Pathogens

12 pages, 455 KiB

Open AccessArticle

Restoring Control: Real-World Success with Imipenem–Relebactam in Critical MDR Infections—A Multicenter Observational Study

by Andrea Marino, Giuseppe Pipitone, Emmanuele Venanzi Rullo, Federica Cosentino, Rita Ippolito, Roberta Costa, Sara Bagarello, Ylenia Russotto, Chiara Iaria, Bruno Cacopardo and Giuseppe Nunnari

Pathogens 2025, 14(7), 685; https://doi.org/10.3390/pathogens14070685 - 11 Jul 2025

Abstract

Background: Multidrug-resistant (MDR) Gram-negative infections, particularly those caused by carbapenem-resistant Enterobacterales (CRE) and difficult-to-treat Pseudomonas aeruginosa (DTR-Pa), present a growing global healthcare challenge, especially in critically ill populations. Imipenem–relebactam (I/R), a novel β-lactam/β-lactamase inhibitor combination, has shown efficacy in clinical trials, but [...] Read more.

Background: Multidrug-resistant (MDR) Gram-negative infections, particularly those caused by carbapenem-resistant Enterobacterales (CRE) and difficult-to-treat Pseudomonas aeruginosa (DTR-Pa), present a growing global healthcare challenge, especially in critically ill populations. Imipenem–relebactam (I/R), a novel β-lactam/β-lactamase inhibitor combination, has shown efficacy in clinical trials, but real-world data remain limited. Methods: We conducted a multicenter, retrospective–prospective observational study across tertiary-care hospitals in Italy between January 2020 and May 2025. Adult patients (≥18 years) treated with I/R for ≥48 h for suspected or confirmed MDR Gram-negative infections were included. Primary endpoints were clinical success at the end of therapy and 30-day all-cause mortality. Secondary endpoints included microbiological eradication, recurrence, safety, and predictors of treatment failure. Statistical analysis involved descriptive methods and correlation analysis for mortality predictors. Results: Twenty-nine patients were included (median age 66 years; 58.6% ICU admission; 71.4% mechanical ventilation). Clinical success was achieved in 22/29 patients (75.9%), while 30-day mortality was 24.1% (7/29). The most common pathogen was Klebsiella pneumoniae (62.1%), with 41.4% of infections being polymicrobial. Microbiological eradication was confirmed in all the BSIs. Parenteral nutrition (p = 0.016), sepsis at presentation (p = 0.04), candidemia (p = 0.036), and arterial catheter use (p = 0.029) were significantly more frequent in non-survivors. Survivors showed significant reductions in CRP, PCT, and bilirubin at 48 h, while non-survivors did not. Parenteral nutrition (rho = 0.427, p = 0.023), sepsis (rho = 0.378, p = 0.043), and arterial catheter use (rho = 0.384, p = 0.04) were significantly correlated with mortality. Conclusions: In this Italian multicenter cohort of critically ill patients, imipenem–relebactam demonstrated high clinical success and acceptable mortality rates in the treatment of severe MDR Gram-negative infections, particularly those caused by KPC-producing K. pneumoniae. Early biomarker dynamics may aid in monitoring treatment response. Larger prospective studies are needed to confirm these findings and define optimal treatment strategies. Full article

(This article belongs to the Special Issue Drug Resistant Pathogens: Diagnosis, Treatment, and Global Health Implications)

11 pages, 4576 KiB

Open AccessCase Report

First Confirmed Case of Zoonotic Transmission of RR-TB from a Dog to a Human, a Neglected Mode of Mycobacterium tuberculosis Infection—Case Report and Review of the Literature

by Ljiljana Zmak, Marija Gomercic Palcic, Mihaela Obrovac, Ivana Folnozic, Drazen Strelec, Irena Reil, Ana Miljan, Maja Zdelar-Tuk, Sanja Duvnjak, Diana Mihalac, Danka Jovetic and Silvio Spicic

Pathogens 2025, 14(7), 684; https://doi.org/10.3390/pathogens14070684 - 11 Jul 2025

Abstract

Mycobacterium (M.) tuberculosis mostly spreads from active tuberculosis (TB) patients to human contacts, although human-to-animal and animal-to-human transmission has been described. Here, we present a rare case of rifampicin-resistant tuberculosis (RR-TB) transmission from a companion dog to its owner, highlighting the zoonotic potential [...] Read more.

