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Tackling Biofilm-Forming Pathogens: A Challenge to Overcome in the Fight Against Infectious Diseases -
Functional Antibody-Dependent Enhancement as an Immune Assessment Platform: Development, Standardization, and Translational Interpretation in Flavivirus Research -
Diagnostic Pathways and Genotyping of Cases of Echinococcus granulosus from Polish Patients
Journal Description
Pathogens
Pathogens
is an international, peer-reviewed, open access journal on pathogens and pathogen-host interactions published monthly online by MDPI.
- Open Access— free for readers, with article processing charges (APC) paid by authors or their institutions.
- High Visibility: indexed within Scopus, SCIE (Web of Science), PubMed, MEDLINE, PMC, Embase, PubAg, CaPlus / SciFinder, AGRIS, and other databases.
- Journal Rank: JCR - Q2 (Microbiology) / CiteScore - Q1 (Infectious Diseases)
- Rapid Publication: manuscripts are peer-reviewed and a first decision is provided to authors approximately 12.5 days after submission; acceptance to publication is undertaken in 2.8 days (median values for papers published in this journal in the first half of 2026).
- Recognition of Reviewers: reviewers who provide timely, thorough peer-review reports receive vouchers entitling them to a discount on the APC of their next publication in any MDPI journal, in appreciation of the work done.
- Companion journals for Pathogens include: Parasitologia and Bacteria.
- Journal Cluster of Microbiology: Acta Microbiologica Hellenica, Applied Microbiology, Bacteria, Journal of Fungi, Microorganisms, Microbiology Research, Pathogens, Viruses, Fermentation and Germs.
Impact Factor:
3.8 (2025);
5-Year Impact Factor:
3.9 (2025)
Latest Articles
District-Level Risk Mapping of Highly Pathogenic Avian Influenza in Poultry in Kazakhstan Using a Multi-Criteria Decision Model
Pathogens 2026, 15(8), 796; https://doi.org/10.3390/pathogens15080796 - 27 Jul 2026
Abstract
Highly pathogenic avian influenza (HPAI) is a transboundary disease of birds with zoonotic potential. Since 2021, H5 HPAI virus variants have spread globally, with migratory waterfowl playing a key role in transcontinental dissemination. Kazakhstan lies at the intersection of major flyways, creating a
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Highly pathogenic avian influenza (HPAI) is a transboundary disease of birds with zoonotic potential. Since 2021, H5 HPAI virus variants have spread globally, with migratory waterfowl playing a key role in transcontinental dissemination. Kazakhstan lies at the intersection of major flyways, creating a persistent risk of virus introduction into domestic poultry. While HPAI risk in wild birds has been explored in Kazakhstan, the spatial risk for poultry has remained underassessed. Here, we present a risk map for HPAI in poultry across 174 administrative districts of Kazakhstan, using a multi-criteria decision analysis (TOPSIS) that integrates five quantitative risk indicators that relate to wild bird habitat, virus survival in the environment and poultry farm census. The obtained district-level risk index agreed well with historical outbreak data (2005–2025), with an area under the receiver operating characteristic (ROC) curve of 0.899 (95% CI: 0.846–0.952). Most districts (56%) were at negligible risk, whereas medium- and high-risk clusters occurred in the north, central, and southern regions, and near the Caspian Sea. These findings provide an operational, spatially explicit tool for targeting surveillance and biosecurity measures to high-risk areas, ultimately helping to mitigate the impact of one of the most devastating transboundary poultry diseases in Central Asia.
Full article
(This article belongs to the Special Issue New Insights into Viral Infections of Domestic Animals)
Open AccessReview
Prevention and Treatment of Staphylococcus aureus Biofilms Using Promising Agr-QS-Targeting Anti-Biofilm Agents
by
Salma Waheed Sheikh, Ahmad Ali, Asma Ahsan, Fei Shang, Ting Xue and Lauren Gollahon
Pathogens 2026, 15(8), 795; https://doi.org/10.3390/pathogens15080795 - 27 Jul 2026
Abstract
Staphylococcus aureus (S. aureus), a leading cause of nosocomial infections, contributes significantly to increased morbidity and mortality, especially when it forms biofilms on medical devices. This pathogen, specifically methicillin-resistant S. aureus (MRSA), remains a challenge to treat due to its ability
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Staphylococcus aureus (S. aureus), a leading cause of nosocomial infections, contributes significantly to increased morbidity and mortality, especially when it forms biofilms on medical devices. This pathogen, specifically methicillin-resistant S. aureus (MRSA), remains a challenge to treat due to its ability to form biofilms and rapidly develop resistance against antibiotics. Biofilm formation allows bacteria to adhere to biotic and abiotic surfaces, creating a protective matrix that shields them from immune responses and antibiotic therapies. The widespread prevalence of multidrug-resistant S. aureus biofilms poses a significant therapeutic challenge in clinical settings. Several novel therapeutic strategies have been developed to combat S. aureus biofilm-associated infections. Accumulating evidence suggests that natural plants and their derivatives possess antimicrobial and chemo preventive properties that can disrupt established biofilms. Several plant-derived compounds with anti-biofilm activities have been reported to target the regulatory proteins involved in the Agr quorum sensing (Agr-QS) system, underscoring their potential as therapeutic candidates for the prevention and treatment of biofilm-associated infections. However, despite these encouraging findings, clinical validation of these plant-based agents is essential to ensure their efficacy, safety, and optimal application in treating S. aureus biofilm infections. The continued exploration of natural biofilm inhibitors anticipates the urgent need for new treatments to combat biofilm-associated infections and multidrug-resistant pathogens like MRSA. This review provides a detailed overview of preventive and therapeutic interventions to eradicate biofilm-forming S. aureus infections.
