Pathogens

13 pages, 3018 KB

Open AccessArticle

Development of Immunodetection Systems Using a Specific Antibody Against the Recombinant Coat Protein for Detecting Sugarcane Streak Mosaic Virus

by Intan Ria Neliana, Bambang Sugiharto, Rikno Harmoko and Wahyu Indra Duwi Fanata

Pathogens 2025, 14(11), 1106; https://doi.org/10.3390/pathogens14111106 (registering DOI) - 30 Oct 2025

Abstract

Sugarcane streak mosaic virus (SCSMV) is one of the mosaic viruses found in mixed infection with two or more viruses. Infections of mosaic viruses show highly similar mosaic symptoms and are difficult to distinguish. This study aimed to develop a specific antibody against [...] Read more.

Sugarcane streak mosaic virus (SCSMV) is one of the mosaic viruses found in mixed infection with two or more viruses. Infections of mosaic viruses show highly similar mosaic symptoms and are difficult to distinguish. This study aimed to develop a specific antibody against SCSMV that could potentially be utilized to differentiate mosaic virus infections. The cDNA encoding the coat protein (CP) of SCSMV was isolated by RT-PCR from the symptomatic sugarcane leaves. The cDNA was then used for the production of CP for the development of its polyclonal antibody. The nucleotide sequence of the cDNA showed high homology of 94.2–97.3% at the amino acid level with the CP-cDNA of SCSMV isolated from India (AM749403.1), Africa (OR195142.1), and USA (U75456.1). CP was produced as a recombinant protein with a molecular size of 36.5 kDa in Escherichia coli. The injection of recombinant CP into a rabbit resulted in the production of polyclonal antibodies, which were used for the immunodetection of SCSMV in sugarcane. Immunoblot analysis revealed a specific reaction of SCSMV-CP in symptomatic sugarcane leaves. ELISA (Enzyme-Linked Immunosorbent Assay) and IC-RT-PCR (Immunocapture-Reverse Transcription-Polymerase Chain Reaction) using the CP antibody proved successful for detecting SCSMV infection in sugarcane leaves. The results indicate that the SCSMV-CP antibody is suitable for an immunodetection system and exhibits high specificity for SCSMV infection. Full article

(This article belongs to the Section Viral Pathogens)

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15 pages, 7711 KB

Open AccessArticle

Trem2-MICAL1-P-ERK Axis in Macrophages Confers Protection Against Toxoplasma gondii-Induced Adverse Pregnancy Outcomes

by Xiaoyu Geng, Haochen Yang, Zihan Wang, Ziqian Chen, Jinling Chen and Mei Yang

Pathogens 2025, 14(11), 1105; https://doi.org/10.3390/pathogens14111105 (registering DOI) - 30 Oct 2025

Abstract

Toxoplasma gondii (T. gondii) infection during pregnancy can cause severe placental damage and fetal impairment. Although triggering the receptor expressed on myeloid cells 2 (Trem2) confers protection against T. gondii infection, the precise molecular mechanisms underlying this immunoregulatory role remain incompletely understood. [...] Read more.

Toxoplasma gondii (T. gondii) infection during pregnancy can cause severe placental damage and fetal impairment. Although triggering the receptor expressed on myeloid cells 2 (Trem2) confers protection against T. gondii infection, the precise molecular mechanisms underlying this immunoregulatory role remain incompletely understood. Using a mouse model, this study identifies a novel Trem2-MICAL1-P-ERK axis in macrophages that protects against T. gondii-induced adverse pregnancy outcomes (APO). RNA-seq of Trem2-overexpressing macrophages revealed significant upregulation of 1857 genes, with MICAL1 among the most markedly altered, highlighting its potential role in Trem2-mediated signaling. Mechanistically, correlation analysis, molecular docking, fluorescence co-localization, and immunoprecipitation assays demonstrate that Trem2 directly interacts with MICAL1, which modulates downstream phosphorylated ERK (P-ERK) signaling. In a T. gondii-infected murine pregnancy model, genetic ablation of Trem2 exacerbated pathogen-induced suppression of MICAL1 and P-ERK, whereas macrophage-specific overexpression of Trem2-DAP12 restored this signaling axis. Conversely, MICAL1 overexpression rescued P-ERK activation but failed to regulate Trem2 expression. Further studies in bone marrow-derived macrophages (BMDMs) revealed that Trem2 deficiency potentiated the inhibitory effects of soluble T. gondii antigens (TgAg) on MICAL1 and P-ERK. These findings elucidate how T. gondii disrupts placental immunity through targeted suppression of Trem2-mediated signaling and establish the Trem2-MICAL1-P-ERK cascade as a core regulatory pathway in immune homeostasis during pregnancy. Full article

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16 pages, 2123 KB

Open AccessArticle

Low Neutrophil Counts in Milk Are Associated with an Increased Frequency of Antimicrobial Treatments

by Alfonso Zecconi, Valerio Sora, Emanuele Invernizzi, Francesca Zaghen and Viviana Chierici Guido

Pathogens 2025, 14(11), 1104; https://doi.org/10.3390/pathogens14111104 - 29 Oct 2025

Abstract

Several studies have demonstrated an association between impaired innate immunity and metabolic parameters, particularly during the periparturient period. However, to our knowledge, no study has been conducted under field conditions investigating the link between low milk polymorphonuclear neutrophil (PMN) levels and increased disease [...] Read more.

