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Pathogens, Volume 14, Issue 6 (June 2025) – 97 articles

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21 pages, 1074 KiB  
Article
The Epidemiology of Coccidioidomycosis (Valley Fever) and the Disease Ecology of Coccidioides spp. in New Mexico (2006–2023)
by Paris S. Salazar-Hamm, Sarah Shrum Davis, Jovani Catalán-Dibene, Adriana L. Romero-Olivares, Karen Edge, Andrew W. Bartlow, Donald O. Natvig and Morgan E. Gorris
Pathogens 2025, 14(6), 607; https://doi.org/10.3390/pathogens14060607 - 19 Jun 2025
Abstract
Coccidioidomycosis (Valley fever), caused by Coccidioides spp., is a fungal infection endemic to semi-arid regions of the Americas. Despite 80 years of disease recognition in New Mexico, there is limited disease awareness. We incorporated clinical, epidemiological, and ecological datasets to summarize the knowledge [...] Read more.
Coccidioidomycosis (Valley fever), caused by Coccidioides spp., is a fungal infection endemic to semi-arid regions of the Americas. Despite 80 years of disease recognition in New Mexico, there is limited disease awareness. We incorporated clinical, epidemiological, and ecological datasets to summarize the knowledge of Valley fever in New Mexico. We analyzed 1541 human cases from 2006 to 2023. On average, 86 cases were reported each year (4.1 cases per 100,000 population per year). The highest levels of incidence were in southwestern New Mexico. American Indian or Alaska Natives in New Mexico had a 1.9 times higher incidence rate of coccidioidomycosis than White people, and among age groups, older populations in New Mexico had the highest incidence rates. We analyzed 300 soil samples near Las Cruces, New Mexico, for the presence of Coccidioides and reported the first known positive soil samples collected from the state, the majority of which were from grassland-dominated sites and from animal burrows. Sequence analyses in clinical specimens, wild animals, and soil samples confirmed that Coccidioides posadasii is the main causative species of coccidioidomycosis in New Mexico. Environmental surveillance validated that locally acquired infections could occur in, but are not limited to, Catron, Doña Ana, Sierra, and Socorro Counties. Full article
(This article belongs to the Special Issue An Update on Fungal Infections)
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17 pages, 2034 KiB  
Article
Characterization of the Diversity in Host Range of an Extensively Drug-Resistant (XDR) Type IV Secretion System-Encoding Plasmid in Acinetobacter
by Kailey Martz, Dalya Alomar, Marisha Karim, Sara Knezevic and Vanessa M. D’Costa
Pathogens 2025, 14(6), 606; https://doi.org/10.3390/pathogens14060606 - 19 Jun 2025
Abstract
The World Health Organization (WHO) cites antimicrobial resistance as among the greatest threats to human health. The multidrug-resistant pathogen Acinetobacter baumannii, recognized as a priority pathogen for healthcare and research, is responsible for a diverse array of infections including respiratory tract, soft [...] Read more.
The World Health Organization (WHO) cites antimicrobial resistance as among the greatest threats to human health. The multidrug-resistant pathogen Acinetobacter baumannii, recognized as a priority pathogen for healthcare and research, is responsible for a diverse array of infections including respiratory tract, soft tissue and wound, and bloodstream infections. Despite this importance, the mechanisms of its pathogenesis remain poorly understood. Conjugation represents a central mechanism for bacterial adaptation and evolution and is responsible for the spread of genes that promote pathogen survival, antibiotic resistance, virulence, and biofilm formation. Our laboratory recently characterized a large group of almost 120 Type IV Secretion System (T4SS)-encoding plasmids in Acinetobacter, distributed globally across 20 countries spanning four continents, and demonstrated that an XDR A. baumannii plasmid from this family was transmissible to another A. baumannii strain. This research investigated the potential diversity of host strains for this representative member plasmid. Using the GC1 lineage strain A. baumannii AB5075-UW harbouring the XDR plasmid p1AB5075 and a series of previously characterized clinical and environmental Acinetobacter strains, conjugative analyses demonstrated transfer of the XDR plasmid to both A. baumannii strains of more genetically divergent sequence types and to non-baumannii Acinetobacter species both inside and outside the Acinetobacter calcoaceticus–baumannii (ACB) complex. Successful recipients included diverse strains of both clinical and environmental origin within the Acinetobacter genus. Collectively, this research could provide insights into an important genetic element for future surveillance. Full article
13 pages, 1514 KiB  
Article
Establishment of a Sandwich ELISA for Detection of Pan-Merbecoviruses
by Kaixin Li, Misa Katayama, Ayano Ichikawa, Hiromichi Matsugo, Yuta Wakabayashi, Akiko Takenaka-Uema, Wataru Sekine, Taisuke Horimoto and Shin Murakami
Pathogens 2025, 14(6), 605; https://doi.org/10.3390/pathogens14060605 - 19 Jun 2025
Abstract
Merbecovirus, a subgenus of Betacoronavirus, includes MERS-CoV and multiple bat-derived viruses with zoonotic potential. Given the unpredictable emergence of these viruses and their genetic diversity, development of broad-spectrum diagnostic tools is expected. In this study, we established a sandwich ELISA targeting [...] Read more.
Merbecovirus, a subgenus of Betacoronavirus, includes MERS-CoV and multiple bat-derived viruses with zoonotic potential. Given the unpredictable emergence of these viruses and their genetic diversity, development of broad-spectrum diagnostic tools is expected. In this study, we established a sandwich ELISA targeting the nucleocapsid (N) protein of merbecoviruses. We generated monoclonal antibodies (mAbs) using recombinant N protein of a bat merbecovirus, VsCoV-1, and selected cross-reactive clones for other merbecoviruses. Three mAbs showed strong reactivities with multiple merbecoviruses but not with SARS-CoV-2 or endemic human coronaviruses. Pairwise ELISA screening identified 1A8/10H6 mAbs as the optimal combination for detection of N protein from six merbecoviruses—VsCoV-1, EjCoV-3, MERS-CoV, NeoCoV, HKU4, and HKU5—with limits of detection (LODs) below 7.81 ng/mL, including 1.25 ng/mL for VsCoV-1. Infectious bat merbecovirus EjCoV-3 was detected at 1.3 × 103 PFU/mL. No cross-reactivity was observed with non-merbecoviruses, indicating its high specificity. This sandwich ELISA offers a rapid, reproducible, and cost-effective diagnostic platform with potential for high-throughput screening and automation. Moreover, its design is amenable to adaptation into point-of-care formats such as lateral flow assays, highlighting its value for field-based surveillance and pandemic preparedness. Full article
(This article belongs to the Special Issue The Epidemiology and Diagnosis of Acute Respiratory Infections)
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17 pages, 2101 KiB  
Article
CRISPR-Cas Dynamics in Carbapenem-Resistant and Carbapenem-Susceptible Klebsiella pneumoniae Clinical Isolates from a Croatian Tertiary Hospital
by Ivana Jurić, Marko Jelić, Manda Markanović, Lucija Kanižaj, Zrinka Bošnjak, Ana Budimir, Tomislav Kuliš, Arjana Tambić-Andrašević, Ivana Ivančić-Baće and Ivana Mareković
Pathogens 2025, 14(6), 604; https://doi.org/10.3390/pathogens14060604 - 19 Jun 2025
Abstract
(1) Background: CRISPR-Cas systems provide adaptive immunity against mobile genetic elements (MGEs) carrying antimicrobial resistance (AMR) genes. Carbapenem-resistant (CR) Klebsiella pneumoniae is a major public health concern, and the role of CRISPR-Cas in its resistance is understudied. This study explored CRISPR-Cas associations with [...] Read more.