Full article
(This article belongs to the Section Bacterial Pathogens)
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Open AccessArticle
Evaluation of an Automated Cartridge-Based PCR Assay for the Detection of Leishmania spp. DNA in Canine Lymph Node Samples
by
Eva Spada, Francesca Di Gaudio, Germano Castelli, Federica Bruno, Roberta Perego, Luciana Baggiani, Vito Biondi, Fabrizio Vitale, Michela Tognoni and Daniela Proverbio
Pathogens 2026, 15(8), 794; https://doi.org/10.3390/pathogens15080794 - 27 Jul 2026
Abstract
An automated cartridge-based Vcheck M Canine Vector 8 Panel for qualitative detection of Leishmania spp. DNA in canine lymph node aspirates, an off-label specimen type, using laboratory qPCR as the comparator, was evaluated. Fifty-seven residual lymph node aspirate suspensions from dogs investigated for
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An automated cartridge-based Vcheck M Canine Vector 8 Panel for qualitative detection of Leishmania spp. DNA in canine lymph node aspirates, an off-label specimen type, using laboratory qPCR as the comparator, was evaluated. Fifty-seven residual lymph node aspirate suspensions from dogs investigated for suspected canine leishmaniosis (CanL) were tested. Reference qPCR detected L. infantum DNA in 29 samples. Vcheck M was positive in 22/29 qPCR-positive samples and negative in 28/28 qPCR-negative samples, corresponding to positive percent agreement/sensitivity of 75.9% (95% CI, 56.5–89.7) and negative percent agreement/specificity of 100.0% (95% CI, 87.7–100.0). Agreement was substantial (Cohen’s kappa, 0.76), and discordance was asymmetric (McNemar p = 0.016). Vcheck-negative/qPCR-positive results were mainly observed at low qPCR parasite loads: 6/7 discordant samples contained ≤30 parasites/mL, whereas all samples with ≥500 parasites/mL were Vcheck positive. Among Vcheck-positive clinical samples, Vcheck Ct correlated inversely with log10 qPCR parasite load (Spearman rho = −0.77; p < 0.001). In a single-run dilution series, the lowest instrument-positive L. infantum standard was 103 parasites/mL. Purified L. major, L. braziliensis, and L. tropica DNA were also detected. Vcheck M showed high specificity as a rapid rule-in test for Leishmania spp. detection in canine lymph node aspirates. However, negative results should not exclude infection in symptomatic or strongly suspected dogs and should be confirmed by qPCR, particularly when low parasite burden is plausible or when the result is critical for diagnostic or therapeutic decision-making.
Full article
(This article belongs to the Section Parasitic Pathogens)
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Open AccessArticle
Molecular Insights from Differential Proteomic Profiling of Premalignant Cervical Lesions and Cervical Cancer
by
Diana Laura Gonzalez-Tolentino, Olga Lilia Garibay-Cerdenares, Sergio Encarnación-Guevara, Ángel Gabriel Martínez-Batallar, Ramiro Alonso-Bastida, Jeovanis Gil, Jorge Organista-Nava, Luz del Carmen Alarcón-Romero, Marco Antonio Leyva-Vázquez and Berenice Illades-Aguiar
Pathogens 2026, 15(8), 793; https://doi.org/10.3390/pathogens15080793 - 26 Jul 2026
Abstract
Cervical cancer (CC) affects women worldwide, and more than 95% of cases are caused by persistent infection with high-risk human papillomavirus (HR-HPV), such as type 16, which promotes the progression of precancerous lesions to cancer. This study aimed to identify differentially expressed proteins
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Cervical cancer (CC) affects women worldwide, and more than 95% of cases are caused by persistent infection with high-risk human papillomavirus (HR-HPV), such as type 16, which promotes the progression of precancerous lesions to cancer. This study aimed to identify differentially expressed proteins (DEPs) in biopsies from patients with HPV16+ low-grade squamous intraepithelial lesions (LSILs) and from patients with HPV16+ squamous cell carcinoma (SCC) compared with those from HPV-negative normal cervical tissue (NCT HPV−) controls. The samples were analyzed by high-performance liquid chromatography–tandem mass spectrometry (HPLC-MS/MS) using a data-independent acquisition (DIA) approach. Data processing and differential protein expression analysis were performed with the DIA-NN software (Data-Independent Acquisition Neural Networks), followed by bioinformatics analyses, including Venn diagrams, pathway enrichment, functional interactome, The Cancer Genome Atlas (TCGA)-SCC data integration, and Western blot detection. In total, 1607 DEPs associated with cell adhesion and extracellular matrix proteins were identified in LSILs, whereas 1516 DEPs associated with catalytic and transport activities were identified in SCC; the proteins overexpressed in LSILs (332) were enriched in processes such as metabolism, immune response activation, and stress and cell death responses. In contrast, proteins overexpressed in SCC (205) were associated with the cell cycle, DNA damage, drug metabolism, proteasome degradation, methylation, and immune response. Interaction analyses highlighted proteins related to early proteins 1,5,6 and 7 (E1, E5, E6, and E7). In terms of the two DEPs, S100 calcium binding protein A10 (S100A10/p11) and thymidine phosphorylase (TYMP) were detected in patients with LSIL, HSIL, and SCC at the protein level, consistent with their higher transcript levels in public datasets. Given the small, exploratory cohort, these findings are hypothesis-generating, and validation in a larger, balanced, independent cohort is required. In conclusion, this study identified DEPs associated with the progression of premalignant lesions to SCC that may represent candidate biomarkers and therapeutic targets warranting further investigation.
Full article
(This article belongs to the Special Issue Recent Advances in Human Papillomavirus Research)
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Open AccessArticle
Leishmaniasis Incidence and Leishmania Species Distribution in the Apurímac, Ene and Mantaro River Valley, a High-Transmission Endemic Area of Peru
by
Jime Rivera-Villar, Nyshon Rojas-Palomino, José Alarcón-Guerrero, Víctor Cárdenas-López, Rilder Gastelú-Quispe, Aide Sandoval-Juarez and Saúl Chuchón-Martínez
Pathogens 2026, 15(8), 792; https://doi.org/10.3390/pathogens15080792 - 25 Jul 2026
Abstract
Background: Leishmaniasis remains a major public health problem in Peru, and the Apurímac, Ene and Mantaro River Valley (VRAEM) is an endemic area where, despite its epidemiological relevance, information on incidence and circulating Leishmania species remains limited. Methods: We estimated district-level cumulative incidence
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Background: Leishmaniasis remains a major public health problem in Peru, and the Apurímac, Ene and Mantaro River Valley (VRAEM) is an endemic area where, despite its epidemiological relevance, information on incidence and circulating Leishmania species remains limited. Methods: We estimated district-level cumulative incidence of leishmaniasis in the 60 districts of the VRAEM during 2015–2024, using national surveillance data and census population figures. In addition, molecular species typing was performed on 112 Giemsa-stained tissue smears obtained in health facilities located in Ayacucho districts belonging to the VRAEM region. Results: A total of 3589 cases were reported, yielding an estimated annual incidence rate of 80.01 cases per 100,000 inhabitants, which is 4.3-fold higher than the national average. Marked spatial heterogeneity was observed, with districts such as Pangoa, Río Tambo, Llochegua, and Pichari reporting estimated annual incidence rates above 130 cases per 100,000 inhabitants. Furthermore, of 55 samples processed by High-Resolution Melting Analysis, Leishmania braziliensis was found in 54.6% of samples, followed by Leishmania guyanensis in 21.8%. Conclusions: The VRAEM constitutes a high-transmission endemic focus of tegumentary leishmaniasis with marked inter-district heterogeneity and circulation of multiple Leishmania species, with Leishmania braziliensis as the predominant species. These findings support the need for targeted surveillance and species-informed clinical management strategies.