Several studies have demonstrated an association between impaired innate immunity and metabolic parameters, particularly during the periparturient period. However, to our knowledge, no study has been conducted under field conditions investigating the link between low milk polymorphonuclear neutrophil (PMN) levels and increased disease frequency. In an attempt to address this knowledge gap, this study examined 6209 cows from 20 dairy herds in Lombardy that were enrolled in a monthly individual dairy herd improvement milk testing program. Analyses of milk test record samples (MTR) included somatic cell count (SCC) and differential cell count (DSCC). A third variable, PLCC (polymorphonuclear neutrophil leukocyte cell count), was calculated by multiplying SCC × DSCC, thus representing PMN cells/mL. A database including compulsory records of all antimicrobial treatments applied in each herd was used as a proxy for disease frequency. In total, 58,090 valid MTR and 12,014 antimicrobial treatments (AMT) were considered for this study. Statistical analyses showed a significant association between the prevalence of cows with a low number of milk PMN and the prevalence of AMT. These results allow routine identification of whether the number of cows with low PLCC exceeds an alarm level within a herd. This threshold was calculated using an ROC curve with a cut-off point of 6% for AMT. This threshold was estimated at 2%, providing 78% accuracy in identifying herds at risk of an increasing treatment rate. This study confirms that cellular markers measured within MTR systems are useful in identifying herds at risk of impaired cellular immunity, thus paving the way for further studies assessing herd and cow immune status with routine milk sampling. Full article

(This article belongs to the Special Issue Dairy Cattle Health: Mastitis, Milk Quality, and Antimicrobial Resistance from a One Health Perspective)

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13 pages, 699 KB

Open AccessArticle

Impact of Empirical and Definitive Antibiotics on Pediatric Febrile Urinary Tract Infection Caused by ESBL-Producing Enterobacterales

by Jin Lee, Yejin Kim, Ye Ji Kim, Seung Beom Han, Jin-Soon Suh, Soo Young Lee and Jong-Hyun Kim

Pathogens 2025, 14(11), 1103; https://doi.org/10.3390/pathogens14111103 - 29 Oct 2025

Abstract

Although the clinical impact of empirical antibiotic susceptibility on urinary tract infection (UTI) caused by extended-spectrum β-lactamase-producing Enterobacterales (ESBL-PEs) is limited, the role of definitive antibiotics remains unclear. This study evaluated therapeutic outcomes and recurrence in children with febrile UTI caused by ESBL-PE, [...] Read more.

Although the clinical impact of empirical antibiotic susceptibility on urinary tract infection (UTI) caused by extended-spectrum β-lactamase-producing Enterobacterales (ESBL-PEs) is limited, the role of definitive antibiotics remains unclear. This study evaluated therapeutic outcomes and recurrence in children with febrile UTI caused by ESBL-PE, focusing on pathogen susceptibility to empirical and definitive antibiotics. Medical records of 376 UTI episodes caused by ESBL-PE in children aged <10 years were retrospectively reviewed, and 319 were analyzed. Episodes were classified as empirical-susceptible (n = 144) and empirical-non-susceptible (n = 175) groups, and further categorized into four groups based on definitive antibiotic susceptibility and treatment duration. Clinical outcomes were comparable between empirical-susceptible and empirical-non-susceptible groups. Among the four definitive-therapy groups, early clinical response did not differ significantly; however, UTI recurrence showed an increasing trend from 1.0% in those receiving empirical and definitive antibiotics to which pathogens were susceptible for ≥7 days to 6.3% in those receiving empirical antibiotics to which pathogens were non-susceptible and not receiving antibiotics to which pathogens were susceptible ≥7 days (p = 0.045). Despite favorable early response to empirical non-susceptible antibiotics, switching to susceptible antibiotics and maintaining therapy for ≥7 days is recommended for children with ESBL-PE UTIs. Full article

(This article belongs to the Special Issue Current Progress on Bacterial Antimicrobial Resistance)

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17 pages, 3105 KB

Open AccessReview

Novel Therapies for Prosthetic Joint Infections Caused by Methicillin-Resistant Staphylococcus aureus

by Xi Xiang, Xin Jin, Qi Yang, Lili Zou, Yueqing Wang, Tianxu Wang and Xun Sun

Pathogens 2025, 14(11), 1102; https://doi.org/10.3390/pathogens14111102 (registering DOI) - 29 Oct 2025

Abstract

Periprosthetic joint infection (PJI) is a serious complication following total joint replacement, with methicillin-resistant Staphylococcus aureus (MRSA) being the primary pathogen. The treatment challenges posed by MRSA’s antibiotic resistance further highlight the critical importance of research in this field. Current antibiotic therapies for [...] Read more.