(1) Background: CRISPR-Cas systems provide adaptive immunity against mobile genetic elements (MGEs) carrying antimicrobial resistance (AMR) genes. Carbapenem-resistant (CR) Klebsiella pneumoniae is a major public health concern, and the role of CRISPR-Cas in its resistance is understudied. This study explored CRISPR-Cas associations with multidrug resistance in clinical K. pneumoniae. (2) Methods: 400 K. pneumoniae isolates (200 CR and 200 carbapenem susceptible (CS)) were analyzed. Carbapenemase genes (blaOXA-48, blaNDM-1, blaKPC-2), cas1, rpoB, and CRISPR1-3 loci were identified by PCR, while only CRISPR loci were sequenced. Genetic relatedness was assessed via PFGE, MLST, and spacer analysis. Statistical analysis utilized chi-squared and Fisher’s exact tests. (3) Results: CRISPR-Cas was present in 15.8% of isolates, mainly subtypes I-E and I-E* (93.3%), with CRISPR3 loci showing greater activity. Clonal complexes ST14/15/101 (CR) and ST35 (CS) were identified. blaOXA-48 was linked to CRISPR-Cas-negative strains, while blaNDM-1 and blaKPC-2 were more frequent in CRISPR-Cas-positive strains (p < 0.0001). Imipenem/relebactam resistance was higher in CRISPR-Cas-negative isolates. (4) Conclusions: K. pneumoniae CRISPR-Cas systems correlate with specific carbapenemase profiles, suggesting pressure against blaOXA-48 acquisition. The coexistence of I-E and I-E* subtypes highlight synergies in targeting MGEs. CRISPR loci could be tools for subtyping organisms following MLST. Full article
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13 pages, 3015 KiB  
Article
Epidemiological and Clinical Characteristics of Respiratory Syncytial Virus Infection in Children in Hangzhou (2022–2023)
by Qin-Rui Lai, Xiao-Li Chu, Ying-Ying Chen, Wei Li, Ya-Jun Guo and Shi-Qiang Shang
Pathogens 2025, 14(6), 603; https://doi.org/10.3390/pathogens14060603 - 19 Jun 2025
Abstract
Human respiratory syncytial virus (HRSV) is a highly contagious RNA virus that causes respiratory infections, especially in children. This study evaluated the impact of COVID-19 control measure changes on HRSV infection patterns in Hangzhou by comparing epidemiological and clinical characteristics. We conducted a [...] Read more.
Human respiratory syncytial virus (HRSV) is a highly contagious RNA virus that causes respiratory infections, especially in children. This study evaluated the impact of COVID-19 control measure changes on HRSV infection patterns in Hangzhou by comparing epidemiological and clinical characteristics. We conducted a retrospective analysis of 12,993 pediatric nasopharyngeal swab samples from children with acute respiratory infections at The Children’s Hospital of Zhejiang University School of Medicine. These samples, collected between February 2022 and January 2024, were screened for HRSV and 12 other respiratory pathogens using capillary electrophoresis technology. From February 2022 to January 2023, the HRSV positivity rate was 7.06%. In 2023, it increased to 9.26%. The highest positivity rates were in infants aged 0–6 months and children aged 6 months to 1 year. Coinfections were most common with rhinovirus in 2022 and Mycoplasma pneumoniae in 2023. HRSV positivity rates were low from April to September 2022, peaking in December. In 2023, the peak occurred from April to September. Symptoms ranged from mild to severe pneumonia, with higher hospitalization rates in children with underlying conditions. The study revealed significant changes in HRSV infection rates following pandemic restriction relaxations, emphasizing the need for the early identification and prevention of severe cases. Full article
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11 pages, 411 KiB  
Article
The Usefulness of the BD MAX MDR-TB Molecular Test in the Rapid Diagnosis of Multidrug-Resistant Tuberculosis
by Tomasz Bogiel, Edyta Dolska, Małgorzata Zimna, Kornelia Nakonowska, Dorota Krawiecka, Renata Żebracka, Maciej Pochowski and Agnieszka Krawczyk
Pathogens 2025, 14(6), 602; https://doi.org/10.3390/pathogens14060602 - 19 Jun 2025
Abstract
Tuberculosis (TB), primarily caused by Mycobacterium tuberculosis complex (MTBC), remains a global health challenge and can lead to severe pulmonary and extrapulmonary complications. Multidrug-resistant TB (MDR-TB) poses additional challenges, requiring advanced diagnostic and treatment strategies. This study evaluates the BD MAX MDR-TB molecular [...] Read more.
Tuberculosis (TB), primarily caused by Mycobacterium tuberculosis complex (MTBC), remains a global health challenge and can lead to severe pulmonary and extrapulmonary complications. Multidrug-resistant TB (MDR-TB) poses additional challenges, requiring advanced diagnostic and treatment strategies. This study evaluates the BD MAX MDR-TB molecular test for a rapid diagnosis of MDR-TB, detecting resistance to rifampicin (RIF) and isoniazid (INH). The BD MAX MDR-TB test, utilizing real-time PCR, was used to analyze specimens collected from TB-suspected patients, identifying MTB DNA and mutations associated with rifampicin and isoniazid resistance. Results were compared with traditional drug susceptibility testing, and 79 out of 638 samples tested were positive for MTB DNA, with 65 showing a sufficient amount of genetic material for resistance gene identification. The BD MAX test showed a 100% correlation with phenotypic rifampicin resistance, though discrepancies were noted for isoniazid resistance, with a 93% concordance. The BD MAX MDR-TB test is an effective tool for a rapid diagnosis of MDR-TB, especially for rifampicin resistance. However, it may not detect certain mutations related to isoniazid resistance. Complementary tests like Xpert MTB/XDR or whole-genome sequencing could improve diagnostic accuracy and support more effective TB control strategies. Full article
(This article belongs to the Section Emerging Pathogens)
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14 pages, 1202 KiB  
Article
Deploying Metagenomics to Characterize Microbial Pathogens During Outbreak of Acute Febrile Illness Among Children in Tanzania
by Shabani Ramadhani Mziray, George Githinji, Zaydah R. de Laurent, Peter M. Mbelele, Khadija S. Mohammed, Boaz D. Wadugu, Brian S. Grundy, Scott K. Heysell, Stellah G. Mpagama and Jaffu O. Chilongola
Pathogens 2025, 14(6), 601; https://doi.org/10.3390/pathogens14060601 - 19 Jun 2025
Abstract
Outbreaks of infectious diseases contribute significantly to morbidity and mortality in resource-limited settings, yet the capacity to identify their etiology remains limited. We aimed to characterize microbes and antimicrobial resistance (AMR) genes in Tanzanian children affected by an acute febrile illness (AFI) outbreak [...] Read more.
Outbreaks of infectious diseases contribute significantly to morbidity and mortality in resource-limited settings, yet the capacity to identify their etiology remains limited. We aimed to characterize microbes and antimicrobial resistance (AMR) genes in Tanzanian children affected by an acute febrile illness (AFI) outbreak using metagenomic next-generation sequencing (mNGS). A cross-sectional study was conducted on archived blood samples from children who presented with AFI between 2018 and 2019. Total nucleic acids were extracted from 200 µL of blood, and complementary DNA (cDNA), along with enriched pathogenic DNA, was sequenced using the Illumina MiSeq platform. mNGS data were analyzed using CZ-ID Illumina mNGS bioinformatics pipeline v7.0. Results were obtained from 25 participants (mean age: 11.6 years; SD ± 5), of whom 36% had a moderate to high-grade fever. The following five potential microbial causes of AFI were identified: Escherichia coli (n = 19), Paraclostridium bifermentans (n = 2), Pegivirus C (n = 2), Shigella flexneri (n = 1) and Pseudomonas fluorescens (n = 1), with E. coli being the most prevalent. Twelve AMR genes were detected, including mdtC, acrF, mdtF, and emrB. E. coli harbored most of the AMR genes previously associated with resistance to commonly used antibiotics. mNGS offers a promising complementary approach to conventional diagnostics for identifying pathogens and AMR profiles in vulnerable populations. Full article
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9 pages, 194 KiB  
Communication
Neurodevelopmental Outcomes in Children with Neonatal Parechovirus CNS Infections
by Anna Piwowarczyk, Julia Śladowska, Agata Lipiec, Ernest Kuchar and Elżbieta Stawicka
Pathogens 2025, 14(6), 600; https://doi.org/10.3390/pathogens14060600 - 18 Jun 2025
Viewed by 53
Abstract
Human parechoviruses, officially known as Parechovirus A (PeV-A), are more frequently reported as a significant cause of serious infections in newborns and young infants. We aimed to describe the clinical features and neurological outcomes of PeV-A encephalitis cases identified in Warsaw. Infants with [...] Read more.