Full article
(This article belongs to the Special Issue Leishmania & Leishmaniasis)
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Open AccessArticle
Analytical Validation and Preliminary Diagnostic Performance Evaluation of GenoPATHX™ Multiplex qPCR for Quantitative Detection of Key Salmonella Serovars in Poultry Matrices
by
Rejoice Nyarku, Emmanuel Kuufire, Viona Osei, Kingsley E. Bentum, Asmaa Elrefaey, Emmanuel Piiru, Tyric James, Yilkal Woube, Temesgen Samuel and Woubit Abebe
Pathogens 2026, 15(8), 791; https://doi.org/10.3390/pathogens15080791 - 25 Jul 2026
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Rapid detection and quantification of epidemiologically important Salmonella enterica serovars are critical for poultry surveillance, food safety monitoring, and risk-based intervention strategies. This study performed comprehensive analytical validation together with a preliminary field-based diagnostic performance evaluation of GenoPATHX™, a multiplex probe-based qPCR platform
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Rapid detection and quantification of epidemiologically important Salmonella enterica serovars are critical for poultry surveillance, food safety monitoring, and risk-based intervention strategies. This study performed comprehensive analytical validation together with a preliminary field-based diagnostic performance evaluation of GenoPATHX™, a multiplex probe-based qPCR platform designed for the simultaneous detection and quantification of priority Salmonella serovars in poultry-associated matrices. The platform consists of two multiplex panels, designated the Chicken Key Performance Indicator (CKPI) and Turkey Key Performance Indicator (TKPI), each designed to detect priority poultry-associated Salmonella serovars together with a genus-level S. enterica marker. Analytical performance was evaluated for amplification efficiency, linearity, limit of detection (LoD95), limit of quantification (LoQ), repeatability, intermediate precision, analytical specificity (inclusivity/exclusivity), robustness, matrix effects, and performance in artificially inoculated matrices. Diagnostic performance was further assessed using naturally contaminated poultry environmental samples. The assay demonstrated robust amplification performance in both singleplex and multiplex formats, with high linearity (R2 = 0.987–0.999) and LoD95 values ranging from 60 to 545 genome equivalents per reaction. Complete analytical inclusivity and high exclusivity were achieved for the evaluated isolate panel. In field samples, the direct GenoPATHX™ workflow demonstrated 81.0% sensitivity, 91.3% specificity, and substantial agreement with the USDA-FSIS reference culture method (κ = 0.73). Overall, GenoPATHX™ exhibited robust analytical performance and enabled rapid, same-day quantitative detection of priority Salmonella serovars in poultry-associated matrices, supporting its application for poultry surveillance and food safety monitoring.
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Open AccessArticle
Molecular Characterization and Prevalence of Trypanozoon Infection in Livestock on Small-Scale Farms in Thailand: An Integrative Control Approach
by
Apiraya Rudeekiatthamrong, Pairpailin Jhaiaun, Giang Thi Nguyen, Wissanuwat Chimnoi, Darunwan Chuensaengarun, Chattraporn Rungchalermlak, Tanapat Mutchimadilok, Nipa Thammasonthijarern, Thawijit Phannithi, Tawanhathai Apichaimongkonkun, Laphatsararach Apinantanakorn, Kanittha Phetudomsinsuk, Tawin Inpankaew, Burin Nimsuphan, Ruttayaporn Ngasaman, Nuttapon Manojai, Jumnongjit Phasuk and Ketsarin Kamyingkird
Pathogens 2026, 15(8), 790; https://doi.org/10.3390/pathogens15080790 - 24 Jul 2026
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Trypanosomosis (Surra), caused by the hemoflagellate protozoan Trypanosoma evansi, has a negative impact on animal health and the livestock economy in many countries, including Thailand. Molecular epidemiological data and integrative control approaches for small-scale farms remain limited. This study determined the prevalence
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Trypanosomosis (Surra), caused by the hemoflagellate protozoan Trypanosoma evansi, has a negative impact on animal health and the livestock economy in many countries, including Thailand. Molecular epidemiological data and integrative control approaches for small-scale farms remain limited. This study determined the prevalence of Trypanozoon infection and its associated risk factors in Thai livestock under an integrative control approach. Molecular tools were used to characterize T. evansi typing. A total of 947 blood samples were collected from cattle and buffalo across 123 small-scale farms in 16 provinces covering four regions of Thailand, with diagnostic results and control recommendations communicated to farmers within 14 days. The ITS2-positive samples were then subjected to further molecular characterization, together with two dogs, seven horses, and an in vitro T. evansi isolate, comprising 40 Trypanozoon-positive samples. These were characterized using PCR targeting the TBR primer, maxicircle NADH5, VSG RoTat 1.2, and minicircle B, followed by ITS2-based phylogenetic analysis. Risk factors were assessed using Chi-square and Fisher’s exact tests, logistic regression, a generalized linear mixed model, and Firth’s penalized logistic regression. The overall prevalence of Trypanozoon infection was 3.17% (30/947), detected exclusively in beef cattle in the Northern region (15.30%; 30/196), specifically in Lampang (46.15%, 12/26), Tak (23.08%, 3/13), and Phrae (17.24%, 15/87) Provinces. No infections were reported in the Northeastern, Central, or Southern regions. Male sex, age under one year, and residence in the Northern region were significant individual-level risk factors, while farm-level lacrimation and the presence of Stomoxys were significantly associated with infection. Molecular characterization identified T. evansi type A and type non-A/B, with no type B or maxicircle-positive samples detected. Phylogenetic analysis grouped the Thai isolates with previously reported isolates from Thai deer and cattle, Iranian camels, and Colombian dogs. On follow-up, no Trypanozoon infection was detected in revisited farms. These findings confirmed that T. evansi, comprising types A and non-A/B, continues to circulate in Thai livestock, particularly in beef cattle in Northern Thailand. Data from this study can be used for surveillance and future development of more effective diagnostic systems.