Periprosthetic joint infection (PJI) is a serious complication following total joint replacement, with methicillin-resistant Staphylococcus aureus (MRSA) being the primary pathogen. The treatment challenges posed by MRSA’s antibiotic resistance further highlight the critical importance of research in this field. Current antibiotic therapies for periprosthetic joint infection caused by methicillin-resistant Staphylococcus aureus (MRSA-PJI) are limited by considerable side effects, such as high costs and the development of resistance. Therefore, there is an urgent need to explore novel alternative or adjunctive therapies. This review provides a comprehensive overview of several innovative therapeutic strategies. These include monoclonal antibody therapies that target specific bacterial components; phage therapy, which can either independently or synergistically degrade biofilms and enhance antimicrobial efficacy, characterized by its high specificity; antimicrobial peptides, capable of disrupting bacterial membrane integrity and exhibiting dual antibiofilm activity, with a reduced tendency to induce resistance; and nanoparticles and hydrogels, which function as drug delivery systems for sustained release, thereby improving both preventive and therapeutic outcomes. However, these novel therapies also face challenges such as high production costs and limited stability, underscoring the need for further research and optimization. Future efforts should focus on additional studies, clinical trials, and the development of robust regulatory frameworks to fully realize the potential of these treatments for MRSA-PJI. Full article

(This article belongs to the Special Issue Advanced Antimicrobial Agents: Combatting Multi-Drug Resistant Bacterial Infections)

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13 pages, 2848 KB

Open AccessArticle

Coxiella burnetii Strains Elicit Distinct Inflammatory Responses in Human Macrophages

by Madhur Sachan, Amanda Dragan, Het Adhvaryu, Daniel E. Voth and Rahul Raghavan

Pathogens 2025, 14(11), 1101; https://doi.org/10.3390/pathogens14111101 - 29 Oct 2025

Abstract

Coxiella burnetii, the causative agent of human Q fever, subverts macrophage antimicrobial functions to establish an intracellular replicative niche. To better understand host–pathogen interactions, we investigated the transcriptional responses of human alveolar macrophages (hAMs) infected with virulent [NMI, G (Q212)], attenuated (NMII), [...] Read more.

Coxiella burnetii, the causative agent of human Q fever, subverts macrophage antimicrobial functions to establish an intracellular replicative niche. To better understand host–pathogen interactions, we investigated the transcriptional responses of human alveolar macrophages (hAMs) infected with virulent [NMI, G (Q212)], attenuated (NMII), and avirulent (Dugway) strains of C. burnetii. RNA sequencing indicated that all strains activated proinflammatory pathways, particularly IL-17 signaling, though the magnitude and nature of the response varied by strain. Infections with NMI, NMII or G (Q212) resulted in differential expression of roughly the same number of genes, while Dugway infection induced a stronger transcriptional response. Dugway and G (Q212) tended to polarize macrophages toward M1-like states, whereas responses to NMI and NMII were variable. Cytokine assays of NMII-infected THP-1 macrophages suggested the activation of IL-17 signaling, but only at later stages of infection, and single-cell RNA sequencing of NMII-infected THP-1 macrophages indicated heterogeneity in host response to infection, with distinct subpopulations exhibiting M1-like and M2-like inflammatory profiles. These findings highlight the complexity of macrophage response to C. burnetii and underscore the importance of strain-specific and cell-specific factors in shaping host immunity. Understanding these dynamics may inform the development of targeted therapies for Q fever. Full article

(This article belongs to the Special Issue Coxiella burnetii: The Host Immune Response, Host-Pathogen Interaction and Pathogenesis)

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13 pages, 906 KB

Open AccessReview

Alternative Sigma Factors of RNA Polymerase as Master Regulators in the Pathogenic Spirochaete Leptospira interrogans

by Sabina Kędzierska-Mieszkowska and Zbigniew Arent

Pathogens 2025, 14(11), 1100; https://doi.org/10.3390/pathogens14111100 - 29 Oct 2025

Abstract

This review summarizes the current knowledge on the role of alternative σ factors in the highly invasive spirochaete Leptospira interrogans, the causative agent of leptospirosis. This globally distributed zoonosis affects both animals and humans, resulting in substantial public health and economic consequences. [...] Read more.

This review summarizes the current knowledge on the role of alternative σ factors in the highly invasive spirochaete Leptospira interrogans, the causative agent of leptospirosis. This globally distributed zoonosis affects both animals and humans, resulting in substantial public health and economic consequences. Together with the primary σ⁷⁰, alternative σ factors provide transcriptional flexibility essential for bacterial adaptation to environmental changes and host–pathogen interactions. Comparative genomic analyses have revealed that the L. interrogans genome encodes 14 σ factors, including one housekeeping σ⁷⁰-like factor and three types of alternative σ factors: σ⁵⁴, σ²⁸, and 11 predicted extracytoplasmic function (ECF, σᴱ-type) factors. This review discusses the characteristics of these regulators, with particular emphasis on the poorly understood ECF σ factors and their potential roles in gene regulation, adaptive responses, and pathogenicity. Full article

(This article belongs to the Special Issue Emerging and Re-Emerging Bacterial Infections)

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18 pages, 2532 KB

Open AccessArticle

Genome-Based In Silico Analysis of the Structural and Functional Characteristics of the Type Three Secretion System (T3SS) and Core Effector Proteins in Enteropathogenic Escherichia coli (EPEC) Strains Isolated from Food-Producing Animals and Products of Animal Origin

by Refiloe Malesa, Rian Pierneef, Thendo Mafuna, Kudakwashe Magwedere, Emmanuel Seakamela and Itumeleng Matle

Pathogens 2025, 14(11), 1099; https://doi.org/10.3390/pathogens14111099 - 29 Oct 2025

Abstract

Enteropathogenic Escherichia coli (EPEC) is a significant diarrheagenic pathotype responsible for severe gastrointestinal infections, particularly in vulnerable populations. The aim of this study is to utilize genome-based in silico analysis to study the structural and functional characteristics of the Type III Secretion System [...] Read more.

