only 10.5% have been diagnosed, underscoring the urgent need to expand testing to meet the World Health Organization’s HBV elimination targets by 2030. Many HBV diagnostic tests [...] Read more.
Globally, an estimated 254 million people are living with chronic hepatitis B virus (HBV) infection, yet only 10.5% have been diagnosed, underscoring the urgent need to expand testing to meet the World Health Organization’s HBV elimination targets by 2030. Many HBV diagnostic tests remain expensive and inaccessible in resource-limited settings. In this study, we demonstrate how individually sourced, commercially available reagents can be used to develop cost-effective in-house assays for total DNA isolation, HBV viral load quantification by (q)PCR, and qHBsAg and qHBeAg measurement using sandwich ELISA. These assays were validated using known HBV-positive and HBV-negative plasma samples (genotypes A–F) and HepAD38 cells treated with tenofovir disoproxil fumarate (TDF). DNA isolation using a commercial column-based kit was compared to a high-throughput, column-free method, allowing for HBV quantification from 50 µL of plasma with lower limits of detection (LLOD) of 1.8 × 103 and 1.8 × 104 HBV DNA copies IU/mL, respectively. Both commercial and in-house DNA isolation methods yielded comparable half-maximal effective concentration (EC50) values in TDF-treated HepAD38 cells. Additionally, in-house sandwich ELISA assays were developed for quantitative HBsAg and HBeAg detection, with LLOD values of 0.78 IU/mL and 0.38 PEI U/mL (Paul Ehrlich Institute), respectively. The in-house reagents for DNA isolation, molecular testing, and serological detection of HBV were estimated to be at least 10 times more cost-effective than commercially available kits, highlighting their potential for broader application in resource-limited regions. Full article
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15 pages, 2282 KB  
Article
Development and Characterization of Fluorescent Protein-Tagged Vibrio harveyi Strains as a Versatile Tool for Studying Infection Dynamics and Strain Interactions
by Charalampos Chalmoukis, Stavros Droubogiannis, Vassiliki A. Michalopoulou, Adriana Triga, Panagiotis F. Sarris and Pantelis Katharios
Pathogens 2025, 14(3), 247; https://doi.org/10.3390/pathogens14030247 - 3 Mar 2025
Cited by 2 | Viewed by 2140
Abstract
Fluorescent protein-tagged bacterial strains are widely used tools for studying host-pathogen interactions and microbial dynamics. In this study, we developed and characterized Vibrio harveyi strains genetically modified to express green fluorescent protein (GFP) and red fluorescent protein (RFP). These strains were constructed using [...] Read more.
Fluorescent protein-tagged bacterial strains are widely used tools for studying host-pathogen interactions and microbial dynamics. In this study, we developed and characterized Vibrio harveyi strains genetically modified to express green fluorescent protein (GFP) and red fluorescent protein (RFP). These strains were constructed using triparental mating and evaluated for phenotypic, genomic, and virulence attributes. Genomic analyses revealed strain-specific variations, including mutations in key regulatory and metabolic genes, such as luxO and transketolase. While plasmid acquisition imposed metabolic costs, resulting in altered growth and antibiotic sensitivities in certain transconjugants, others demonstrated robust phenotypic stability. Virulence assays using gilthead seabream larvae revealed that most tagged strains retained moderate pathogenicity, with visualization of co-infections highlighting the potential for studying strain-specific interactions. Furthermore, fluorescent microscopy confirmed the successful colonization and localization of tagged bacteria within host tissues. These findings underscore the utility of GFP- and RFP-tagged Vibrio harveyi as versatile tools for infection dynamics, offering a foundation for future research on strain interactions and pathogen-host relationships. Full article
(This article belongs to the Special Issue Fish Pathogens: An Ongoing Challenge for Aquaculture)
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21 pages, 702 KB  
Review
Latency Reversing Agents and the Road to an HIV Cure
by Louis Tioka, Rafael Ceña Diez, Anders Sönnerborg and Maarten A. A. van de Klundert
Pathogens 2025, 14(3), 232; https://doi.org/10.3390/pathogens14030232 - 27 Feb 2025
Cited by 1 | Viewed by 5400
Abstract
HIV-1 infection cannot be cured due to the presence of HIV-1 latently infected cells. These cells do not produce the virus, but they can resume virus production at any time in the absence of antiretroviral therapy. Therefore, people living with HIV (PLWH) need [...] Read more.