Human parechoviruses, officially known as Parechovirus A (PeV-A), are more frequently reported as a significant cause of serious infections in newborns and young infants. We aimed to describe the clinical features and neurological outcomes of PeV-A encephalitis cases identified in Warsaw. Infants with suspected encephalitis were retrospectively identified in three hospitals in the summer of 2022. Cases of confirmed PeV-A infection had their comprehensive demographic, clinical, laboratory, imaging, and outcome data reviewed. The psychomotor development of the children up to the age of 2 years was assessed by using the standardized tools. We identified 18 cases of confirmed encephalitis with a PeV-A infection. Their median age was 16 days. Fourteen cases were included in the analysis, while one patient dropped out after the first visit. Most were boys (9/14), and one patient was born preterm. Three patients had white matter alterations on brain MRI at discharge. No significant neurologic sequelae were observed after acute illness. At the 24-month follow-up, based on the Bayley Scales of Infant and Toddler Development (BSID-IV) and the Brunet–Lézine Scale, the children showed no neurodevelopmental sequelae. Brain MRIs were obtained in all of the participants up to 12 months of age and revealed no significant lesions. Neurodevelopmental complications are not frequent in children after PeV-A encephalitis at 24 months of age. Continued follow-up in larger cohorts is needed to explore the predictors of long-term morbidity. Full article
11 pages, 822 KiB  
Article
Bat Influenza M2 Shows Functions Similar to Those of Classical Influenza A Viruses
by Wenyu Yang, Liping Wang, Lei Shi, Jialin Zhang, Heidi Liu, Jun Wang and Wenjun Ma
Pathogens 2025, 14(6), 599; https://doi.org/10.3390/pathogens14060599 - 18 Jun 2025
Viewed by 90
Abstract
Novel bat influenza viruses show different features in contrast to classical influenza A viruses (IAVs). The M2 of IAVs functions as an ion channel that plays an important role in virus entry, viral assembly, and release and also serves as the antiviral target. [...] Read more.
Novel bat influenza viruses show different features in contrast to classical influenza A viruses (IAVs). The M2 of IAVs functions as an ion channel that plays an important role in virus entry, viral assembly, and release and also serves as the antiviral target. To date, whether bat influenza M2 functions as the ion channel like classical IAV M2 remains unknown. Here, we show that the bat influenza M2 amino acid at position 31 (N/S) is critical for sensitivity to antivirals targeting the ion channel such as amantadine and other tested antivirals and that the amino acids at position 37 (H/G) and 41 (W/A) are crucial for virus replication and survival. The results indicate that bat influenza M2 functions similarly to conventional IAVs despite the low identity between the two. Full article
(This article belongs to the Section Viral Pathogens)
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13 pages, 244 KiB  
Article
Evaluation of HIV Late Presentation Trends: A Cross-Sectional Analysis from a Leading Ecuadorian Public Hospital
by Adriana D. Suarez-Vizcaino, Nicole C. Bustamante-Pancho, Juan S. Izquierdo-Condoy, Hugo Pereira-Olmos, I. Alberto Castillo and Esteban Ortiz-Prado
Pathogens 2025, 14(6), 598; https://doi.org/10.3390/pathogens14060598 - 18 Jun 2025
Viewed by 70
Abstract
The global impact of HIV is especially significant when diagnoses are made in advanced stages. While strategies exist to mitigate late presentations, Ecuador’s 2018–2022 strategic plan has not yet been evaluated. This study assesses the prevalence and implications of late and advanced HIV [...] Read more.
The global impact of HIV is especially significant when diagnoses are made in advanced stages. While strategies exist to mitigate late presentations, Ecuador’s 2018–2022 strategic plan has not yet been evaluated. This study assesses the prevalence and implications of late and advanced HIV presentations in Ecuador, using data from a reference hospital in Quito. A cross-sectional analysis of 436 medical records of people living with HIV from the “Hospital de Especialidades Eugenio Espejo” was conducted between November 2015 and February 2020. The data were divided into “Pre-Plan” and “Post-Plan” periods for comparative analysis. The mean CD4 T count showed a non-statistically significant increase in the post-plan period (January 2018–February 2020). Notably, 65.1% of patients presented late, and 39.4% had advanced disease. Demographic data indicated that 89.9% were men, and 54.1% were under 30 years of age. No characteristics were identified that were associated with advanced late presentation of HIV infection. Sexual orientation data revealed that 69.1% identified as homosexual or bisexual. A predominance of late and advanced presenters was identified in the post-plan period, associated with being employed (p < 0.05) and being drug users (p < 0.001). There was also a greater incidence of late presenters among immigrants in the post-plan period (p = 0.045). Despite the implementation of Ecuador’s 2018–2022 strategic plan for HIV, substantial challenges in reducing late presentations remain. This study suggests that early diagnoses have not significantly improved. Employed patients and drug users were more likely to present late, with drug users also accounting for many advanced cases. This study highlights the need for more focused and targeted strategies to supplement the existing plan. Full article
11 pages, 2217 KiB  
Article
Molecular and Epidemiological Investigation of Cryptosporidium Infection in Goat Population from Bouira Province, Algeria
by Samia Bedjaoui, Djamel Baroudi, Karim Tarik Adjou, Bernard Davoust and Younes Laidoudi
Pathogens 2025, 14(6), 597; https://doi.org/10.3390/pathogens14060597 - 18 Jun 2025
Viewed by 104
Abstract
Cryptosporidiosis is a gastrointestinal disease affecting terrestrial and aquatic vertebrates worldwide. This study investigated molecularly and microscopically the prevalence and the diversity of Cryptosporidium spp. in goats across the Bouira communes, Algeria. A total of 559 fecal samples were collected from 70 farms, [...] Read more.
Cryptosporidiosis is a gastrointestinal disease affecting terrestrial and aquatic vertebrates worldwide. This study investigated molecularly and microscopically the prevalence and the diversity of Cryptosporidium spp. in goats across the Bouira communes, Algeria. A total of 559 fecal samples were collected from 70 farms, representing 16.6% of the regional goat population. Samples were analyzed using microscopy (modified Ziehl-Neelsen staining) and molecular methods (i.e., qPCR and nested PCR targeting the 18S rRNA gene, followed by sequencing). Microscopy detected Cryptosporidium in 6.1% of samples, while qPCR revealed a significantly higher prevalence of 13.6% (p < 0.00001), confirming the superior sensitivity of molecular diagnostics. Spatial analysis identified significant clustering (Moran’s I = 0.330, p = 0.0003), with communes-level prevalence ranging from 6.7% to 45.7%. Infection rates correlated positively with humidity and rainfall but negatively with temperature. Phylogenetic analysis confirmed Cryptosporidium xiaoi as the sole species circulating, showing 100% genetic similarity to global caprine isolates. Despite C. xiaoi’s host adaptation, a GenBank review highlighted six other zoonotic species infecting goats worldwide, underscoring potential cross-species transmission risks. The study emphasizes the need for PCR-based surveillance to assess true prevalence and zoonotic threats, while climatic findings support targeted interventions in high-risk areas. Full article
(This article belongs to the Special Issue Biology, Epidemiology and Interactions of Parasitic Diseases)
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13 pages, 1427 KiB  
Article
Immunohistochemical Profiling of Immune Checkpoints in Chronic Hepatitis B Liver Tissue
by João Panão-Costa, Rui Caetano Oliveira, Paulo Teixeira, Francisco Caramelo, Maria Augusta Cipriano, Olga Borges and Armando Carvalho
Pathogens 2025, 14(6), 596; https://doi.org/10.3390/pathogens14060596 - 18 Jun 2025
Viewed by 87
Abstract
Chronic hepatitis B (CHB) remains a significant global health concern due to complications like cirrhosis and liver cancer. Immune cell exhaustion, characterized by increased suppressive molecules and inhibitory receptors, represents a critical feature of CHB. Understanding the mechanisms of hepatic immune exhaustion in [...] Read more.