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Open AccessArticle
First Isolation, Identification, and Whole-Genome Sequencing of a Multidrug-Resistant Bovine-Derived Providencia stuartii in China
by
Rong-Jun Gong, Xue-Li Ge, Jia-Min Ma, Yang-Sini Fu, Jia-Hao Chen, Man-Ting Li, Yong-Xiang Zhao, Liang Zhu, Qing-Hong Guo, Xin-Chao Liu and Wen-Chao Li
Pathogens 2026, 15(8), 789; https://doi.org/10.3390/pathogens15080789 - 24 Jul 2026
Abstract
Background: Providencia stuartii is an opportunistic pathogen associated with multidrug resistance. However, its occurrence, genomic characteristics, antimicrobial resistance profiles, and pathogenic potential in cattle-associated isolates remain poorly understood. Methods: A P. stuartii strain was isolated from a rectal swab of a diarrheic beef
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Background: Providencia stuartii is an opportunistic pathogen associated with multidrug resistance. However, its occurrence, genomic characteristics, antimicrobial resistance profiles, and pathogenic potential in cattle-associated isolates remain poorly understood. Methods: A P. stuartii strain was isolated from a rectal swab of a diarrheic beef cattle individual during bacterial investigation on a commercial farm in Anhui Province, China. The strain was identified by Gram staining, biochemical tests, 16S rRNA sequencing, and whole-genome sequencing. Antimicrobial susceptibility was assessed using the Kirby–Bauer method, and resistance and virulence genes were detected by PCR and genomic analysis. Pathogenicity was evaluated in a murine infection model. Results: The isolate was confirmed as P. stuartii, with a 4.22 Mb genome and 41.28% GC content. It showed multidrug resistance, including resistance to eight antimicrobial agents. Genomic analysis revealed multiple resistance and virulence determinants, genomic islands, prophages, and CRISPR regions. Under high-dose intraperitoneal challenge conditions, the isolate caused dose-dependent mortality, systemic recovery from major organs, and histopathological lesions in mice. Conclusions: This study reports the genomic and antimicrobial resistance characteristics of a multidrug-resistant P. stuartii strain recovered from a rectal swab of diarrheic beef cattle in China. High-dose intraperitoneal inoculation demonstrated that the isolate possessed pathogenic potential in mice under experimental conditions. However, the murine model did not reproduce natural exposure in cattle, and the findings do not establish bovine tissue invasion, clinical pathogenicity, or a causal role in diarrhea. Further epidemiological investigations and cattle-relevant experimental studies are required to determine the clinical significance of this organism in bovine populations.
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(This article belongs to the Section Bacterial Pathogens)
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Open AccessArticle
A Novel Real-Time SYBR Green PCR Assay for Integron Profiling in Pseudomonas aeruginosa: Unveiling the Resistance Nexus Between Food and Clinical Isolates
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Hend Emad, Mahmoud M. Amer, Munirah M. Alhammadi, Mona Alharbi, Yasmen F. Mahran, Attia A. Attia, Khalid Maniah and Azza SalahEldin El-Demerdash
Pathogens 2026, 15(8), 788; https://doi.org/10.3390/pathogens15080788 - 24 Jul 2026
Abstract
Background: The dissemination of extensively drug-resistant Pseudomonas aeruginosa strains across the human–food interface represents a critical threat to global public health, complicating empirical therapeutic choices and challenging traditional nosocomial restriction paradigms. Objective: This study investigated the prevalence, phenotypic resistome landscapes, and underlying integron-mediated
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Background: The dissemination of extensively drug-resistant Pseudomonas aeruginosa strains across the human–food interface represents a critical threat to global public health, complicating empirical therapeutic choices and challenging traditional nosocomial restriction paradigms. Objective: This study investigated the prevalence, phenotypic resistome landscapes, and underlying integron-mediated mobilization mechanics of P. aeruginosa across the One Health continuum using an optimized real-time quantitative PCR (qPCR) platform. Methods: A total of 116 samples comprising human clinical specimens (n = 61) and retail food matrices (n = 55) were screened. Phenotypic resistance was profiled against 16 antipseudomonal agents, with polymyxin and colistin resistance validated via reference broth microdilution. A novel real-time SYBR Green qPCR assay backed by derivative melting curve analysis was developed to track Class 1 (intI1) and Class 2 (intI2) integrons. Results: Pseudomonas aeruginosa was recovered from 36.2% (n = 42/116) of total samples, exhibiting a homogeneous distribution across clinical (32.8%) and retail food matrices (40.0%, p = 0.445). Antibiogram characterization revealed a critical resistance burden: 100% of isolates fell into extreme epidemiological tiers, with 71.4% (n = 30) exhibiting an extensively drug-resistant (XDR) phenotype and 28.6% (n = 12) reaching absolute pandrug-resistant (PDR) status. Absolute phenotypic resistance (100%) was recorded across both cohorts for meropenem, ceftazidime, aztreonam, tobramycin, polymyxins, and novel combination therapeutics (ceftazidime–avibactam and ceftolozane–tazobactam). Pairwise tracking demonstrated an exceptionally strong positive correlation in resistance distribution patterns between human and food isolates (Spearman’s rho = 0.948, p < 0.001). Analytical validation of the qPCR assay demonstrated tight, reproducible specific derivative melting peaks (Tm) at 83.2 °C for intI1 and 83.8 °C for intI2. Genotypic screening revealed universal conservation of plasmid-borne intI1 (100%), whereas plasmidic intI2 was variably distributed (61.9%). Crucially, intI2 carriage served as a definitive marker for the transition to complete pandrug resistance, being detected in 100% (n = 12/12) of PDR isolates compared to 46.7% (n = 14/30) of XDR strains (p = 0.001), supported by a strong positive correlation with the PDR phenotype (rho = 0.496, p = 0.0008). Conclusions: These findings reveal an overlapping, extreme resistome across the food-clinical interface and identify intI2 as a genetic tipping point for absolute pandrug resistance. The validated real-time assay provides a highly sensitive, proactive molecular surveillance framework capable of mapping high-risk mobile genetic platforms before they manifest as catastrophic empirical treatment failures.
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(This article belongs to the Section Bacterial Pathogens)
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Open AccessCommunication
Establishment of an HSV-1 Mouse Model with Cutaneous Lesions
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Hye-Myung Ryu, Bushra Riaz and Seonghyang Sohn
Pathogens 2026, 15(8), 787; https://doi.org/10.3390/pathogens15080787 - 24 Jul 2026
Abstract
Herpes simplex virus type 1 (HSV-1) causes recurrent mucocutaneous lesions, yet existing animal models incompletely recapitulate characteristic skin manifestations. Here, we established a mouse model of cutaneous HSV-1 infection by combining epithelial barrier disruption with localized viral inoculation. Superficial scarification of the ear
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Herpes simplex virus type 1 (HSV-1) causes recurrent mucocutaneous lesions, yet existing animal models incompletely recapitulate characteristic skin manifestations. Here, we established a mouse model of cutaneous HSV-1 infection by combining epithelial barrier disruption with localized viral inoculation. Superficial scarification of the ear followed by HSV-1 exposure resulted in consistent lesion formation. Administration of hydrocortisone further increased the incidence of lesions, highlighting the critical roles of epithelial integrity and host immune regulation in the pathogenesis of HSV-1 infection. Using this model, we evaluated the antiviral efficacy of Acyclovir. Treatment significantly reduced lesion severity, lesion size, and viral gene expression, indicating partial suppression of viral replication. Notably, acyclovir treatment was associated with increased expression of T-bet and Foxp3 in lymphoid tissues, suggesting modulation of both effector and regulatory immune responses. Collectively, this model successfully reproduces localized HSV-1 skin lesions, serving as a useful platform for investigating viral pathogenesis and evaluating antiviral therapies. However, further studies, including direct viral quantification and histopathological analysis, are required.