Mycobacterium (M.) tuberculosis mostly spreads from active tuberculosis (TB) patients to human contacts, although human-to-animal and animal-to-human transmission has been described. Here, we present a rare case of rifampicin-resistant tuberculosis (RR-TB) transmission from a companion dog to its owner, highlighting the zoonotic potential of the pathogen. Namely, a 37-year-old Croatian man was diagnosed with RR-TB, with whole-genome sequencing analysis revealing a close genetic link to the strain isolated from his dog, which had died of miliary TB six years earlier. This case emphasizes the complexity of TB transmission dynamics, particularly involving companion animals, and underlines the importance of integrated “One Health” approaches for TB control. Awareness of zoonotic TB risks is essential for the early detection and prevention of cross-species transmission, especially in vulnerable populations and households with close human–animal contact. Full article

(This article belongs to the Special Issue Emerging and Neglected Pathogens in the Balkans)

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25 pages, 5421 KiB

Open AccessArticle

NOD2 (Nucleotide-Binding Oligomerization Domain-Containing Protein 2)-Mediated Modulation of the Immune Response Induced by BCG (Bacillus Calmette-Guérin) Bacilli

by Magdalena Jurczak, Joanna Kaczmarek, Magdalena Kowalewska-Pietrzak, Paulina Stelmach and Magdalena Druszczynska

Pathogens 2025, 14(7), 683; https://doi.org/10.3390/pathogens14070683 - 11 Jul 2025

Abstract

The Bacillus Calmette-Guérin (BCG) vaccine confers broad, non-specific immunity that may bolster defenses against respiratory viruses. While NOD2 (nucleotide-binding oligomerization domain-containing protein 2)-driven pathways are central to innate immune responses, the contribution of surface receptor modulation on monocytes to shaping these responses remains [...] Read more.

The Bacillus Calmette-Guérin (BCG) vaccine confers broad, non-specific immunity that may bolster defenses against respiratory viruses. While NOD2 (nucleotide-binding oligomerization domain-containing protein 2)-driven pathways are central to innate immune responses, the contribution of surface receptor modulation on monocytes to shaping these responses remains underexplored. We analyzed whole-blood cultures from BCG-vaccinated Polish children, stratified by serostatus to SARS-CoV-2 and RSV, and stimulated for 48 h with live BCG, purified viral antigens, or both. RT-qPCR quantified mRNA levels of NOD2 and key cytokines (IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, TNF), while flow cytometry assessed CD14, HLA-DR, CD11b, and CD206 expression. Co-stimulation with BCG + RSV elicited the strongest transcriptional response, notably a 2–4-fold upregulation of NOD2, IL-1β, and IL-6 versus RSV alone. In SARS-CoV-2(+) donors, RSV alone induced higher NOD2 expression than BCG or BCG + RSV, while IL-2 peaked following BCG + SARS-CoV-2. Across conditions, NOD2 positively correlated with IL-4 and IL-6 but negatively correlated with IL-1β in SARS-CoV-2 cultures. Viral antigens increased CD14 and HLA-DR on monocytes, suggesting activation; CD206 rose only in dual-seropositive children. Our findings indicate that BCG stimulation affects pediatric antiviral immunity through NOD2-related cytokine production and induction of a CD14⁺HLA-DR⁺ phenotype, supporting its potential role in boosting innate defenses against respiratory pathogens. Full article

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16 pages, 613 KiB

Open AccessArticle

Isolation and Molecular Characterization of Antimicrobial-Resistant Bacteria from Vegetable Foods

by Annamaria Castello, Chiara Massaro, Erine Seghers, Clelia Ferraro, Antonella Costa, Rosa Alduina and Cinzia Cardamone

Pathogens 2025, 14(7), 682; https://doi.org/10.3390/pathogens14070682 - 10 Jul 2025

Abstract

Antimicrobial resistance (AMR) poses a growing threat to global health, and its spread through the food chain is gaining increasing attention. While AMR in food of animal origin has been extensively studied, less is known about its prevalence in plant-based foods, particularly fresh [...] Read more.

Antimicrobial resistance (AMR) poses a growing threat to global health, and its spread through the food chain is gaining increasing attention. While AMR in food of animal origin has been extensively studied, less is known about its prevalence in plant-based foods, particularly fresh and ready-to-eat (RTE) vegetables. This study investigated the occurrence of antimicrobial-resistant bacteria in fresh and RTE vegetables. Isolates were subjected to antimicrobial susceptibility testing and molecular analyses for the characterization of antimicrobial resistance genes (ARGs). A significant proportion of samples were found to harbor antimicrobial-resistant bacteria, including multidrug-resistant strains. Several ARGs, including those encoding extended-spectrum