Chronic hepatitis B (CHB) remains a significant global health concern due to complications like cirrhosis and liver cancer. Immune cell exhaustion, characterized by increased suppressive molecules and inhibitory receptors, represents a critical feature of CHB. Understanding the mechanisms of hepatic immune exhaustion in CHB patients is imperative for the development of effective therapeutic interventions. In this study, we investigated the expression levels and histological distribution of various immune checkpoint receptors and ligands in liver biopsies obtained from CHB patients. Additionally, we aimed to evaluate potential concurrent overexpression of specific receptors and their association with clinical parameters such as ALT levels. Our analysis revealed that PD-1, PD-L1, CTLA-4, TIM-3, GAL-9, CD272, TIGIT, and 2B4 exhibited predominant localization in portal tracts and sinusoids. Furthermore, we observed a correlation between the expression of PD-1, TIM-3, and GAL-9 with ALT levels in CHB patients. Additionally, a strong relationship was identified between the expression of CD272 and TIGIT, as well as between GAL-9 and CTLA-4 within the studied population. Our findings underscore the significance of the TIM-3:GAL-9 pathway in the immunopathogenesis of CHB. This detailed analysis sets the stage for future combined immunotherapy strategies aimed at leveraging checkpoint receptors to enhance clinical outcomes. Full article
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20 pages, 2180 KiB  
Article
Insights into the Regulatory Roles of miRNAs in the Salivary Glands of the Soft Ticks Ornithodoros moubata and Ornithodoros erraticus
by Ana Laura Cano-Argüelles, Ricardo Pérez-Sánchez, Cristian Gallardo-Escárate, Rocío Vizcaíno-Marín, María González-Sánchez and Ana Oleaga
Pathogens 2025, 14(6), 595; https://doi.org/10.3390/pathogens14060595 - 17 Jun 2025
Viewed by 45
Abstract
MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression by inhibiting or degrading messenger RNAs (mRNAs). In ticks, salivary miRNAs are proposed to play key roles in modulating host–vector interactions during blood feeding. Previously, we identified salivary miRNAs in Ornithodoros moubata and [...] Read more.
MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression by inhibiting or degrading messenger RNAs (mRNAs). In ticks, salivary miRNAs are proposed to play key roles in modulating host–vector interactions during blood feeding. Previously, we identified salivary miRNAs in Ornithodoros moubata and Ornithodoros erraticus, major vectors of African swine fever and tick-borne human relapsing fever. In this study, we investigated the regulatory roles of salivary miRNAs in tick biology. Salivary miRNA datasets were re-analysed to identify conserved miRNAs, and putative target genes were predicted using the sialotranscriptomes of both species. In silico predictions were validated through experimental inhibition of specific miRNAs using antagomirs. Knockdown of miR-375 and miR-1 significantly reduced blood intake, oviposition, and fertility, indicating their involvement in feeding and reproductive processes. Silencing miR-252b in O. moubata led to increased mortality, suggesting a critical role in survival. Notably, Metis1 was identified as a likely target of miR-252b, and its dysregulation may underlie the observed lethality in miR-252b-silenced ticks. These findings highlight the functional relevance of salivary miRNAs in tick physiology and host interaction, offering new perspectives for the development of innovative tick control strategies. Full article
(This article belongs to the Section Ticks)
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15 pages, 2127 KiB  
Article
The Incidence and Trends of Yellow Fever from 1990 to 2021 in Major Endemic Regions: A Systematic Analysis Based on the 2021 Global Burden of Disease Study
by Xinwei Wang, Bin Li, Baoren He, Xipeng Yan, Linbin Huang, Jinlian Li, Rongji Lai, Mingshuang Lai, He Xie, Qiuhong Mo and Limin Chen
Pathogens 2025, 14(6), 594; https://doi.org/10.3390/pathogens14060594 - 16 Jun 2025
Viewed by 132
Abstract
As a re-emerging disease, the worldwide burden and trends of yellow fever (YF) remain inadequately quantified. This study aims to assess the incidence of YF both globally and in major endemic regions from 1990 to 2021. Utilizing data from the Global Burden of [...] Read more.
As a re-emerging disease, the worldwide burden and trends of yellow fever (YF) remain inadequately quantified. This study aims to assess the incidence of YF both globally and in major endemic regions from 1990 to 2021. Utilizing data from the Global Burden of Disease (GBD) database, we evaluated the burden of YF. We employed an age–period–cohort model to assess the influence of age, period, and cohort on the incidence of YF from 1992 to 2021. A secondary data analysis based on GBD database showed the following: in 2021, there were 86,509 incident cases of YF. Between 1990 and 2021, the number of incident cases decreased by 74.7%, while the age-standardized incidence rate (ASIR) declined at an EAPC of −4.76% (95% confidence interval: −5.10 to −4.42). In 2021, the highest ASIRs of YF were observed in Western Sub-Saharan Africa, Central Sub-Saharan Africa, and Eastern Sub-Saharan Africa. The analysis of age effects indicates that children aged 5–10 years old exhibit the highest incidence rate. Both period and cohort effects demonstrated a decline in morbidity risk. The decomposition analysis identified epidemiological changes as the primary factor contributing to the global reduction in the YF burden. Despite considerable reduction in incidence, YF remains a significant public health threat in Sub-Saharan Africa. Full article
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14 pages, 1855 KiB  
Article
Oxidative Stress and Apoptotic Markers in Goats Naturally Infected with Mycobacterium avium subsp. paratuberculosis
by Merve Ozturk, Muhammet Bahaeddin Dortbudak, Bayram Bekmez, Lucia Biagini, Nuri Altuğ, Giacomo Rossi, Yasin Ozturk and Alessandro Di Cerbo
Pathogens 2025, 14(6), 593; https://doi.org/10.3390/pathogens14060593 - 16 Jun 2025
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Abstract
Paratuberculosis, caused by Mycobacterium avium subspecies paratuberculosis (MAP), is a chronic granulomatous enteritis with significant implications for ruminant health, economic productivity, and potential zoonotic risk. This study investigated the expression of biomarkers of oxidative stress and apoptosis in goats naturally infected with MAP, [...] Read more.
Paratuberculosis, caused by Mycobacterium avium subspecies paratuberculosis (MAP), is a chronic granulomatous enteritis with significant implications for ruminant health, economic productivity, and potential zoonotic risk. This study investigated the expression of biomarkers of oxidative stress and apoptosis in goats naturally infected with MAP, focusing on three biological matrices: serum, intestinal mucosa, and mesenteric lymph nodes. Twenty MAP-positive goats and ten healthy controls were included. Serum and tissue levels of malondialdehyde (MDA), glutathione S-transferase (GST), glutathione peroxidase (GPX), superoxide dismutase (SOD), glutathione reductase (GSR), and caspase-3 were quantitatively assessed using ELISA tests. Gross and histopathological analyses confirmed MAP infection. Infected animals showed significantly elevated serum levels of MDA and caspase-3 (p < 0.001), along with decreased antioxidant enzyme activities (GSR, GST, GPX, SOD). Tissue analysis revealed increased MDA and caspase-3 levels, particularly in the intestinal mucosa compared to mesenteric lymph nodes, suggesting localized oxidative damage and apoptosis. Conversely, antioxidant enzyme activity was higher in mesenteric lymph nodes, indicating a compensatory response and a pronounced involvement of the intestinal tract. These findings demonstrate that MAP infection induces marked oxidative stress and apoptotic processes, especially in the intestinal mucosa. The imbalance between pro-oxidant and antioxidant systems may play a key role in the pathogenesis and chronic progression of the disease. Caspase-3 and MDA, in particular, have been identified as promising diagnostic or prognostic biomarkers for MAP infection. This study highlights the importance of developing improved diagnostic tools and therapeutic strategies targeting oxidative stress pathways in paratuberculosis. Full article
(This article belongs to the Special Issue Biology of Mycobacterial Pathogens)
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16 pages, 876 KiB  
Article
M72 Fusion Proteins in Nanocapsules Enhance BCG Efficacy Against Bovine Tuberculosis in a Mouse Model
by Federico Carlos Blanco, Renée Onnainty, María Rocío Marini, Laura Inés Klepp, Elizabeth Andrea García, Cristina Lourdes Vazquez, Ana Canal, Gladys Granero and Fabiana Bigi
Pathogens 2025, 14(6), 592; https://doi.org/10.3390/pathogens14060592 - 16 Jun 2025
Viewed by 162
Abstract
Mycobacterium bovis is the causative pathogen of bovine tuberculosis (bTB), a disease that affects cattle and other mammals, including humans. Currently, there is no efficient vaccine against bTB, underscoring the need for novel immunization strategies. The M72 fusion protein, composed of three polypeptides derived [...] Read more.