Enteropathogenic Escherichia coli (EPEC) is a significant diarrheagenic pathotype responsible for severe gastrointestinal infections, particularly in vulnerable populations. The aim of this study is to utilize genome-based in silico analysis to study the structural and functional characteristics of the Type III Secretion System (T3SS) and its core effector proteins in EPEC strains. Representative proteins were selected, with particular interest placed on EscV and EscD as major parts of the export apparatus and the basal body, while the EspA effector protein forms the filamentous structure. Several in silico-based techniques were employed, revealing key structural proteins, core effectors, and adhesion-related proteins among the sequenced isolates. Of the 27 isolates analyzed, only 3 (11%) were found to carry LEE-encoded proteins associated with T3SS structural components (escV, escN, escD, and escU) and core effector proteins (espA, espD, espG, and eae). Structural predictions and Ramachandran plot validations suggested stability and potential functional conservation of T3SS proteins, with EscV and EspA selected for detailed 3D structural modelling. Insights into transmembrane domains, protein–protein interaction, and secondary structures were obtained, providing a comprehensive understanding of T3SS assembly and function. These findings suggest that the T3SS in EPEC consists of stable proteins that enable the system to remain functional. The structural and functional properties of the LEE genes encoding the T3SS in the EPEC pathotype represent promising targets for developing virulence blockers to disrupt the pathogenesis of a broad range of bacteria. This study is the first to report EPEC strains with functional T3SS in South Africa, emphasizing the importance of continued surveillance and molecular characterization of EPEC strains. The findings contribute to the development of targeted interventions to mitigate foodborne infections and improve public health. Full article

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11 pages, 843 KB

Open AccessArticle

Eighteen Years of Human Rhinovirus Surveillance in the Republic of Korea (2007–2024): Age- and Season-Specific Trends from a Single-Center Study with Public Health Implications

by Yu Jeong Kim, Jeong Su Han, Sung Hun Jang, Jae-Sik Jeon and Jae Kyung Kim

Pathogens 2025, 14(11), 1098; https://doi.org/10.3390/pathogens14111098 - 28 Oct 2025

Abstract

Human rhinovirus (HRV) is the most common cause of upper respiratory tract infections and can cause substantial morbidity in children. Because its clinical features are nonspecific, differentiation from influenza virus and respiratory syncytial virus is often difficult, underscoring the diagnostic importance of real-time [...] Read more.

Human rhinovirus (HRV) is the most common cause of upper respiratory tract infections and can cause substantial morbidity in children. Because its clinical features are nonspecific, differentiation from influenza virus and respiratory syncytial virus is often difficult, underscoring the diagnostic importance of real-time reverse transcriptase polymerase chain reaction (Real-Time RT-PCR)-based detection. This study aimed to characterize long-term epidemiological patterns of HRV in the Republic of Korea and assess their clinical and public health implications. We retrospectively analyzed 23,284 nasopharyngeal swab specimens collected between 2007 and 2024 from outpatients and inpatients presenting with influenza-like illness at a tertiary care hospital. HRV RNA was detected by Real-Time RT-PCR, and positivity rates were compared by year, month, and age group. Annual detection peaked in 2015 (31.3%) and 2016 (28.6%), then dropped sharply during the COVID-19 pandemic (2020–2021, 4.2–11.0%) and remained low through 2024. Seasonally, rates were highest in July (24.4%) and September (24.1%) and lowest in January (6.9%). Age-specific analysis showed peak positivity in children (26.1%) and infants (20.3%), with lower rates in adults (3.9%) and older adults (3.3%). These findings underscore the diagnostic value of HRV detection and provide evidence for pediatric-focused prevention, outbreak preparedness, and climate-informed surveillance strategies. Full article

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17 pages, 15673 KB

Open AccessArticle

Mitochondrial Network Fragmentation Leads to Dysfunction of Macrophages During Echinococcus multilocularis Protoscoleces Infection

by Zihan Yang, Yaogang Zhang, Tao Zhang, Jing Hou, Meiyuan Tian, Dengliang Huang, Yuan Jiang, Li Sun, Panlong Wei and Yanyan Ma

Pathogens 2025, 14(11), 1097; https://doi.org/10.3390/pathogens14111097 - 28 Oct 2025

Abstract

Alveolar echinococcosis (AE) is a fatal foodborne parasitic disease caused by the larvae of Echinococcus multilocularis. The disease primarily affects the liver. Previous studies have found that Kupffer cells have an immune protective effect, but in the late stages of AE, they [...] Read more.