β

-lactamases (ESBLs) and resistance to critically important antimicrobials, were detected. The findings point to environmental contamination—potentially originating from wastewater reuse and agricultural practices—as a likely contributor to AMR dissemination in vegetables. The presence of antimicrobial-resistant bacteria and ARGs in fresh produce raises concerns about food safety and public health. The current regulatory framework lacks specific criteria for monitoring AMR in vegetables, highlighting the urgent need for surveillance programs and risk mitigation strategies. This study contributes to a better understanding of AMR in the plant-based food sector and supports the implementation of a One Health approach to address this issue. Full article

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26 pages, 888 KiB

Open AccessReview

Current Trends in Approaches to Prevent and Control Antimicrobial Resistance in Aquatic Veterinary Medicine

by Dongqing Zhao, Konrad Wojnarowski, Paulina Cholewińska and Dušan Palić

Pathogens 2025, 14(7), 681; https://doi.org/10.3390/pathogens14070681 - 10 Jul 2025

Abstract

The growth of aquaculture production in recent years has revealed multiple challenges, including the rise of antimicrobial resistance (AMR) in aquatic animal production, which is currently attracting significant attention from multiple one-health stakeholders. While antibiotics have played a major role in the treatment [...] Read more.

The growth of aquaculture production in recent years has revealed multiple challenges, including the rise of antimicrobial resistance (AMR) in aquatic animal production, which is currently attracting significant attention from multiple one-health stakeholders. While antibiotics have played a major role in the treatment of bacterial infections for almost a century, a major consequence of their use is the increase in AMR, including the emergence of AMR in aquaculture. The AMR phenomenon creates a situation where antibiotic use in one system (e.g., aquaculture) may impact another system (e.g., terrestrial–human). Non-prudent use of antibiotics in aquaculture and animal farming increases the risk of AMR emergence, since bacteria harboring antibiotic resistance genes can cross between compartments such as wastewater or other effluents to aquatic environments, including intensive aquaculture. Transferable antimicrobial resistance gene (AMG) elements (plasmids, transposons, integrons, etc.) have already been detected in varying degrees from pathogenic bacteria that are often causing infections in farmed fish (Aeromonas, Vibrio, Streptococcus, Pseudomonas, Edwardsiella, etc.). This review of current veterinary approaches for the prevention and control of AMR emergence in aquaculture focuses on the feasibility of alternatives to antimicrobials and supplemental treatment applications during on-farm bacterial disease control and prevention. The use of vaccines, bacteriophages, biosurfactants, probiotics, bacteriocins, and antimicrobial peptides is discussed. Full article

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13 pages, 902 KiB

Open AccessArticle

Mitochondrial Genomes of the Blood Flukes Cardicola forsteri and Cardicola orientalis (Trematoda: Aporocotylidae)

by Jemma Hudson, Sunita B. Sumanam, Bronwyn E. Campbell, Lachlan Coff, Barbara F. Nowak, Paul A. Ramsland, Neil D. Young and Nathan J. Bott

Pathogens 2025, 14(7), 680; https://doi.org/10.3390/pathogens14070680 - 10 Jul 2025

Abstract

Parasitic diseases can be a significant constraint on aquaculture industries, which continue to develop in response to the rise in global demand for sustainable protein sources. Blood flukes, Cardicola forsteri and Cardicola orientalis, are economically significant parasites of Southern bluefin tuna (Australia), [...] Read more.

Parasitic diseases can be a significant constraint on aquaculture industries, which continue to develop in response to the rise in global demand for sustainable protein sources. Blood flukes, Cardicola forsteri and Cardicola orientalis, are economically significant parasites of Southern bluefin tuna (Australia), Pacific bluefin tuna (Japan), and Atlantic bluefin tuna (Mediterranean) as they are responsible for blood vessel obstruction in the gills leading to branchitis and mortalities when untreated. Here, we have defined the mitochondrial genomes for these species—the first for any aporocotylids. Oxford nanopore long-read sequencing was used to sequence C. orientalis from a single individual. The mitochondrial genome of C. forsteri was assembled and curated from available sequence data. Both Cardicola spp. mitogenomes contained 12 protein coding, 2 ribosomal and 22 tRNA genes, with the gene order matching that of Asian schistosomes. A control region was identified for each species which contained long and short repeats; the region for C. forsteri was longest, and the overall pattern differed between the two species. A surprisingly high nucleotide diversity was observed between the two species, generating interest into the mitochondrial genes of related species. This paper provides a useful resource for future genetics-based research of aporocotylids and other flatworm parasites of socioeconomic significance. Full article