HIV-1 infection cannot be cured due to the presence of HIV-1 latently infected cells. These cells do not produce the virus, but they can resume virus production at any time in the absence of antiretroviral therapy. Therefore, people living with HIV (PLWH) need to take lifelong therapy. Strategies have been coined to eradicate the viral reservoir by reactivating HIV-1 latently infected cells and subsequently killing them. Various latency reversing agents (LRAs) that can reactivate HIV-1 in vitro and ex vivo have been identified. The most potent LRAs also strongly activate T cells and therefore cannot be applied in vivo. Many LRAs that reactivate HIV in the absence of general T cell activation have been identified and have been tested in clinical trials. Although some LRAs could reduce the reservoir size in clinical trials, so far, they have failed to eradicate the reservoir. More recently, immune modulators have been applied in PLWH, and the first results seem to indicate that these may reduce the reservoir and possibly improve immunological control after therapy interruption. Potentially, combinations of LRAs and immune modulators could reduce the reservoir size, and in the future, immunological control may enable PLWH to live without developing HIV-related disease in the absence of therapy. Full article
(This article belongs to the Section Viral Pathogens)
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20 pages, 3172 KB  
Article
Integrated One Health Surveillance of West Nile Virus and Usutu Virus in the Veneto Region, Northeastern Italy, from 2022 to 2023
by Federica Gobbo, Giulia Chiarello, Sofia Sgubin, Federica Toniolo, Francesco Gradoni, Lidia Iustina Danca, Sara Carlin, Katia Capello, Giacomo De Conti, Alessio Bortolami, Maria Varotto, Laura Favero, Michele Brichese, Francesca Russo, Franco Mutinelli, Stefania Vogiatzis, Monia Pacenti, Luisa Barzon and Fabrizio Montarsi
Pathogens 2025, 14(3), 227; https://doi.org/10.3390/pathogens14030227 - 25 Feb 2025
Cited by 1 | Viewed by 2913
Abstract
West Nile virus (WNV) and Usutu virus (USUV) are neurotropic mosquito-borne orthoflaviviruses maintained in an enzootic cycle, in which birds are amplifying/reservoir hosts, while humans and equids are dead-end hosts. As northern Italy, especially the Veneto Region, is considered an endemic area for [...] Read more.
West Nile virus (WNV) and Usutu virus (USUV) are neurotropic mosquito-borne orthoflaviviruses maintained in an enzootic cycle, in which birds are amplifying/reservoir hosts, while humans and equids are dead-end hosts. As northern Italy, especially the Veneto Region, is considered an endemic area for WNV and USUV circulation, a surveillance plan based on a One Health approach has been implemented since 2008. This work reports the results of entomological, veterinary and human surveillances for WNV and USUV in the Veneto Region in 2022 and 2023, through virological and/or serological examinations. In 2022, 531 human WNV infections were recorded, and 93,213 mosquitoes and 2193 birds were virologically tested, showing infection rates (IRs) of 4.85% and 8.30%, respectively. The surveillance effort in 2023 provided these results: 56 human WNV infections were confirmed, and 133,648 mosquitoes and 1812 birds were virologically tested, showing IRs of 1.78% and 4.69%, respectively. This work highlights the exceptional circulation of WNV in the Veneto Region, due to the new re-introduction of WNV lineage 1 and co-circulation with WNV lineage 2. This paper confirms the efficacy of integrated surveillance for early warning of viral circulation and gives new insights about avian hosts involved in the enzootic cycle of orthoflavivirus in the endemic region of Italy. Full article
(This article belongs to the Section Viral Pathogens)
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42 pages, 5461 KB  
Review
Anisakidae and Anisakidosis: A Public Health Perspective
by Diana Nonković, Vanja Tešić, Vida Šimat, Svjetlana Karabuva, Alan Medić and Jerko Hrabar
Pathogens 2025, 14(3), 217; https://doi.org/10.3390/pathogens14030217 - 22 Feb 2025
Cited by 7 | Viewed by 4732
Abstract
Fish and seafood are increasingly recognised as safe and nutritiously valuable foods of animal origin, being a source of about 17% of animal protein globally. Novel culinary trends encourage the consumption of raw or thermally lightly processed fishery products. At the same time, [...] Read more.