Alveolar echinococcosis (AE) is a fatal foodborne parasitic disease caused by the larvae of Echinococcus multilocularis. The disease primarily affects the liver. Previous studies have found that Kupffer cells have an immune protective effect, but in the late stages of AE, they are associated with parasite immune escape. The present study analyzed the effects of Echinococcus multilocularis protoscoleces (PSCs) infection on the mitochondrial morphology and function of macrophages, as well as their phagocytic function and apoptosis. Infection with PSCs has been shown to result in the fragmentation of the macrophage mitochondrial network, the impairment of mitochondrial membrane potential, the elevation of mitochondrial reactive oxygen species, and the reduction in mitochondrial DNA copy number. This cascade of events, consequent to the infection, has been demonstrated to promote the apoptosis of macrophages and impair their phagocytic function. Inhibiting mitochondrial fission during PSCs infection has been shown to mitigate mitochondrial dysfunction, suppress macrophage apoptosis, and enhance macrophage phagocytic function. This discovery provides insights into improving macrophage function during the progression of AE. Full article

(This article belongs to the Section Parasitic Pathogens)

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17 pages, 4755 KB

Open AccessArticle

Experimental Evaluation of the Efficacy of Air-Sanitizing Equipment in Neutralizing Airborne SARS-CoV-2 Virus

by Alberto Izzotti, Nicolò Ruzzarin, Oriana Ferrante and Alessandra Pulliero

Pathogens 2025, 14(11), 1096; https://doi.org/10.3390/pathogens14111096 - 28 Oct 2025

Abstract

Air-sanitizing equipment is a collection of protective devices using filtration and/or UV irradiation to entrap aerosol and kill viruses, to prevent the spread of airborne infective diseases in indoor environments. The aim of the herein reported experimental study was to evaluate the possibility [...] Read more.

Air-sanitizing equipment is a collection of protective devices using filtration and/or UV irradiation to entrap aerosol and kill viruses, to prevent the spread of airborne infective diseases in indoor environments. The aim of the herein reported experimental study was to evaluate the possibility of attenuating the environmental spread of the SARS-CoV-2 virus by sanitizing indoor air. Aerosols were generated from human throat swab samples containing viable wild-type SARS-CoV-2. These samples were introduced into a controlled airflow channel and collected in buffered saline, with or without air sanitization. The viral presence was evaluated by antigenic test and qPCR. 34 different types of air- sanitizers were tested for their ability to neutralize viral aerosols. All devices neutralized viral infectivity as evaluated by a antigen test, qPCR, and cell infectivity, except for the unit without filtration and using LED-UV instead of bulbs, which was ineffective at 5 min but effective after 10 min of treatment. The obtained results provide evidence that 97% of the tested sanitizing devices are effective in breaking down the airborne viral load of wild human SARS-CoV-2 virus, even at a very high concentration, with a single passage of air. These results provide evidence that high-quality air-sanitizing devices may be used as a preventive tool to prevent the risk of airborne infections in indoor environments. Full article

(This article belongs to the Special Issue Airborne Transmission of Pathogens)

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17 pages, 711 KB

Open AccessArticle

Toxocariasis as an Elderly Zoonosis: Seroprevalence, Neurocognitive Assessment, and Associated Risk Factors in Persons 50 Years and Older

by Gabriela Geraldi da Silva Rapchan, Isabella Braghin Ferreira, Viviane dos Santos Vaccaro Lima, Susana Angélica Zevallos Lescano, Giovanna Ribelatto Monteiro, Gustavo Cardoso dos Santos, Larissa Sapucaia Ferreira Esteves, Fabiano Borges Figueiredo, Louise Bach Kmetiuk, Alexander Welker Biondo, Rogerio Giuffrida and Vamilton Alvares Santarem

Pathogens 2025, 14(11), 1095; https://doi.org/10.3390/pathogens14111095 - 28 Oct 2025

Abstract

Toxocariasis, a geohelminthiasis caused by Toxocara canis and Toxocara cati nematodes, has an estimated 19% seroprevalence worldwide. Although children have been considered more prone to infection, adults and the elderly may also be considered at risk. Accordingly, the present study aimed to assess [...] Read more.

Toxocariasis, a geohelminthiasis caused by Toxocara canis and Toxocara cati nematodes, has an estimated 19% seroprevalence worldwide. Although children have been considered more prone to infection, adults and the elderly may also be considered at risk. Accordingly, the present study aimed to assess the associated risk factors for Toxocara spp. seropositivity in a population of 290 individuals over 50 years old, attended by the Public Health System in western São Paulo state. Socioepidemiological information was obtained by a semi-structured questionnaire, a blood (serum) sample, and a neurocognitive function assessment by the 10-point cognitive screener (10-CS). Overall, 89/290 (30.7%; 95% CI: 25.7–36.2%) individuals presented anti-Toxocara spp. IgG antibodies by ELISA. Multivariate analysis revealed that individuals raising both dogs and cats were 3-fold more likely to be seropositive than the ones without any pet (p = 0.002), while educational level resulted in an associated protective factor (p = 0.001). No seropositivity influence was observed for the other evaluated variables, including age, gender, monthly income, consumption of drinkable water or raw/undercooked meat, having a dirty floor at home, contact with soil, washing vegetables and hands before meals, and onychophagy. Although seropositivity was not statistically associated with dementia (p = 0.198) and neuropsychiatric disorder (p = 0.440), results herein indicated toxocariasis as a likely under-reported and neglected infection in the older human population. As an immunosenescence risk group that should be continuously monitored, elderly pet owners may be at risk and should be extra careful with self-hygiene and pet deworming, particularly when owning multiple pet species, to reduce the risk of toxocariasis infection. Full article

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41 pages, 2985 KB

Open AccessPerspective

Pathogens That Rewrite the Rules: Ascoviruses, Elegant Manipulators of Cell Death Pathways and Architects of the Extracellular Viral Paradigm

by Sarah R. Rudd, Leticia S. Miranda, Sharon J. Asariah, Chloe S. Rodgers, Jenive T. Estrada, Michael A. Alonzo and Dennis K. Bideshi

Pathogens 2025, 14(11), 1094; https://doi.org/10.3390/pathogens14111094 - 27 Oct 2025

Abstract

Ascoviruses (AVs) are obligate intracellular pathogens that target the larval and pupal stages of lepidopteran insects, specifically moth caterpillars. AVs are unique among viruses in their (i) transmission mode, (ii) gross pathology, (iii) virion ultrastructure, (iv) genomic architecture featuring a remarkable combination of [...] Read more.