Full article
(This article belongs to the Special Issue Pathogen–Host Interactions: Death, Defense, and Disease)
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Open AccessArticle
Real-World Use of Cefiderocol for Multidrug-Resistant Gram-Negative Infections in Critically Ill ICU Patients: A Single-Center Case Series
by
Stelian Adrian Ritiu, Adelina Baloi, Marius Păpurică, Dorel Sandesc, Daiana Toma, Claudiu Rafael Bârsac, Sonia Elena Popovici, Madalina Butaș, Ana-Maria-Ionela Botoaca and Ovidiu Bedreag
Pathogens 2026, 15(8), 786; https://doi.org/10.3390/pathogens15080786 - 24 Jul 2026
Abstract
Background/Objectives: Carbapenem-resistant Gram-negative (CR-GN) infections are associated with high mortality in intensive care units (ICUs), with limited therapeutic options. We aimed to evaluate microbiological and clinical outcomes associated with cefiderocol in critically ill patients with multidrug-resistant (MDR) Gram-negative infections. Methods: This retrospective, single-center
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Background/Objectives: Carbapenem-resistant Gram-negative (CR-GN) infections are associated with high mortality in intensive care units (ICUs), with limited therapeutic options. We aimed to evaluate microbiological and clinical outcomes associated with cefiderocol in critically ill patients with multidrug-resistant (MDR) Gram-negative infections. Methods: This retrospective, single-center case series included 25 critically ill patients with carbapenem-resistant or multidrug-resistant Gram-negative infections treated with cefiderocol between May 2024 and October 2025 in a mixed ICU of a tertiary hospital in Romania. Microbiological cure was defined as eradication of the designated target pathogen at the end of therapy. Clinical evolution was assessed using the Acute Physiology and Chronic Health Evaluation II (APACHE II) and Sequential Organ Failure Assessment (SOFA) scores, as well as inflammatory biomarkers including white blood cell count (WBC), C-reactive protein (CRP), and procalcitonin (PCT). Results: Klebsiella pneumoniae was the predominant pathogen, identified in 23 of 25 patients (92%), followed by Acinetobacter baumannii (11/25, 44%) and Pseudomonas aeruginosa (6/25, 24%). As multiple pathogens were co-isolated in 16 patients (64%), percentages exceed 100% and are reported as proportions of patients rather than of total isolates. Microbiological cure was achieved in 68% of patients. Mortality was markedly higher in patients without microbiological eradication (88% vs. 35%). Higher baseline APACHE II (HR 1.18, 95% CI 1.02–1.37) and SOFA scores (HR 1.33, 95% CI 1.07–1.65) were associated with increased mortality. Early initiation of cefiderocol (≤10 days from ICU admission) was associated with improved microbiological and clinical outcomes compared to delayed treatment. Reductions in severity scores and inflammatory markers, including C-reactive protein (CRP) and procalcitonin (PCT), were observed during therapy. Pathogen type and combination therapy were not clearly associated with outcomes in this cohort. Conclusions: Cefiderocol was observed in association with clinically relevant rates of microbiological eradication and improvements in clinical parameters in critically ill patients with MDR Gram-negative infections. Outcomes appeared to be primarily determined by baseline disease severity, while earlier initiation of therapy was associated with more favourable microbiological and clinical outcomes, although causality cannot be established given the observational design and absence of a comparator group. These findings are hypothesis-generating and supportive of a potential role for cefiderocol as a salvage option in high-risk ICU populations, pending validation in larger, prospective, controlled studies.
Full article
(This article belongs to the Special Issue Advanced Antimicrobial Agents: Combatting Multi-Drug Resistant Bacterial Infections)
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Open AccessArticle
Compact Gut Microbial Dysbiosis Signature Associated with Necrotizing Enterocolitis and Altered Early Microbiota Development
by
Ying Xiang, Zhuoqi Zhao, Zhong Feng, Jialu Zhuang, Yu Chen, Yansong Chen, Shumin Feng, Liya Pan, Zhilong Yan and Li Hong
Pathogens 2026, 15(8), 785; https://doi.org/10.3390/pathogens15080785 - 24 Jul 2026
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Background: Necrotizing enterocolitis (NEC) is a life-threatening intestinal disorder in preterm infants and is strongly associated with gut microbial dysbiosis. However, whether recurrent genus-level dysbiosis patterns can be observed across heterogeneous NEC cohorts and summarized as a compact, interpretable microbial signature remains unclear.
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Background: Necrotizing enterocolitis (NEC) is a life-threatening intestinal disorder in preterm infants and is strongly associated with gut microbial dysbiosis. However, whether recurrent genus-level dysbiosis patterns can be observed across heterogeneous NEC cohorts and summarized as a compact, interpretable microbial signature remains unclear. Methods: We analyzed two public neonatal gut microbiome cohorts and an independent real-world cohort within a three-stage framework of discovery, contextual analysis, and external validation. Community composition, alpha diversity, beta diversity, and differential genera were evaluated using harmonized genus-level features within each cohort. Machine learning feature prioritization was used to derive a reduced NEC-associated microbial signature. A four-group cohort was then used to examine this signature in relation to disease status, antibiotic exposure, and early postnatal development. The reduced signature was finally examined in our collected samples. Results: NEC-related microbial alterations showed marked heterogeneity at the whole-community level across cohorts, whereas several genus-level directional patterns recurred across datasets. Across datasets, NEC was repeatedly associated with enrichment of several opportunistic genera, including Enterobacter, Klebsiella, Serratia, and Escherichia–Shigella, and depletion of commensal or probiotic-associated taxa such as Bifidobacterium, Lactobacillus, and Pediococcus. Feature prioritization yielded a compact microbial signature that improved discrimination over the unfiltered abundance profile in the discovery cohort (AUC 0.674 vs. 0.59). In the four-group cohort, the NEC_ABT group remained distinct from healthy controls, supporting partial modification rather than normalization of the NEC-associated pattern. In our collected samples, the signature retained directional consistency but showed only modest external discrimination (AUC 0.618). Conclusions: These findings identify recurrent genus-level dysbiosis features associated with NEC across clinically heterogeneous settings. The compact microbial signature may serve as an exploratory biomarker for longitudinal and mechanistic validation, but not as a standalone diagnostic tool.