Fish and seafood are increasingly recognised as safe and nutritiously valuable foods of animal origin, being a source of about 17% of animal protein globally. Novel culinary trends encourage the consumption of raw or thermally lightly processed fishery products. At the same time, consumers prefer wild, fresh and whole fish over farmed or processed fish. However, the consumption of raw or undercooked fish and other marine organisms poses a risk of contracting parasitic infections, potentially representing a public health risk. Among the most common seafoodborne parasites are members of the Anisakidae family, especially the genus Anisakis, which can cause potentially detrimental effects to human health. These parasites are the causative agent of a zoonosis termed anisakidosis that is prevalent in countries with high per capita fish consumption. Although the number of annual clinical cases varies among countries and regions and is generally not high, sensitisation to this parasite in the general population seems to be considerably higher. Therefore, anisakidosis is still significantly underreported and misdiagnosed globally, making it a disease of rising public health concerns. To prevent infection and mitigate potential negative effects on human health, proper preventive measures such as gutting the fish, freezing or thermal processing are needed. Moreover, a holistic approach implementing One Health principles together with educational campaigns towards the general public and primary care physicians can extend the knowledge on the occurrence of these parasites in their natural hosts and the diagnosis and incidence of anisakidosis, with a final goal to minimize risks for human health and reducing costs for health systems. Full article
(This article belongs to the Section Parasitic Pathogens)
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18 pages, 10968 KB  
Article
The Historical Baseline of Hard Tick Records in Spain (1985–2024)
by Agustín Estrada-Peña, Miguel A. Habela Martínez-Estéllez, Carlos Pradera and Joaquim Castellà
Pathogens 2025, 14(2), 173; https://doi.org/10.3390/pathogens14020173 - 9 Feb 2025
Cited by 1 | Viewed by 3009
Abstract
Ticks are important vectors of pathogens, producing diseases in animals and humans. The planning of information campaigns or prevention programs is heavily based on the knowledge of highly detailed data on tick distribution. This study reports unpublished data on the distribution of more [...] Read more.
Ticks are important vectors of pathogens, producing diseases in animals and humans. The planning of information campaigns or prevention programs is heavily based on the knowledge of highly detailed data on tick distribution. This study reports unpublished data on the distribution of more than 30,000 tick specimens, collected by active surveys in the years 1985–2024 in Spain, from 2285 surveys in 1636 unique sites, providing coordinates with variable accuracy. The report covers new records of Dermacentor marginatus, Dermacentor reticulatus, Haemaphysalis punctata, Haemaphysalis sulcata, Hyalomma marginatum, Hyalomma lusitanicum, Ixodes ricinus, Rhipicephalus bursa, Rhipicephalus hibericus, and Rhipicephalus sanguineus (either s.s. or s.l.). Other species were sporadically collected. Many specimens have been re-examined for accurate identification according to current taxonomic views, mainly in the genus Rhipicephalus. We summarized this newly available information using the Köppen–Geiger climate classification. This compilation of unpublished tick records pinpoints the importance of the systematic monitoring of ticks. It is intended as the baseline over which the ongoing national tick collection program is built in order to track the long-term changes of tick distribution in Spain, because of the land use changes, the spread of invasive vertebrates, or the climate trends. Full article
(This article belongs to the Section Ticks)
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13 pages, 1026 KB  
Article
Molecular and Clinical Characterization of Invasive Streptococcus pyogenes Isolates: Insights from Two Northern-Italy Centers
by Carmelo Bonomo, Eva Mannino, Dafne Bongiorno, Caterina Vocale, Armando Amicucci, Dalida Bivona, Davide Guariglia, Emanuele Nicitra, Grete Francesca Privitera, Giuseppe Sangiorgio, Stefania Stefani and Simone Ambretti
Pathogens 2025, 14(2), 152; https://doi.org/10.3390/pathogens14020152 - 5 Feb 2025
Cited by 3 | Viewed by 2481
Abstract
Streptococcus pyogenes (Group A Streptococcus, GAS) is a Gram-positive pathogen responsible for both superficial and invasive infections (iGAS), with increasing global incidence in recent years. This study aims to characterize the molecular and clinical features of iGAS cases in Bologna and Imola (Italy) [...] Read more.