Ascoviruses (AVs) are obligate intracellular pathogens that target the larval and pupal stages of lepidopteran insects, specifically moth caterpillars. AVs are unique among viruses in their (i) transmission mode, (ii) gross pathology, (iii) virion ultrastructure, (iv) genomic architecture featuring a remarkable combination of genes, and (v) ability to reprogram host cell death and lipid biosynthetic pathways to generate virion-containing vesicles (VCVs). The metabolically active acellular VCVs are repurposed to complete virogenesis and to facilitate dissemination by endoparasitoid wasps. Since their discovery in the late 20th century, research has focused on these distinctive traits and, to a lesser extent, their potential for biological control. Among AV proteins are the large DNA-binding P64 family, inhibitor of apoptosis-like proteins (IAPs), executioner caspase and caspase-like proteins, and lipid-modifying enzymes, which together drive their novel cytopathology. This review synthesizes current knowledge of AV biology and proposes a framework for understanding VCV formation by integrating predicted viral protein functions with host cellular pathways, including the ceramide–sphingosine-1-phosphate rheostat and apoptosis. Full article

(This article belongs to the Section Viral Pathogens)

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10 pages, 213 KB

Open AccessBrief Report

Standardized Diagnostic Assays for Omsk Hemorrhagic Fever Virus

by Jeong-Hyun Lee, Sunyoung Jung, Hwajung Yi and Yoon-Seok Chung

Pathogens 2025, 14(11), 1093; https://doi.org/10.3390/pathogens14111093 - 27 Oct 2025

Abstract

Omsk hemorrhagic fever is an acute zoonotic disease caused by Omsk hemorrhagic fever virus, a member of the genus Flavivirus (family Flaviviridae), with a reported case-fatality rate of approximately 3%. Historically confined to southwestern Siberia, ecological changes raise concerns about possible spread to [...] Read more.

Omsk hemorrhagic fever is an acute zoonotic disease caused by Omsk hemorrhagic fever virus, a member of the genus Flavivirus (family Flaviviridae), with a reported case-fatality rate of approximately 3%. Historically confined to southwestern Siberia, ecological changes raise concerns about possible spread to non-endemic regions. Although no Omsk hemorrhagic fever cases have been reported in the Republic of Korea, the risk of accidental importation highlights the importance of establishing a reliable diagnostic protocol. We established and validated an institutionally developed diagnostic protocol employing real-time reverse transcription polymerase chain reaction targeting the NS2A and C genes of Omsk hemorrhagic fever virus. Primers and probes were designed from all available genomes to ensure broad strain coverage. Human ribonuclease P was used as an internal control to verify nucleic acid extraction and amplification. Using synthetic deoxyribonucleic acid fragments and in vitro-transcribed ribonucleic acid, assay performance was optimized, and analytical sensitivity was determined using probit analysis. The limits of detection were 74.50 copies/µL (threshold cycle 32.99) for NS2A and 70.41 copies/µL (threshold cycle 35.38) for C. Specificity testing using representative flaviviruses (West Nile virus, Yellow fever virus, Zika virus, St. Louis encephalitis virus, and Tick-borne encephalitis virus) and an alphavirus (Venezuelan equine encephalitis virus) demonstrated no cross-reactivity. The assay demonstrated high sensitivity, specificity, and reproducibility, supporting its potential application in national and international Omsk hemorrhagic fever virus surveillance systems. Full article

(This article belongs to the Special Issue Emerging Viral Pathogens: Diagnosis, Treatment, and Public Health Interventions)

12 pages, 1373 KB

Open AccessArticle

Genomic Surveillance of Plasmodium falciparum Drug Resistance Markers Between October 2021 and June 2023 in Kigali, Rwanda

by Sandra Noukimi Fankem, Jean-Bosco Mbonimpa, Edgar Mutebwa Kalimba, Mariama Telly Diallo, Mary Efeti Teke and Jacob Souopgui

Pathogens 2025, 14(11), 1092; https://doi.org/10.3390/pathogens14111092 - 27 Oct 2025

Abstract

Artemisinin-based combination therapies (ACTs) remain the cornerstone of malaria treatment in Rwanda, but the emergence of drug resistance threatens their efficacy. This study conducted genomic surveillance of Plasmodium falciparum isolates collected in Kigali between October 2021 and June 2023 to assess resistance markers. [...] Read more.