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Open AccessArticle
Arthrobotrys thaumasia and A. musiformis (Fungi: Orbiliales) as Natural Potential Enemies of Haemonchus contortus (Nematoda: Trichostrongylidae)
by
Mariana Cervantes-Martínez, Rosa Isabel Higuera-Piedrahita, Héctor Alejandro de la Crúz-Crúz, Gustavo Pérez-Anzúrez, María Eugenia López-Arellano, Agustín Olmedo-Juárez, Edgar Jesús Delgado-Núñez, Ana Yuridia Ocampo-Gutiérrez and Pedro Mendoza-de Gives
Pathogens 2026, 15(8), 784; https://doi.org/10.3390/pathogens15080784 - 23 Jul 2026
Abstract
Haemonchus contortus (Hc) is a parasitic nematode affecting sheep. Arthrobotrys is a group of nematophagous fungi (NF) that capture nematodes with specialized traps. This study evaluated the in vitro predatory ability of Arthrobotrys thaumasia (At) and A. musiformis (
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Haemonchus contortus (Hc) is a parasitic nematode affecting sheep. Arthrobotrys is a group of nematophagous fungi (NF) that capture nematodes with specialized traps. This study evaluated the in vitro predatory ability of Arthrobotrys thaumasia (At) and A. musiformis (Am), as well as the nematocidal activity of their liquid culture filtrates against Hc. NF isolates were obtained by sprinkling soil onto agar plates containing nematodes. Isolates were identified morphologically and molecularly. The predatory activity (PA) was tested on agar plates inoculated with nematodes and fungi; plates inoculated only with nematodes served as controls. After 10 days at 18 °C and 28 °C, larvae were counted. For nematocidal activity (NA) of LCFs, fungi were grown in Czapek–Dox (CzDB) and potato dextrose broths. Two concentrations (100 mg/mL and 130 mg/mL) were tested. Nematodes and LCFs were incubated for 48 or 72 h. PA was analyzed using unpaired Student’s t-tests, and LCFs were evaluated by ANOVA with Tukey’s post hoc test. Taxonomic procedures confirmed the isolates as Am and At. For PA, effectiveness was 74.64% for Am and 83.53% for At. The highest NA of LCF of Am in CzDB was 51% at 100 mg/mL and 76.7% at 130 mg/mL. That of the LCF of At was 22% at the highest concentration. Both isolates show strong potential as biocontrol agents against haemonchosis.
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(This article belongs to the Section Parasitic Pathogens)
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Open AccessReview
Clinical Guidelines for Hepatitis E Vaccination in India: An Expert Panel Consensus Report on the Recombinant Hepatitis E Vaccine, HEV 239
by
Mohammad Sultan Khuroo and Naira S. Khuroo
Pathogens 2026, 15(8), 783; https://doi.org/10.3390/pathogens15080783 - 23 Jul 2026
Abstract
(1) Background: Hepatitis E remains a major public health challenge in India. (2) Methods: In August 2025, the recombinant HEV 239 vaccine was approved in India for adults aged 18 to 65 years. To establish clinical guidelines tailored to the Indian setting, an
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(1) Background: Hepatitis E remains a major public health challenge in India. (2) Methods: In August 2025, the recombinant HEV 239 vaccine was approved in India for adults aged 18 to 65 years. To establish clinical guidelines tailored to the Indian setting, an expert panel consensus was conducted using a modified Delphi process in accordance with the ACCORD reporting guidelines. (3) Results: A steering committee put forth 18 statements covering vaccine safety, efficacy, and clinical indications, which were independently evaluated by 33 senior Indian hepatologists and epidemiologists. Consensus was assessed using the GRADE framework for level of evidence, balance of benefits and harms, and strength of recommendations. Of the 18 statements, 12 reached the 70% consensus threshold. The panel concluded that the vaccine, which is administered on a standard three-dose schedule, is safe and highly effective in healthy adults, providing protection for up to 10 years. Targeted vaccination was recommended for five high-risk populations: outbreak-affected groups, hyperendemic pockets, women of childbearing age, patients with CLD, and solid organ transplant recipients. Although derived from HEV genotype 1, the vaccine demonstrated cross-protective efficacy against HEV genotype 4. (4) Conclusions: This consensus report provides a framework for deploying the HEV vaccine to mitigate disease burden in India while emphasizing the need for real-world effectiveness and safety data from India.
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(This article belongs to the Special Issue Hepatitis E: Virus, Disease and Vaccine)
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Open AccessArticle
Implementation of a Composite Reference Standard Methodology in the Greek National Malaria Reference Center, Insights from a Seven-Year Experience
by
Nikolaos Tegos, Leonidas Georgalis, Anastasia Bimpa, Anastasia Panagopoulou, Stavroula Beleri, Vasilios Papavasilopoulos, Annita Vakali, Chrisovaladou Kefaloudi, Danai Pervanidou, Christos Hatzichristodoulou and Eleni Patsoula
Pathogens 2026, 15(8), 782; https://doi.org/10.3390/pathogens15080782 - 23 Jul 2026
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Background: Malaria is currently ranked as an important public health issue in Greece despite its initial eradication back in 1974. Given that the clinical picture is often nonspecific, the need for the implementation of targeted laboratory methodologies is imperative to ensure prompt Plasmodium
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Background: Malaria is currently ranked as an important public health issue in Greece despite its initial eradication back in 1974. Given that the clinical picture is often nonspecific, the need for the implementation of targeted laboratory methodologies is imperative to ensure prompt Plasmodium identification and diagnosis. Methods: A total of 780 clinical samples from 763 suspected malaria cases from the National Malaria Reference Center were analyzed within a seven-year period (2017–2023). A composite laboratory reference standard methodology, including rapid diagnostic testing, microscopy, and polymerase chain reactions, was implemented for the purposes set by the current study in a group of 532 samples. A multivariable statistical analysis was implemented for the interpretation of the extracted data. Results: Microscopy and PCR successfully identified P. falciparum (111 vs. 131) and P. vivax (95 vs. 95) clinical samples. In addition, microscopy successfully identified selected cases of P. ovale (16) and P. malariae (nine), with current cases being confirmed by targeted, species-level real-time PCR protocols. Conclusions: The current study highlighted the importance of microscopic examination in malaria diagnostic procedures for Plasmodium species identification and typing. The combination of microscopy and PCR ensures a credible laboratory approach for submicroscopic clinical cases and patients with prior antimalarial treatment. This composite laboratory reference standard methodology is recommended by the Greek Malaria Reference Center for prompt identification of Plasmodium species in suspected malaria clinical cases.