Streptococcus pyogenes (Group A Streptococcus, GAS) is a Gram-positive pathogen responsible for both superficial and invasive infections (iGAS), with increasing global incidence in recent years. This study aims to characterize the molecular and clinical features of iGAS cases in Bologna and Imola (Italy) between 2022 and 2024. Thirty-five invasive isolates were analyzed through whole-genome sequencing (WGS) to investigate the distribution of emm types, antimicrobial resistance (AMR) genes, and virulence factors. Clinical and epidemiological data were retrospectively collected and analyzed. The majority of cases (80%) were recorded in 2023, predominantly among patients aged over 65 (60%). Bloodstream infections were present in 97.1% of cases, and comorbidities such as diabetes and immunosuppression were common. Empirical antibiotic therapy often involved penicillin/β-lactam inhibitors, while oxazolidinones were the most frequently used in targeted regimens. The in-hospital mortality rate was 20%. Genomic analysis identified emm1, emm12, and emm89 as the most prevalent types, associated with specific virulence profiles and resistance determinants. This study highlights the critical role of emm typing and genomic characterization in understanding the pathogenicity of GAS. These findings contribute to the identification of risk factors for severe outcomes and underscore the need for targeted prevention and treatment strategies in vulnerable populations. Full article
(This article belongs to the Section Bacterial Pathogens)
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23 pages, 3056 KB  
Article
Putrescine Depletion in Leishmania donovani Parasites Causes Immediate Proliferation Arrest Followed by an Apoptosis-like Cell Death
by Julia Johnston, Jonathan Taylor, Surbhi Nahata, Angelica Gatica-Gomez, Yvette L. Anderson, Sophia Kiger, Thong Pham, Kayhan Karimi, Jasmin-Faith Lacar, Nicola S. Carter and Sigrid C. Roberts
Pathogens 2025, 14(2), 137; https://doi.org/10.3390/pathogens14020137 - 2 Feb 2025
Cited by 1 | Viewed by 2037
Abstract
The polyamine pathway in Leishmania parasites has emerged as a promising target for therapeutic intervention, yet the functions of polyamines in parasites remain largely unexplored. Ornithine decarboxylase (ODC) and spermidine synthase (SPDSYN) catalyze the sequential conversion of ornithine to putrescine and spermidine. We [...] Read more.