Artemisinin-based combination therapies (ACTs) remain the cornerstone of malaria treatment in Rwanda, but the emergence of drug resistance threatens their efficacy. This study conducted genomic surveillance of Plasmodium falciparum isolates collected in Kigali between October 2021 and June 2023 to assess resistance markers. Using Oxford Nanopore Technology and Sanger sequencing methods, we analyzed 250 clinical isolates focusing on mutations in the pfcrt, pfmdr1, pfdhfr, pfdhps, and Pfkelch13 genes. Resistance-associated mutations were highly prevalent: pfcrt 76T (26%) and pfmdr1 184F (72.8%) were common, indicating continued lumefantrine pressure. All isolates carried mutations in pfdhfr and pfdhps, with the IRNI-SAEAA and IRNI-SAEGA haplotypes found in 45.6% and 24.8% of samples, respectively, suggesting sustained antifolate resistance. Pfkelch13 mutations were present in 50.4% of isolates, including validated R561H (25.6%), A675V and candidate P441L mutations. Novel haplotypes, including K189T + R561H (24.8%), were identified for the first time in Rwanda. The BTB/POZ domain mutation H384R was observed in 6.4% of isolates, raising questions about its potential functional role. These findings highlight complex and evolving resistance patterns and emphasize the urgent need for continued molecular surveillance and functional validation to inform malaria control strategies in Rwanda. Full article

(This article belongs to the Topic Multidrug Resistance Across Pathogens: Fungi, Bacteria, Parasites, and Viruses)

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13 pages, 2384 KB

Open AccessArticle

Phylodynamics of SARS-CoV-2 Lineages B.1.1.7, B.1.1.529 and B.1.617.2 in Nigeria Suggests Divergent Evolutionary Trajectories

by Babatunde O. Motayo, Olukunle O. Oluwasemowo, Anyebe B. Onoja, Paul A. Akinduti and Adedayo O. Faneye

Pathogens 2025, 14(11), 1091; https://doi.org/10.3390/pathogens14111091 - 26 Oct 2025

Abstract

Background: The early months of the COVID-19 pandemic were characterized by high transmission rates and mortality, compounded by the emergence of multiple SARS-CoV-2 lineages, including Variants of Concern (VOCs). This study investigates the phylodynamic and spatio-temporal trends of VOCs during the peak of [...] Read more.

Background: The early months of the COVID-19 pandemic were characterized by high transmission rates and mortality, compounded by the emergence of multiple SARS-CoV-2 lineages, including Variants of Concern (VOCs). This study investigates the phylodynamic and spatio-temporal trends of VOCs during the peak of the pandemic in Nigeria. Methods: Whole-genome sequencing (WGS) data from three major VOCs circulating in Nigeria, B.1.1.7 (Alpha), B.1.617.2 (Delta), and B.1.1.529 (Omicron), were analyzed using tools such as Nextclade, R Studio v 4.2.3, and BEAST X v 10.5.0. The spatial distribution, evolutionary history, viral ancestral introductions, and geographic dispersal patterns were characterized. Results: Three major lineages following WHO nomenclature were identified: Alpha, Delta, and Omicron. The Delta variant exhibited the widest geographic spread, detected in 14 states, while the Alpha variant was the least distributed, identified in only eight states but present across most epidemiological weeks studied. Evolutionary rates varied slightly, with Alpha exhibiting the slowest rate (2.66 × 10⁻⁴ substitutions/site/year). Viral population analyses showed distinct patterns: Omicron sustained elevated population growth over time, while Delta declined after initial expansion. The earliest Times to Most Recent Common Ancestor (TMRCA) were consistent with the earliest outbreaks of SARS-CoV-2 globally. Geographic transmission analysis indicated a predominant coastal-to-inland spread for all variants, with Omicron showing the most diffuse dispersal, highlighting commercial routes as significant drivers of viral diffusion. Conclusion: The SARS-CoV-2 epidemic in Nigeria was characterized by multiple variant introductions and a dominant coastal-to-inland spread, emphasizing that despite lockdown measures, commercial trade routes played a critical role in viral dissemination. These findings provide insights into pandemic control strategies and future outbreak preparedness. Full article

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15 pages, 608 KB

Open AccessReview

A Comprehensive Review of Nanoparticles in the Fight Against Antimicrobial Resistance

by Antonios Mouzakis, Periklis Panagopoulos, Dimitrios Papazoglou and Vasileios Petrakis

Pathogens 2025, 14(11), 1090; https://doi.org/10.3390/pathogens14111090 - 26 Oct 2025

Abstract

(1) Background: The escalating crisis of multidrug-resistant (MDR) bacteria presents a formidable threat to global public health, necessitating the urgent development of alternative antimicrobial strategies. Nanoparticles (NPs) have emerged as a promising frontier in this effort, leveraging their unique physicochemical properties and multi-modal [...] Read more.