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Open AccessArticle
Effects of Essential Oil Treatment on Clostridium perfringens Cells
by
Safa Q. Alfattani and Mahfuzur R. Sarker
Pathogens 2026, 15(8), 781; https://doi.org/10.3390/pathogens15080781 - 23 Jul 2026
Abstract
Clostridium perfringens type F is a Gram-positive, spore-forming bacterium that causes food poisoning (FP) in humans. Our previous findings demonstrated that essential oils (EOs), including clove, rosemary, and peppermint oils, significantly inhibited the germination, outgrowth, and vegetative growth of C. perfringens spores. We
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Clostridium perfringens type F is a Gram-positive, spore-forming bacterium that causes food poisoning (FP) in humans. Our previous findings demonstrated that essential oils (EOs), including clove, rosemary, and peppermint oils, significantly inhibited the germination, outgrowth, and vegetative growth of C. perfringens spores. We hypothesized that cytoplasmic membrane damage could be one mechanism by which EOs exert their inhibitory effects against C. perfringens vegetative cells. To test this hypothesis, vegetative cells of two clinical FP strains of C. perfringens, SM101 and E13, were treated with EOs, and the leakage of intracellular molecules was compared between EO-treated and untreated cells. Our results demonstrated increased leakage of intracellular molecules, including ATP, 260-nm-absorbing materials, and proteins, from EO-treated cells relative to untreated cells. In addition, EO-treated cells exhibited greater crystal violet uptake than untreated cells. These results suggest that EOs induced cytoplasmic membrane damage, thereby promoting the leakage of intracellular molecules and the uptake of extracellular crystal violet. Scanning electron microscopy further confirmed cytoplasmic membrane damage by revealing substantially greater membrane disruption in EO-treated cells than in untreated cells. In conclusion, EOs may exert their anti-C. perfringens effects by damaging the cytoplasmic membrane, thereby disrupting membrane permeability and suppressing cell multiplication.
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(This article belongs to the Special Issue Fighting Pathogens with Natural Antimicrobials)
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Open AccessArticle
Epidemiological Trends, Statistical Correlations, and Forecasting of Notifiable Infectious Diseases in South Korea, 2001–2024: A Regression and Time Series Analysis with Projections to 2028
by
Hyeran Jung and Minsun Jung
Pathogens 2026, 15(8), 780; https://doi.org/10.3390/pathogens15080780 - 23 Jul 2026
Abstract
Background: Notifiable infectious diseases impose a substantial, evolving burden on public health systems. This study characterized long-term epidemiological trends, inter-class associations, and age-stratified patterns of notifiable infectious diseases in South Korea and projected future burden through 2028. Methods: We analyzed aggregate national surveillance
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Background: Notifiable infectious diseases impose a substantial, evolving burden on public health systems. This study characterized long-term epidemiological trends, inter-class associations, and age-stratified patterns of notifiable infectious diseases in South Korea and projected future burden through 2028. Methods: We analyzed aggregate national surveillance data (2001–2024) from the Korea Disease Control and Prevention Agency (KDCA) via the Korean Statistical Information Service (KOSIS). Because the statutory classification changed in 2020 from a four-group (Class 1–4, gun) to a four-grade (geup) system, all inferential trend, correlation, and forecasting analyses were restricted to the internally consistent 2001–2019 group-system period, and 2020–2024 grade-system data were used descriptively. Secular trends were estimated with ordinary least squares and, for count-specific inference, negative-binomial log-linear models (annual percent change, APC); residual autocorrelation was assessed (Durbin–Watson, Ljung–Box). Structural breaks were estimated objectively (Bai–Perron least-squares). Inter-class associations were tested with Pearson/Spearman correlations and re-evaluated with partial correlation (controlling for calendar year) and first-difference detrending to guard against spurious co-trending. Forecasts used a random-walk ARIMA (identified by ADF stationarity testing, ACF/PACF, and AICc) and a damped Holt–Winters model on the H1N1-adjusted series, combined into an ensemble; performance was validated by rolling-origin cross-validation (RMSE, MAE, MAPE). Results: After handling the 2009 H1N1 pandemic outlier, total notifications rose significantly over 2001–2019 (slope = 8344 cases/year; 95% CI 6603–10,084; R2 = 0.866; p < 0.001). Class 2 (respiratory/vaccine-preventable) diseases showed the strongest trend (APC = 17.2%/year; 95% CI 8.0–27.3), with an objectively estimated structural break at 2004 (supF = 10.5) separating an early variable phase from sustained growth (2005–2019: +7418 cases/year; R2 = 0.94). Raw Class 1–Class 2 correlation (Pearson r = 0.569, p = 0.011) did not survive temporal adjustment (partial r = 0.13, p = 0.59; detrended r = −0.31, p = 0.21), indicating shared secular co-trending rather than a direct epidemiological association; only Class 2–Class 3 remained associated after detrending (r = 0.52, p = 0.03). Young working-age adults (30–49 years) showed the steepest Class 1 increases. Ensemble projections for the pre-COVID trajectory were 238,000 (2025) to 269,000 (2028) annual notifications, with wide prediction intervals reflecting substantial uncertainty. Conclusions: South Korean notifiable-disease epidemiology was dominated by a genuine upward Class 2 respiratory trend; several previously reported inter-class correlations were attributable to common temporal trends rather than shared transmission. Post-COVID normalization monitoring and respiratory-disease surveillance investment are priority actions.