The polyamine pathway in Leishmania parasites has emerged as a promising target for therapeutic intervention, yet the functions of polyamines in parasites remain largely unexplored. Ornithine decarboxylase (ODC) and spermidine synthase (SPDSYN) catalyze the sequential conversion of ornithine to putrescine and spermidine. We previously found that Leishmania donovani Δodc and Δspdsyn mutants exhibit markedly reduced growth in vitro and diminished infectivity in mice, with the effect being most pronounced in putrescine-depleted Δodc mutants. Here, we report that, in polyamine-free media, ∆odc mutants arrested proliferation and replication, while ∆spdsyn mutants showed a slow growth and replication phenotype. Starved ∆odc parasites also exhibited a marked reduction in metabolism, which was not observed in the starved ∆spdsyn cells. In contrast, both mutants displayed mitochondrial membrane hyperpolarization. Hallmarks of apoptosis, specifically DNA fragmentation and membrane modifications, were observed in Δodc mutants incubated in polyamine-free media. These results show that putrescine depletion had an immediate detrimental effect on cell growth, replication, and mitochondrial metabolism and caused an apoptosis-like death phenotype. Our findings establish ODC as the most promising therapeutic target within the polyamine biosynthetic pathway for treating leishmaniasis. Full article
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14 pages, 698 KB  
Article
Long-Term Surveillance of Food Products of Diverse Origins: A Five-Year Survey of Hepatitis A and Norovirus in Greece, 2019–2024
by Rafail Fokas, Zoi Anastopoulou, Kalypso-Angeliki Koukouvini, Maria-Eleni Dimitrakopoulou, Zoi Kotsiri, Eleftheria Chorti-Tripsa, Chrysoula Kotsalou, Dimosthenis Tzimotoudis and Apostolos Vantarakis
Pathogens 2025, 14(2), 135; https://doi.org/10.3390/pathogens14020135 - 2 Feb 2025
Cited by 2 | Viewed by 1797
Abstract
This study examines at the prevalence and spread of Hepatitis A Virus (HAV) and norovirus GI/GII in local and imported food products in Greece over a five-year period (2019–2024). A total of two hundred sixty-six food samples were evaluated using obligatory inspections and [...] Read more.
This study examines at the prevalence and spread of Hepatitis A Virus (HAV) and norovirus GI/GII in local and imported food products in Greece over a five-year period (2019–2024). A total of two hundred sixty-six food samples were evaluated using obligatory inspections and virus detection procedures, including 202 for Hepatitis A and 64 for Norovirus. High-risk categories analyzed were vegetables [138 (HAV), 17 (NoV)], fruits [16 (HAV), 7 (NoV)], soft fruits/berries [37 (HAV), 31 (NoV)], processed meals [4 (HAV), 4 (NoV)], and animal-based products [1 (HAV), 5 (NoV)]. Viral RNA was isolated using QIAamp Viral RNA Mini Kit and detected using established RT-qPCR procedures that met ISO requirements for high sensitivity and reproducibility. The results demonstrated HAV contamination mostly in vegetables (4.35% positive rate), with sporadic findings in other categories. Norovirus GI/GII was detected primarily in soft fruits/berries, with a category-specific positive rate of 6.45%. A temporal study revealed that HAV peaks in 2020, while Norovirus contaminations were detected in 2021 and 2024. The findings highlight the important need to incorporate viral testing into routine food safety procedures, especially for high-risk product categories. This study establishes a basic framework for public health initiatives that address gaps in foodborne virus surveillance in Greece. The study’s ramifications extend to global efforts to monitor and reduce foodborne virus contamination, pushing for higher regulatory requirements and targeted preventative actions. Full article
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15 pages, 2841 KB  
Article
Biocontrol Potential of Trichoderma spp. Against Phytophthora ramorum
by Elisa Becker, Nirusan Rajakulendran and Simon Francis Shamoun
Pathogens 2025, 14(2), 136; https://doi.org/10.3390/pathogens14020136 - 2 Feb 2025
Cited by 3 | Viewed by 2357
Abstract
Phytophthora ramorum, the cause of Sudden Oak Death and related diseases, threatens over 130 tree and shrub species. We evaluated the biocontrol potential of isolates from nine Trichoderma species against P. ramorum using growth-rate studies, dual-culture assays, and culture-filtrate assays. Results showed [...] Read more.