(1) Background: The escalating crisis of multidrug-resistant (MDR) bacteria presents a formidable threat to global public health, necessitating the urgent development of alternative antimicrobial strategies. Nanoparticles (NPs) have emerged as a promising frontier in this effort, leveraging their unique physicochemical properties and multi-modal mechanisms of action to combat bacterial infections. This systematic review aims to comprehensively evaluate the current body of evidence on the dynamic interplay between nanoparticles and bacterial resistance. (2) Methods: A comprehensive search of electronic databases, including PubMed, Scopus, and Web of Science, was performed using a combination of keywords and Medical Subject Headings (MeSH) terms to identify relevant primary research articles. Eligibility criteria focused on studies evaluating the antimicrobial effects of nanoparticles on MDR bacterial strains, reporting on mechanisms of action, efficacy, or resistance development. (3) Results: The synthesis of findings revealed that nanoparticles exert their antimicrobial effects through multiple pathways, including the generation of reactive oxygen species (ROS), direct disruption of bacterial membranes, and the release of toxic ions. However, the analysis also confirmed that bacteria have evolved sophisticated defense mechanisms against nanoparticles, including surface modifications that prevent adhesion, upregulation of efflux pumps, and chemical neutralization of toxic ions. (4) Conclusions: Nanoparticles represent a potent and versatile tool in the global effort to combat antimicrobial resistance. Their long-term efficacy is not guaranteed, as bacteria have shown a remarkable capacity for adaptation. The future of this field lies in the development of rationally designed nanoparticle systems that not only possess intrinsic antimicrobial activity but also actively disarm bacterial resistance mechanisms. This includes the strategic use of synergistic combinations with conventional antibiotics and the exploration of resistance-agnostic approaches like nanotoxoid vaccines. Full article

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13 pages, 1624 KB

Open AccessArticle

From Hyperendemic to Low Endemicity: The Effect of Hepatitis B Vaccination on HBV and HDV Prevalence in the Brazilian Amazon

by Andreza Pinheiro Malheiros, Michele Soares Gomes-Gouvêa, Leidiane Barbosa Ribeiro, Alex Junior Souza de Souza, Raymundo Soares Azevedo, Dickson Ciro Nascimento de Brito, Candida Maria Abrahão de Oliveira, Heloisa Marceliano Nunes and João Renato Rebello Pinho

Pathogens 2025, 14(11), 1089; https://doi.org/10.3390/pathogens14111089 - 25 Oct 2025

Abstract

The Amazon Basin was historically hyperendemic for HBV and HDV, associated with severe outcomes like fulminant hepatitis. Brazil initiated its hepatitis B vaccination in 1989. This study assessed the current prevalence in this endemic region to evaluate the impact of vaccination. A cross-sectional [...] Read more.

The Amazon Basin was historically hyperendemic for HBV and HDV, associated with severe outcomes like fulminant hepatitis. Brazil initiated its hepatitis B vaccination in 1989. This study assessed the current prevalence in this endemic region to evaluate the impact of vaccination. A cross-sectional population-based survey enrolled 1100 urban and rural residents. HBsAg prevalence was 1.5%, with no cases in individuals under 20 years, demonstrating interrupted vertical and horizontal transmission. Anti-HBc positivity (30.9%) indicated past exposure, predominantly in those over 30 years. Isolated anti-HBc (10.3%) included two occult HBV infections. HDV coinfection occurred in 25% of HBsAg-positive cases, with HDV RNA detected in two. Anti-HDV positivity was exclusive to adults over 30. Vaccination coverage was poorly documented, but 23.7% had protective anti-HBs titers. HBV vaccination has reduced HBsAg prevalence from high to low endemicity in the region, eliminating chronic infections in younger generations. Persistent HDV in older age groups underscores the need for targeted screening. Despite vaccination record gaps, the findings highlight the program’s success in interrupting transmission and support continued efforts toward HBV/HDV elimination. Full article

(This article belongs to the Special Issue Viral Hepatitis Among Specific Populations: Epidemiology, Transmission, Treatment, and Prevention)

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16 pages, 2325 KB

Open AccessArticle

An Update of Epidemiological Trends in Enzootic Bovine Leukosis in Italy and an Analysis of Risk Factors Associated with Infection Persistence

by Cecilia Righi, Carmen Iscaro, Stefano Petrini, Eleonora Scoccia, Silvia Pirani, Alessandro Fiorucci, Roberto Lomolino and Francesco Feliziani

Pathogens 2025, 14(11), 1088; https://doi.org/10.3390/pathogens14111088 - 24 Oct 2025

Abstract

In 2017, the Commission Implementing Decision (European Union [EU]) 2017/1910 officially declared Italy to be enzootic bovine leukosis (EBL)-free. The Commission Delegated Regulation (EU) 2020/689 laid criteria to maintain an official disease-free status. While some infection clusters persist in restricted areas, specific measures [...] Read more.

In 2017, the Commission Implementing Decision (European Union [EU]) 2017/1910 officially declared Italy to be enzootic bovine leukosis (EBL)-free. The Commission Delegated Regulation (EU) 2020/689 laid criteria to maintain an official disease-free status. While some infection clusters persist in restricted areas, specific measures are being implemented to eliminate pockets of viral persistence. This updated analysis of current data, concerning epidemiological trends of EBL in Italy from January 2022 to December 2024, aimed to analyze the status of infection clusters in terms of risk factors associated with bovine leukemia virus (BLV) seropositivity to evaluate the effectiveness of the eradication measures. Our findings highlighted an improvement in EBL eradication; however, the Latium region lags behind in terms of disease eradication while the situation in Apulia is being resolved. Campania, which has implemented restrictive and consistent health measures, has the lowest prevalence and incidence rates compared with previous years. Identifying and assessing risk factors that favor EBL persistence in infection clusters is essential as is implementing specific measures to eliminate such clusters, thereby enabling disease eradication and the adoption of targeted prevention strategies. Full article

(This article belongs to the Section Epidemiology of Infectious Diseases)

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