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(This article belongs to the Special Issue Epidemiology of Vector-Borne Pathogens)
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Open AccessBrief Report
High Burden of Asymptomatic Malaria Among Schoolchildren in Nigeria: Diagnosis Challenges and pfhrp2 Deletions
by
Irene Molina-de la Fuente, Akeem Abiodun Akindele, Thuy-Huong Ta-Tang, Alexandra Martin-Ramírez, Vicenta Gonzalez, Ibukun Temitope Sossou, Samuel Adeyinka Adedokun, Raquel Capote-Morales, Agustín Benito, Sulaiman Adebayo Nassar and Pedro Berzosa
Pathogens 2026, 15(8), 779; https://doi.org/10.3390/pathogens15080779 - 23 Jul 2026
Abstract
Background: Nigeria has a high prevalence of malaria, with asymptomatic patients being one of the possible reservoirs. In this context, accurate diagnosis is essential for malaria control. The efficacy of rapid diagnostic tests (RDTs) is being threatened by false negatives due to pfhrp2
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Background: Nigeria has a high prevalence of malaria, with asymptomatic patients being one of the possible reservoirs. In this context, accurate diagnosis is essential for malaria control. The efficacy of rapid diagnostic tests (RDTs) is being threatened by false negatives due to pfhrp2 and pfhrp3 deletions. This study aims to describe malaria in asymptomatic schoolchildren in Osun State, Southwestern Nigeria and to assess the performance of malaria diagnosis, including the characterisation of pfhrp2 and pfhrp3 deletions. Methods: A total of 350 dried blood spot (DBS) samples from schoolchildren were used for malaria diagnosis using microscopy, RDTs, and polymerase chain reaction (PCR). Sensitivity and specificity were calculated for microscopy and RDTs, using PCR as the gold standard. Pfhrp2 and pfhrp3 deletions were analysed for all P. falciparum-positive samples using multiplex qPCR. Results: The P. falciparum infection frequency among asymptomatic schoolchildren in Osun State was 89%. Considering PCR as the gold standard, microscopy presented better sensitivity at 100% (86% for RDTs), but RDTs had better specificity at 76% (60% for microscopy). The deletion frequency of the pfhrp2 gene among positive samples was 16.6%, while it was 1.3% for pfhrp3 and 0.9% for double deletion. However, considering all positive samples, only 3% of samples yielded false negatives by RDT with deletion in pfhrp2. Conclusions: P. falciparum infection has a high frequency in asymptomatic schoolchildren, being a potential hotspot for malaria. The combination of RDTs and microscopy could increase the accuracy of malaria diagnosis. Deletions in pfhrp2 and pfhrp3 are highly common in Osun State, but their impact on RDT results is still limited. More surveillance studies are recommended to assess the contribution of asymptomatic children to malaria transmission and the impact of deletions.
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(This article belongs to the Special Issue Molecular Epidemiology and Host-Pathogen Interactions in Infectious Diseases)
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Open AccessArticle
Effect of Methyl Gallate on Physiological and Gut Microbiota-Associated Responses Following a Single Intramuscular Administration of Tylosin in Weaned Piglets
by
Hae-Yeon Cho, Syed Al Jawad Sayem, Ga-Yeong Lee, Jonghyun Park, Eon-Bee Lee, Mi-Hyang Hwangbo and Seung-Chun Park
Pathogens 2026, 15(7), 778; https://doi.org/10.3390/pathogens15070778 - 22 Jul 2026
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Tylosin (TYL), a macrolide antibiotic, is commonly used to treat respiratory infections in weaned piglets. Despite its utility, TYL administration is associated with adverse side effects, including immune dysregulation and gut microbiota imbalance. This study evaluated whether methyl gallate (MG), a natural polyphenolic
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Tylosin (TYL), a macrolide antibiotic, is commonly used to treat respiratory infections in weaned piglets. Despite its utility, TYL administration is associated with adverse side effects, including immune dysregulation and gut microbiota imbalance. This study evaluated whether methyl gallate (MG), a natural polyphenolic compound with antimicrobial and immunomodulatory properties, could mitigate these side effects of TYL. Forty weaned piglets were randomly assigned to a saline control group, a TYL (10 mg/kg) group, an MG (10 mg/kg) group, and a TYL + MG (10 mg/kg each) group. Changes in body weight, hematological parameters, plasma cytokines, culturable fecal bacterial populations, and 16S rRNA-based gut microbiota composition were evaluated in all weaned piglets. There were no significant differences (p > 0.05) in ADG, ADFI, or FE among the treatment groups. Hematological analysis revealed significant changes in neutrophil and eosinophil counts, showing significant fluctuations in the MG-treated group compared with the control. TYL administration significantly decreased TNF-α and IL-1β levels while increasing IL-10 and IL-1ra concentrations, indicating immunomodulatory effects. In contrast, MG significantly increased adaptive immune-associated cytokines, including IL-2, IL-4, IL-6, and IL-12. Culture-based fecal microbiological analysis revealed that TYL reduced Lactobacillus abundance and lowered the culture-based Lactobacillus/Enterobacteriaceae ratio, suggesting acute dysbiosis. Co-administration of MG partially attenuated these microbiota-associated alterations and promoted the recovery of bacterial populations. Moreover, 16S rRNA sequencing demonstrated that similar relative abundances of Proteobacteria were observed in the TYL and TYL + MG groups. Therefore, no clear reduction in relative abundance of Proteobacteria was demonstrated under the present experimental conditions. These findings support the possibility that MG may partially attenuate selected TYL-associated immunological and gut microbiota-related changes under the conditions of the present exploratory study. A graphical summary of the study design and the principal findings is presented in the Graphical Abstract.
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Open AccessArticle
Two Diagnostic Challenges in Hepatitis B Serology: Low HBsAg S/CO Values and Isolated Anti-HBc Positivity
by
Şerife Yılmaz Gürbüz, Oğuzhan Yağdı and Erhan Başar
Pathogens 2026, 15(7), 777; https://doi.org/10.3390/pathogens15070777 - 22 Jul 2026
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Hepatitis B surface antigen (HBsAg) signal-to-cutoff (S/CO) values and isolated anti-HBc positivity represent two significant diagnostic challenges in hepatitis B virus (HBV) serology. This retrospective study, conducted in Karabuk between 2021 and 2025, aimed to evaluate the correlation between these profiles and HBV
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Hepatitis B surface antigen (HBsAg) signal-to-cutoff (S/CO) values and isolated anti-HBc positivity represent two significant diagnostic challenges in hepatitis B virus (HBV) serology. This retrospective study, conducted in Karabuk between 2021 and 2025, aimed to evaluate the correlation between these profiles and HBV DNA positivity to determine the optimal S/CO thresholds for confirmatory testing and clarify the clinical relevance of isolated anti-HBc reactivity. Among 17,356 patients screened for anti-HBc, isolated anti-HBc positivity was identified in 506 patients (2.9%), of whom 3.2% had detectable HBV DNA. Positivity was significantly higher in patients older than 50 years and in the Gastroenterology department. Separately, 213 specimens with HBsAg S/CO values between 1.0 and 10.0 underwent HBV DNA confirmation testing. ROC analysis yielded an excellent area under the curve of 0.937, with an S/CO threshold of ≥3.04, achieving 100% sensitivity for HBV DNA detection. All specimens with S/CO values between 1.0 and 3.0 were HBV DNA-negative. In this study population, no serum HBV DNA positivity was detected among samples with S/CO values below 3.04, suggesting that HBV DNA testing may help avoid unnecessary confirmatory work-up in this range. Anti-HBc screening should be prioritized in older patients. S/CO-based confirmatory algorithm using HBV DNA testing may improve diagnostic accuracy and reduce unnecessary clinical interventions in routine hepatitis B screening programs.
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