Phytophthora ramorum, the cause of Sudden Oak Death and related diseases, threatens over 130 tree and shrub species. We evaluated the biocontrol potential of isolates from nine Trichoderma species against P. ramorum using growth-rate studies, dual-culture assays, and culture-filtrate assays. Results showed significant variation in Trichoderma growth rates and biocontrol potential. Some isolates exhibited rapid growth, effective overgrowth, and lethal effects against P. ramorum and produced potent antagonistic metabolites. Faster growth rates only partially correlated with biocontrol efficacy, indicating that factors beyond growth, such as metabolite production, play significant roles. Notably, isolates of T. koningii, T. viride, and the commercial product SoilGard™ (T. virens) showed promising efficacy. We calculated a combined biocontrol variable to rank isolates based on vigour and efficacy to aid in identifying promising candidates. Our findings support the use of Trichoderma spp. as biocontrol agents against P. ramorum and underscore the need for a multifaceted approach to selecting and optimizing isolates. Our evaluation demonstrated the importance of using different assays to assess specific mechanisms of action of biocontrol candidates. Future research should further explore these interactions to enhance the sustainable management of P. ramorum. Full article
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12 pages, 258 KB  
Review
Novel Strategies for Preventing Fungal Infections—Outline
by Damilola J. Agbadamashi and Claire L. Price
Pathogens 2025, 14(2), 126; https://doi.org/10.3390/pathogens14020126 - 1 Feb 2025
Cited by 7 | Viewed by 2783
Abstract
Fungal infections are a significant global health challenge, causing approximately 3.8 million deaths annually, with immunocompromised populations particularly at risk. Traditional antifungal therapies, including azoles, echinocandins, and polyenes, face limitations due to rising antifungal resistance, toxicity, and inadequate treatment options. This review explores [...] Read more.
Fungal infections are a significant global health challenge, causing approximately 3.8 million deaths annually, with immunocompromised populations particularly at risk. Traditional antifungal therapies, including azoles, echinocandins, and polyenes, face limitations due to rising antifungal resistance, toxicity, and inadequate treatment options. This review explores innovative strategies for preventing and managing fungal infections, such as vaccines, antifungal peptides, nanotechnology, probiotics, and immunotherapy. Vaccines offer promising avenues for long-term protection, despite difficulties in their development due to fungal complexity and immune evasion mechanisms. Antifungal peptides provide a novel class of agents with broad-spectrum activity and reduced resistance risk, whilst nanotechnology enables targeted, effective drug delivery systems. Probiotics show potential in preventing fungal infections, particularly vulvovaginal candidiasis, by maintaining microbial balance. Immunotherapy leverages immune system modulation to enhance antifungal defenses, and omics technologies deliver comprehensive insights into fungal biology, paving the way for novel therapeutic and vaccine targets. While these approaches hold immense promise, challenges such as cost, accessibility, and translational barriers remain. A coordinated effort among researchers, clinicians, and policymakers is critical to advancing these strategies and addressing the global burden of fungal infections effectively. Full article
(This article belongs to the Section Fungal Pathogens)
13 pages, 271 KB  
Review
An Update on Vaccines Against Trypanosoma cruzi and Chagas Disease
by Nisha J. Garg
Pathogens 2025, 14(2), 124; https://doi.org/10.3390/pathogens14020124 - 30 Jan 2025
Cited by 2 | Viewed by 2348
Abstract
Chagas disease (CD) is a global health concern, with no existing therapies to prophylactically treat adults traveling to endemic countries or those who may already be infected with Trypanosoma cruzi. The economic burden of Chagas cardiomyopathy and heart failure, due to healthcare [...] Read more.
Chagas disease (CD) is a global health concern, with no existing therapies to prophylactically treat adults traveling to endemic countries or those who may already be infected with Trypanosoma cruzi. The economic burden of Chagas cardiomyopathy and heart failure, due to healthcare costs and lost productivity from premature deaths, provides a strong rationale for investment in the development of immune therapies against CD. Vaccine efficacy is proposed to depend heavily on the induction of a robust Th1 response for the clearance of intracellular pathogens like T. cruzi. In this review, updated information on the efforts for vaccine development against CD is provided. Full article
18 pages, 14380 KB  
Article
Optimization of Enterovirus-like Particle Production and Purification Using Design of Experiments
by Louis Kuijpers, Wouter J. P. van den Braak, Abbas Freydoonian, Nynke H. Dekker and Leo A. van der Pol
Pathogens 2025, 14(2), 118; https://doi.org/10.3390/pathogens14020118 - 27 Jan 2025
Cited by 1 | Viewed by 2168
Abstract
Hand, foot, and mouth disease (HFMD) represents an emerging health concern whose main causative agents are Coxsackievirus A6 (CVA6) and enterovirus A71 (EV71). The lack of a CVA6 vaccine and the rise of new HFMD-causing strains due to the containment of established HFMD-causing [...] Read more.