Mycobacterium bovis is the causative pathogen of bovine tuberculosis (bTB), a disease that affects cattle and other mammals, including humans. Currently, there is no efficient vaccine against bTB, underscoring the need for novel immunization strategies. The M72 fusion protein, composed of three polypeptides derived from Mycobacterium tuberculosis and M. bovis, has demonstrated protective efficacy against M. tuberculosis in clinical trials when combined with the AS01E adjuvant. Given the established efficacy of nanocapsule formulations as vaccine delivery systems, this study evaluated a novel immunization strategy combining BCG with either full-length M72 or a truncated M72 fused to a streptococcal albumin-binding domain (ABDsM72). Both antigens were encapsulated in chitosan/alginate nanocapsules and assessed in a murine M. bovis challenge model. Priming with BCG followed by an M72 boost significantly improved splenic protection compared to BCG alone, but it did not enhance pulmonary protection. Notably, boosting with ABDsM72 further increased the proportion of CD4+KLRG1-CXCR3+ T cells in the lungs of M. bovis-challenged mice, a key correlate of protective immunity. These findings demonstrate that chitosan/alginate-encapsulated antigens enhance BCG-induced immunity, supporting their potential as next-generation vaccine candidates for bTB control. Full article
(This article belongs to the Special Issue Mycobacterial Infection: Pathogenesis and Drug Development)
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20 pages, 4293 KiB  
Article
Novel Antischistosomal Drug Targets: Identification of Alkaloid Inhibitors of SmTGR via Integrated In Silico Methods
by Valéria V. M. Paixão, Yria J. A. Santos, Adriana O. Fernandes, Elaine S. Conceição, Ricardo P. Rodrigues, Daniela A. Chagas-Paula, Silvio S. Dolabella and Tiago B. Oliveira
Pathogens 2025, 14(6), 591; https://doi.org/10.3390/pathogens14060591 - 15 Jun 2025
Viewed by 299
Abstract
Schistosomiasis mansoni is a neglected tropical disease caused by the parasite Schistosoma mansoni, affecting approximately 200 million people annually. Currently, treatment relies primarily on a single drug, praziquantel (PZQ), which shows limited efficacy against the parasite’s immature forms. As a result, Thioredoxin [...] Read more.
Schistosomiasis mansoni is a neglected tropical disease caused by the parasite Schistosoma mansoni, affecting approximately 200 million people annually. Currently, treatment relies primarily on a single drug, praziquantel (PZQ), which shows limited efficacy against the parasite’s immature forms. As a result, Thioredoxin Glutathione Reductase from S. mansoni (SmTGR) has emerged as a promising target for novel drug development. This study presents the development of integrated in silico methods to identify alkaloids from medicinal plants with potential activity against S. mansoni. Fourteen alkaloids were identified, with predicted activity ranging from 61.3 to 85.2%. Among these, lindoldhamine and daibucarboline A demonstrated, for the first time, potential SmTGR inhibition, with probabilities of 85.2% and 75.8%, respectively. These findings highlight the potential of these alkaloids as promising candidates for the development of new therapies against schistosomiasis. Full article
(This article belongs to the Section Parasitic Pathogens)
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12 pages, 2254 KiB  
Article
Latency-Associated Nuclear Antigen (LANA) Promotes Ferroptosis by Suppressing Nrf2/GPX4 and Upregulating MDM2
by Yuejia Cao, Shihan Shao, Yingying Zhang, Dandan Song, Fei Gui, Xinyi Chen, Yu Hong, Rong Chen, Yang Song, Dongmei Li, Xiaohua Tan and Chunhong Di
Pathogens 2025, 14(6), 590; https://doi.org/10.3390/pathogens14060590 - 15 Jun 2025
Viewed by 196
Abstract
Ferroptosis, an iron-dependent cell death driven by lipid peroxidation, is regulated by key mediators including glutathione peroxidase 4 (GPX4) and nuclear factor erythroid 2-related factor 2 (Nrf2). Kaposi’s sarcoma-associated herpesvirus (KSHV) encodes latency-associated nuclear antigen (LANA), a multifunctional protein critical for viral persistence. [...] Read more.
Ferroptosis, an iron-dependent cell death driven by lipid peroxidation, is regulated by key mediators including glutathione peroxidase 4 (GPX4) and nuclear factor erythroid 2-related factor 2 (Nrf2). Kaposi’s sarcoma-associated herpesvirus (KSHV) encodes latency-associated nuclear antigen (LANA), a multifunctional protein critical for viral persistence. Although studies reported that KSHV infection enhanced cellular resistance to ferroptosis, the specific role of LANA in this process remains unexplored. Here, we demonstrate that LANA unexpectedly promotes ferroptosis. In KSHV-positive iSLK.219 cells, LANA knockdown significantly attenuated RSL-3-induced ferroptosis, whereas LANA overexpression sensitized HeLa cells to ferroptotic death. Quantitative analysis revealed that LANA-depleted cells exhibited significantly elevated ROS accumulation (p < 0.01), whereas LANA-overexpressing cells maintained reduced ROS levels during challenge with the ferroptosis inducer RSl-3. Mechanistically, LANA suppressed glutathione peroxidase 4 (GPX4) expression, reduced nuclear factor erythroid 2-related factor 2 (Nrf2) expression and impaired its nuclear translocation, and upregulated mouse double minute 2 homolog (MDM2) expression. Pharmacological inhibition of Nrf2 (ML385) or MDM2 (nutlin3a) reversed the ferroptotic effects of LANA knockdown or overexpression, respectively. These findings reveal a pro-ferroptotic role of LANA via Nrf2/GPX4 suppression and MDM2 activation. Full article
(This article belongs to the Special Issue Herpesvirus Latency and Reactivation)
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29 pages, 2576 KiB  
Review
A Comprehensive Review of the Mechanisms of Human Q Fever: Pathogenesis and Pathophysiology
by José-Luis Pérez-Arellano, Jose Curbelo and Cristina Carranza-Rodriguez
Pathogens 2025, 14(6), 589; https://doi.org/10.3390/pathogens14060589 - 14 Jun 2025
Viewed by 396
Abstract
Coxiella burnetii infection has a worldwide distribution, although the incidence and clinical manifestations vary between and within countries. There are the following four basic forms: asymptomatic infection, acute Q fever, chronic Q fever, and post-Q fever fatigue syndrome. The aim of this review [...] Read more.
Coxiella burnetii infection has a worldwide distribution, although the incidence and clinical manifestations vary between and within countries. There are the following four basic forms: asymptomatic infection, acute Q fever, chronic Q fever, and post-Q fever fatigue syndrome. The aim of this review is to provide a comprehensive overview of the important aspects of its pathogenesis and pathophysiology. First, we provide a brief update of the taxonomic aspects, basic structures, and genotypes of C. burnetii necessary for the proper interpretation of the following sections. Routes of infection, different stages of pathogenesis (respiratory entry of C. burnetii; penetration into alveolar macrophages, life cycle, and effects; systemic dissemination), and innate, acquired humoral and cell-mediated immune responses in different forms of infection are described in detail. The pathophysiology and clinical manifestations of Q fever, such as the main mechanisms of injury, in isolation and in combination, are reviewed. The clinical and biological manifestations of the two main forms of Q fever (acute and chronic) are outlined, with a brief definition and mention of the mechanisms of post-Q fever fatigue syndrome. Full article
(This article belongs to the Section Bacterial Pathogens)
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19 pages, 266 KiB  
Article
Characteristics of Neonates with Sepsis Associated with Antimicrobial Resistance and Mortality in a Tertiary Hospital in Mexico: A Retrospective Observational Study
by Uriel A. Angulo-Zamudio, Maria Luisa Velazquez-Meza, Jesus J. Martinez-Garcia, Nidia Leon-Sicairos, Jorge Velazquez-Roman, Hector Flores-Villaseñor, Claudia Leon-Sicairos, Francisco A. Martínez-Villa and Adrian Canizalez-Roman
Pathogens 2025, 14(6), 588; https://doi.org/10.3390/pathogens14060588 - 14 Jun 2025
Viewed by 238
Abstract
The objective of this study was to determine the epidemiological, clinical, and laboratory characteristics of newborns with sepsis in northwestern Mexico, identify the microorganisms causing early- and late-onset sepsis, and assess antimicrobial resistance. Additionally, it sought to associate neonatal characteristics with antimicrobial resistance [...] Read more.