(This article belongs to the Special Issue Genomic Insights into Parasites of Fishes)

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19 pages, 1713 KiB

Open AccessArticle

Potential for Duplexed, In-Tandem gRNA-Mediated Suppression of Two Essential Genes of Tomato Leaf Curl New Delhi Virus in Crop Plants

by Saher Naveed, Judith K. Brown, Muhammad Mubin, Nazir Javed and Muhammad Shah Nawaz-ul-Rehman

Pathogens 2025, 14(7), 679; https://doi.org/10.3390/pathogens14070679 - 10 Jul 2025

Abstract

Tomato leaf curl New Delhi virus (ToLCNDV) is among the most prevalent and widely distributed begomovirus infecting chili pepper (Capsicum annuum) and tomato in the Indian subcontinent. In this study, a guide RNA (gRNA) sequence-CRISPR-Cas9 approach was used to target and [...] Read more.

Tomato leaf curl New Delhi virus (ToLCNDV) is among the most prevalent and widely distributed begomovirus infecting chili pepper (Capsicum annuum) and tomato in the Indian subcontinent. In this study, a guide RNA (gRNA) sequence-CRISPR-Cas9 approach was used to target and cleave two essential coding regions in the begomovirus genome. The gRNAs were designed to target conserved regions of the ToLCNDV replication-associated protein (rep) gene or ORF AC1, and/or the coat protein (cp) gene or AV1 ORF, respectively. Based on an alignment of 346 representative ToLCNDV genome sequences, all predicted single nucleotide polymorphisms off-target sites were identified and eliminated as potential gRNA targets. Based on the remaining genome regions, four candidate gRNAs were designed and used to build gRNA-Cas9 duplexed constructs, e.g., containing two gRNAs cloned in tandem, in different combinations (1–4). Two contained two gRNAs that targeted the coat protein gene (cp; AV1 ORF), while the other two constructs targeted both the cp and replication-associated protein gene (rep; AC1 ORF). These constructs were evaluated for the potential to suppress ToLCNDV infection in Nicotiana benthamiana plants in a transient expression-transfection assay. Among the plants inoculated with the duplexed gRNA construct designed to cleave ToLCNDV-AV1 or AC1-specific nucleotides, the construct designed to target both the cp (293–993 nt) and rep (1561–2324) showed the greatest reduction in virus accumulation, based on real-time quantitative PCR amplification, and attenuated disease symptoms, compared to plants inoculated with the DNA-A component alone or mock-inoculated, e.g., with buffer. The results demonstrate the potential for gRNA-mediated suppression of ToLCNDV infection in plants by targeting at least two viral coding regions, underscoring the great potential of CRISPR-Cas-mediated abatement of begomovirus infection in numerous crop species. Full article

(This article belongs to the Section Viral Pathogens)

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16 pages, 1969 KiB

Open AccessArticle

Thirteen-Year Sequelae of Marburg Virus Disease Survival: Persistent Cardiometabolic, Immunometabolic, and Haematological Alterations in the Absence of Psychological Morbidity

by Jennifer Serwanga, Raymond Ernest Kaweesa, Joseph Katende Ssebwana, Goeffrey Odoch, Raymond Reuel Wayesu, Anne Daphine Ntabadde, Deborah Mukisa, Peter Ejou, FiloStudy Team, Julius Julian Lutwama and Pontiano Kaleebu

Pathogens 2025, 14(7), 678; https://doi.org/10.3390/pathogens14070678 - 9 Jul 2025

Abstract

Background: Marburg virus disease (MVD) is a highly lethal filoviral infection, yet its long-term health consequences remain poorly understood. We present one of the most temporally distant evaluations of MVD survivors, conducted 13 years post-outbreak in Uganda, offering novel insights into chronic [...] Read more.