Hand, foot, and mouth disease (HFMD) represents an emerging health concern whose main causative agents are Coxsackievirus A6 (CVA6) and enterovirus A71 (EV71). The lack of a CVA6 vaccine and the rise of new HFMD-causing strains due to the containment of established HFMD-causing viruses necessitates the search for alternative vaccine technologies, including virus-like particle (VLP) vaccine candidates. While studies have demonstrated that production of enterovirus-like particles in various organisms can be achieved by expression of the viral P1 structural proteins and the 3CD protease, optimization based on the interplay between the three most commonly altered infection parameters (multiplicity of infection (MOI), viable cell density at the time of infection (VCD), and the infection period) is often not investigated. To address this challenge, we have performed Design of Experiments (DoE) to optimize the production of CVA6 and EV71 VLPs. Our results indicate that CVA6 VLP production peaks at high MOI, high VCD, and long infection periods. Our subsequent downstream purification processes yielded 38 mg and 158 mg of purified CVA6 and EV71 VLPs from 1 L crude harvest, respectively. This translates into thousands of potential vaccine doses and highlights the economic potential of enterovirus-like particles for vaccine purposes. Full article
(This article belongs to the Special Issue Hand–Foot–Mouth Disease)
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11 pages, 839 KB  
Article
Prevalence of T. rubrum and T. interdigitale Exhibiting High MICs to Terbinafine in Clinical Samples Analyzed in the Portuguese Mycology Reference Laboratory
by Helena Schirmer, Camila Henriques, Helena Simões, Cristina Veríssimo and Raquel Sabino
Pathogens 2025, 14(2), 115; https://doi.org/10.3390/pathogens14020115 - 25 Jan 2025
Cited by 5 | Viewed by 2078
Abstract
Cutaneous fungal infections represent a significant burden worldwide with a high impact on public health. Accurate identification of dermatophyte species causing these infections is vital for an appropriate treatment. Terbinafine is the primary agent against Trichophyton species due to its clinical efficacy; however, [...] Read more.
Cutaneous fungal infections represent a significant burden worldwide with a high impact on public health. Accurate identification of dermatophyte species causing these infections is vital for an appropriate treatment. Terbinafine is the primary agent against Trichophyton species due to its clinical efficacy; however, cases of elevated minimum inhibitory concentration (MIC) have been reported, raising clinical and epidemiological concerns. Herein, we aimed to detect Trichophyton rubrum and Trichophyton interdigitale isolates collected from clinical samples with terbinafine-high MICs (TRB-hMIC). A total of 168 isolates, recovered from 2017 to 2023, were identified as T. rubrum complex (140/83.4%) or T. interdigitale (28/16.7%) and further screened regarding their terbinafine susceptibility. Four isolates with capacity to grow in terbinafine media were detected by screening, and these and a further sixteen random isolates were submitted to the broth microdilution method. This methodology confirmed the four (2.4%) isolates as TRB-hMIC. One T. rubrum and three T. interdigitale showed a minimum inhibitory concentration (MIC) higher than 1 mg/L. Partial sequencing of the SQLE gene identified point mutations in T. rubrum (Phe397Iso) and in one T. interdigitale (Phe397Leu) isolate. Notably, in the other two T. interdigitale isolates with TRB-hMIC, no point mutations in the SQLE gene were identified. In conclusion, TRB-hMIC isolates (T. rubrum and T. interdigitale) were identified in clinical samples analyzed in Portugal, as antifungal susceptibility testing is a crucial routine for identifying treatment failures and also for epidemiological purposes aiming to monitor the dynamics of terbinafine resistance. Full article
(This article belongs to the Special Issue An Update on Fungal Infections)
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