The objective of this study was to determine the epidemiological, clinical, and laboratory characteristics of newborns with sepsis in northwestern Mexico, identify the microorganisms causing early- and late-onset sepsis, and assess antimicrobial resistance. Additionally, it sought to associate neonatal characteristics with antimicrobial resistance or mortality. A retrospective study was conducted from August 2021 to April 2023, during which 8382 neonatal clinical records were analyzed to collect epidemiological, clinical, and laboratory characteristics, as well as microorganisms isolated from neonates and their antimicrobial resistance profiles. Of these, 314 neonates with sepsis were included. The incidence of neonatal sepsis was 4% (314/8382), and the mortality was 12.7% (40/314); late-onset sepsis (65.3%) was more frequent than early-onset sepsis (34.7%). Staphylococcus epidermidis was the most frequently isolated bacterium in neonates with sepsis (both early- and late-onset). Gram-positive bacteria, such as Staphylococcus hominis and Enterococcus faecium, were associated with early-onset sepsis, whereas fungi, particularly Candida albicans, were associated with late-onset sepsis. Of the microorganisms, 52.6% were multidrug resistant (MDR), 10.8% were extensively drug resistant (XDR), and 5.5% were pan-drug resistant (PDR). Low birth weight, prematurity, cesarean section, mechanical ventilation, tachycardia, and low hemoglobin and platelet levels, among others, were associated with XDR or MDR microorganisms. In contrast, low birth weight, mechanical ventilation, stroke, unexpected delivery, respiratory distress, tachycardia, convulsive crisis, high procalcitonin, urea, and AST/TGO levels, among others, were associated with mortality. The incidence, types of sepsis, antimicrobial resistance, and associations identified in this study will aid in diagnosing neonatal sepsis earlier and may reduce mortality in our region. Full article
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15 pages, 1423 KiB  
Article
First Outbreak of Aeromoniasis, Caused by Aeromonas veronii, in Farmed European Seabass (Dicentrarchus labrax) in the Ionian Sea, Greece
by Eirini Lampou, Eleni Psychari, Konstantina Louka, Charalampos Kotzamanidis, Andigoni Malousi, Ioannis Petropoulos, Markos N. Kolygas, Dimitrios Doukas and Konstantina Bitchava
Pathogens 2025, 14(6), 587; https://doi.org/10.3390/pathogens14060587 - 14 Jun 2025
Viewed by 200
Abstract
This article documents the first outbreak of aeromoniasis caused by Aeromonas veronii in farmed European seabass (Dicentrarchus labrax) along the Greek Ionian Sea coast. In late spring 2024, commercially sized fish exhibited anorexia, hemorrhages, and ulcers on the skin, accompanied by [...] Read more.
This article documents the first outbreak of aeromoniasis caused by Aeromonas veronii in farmed European seabass (Dicentrarchus labrax) along the Greek Ionian Sea coast. In late spring 2024, commercially sized fish exhibited anorexia, hemorrhages, and ulcers on the skin, accompanied by elevated morbidity and mortality rates. The outbreak spread rapidly across local farms in Sagiada Bay, reaching its peak in late summer, and extending into the Astakos Gulf, southern in the Ionian Sea. The postmortem examination revealed hemorrhages, organomegaly, abscess formation, and granulomatous inflammation. Aeromonas veronii was isolated from all examined individuals in nutrient media and confirmed by biochemical and molecular methods. Whole genome sequencing and phylogenetic analysis demonstrated genetic homogeneity among two strains from two different areas along the Ionian Sea and a close evolutionary relationship with other Aeromonas veronii strains from the Aegean Sea. Although genetically similar, the isolates exhibited differences in phenotypic and biochemical characteristics, indicating regional variability. The present study provides an overview of the pathology, clinical characteristics and progression of aeromoniasis in Ionian Sea aquaculture, highlighting the need for continued monitoring, in-depth genomic and phenotypic assessment, and the design of region-specific preventive strategies, including autogenous vaccines, for effective disease management. Full article
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65 pages, 6483 KiB  
Review
The Challenge of Lyssavirus Infections in Domestic and Other Animals: A Mix of Virological Confusion, Consternation, Chagrin, and Curiosity
by Charles E. Rupprecht, Aniruddha V. Belsare, Florence Cliquet, Philip P. Mshelbwala, Janine F. R. Seetahal and Vaughn V. Wicker
Pathogens 2025, 14(6), 586; https://doi.org/10.3390/pathogens14060586 - 13 Jun 2025
Viewed by 329
Abstract
Lyssaviruses are RNA viruses in the Family Rhabdoviridae, Genus Lyssavirus. They represent the causative agents of acute, progressive encephalitis, known historically as rabies. Regardless of specific etiology, their collective viral morphology, biochemistry, pathobiology, associated clinical signs, diagnosis, epizootiology, and management are essentially [...] Read more.
Lyssaviruses are RNA viruses in the Family Rhabdoviridae, Genus Lyssavirus. They represent the causative agents of acute, progressive encephalitis, known historically as rabies. Regardless of specific etiology, their collective viral morphology, biochemistry, pathobiology, associated clinical signs, diagnosis, epizootiology, and management are essentially the same. Despite centuries of clinical recognition, these quintessential neurotropic agents remain significant pathogens today, with substantive consequences to agriculture, public health, and conservation biology. Notably, the singular morbidity caused by lyssaviruses is incurable and constitutes the highest case fatality of any viral disease. All warm-blooded vertebrates are believed to be susceptible. The dog is the only domestic animal that serves as a reservoir, vector, and victim. In contrast, felids are effective vectors, but not reservoirs. All other rabid domestic species, such as livestock, constitute spillover infections, as a bellwether to local lyssavirus activity. Frequently, professional confusion abounds among the veterinary community, because although the viral species Lyssavirus rabies is inarguably the best-known representative in the Genus, at least 20 other recognized or putative members of this monophyletic group are known. Frequently, this is simply overlooked. Moreover, often the ‘taxonomic etiology’ (i.e., ‘Lyssavirus x’) is mistakenly referenced in a biopolitcal context, instead of the obvious clinical illness (i.e., ‘rabies’). Global consternation persists, if localities believe they are ‘disease-free’, when documented lyssaviruses circulate or laboratory-based surveillance is inadequate to support such claims. Understandably, professional chagrin develops when individuals mistake the epidemiological terminology of control, prevention, elimination, etc. Management is not simple, given that the only licensed veterinary and human vaccines are against rabies virus, sensu lato. There are no adequate antiviral drugs for any lyssaviruses or cross-reactive biologics developed against more distantly related viral members. While representative taxa among the mammalian Orders Chiroptera, Carnivora, and Primates exemplify the major global reservoirs, which mammalian species are responsible for the perpetuation of other lyssaviruses remains a seemingly academic curiosity. This zoonosis is neglected. Clearly, with such underlying characteristics as a fundamental ‘disease of nature’, rabies, unlike smallpox and rinderpest, is not a candidate for eradication. With the worldwide zeal to drive human fatalities from canine rabies viruses to zero by the rapidly approaching year 2030, enhanced surveillance and greater introspection of the poorly appreciated burden posed by rabies virus and diverse other lyssaviruses may manifest as an epidemiological luxury to the overall global program of the future. Full article
(This article belongs to the Special Issue Current Challenges in Veterinary Virology)
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19 pages, 9515 KiB  
Article
Survey of Piroplasmids in Wild Mammals, Unconventional Pets, and Ticks from Goiás State, Midwestern Brazil
by Raphaela Bueno Mendes Bittencourt, Ana Cláudia Calchi, Lucianne Cardoso Neves, Nicolas Jalowitzki de Lima, Gabriel Cândido dos Santos, Ennya Rafaella Neves Cardoso, Warley Vieira de Freitas Paula, Luciana Batalha de Miranda Araújo, Jessica Rocha Gonçalves, Elisângela de Albuquerque Sobreira, Luiz Alfredo Martins Lopes Baptista, Hermes Ribeiro Luz, Marcos Rogério André, Filipe Dantas-Torres and Felipe da Silva Krawczak
Pathogens 2025, 14(6), 585; https://doi.org/10.3390/pathogens14060585 - 12 Jun 2025
Viewed by 343
Abstract
Tick-borne piroplasmids are apicomplexan protozoa that infect a wide range of vertebrate hosts, with significant implications for animal and human health. This study investigated the occurrence and genetic diversity of piroplasmids in wild mammals, unconventional pets, and associated ticks in Goiás state, midwestern [...] Read more.
Tick-borne piroplasmids are apicomplexan protozoa that infect a wide range of vertebrate hosts, with significant implications for animal and human health. This study investigated the occurrence and genetic diversity of piroplasmids in wild mammals, unconventional pets, and associated ticks in Goiás state, midwestern Brazil. Between April 2023 and January 2024, 105 blood samples, 22 tissue samples, and 300 ticks were collected from 21 mammalian species housed in wildlife screening centers, zoos, and veterinary clinics. Molecular screening targeting the 18S rRNA gene of piroplasmids detected a 25.7% (27/105) overall positivity, with gray brockets (Subulo gouazoubira) and South American tapirs (Tapirus terrestris) showing the highest infection rates. Three tick samples tested positive, including two Amblyomma sculptum nymphs and a male of Amblyomma dubitatum collected from a tapir and capybara (Hydrochoerus hydrochaeris). Cytauxzoon brasiliensis was reported, for the first time, in cougars (Puma concolor) from Goiás state, midwestern Brazil, indicating the role of this feline as a host of this parasite. Babesia goianiaensis was confirmed in a capybara, and Theileria terrestris in tapirs. Phylogenetic analyses clustered gray brockets-associated Theileria sequences with Theileria sp. previously detected in Neotropical deer from Brazil and Theileria cervi. While the phylogenetic analysis of amino acid sequences of the cytochrome c oxidase subunit III separated Theileria genotypes detected in S. gouazoubira from T. cervi, hsp70-based phylogenetic inferences clustered the genotypes detected in Tapirus terrestris with Theileria terrestris, suggesting host-specific evolutionary lineages. These findings contribute to the understanding of Piroplasmida diversity and circulation in South American wild mammals, emphasizing the need for enhanced molecular surveillance to elucidate transmission dynamics, assess potential health risks, and contribute to the establishment of wildlife conservation and One Health strategies. Full article
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16 pages, 1357 KiB  
Article
EmsB Microsatellite Analysis of Echinococcus multilocularis Specimens Isolated from Belgian Patients with Alveolar Echinococcosis and from Animal Hosts
by Sabrina Egrek, Jenny Knapp, Rosalie Sacheli, Khalid El Moussaoui, Philippe Léonard, Eva Larranaga Lapique, Laurence Millon, Sara Engelskirchen, Olivier Detry, Annick Linden and Marie-Pierre Hayette
Pathogens 2025, 14(6), 584; https://doi.org/10.3390/pathogens14060584 - 12 Jun 2025
Viewed by 238
Abstract
Alveolar echinococcosis (AE), caused by Echinococcus multilocularis (E. multilocularis), is a severe parasitic zoonosis that is potentially fatal for humans. The parasite is primarily transmitted by wildlife, with red foxes acting as definitive hosts and rodents as intermediate hosts, while humans [...] Read more.
Alveolar echinococcosis (AE), caused by Echinococcus multilocularis (E. multilocularis), is a severe parasitic zoonosis that is potentially fatal for humans. The parasite is primarily transmitted by wildlife, with red foxes acting as definitive hosts and rodents as intermediate hosts, while humans can become accidental but dead-end hosts. The aim of this study is to use EmsB typing on E. multilocularis isolates from human AE cases and local animals such as foxes and rodents. In this study, retrospective EmsB typing was performed on 39 samples, including 11 tissue samples from 10 patients, 18 fecal swabs from foxes, and 10 tissue samples from rodents. A dendrogram was created to determine the EmsB profiles present. The results showed that all the rodent samples were associated with the EmsB P1 profile (10/10), while the human and fox samples shared the EmsB profile P1 (5/11 humans and 8/18 foxes), a profile near P4 (2/11 humans and 3 foxes), and a profile near P8 (1/11 humans and 1/18 foxes). The study demonstrates that the same EmsB profiles circulate among humans and animals, confirming that wildlife reservoirs play a key role in transmission. Full article
(This article belongs to the Section Parasitic Pathogens)
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7 pages, 315 KiB  
Brief Report
Implications of Intravenous and Inhaled Amikacin Breakpoint Reporting for Mycobacterium avium Complex Pulmonary Isolates
by Christian M. Gill, Robin Chamberland and Getahun Abate
Pathogens 2025, 14(6), 583; https://doi.org/10.3390/pathogens14060583 - 12 Jun 2025
Viewed by 251
Abstract
The treatment of Mycobacterium avium complex (MAC) remains a clinical challenge as multidrug regimens are needed and may be limited by treatment-related toxicity. The Clinical and Laboratory Standards Institute (CLSI) endorses breakpoints for several agents used for MAC infection treatment. Amikacin has distinct [...] Read more.
The treatment of Mycobacterium avium complex (MAC) remains a clinical challenge as multidrug regimens are needed and may be limited by treatment-related toxicity. The Clinical and Laboratory Standards Institute (CLSI) endorses breakpoints for several agents used for MAC infection treatment. Amikacin has distinct breakpoints for intravenous (IV) therapy and inhaled therapy using amikacin liposome inhalation suspension (ALIS) for MAC pulmonary disease. The purpose of the present retrospective cohort study of MAC pulmonary isolates was to assess the number of amikacin non-susceptible isolates by the IV breakpoints that remain susceptible to the inhaled breakpoints. One isolate per patient per year was assessed and susceptibility was described for amikacin IV, amikacin inhaled, clarithromycin, moxifloxacin, and linezolid per the CLSI. Of the 218 isolates, 94% [204/218] tested as susceptible to amikacin per the IV breakpoints compared with 99.5% [217/218] to the inhaled breakpoints. Of the amikacin IV non-susceptible isolates, 93% [13/14] were susceptible by the inhaled breakpoints. For comparison, clarithromycin was the next most active agent followed by moxifloxacin and linezolid with 97% [211/218], 82% [178/218], and 66% [143/218] of isolates testing as susceptible to each, respectively. These data highlight the importance of laboratories to report both the IV and inhaled amikacin interpretive criteria so that clinicians do not disregard potential therapeutic options for the treatment of MAC pulmonary disease. Full article
(This article belongs to the Section Bacterial Pathogens)
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21 pages, 3837 KiB  
Article
Evaluation of the In Vitro Activity of Bedaquiline, Delamanid, and Clofazimine Against Mycobacterium abscessus Complex and Their Antibiofilm Potential
by Katarzyna Kania, Katarzyna Wójcik, Alicja Skórkowska and Karolina Klesiewicz
Pathogens 2025, 14(6), 582; https://doi.org/10.3390/pathogens14060582 - 11 Jun 2025
Viewed by 305
Abstract
Mycobacterium abscessus complex (MABc) poses a major therapeutic challenge due to its intrinsic multidrug resistance and ability to form biofilms. This study evaluated the in vitro activity of three antimycobacterial agents—bedaquiline, delamanid, and clofazimine—on 20 clinical MABc isolates, including M. abscessus subsp. abscessus [...] Read more.
Mycobacterium abscessus complex (MABc) poses a major therapeutic challenge due to its intrinsic multidrug resistance and ability to form biofilms. This study evaluated the in vitro activity of three antimycobacterial agents—bedaquiline, delamanid, and clofazimine—on 20 clinical MABc isolates, including M. abscessus subsp. abscessus, massiliense, and bolletii, with a focus on biofilm-forming phenotypes. Biofilm analysis showed that the rough colony morphotypes were mostly weak biofilm formers, while the smooth and mixed morphotypes were predominantly moderate or strong biofilm formers. A statistically significant association was observed between the mixed colony morphology and strong biofilm formation (p = 0.032). Importantly, bedaquiline exhibited potent and consistent activity across all isolates, regardless of the biofilm-forming ability, with MIC values ranging from 0.125 to 1 µg/mL. In contrast, delamanid and clofazimine showed limited efficacy, with MIC values exceeding 16 µg/mL and 8 µg/mL, respectively. These findings strongly support the role of bedaquiline as a promising core agent for future combination therapies targeting drug-resistant MABc infections, including biofilm-associated infections. Our results, among the first from Poland, highlight the critical need for incorporating novel agents such as bedaquiline into therapeutic strategies against this difficult-to-treat pathogen. Full article
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16 pages, 1065 KiB  
Article
First Serologic Analysis of Antibodies Against African Swine Fever Virus Detected in Domestic Pig Farms in South Korea from 2019 to 2024
by Seong-Keun Hong, Mugyeom Moon, Ki-Hyun Cho, Hae-Eun Kang, Jong-Soo Lee and Yeon-Hee Kim
Pathogens 2025, 14(6), 581; https://doi.org/10.3390/pathogens14060581 - 11 Jun 2025
Viewed by 360
Abstract
Background: African swine fever (ASF) is a crucial socioeconomic setback to South Korea’s swine industry. This study aimed to determine seropositivity for ASF virus (ASFV) in pigs that appeared to be infected on farms with reported ASF outbreaks. Methods: A total of 2232 [...] Read more.
Background: African swine fever (ASF) is a crucial socioeconomic setback to South Korea’s swine industry. This study aimed to determine seropositivity for ASF virus (ASFV) in pigs that appeared to be infected on farms with reported ASF outbreaks. Methods: A total of 2232 sera from ASF outbreaks (2019–2024) in South Korea were collected. Two enzyme-linked immunosorbent assay (ELISA) kits were used to detect ASFV antibodies, and an immunoperoxidase test (IPT) was used as a confirmatory test following the method recommended by the World Organisation for Animal Health in the Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. Also, spatial clustering was identified using the Density-Based Spatial Clustering of Applications with Noise (DBSCAN) model to understand ASF hotspots in the wild boar population and assess the spatial relationship between the hotspots and ASF antibody-positive domestic pig farms. Results: Antibodies were first detected in Hwacheon in 2020, but by 2024, only 1.43% of pigs had detectable antibodies against ASFV. Although this percentage is still low, the number of antibody-positive pigs is gradually increasing. Additionally, 32 positive samples were found from nine pig farms with outbreaks, and these samples were confirmed positive in both the two ELISA tests and the IPT. The highest seropositivity was recorded at the finishing stage of pig production. When compared to the confirmatory IPT, both blocking and competition ELISA demonstrated high diagnostic sensitivities. The statistical association between ASF antibody-positive farms and wild boars were analyzed using Fisher’s exact test, yielding a significant p-value of 0.007. This indicates a strong correlation, as eight out of nine ASF-seropositive farms were located within hotspots that were significantly associated. Conclusions: Our findings provide valuable insights into ASFV antibody detection in South Korea and demonstrate a statistical association between farms housing pigs with ASFV antibodies and hotspots of ASFV-infected wild boars. Confirmatory tests, such as the IPT, are needed. These insights will contribute to the improvement of surveillance and biosecurity measures for swine farms. Full article
(This article belongs to the Special Issue Diagnostics of Emerging and Re-Emerging Pathogens)
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16 pages, 1381 KiB  
Article
Characterization of Microbial Growth, Pathogen Presence, and Histamine Accumulation in Chilled Rainbow Trout and Mackerel Samples Collected from Romanian Markets
by Vida Silviu, Alexandra Tabaran, Oana Lucia Crişan Reget, Mihaela Niculina Duma, Luciana Cătălina Panait and Sorin Daniel Dan
Pathogens 2025, 14(6), 580; https://doi.org/10.3390/pathogens14060580 - 11 Jun 2025
Viewed by 341
Abstract
This study aimed to evaluate microbial growth, pathogen presence, and histamine production in rainbow trout and mackerel stored on flaked ice over their shelf life. A total of 72 fish samples (rainbow trout and mackerel) were analyzed across four storage intervals (day 1, [...] Read more.
This study aimed to evaluate microbial growth, pathogen presence, and histamine production in rainbow trout and mackerel stored on flaked ice over their shelf life. A total of 72 fish samples (rainbow trout and mackerel) were analyzed across four storage intervals (day 1, 3, 9, 12/11) on flaked ice. TVC increased from 2.59 to 5.04 log cfu/g in rainbow trout and from 3.18 to 4.88 log cfu/g in mackerel over the storage period. Significant increases were observed in Pseudomonas, Aeromonas, and Enterobacteriaceae populations, especially after the ninth day. Microbial identification revealed spoilage-associated bacteria, such as Pseudomonas fluorescens and Aeromonas salmonicida, as well as opportunistic pathogens, including Francisella tularensis, Yersinia spp., and Chromobacterium violaceum. Histamine levels rose with storage time but remained below toxic thresholds (<200 mg/kg), peaking at 1.56 mg/kg in trout and 1.87 mg/kg in mackerel. A strong positive correlation was found between TVC and histamine levels (Pearson’s r = 0.85 for trout, 0.82 for mackerel). Proper hygiene and storage are crucial, and consumption is recommended before day 9 of storage on flaked ice. Hygiene measures remain essential to minimize contamination risks and preserve product safety. Full article
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20 pages, 2181 KiB  
Article
Genetic Identification of Brazilian Mammalian Hosts of Trypanosoma cruzi: Improving Blood Meal Source Discrimination in Vector-Borne Transmission
by Quezia Moura Oliveira, Thaíla Santos Pessanha and Alena Mayo Iñiguez
Pathogens 2025, 14(6), 579; https://doi.org/10.3390/pathogens14060579 - 10 Jun 2025
Viewed by 400
Abstract
The detection of food sources of blood-sucking vectors is essential for a better understanding of the hosts, reservoirs, and other fauna that participate in the transmission web of hemoparasites. The molecular identification of triatomine blood meal sources (BMSs) has been shown to be [...] Read more.
The detection of food sources of blood-sucking vectors is essential for a better understanding of the hosts, reservoirs, and other fauna that participate in the transmission web of hemoparasites. The molecular identification of triatomine blood meal sources (BMSs) has been shown to be highly sensitive and taxonomically specific when compared to the immunological method. The application of molecular cloning makes it possible to identify multiple BMS species and/or different individuals/haplotypes of the same vertebrate species in a single triatomine specimen. In Brazil, the molecular detection of BMSs is incipient, with insufficient genetic information on the species of animals involved in the transmission of Trypanosoma cruzi. In this work, we evaluated the sensitivity and specificity of a molecular approach using molecular cloning for the detection of multiple Brazilian mammalian species. The DNA was extracted from blood clots of 13 species of canids, bats, xenarthral, marsupials, and rodents. Serial proportions were used to formulate mixtures combining taxonomically close (belonging to the same family or order) and taxonomically distant (different families) species. The results showed that GenBank lacks reference sequences for some native species tested, such as the sylvatic rodent, Necromys lasiurus, and the wild canid, Lycalopex gymnocercus, for cytb and 12S rDNA, and the rodent Oecomys cleberi for 12S rDNA. The study also demonstrated that it is possible to detect multiple different species, even for those that are taxonomically close. This approach was proven to be efficient for the detection of species in equal and even in disparate unequal proportions, which could represent complementary information about the diversity of potential hosts of T. cruzi. The detection of multiple BMS species in mixed samples provides a more comprehensive and accurate landscape of T. cruzi transmission in nature. Full article
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13 pages, 602 KiB  
Systematic Review
An Umbrella Review of E-Cigarettes’ Impact on Oral Microbiota and Biofilm Buildup
by Beatriz Panariello, Fabrízio Dias Panariello, Ashminie Misir and Eliane Porto Barboza
Pathogens 2025, 14(6), 578; https://doi.org/10.3390/pathogens14060578 - 10 Jun 2025
Viewed by 317
Abstract
E-cigarettes, a form of electronic nicotine delivery system (ENDS), have gained significant popularity, particularly among adolescents who often view vaping as a “cool” lifestyle choice. This growing trend has spurred extensive research on the effects of ENDS on both oral and systemic health. [...] Read more.
E-cigarettes, a form of electronic nicotine delivery system (ENDS), have gained significant popularity, particularly among adolescents who often view vaping as a “cool” lifestyle choice. This growing trend has spurred extensive research on the effects of ENDS on both oral and systemic health. By synthesizing data from systematic reviews and meta-analyses, this umbrella review offers a comprehensive evaluation of the impact of e-cigarettes on oral biofilm accumulation and microbiota composition. A systematic search was conducted up to 12 March 2025, across PubMed/MEDLINE, Google Scholar, Cochrane Library, and Scopus for studies published between 2015 and 2025. Ten studies met the eligibility criteria. The quality of the selected papers, as assessed using the AMSTAR 2 tool, ranged from moderate to high. The findings of this review suggest that e-cigarette use may contribute to dysbiosis in the oral microbiota and foster biofilm accumulation, thereby increasing the risk of oral diseases such as periodontitis, peri-implantitis, oral candidiasis, and caries. The findings also highlight the need for further research into the long-term effects of e-cigarette use on oral health. This review is registered with PROSPERO (CRD420251025639). Full article
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