Background: Marburg virus disease (MVD) is a highly lethal filoviral infection, yet its long-term health consequences remain poorly understood. We present one of the most temporally distant evaluations of MVD survivors, conducted 13 years post-outbreak in Uganda, offering novel insights into chronic physiological, biochemical, haematological, and psychosocial outcomes. Methods: A cross-sectional, community-based study compared ten MVD survivors with nineteen age- and sex-matched unexposed controls. Clinical evaluations included vital signs, anthropometry, mental health screening, and symptom reporting. Laboratory analyses covered electrolytes, inflammatory markers, renal and liver function tests, haematology, and urinalysis. Standardised psychological assessments measured anxiety, depression, perceived stigma, and social support. Findings: Survivors exhibited an elevated body mass index (BMI), higher systolic and diastolic blood pressure, and lower respiratory rates compared to controls, indicating ongoing cardiometabolic and autonomic changes. These trends may reflect persistent cardiometabolic stress and potential alterations in autonomic regulation, warranting further investigation. Biochemically, survivors exhibited disruptions in serum chloride, bilirubin, and total protein levels, suggesting subclinical hepatic and renal stress. Haematological analysis revealed persistent reticulocytosis despite normal haemoglobin levels, indicating long-term erythropoietic modulation. Despite these physiological changes, survivors reported minimal psychological morbidity, sharply contrasting with the post-recovery profiles of other viral haemorrhagic fevers. Stigma was prevalent during the outbreak; however, strong family support alleviated long-term psychosocial distress. Interpretation: Thirteen years post-infection, MVD survivors demonstrate multisystem physiological perturbations without marked psychological sequelae. These findings challenge assumptions of universal post-viral trauma and highlight the necessity for tailored survivor care models. Future longitudinal studies should investigate the mechanistic pathways underlying cardiometabolic and haematological reprogramming to inform intervention strategies in resource-limited settings. Full article

(This article belongs to the Special Issue Marburg Virus)

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11 pages, 1596 KiB

Open AccessArticle

SNX11 Deletion Inhibits Dabie bandavirus Infection by Interfering with the Assembly of V-ATPase

by Tiezhu Liu, Xueqi Wang, Yang Fang, Ping Zhang, Qiang Sun, Jiandong Li and Shiwen Wang

Pathogens 2025, 14(7), 677; https://doi.org/10.3390/pathogens14070677 - 9 Jul 2025

Abstract

SNX11, a sorting nexin protein localized on the endosomal membrane, is an important protein closely related to protein sorting and endosomal trafficking. Previously, through a genome-wide CRISPR screening, we identified SNX11 as a critical protein for the entry of Dabie bandavirus. SNX11 deletion [...] Read more.

SNX11, a sorting nexin protein localized on the endosomal membrane, is an important protein closely related to protein sorting and endosomal trafficking. Previously, through a genome-wide CRISPR screening, we identified SNX11 as a critical protein for the entry of Dabie bandavirus. SNX11 deletion significantly inhibits the replication of Dabie bandavirus. We further discovered that the loss of SNX11 alters endosomal pH, potentially affecting the release process of Dabie bandavirus from endosomes to the cytoplasm. However, the mechanism by which SNX11 modulates endosomal pH and whether SNX11 deletion similarly inhibits other viruses remain to be elucidated. This study reveals that SNX11 can interact with the V1 subunit of the endosomal proton pump V-ATPase, affecting the expression level of this subunit on the endosomal membrane and thereby disrupting the assembly of V-ATPase. Additionally, we found that SNX11 deletion significantly inhibits the replication of dengue virus, hantavirus, and influenza virus. These findings suggest that SNX11 may be a key protein in the process of viral infection and could serve as a broad-spectrum antiviral target. Full article

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10 pages, 671 KiB

Open AccessArticle

Temporal Parasitemia Trends Predict Risk and Timing of Experimental Cerebral Malaria in Mice Infected by Plasmodium berghei ANKA

by Peyton J. Murin, Cláudio Tadeu Daniel-Ribeiro, Leonardo José Moura Carvalho and Yuri Chaves Martins

Pathogens 2025, 14(7), 676; https://doi.org/10.3390/pathogens14070676 - 9 Jul 2025

Abstract

Background: Experimental models using Plasmodium berghei ANKA (PbA)-infected mice have been essential for uncovering cerebral malaria (CM) pathogenesis. However, variability in experimental CM (ECM) incidence, onset, and mortality introduce challenges when analyses rely solely on infection day, which may reflect different disease stages [...] Read more.

Background: Experimental models using Plasmodium berghei ANKA (PbA)-infected mice have been essential for uncovering cerebral malaria (CM) pathogenesis. However, variability in experimental CM (ECM) incidence, onset, and mortality introduce challenges when analyses rely solely on infection day, which may reflect different disease stages among animals. Methods: We applied machine learning to predict ECM risk and onset in a cohort of 153 C57BL/6, 164 CBA, and 53 Swiss Webster mice. First, we fitted a logistic regression model to estimate the risk of ECM at any day using parasitemia data from day 1 to day 4. Next, we developed and trained a Random Forest Regressor model to predict the exact day of symptom onset. Results: A total of 64.5% of the cohort developed ECM, with onset ranging between 5 and 11 days. Early increases in parasitemia were strong predictors for the development of ECM, with an increase in parasitemia equal to or greater than 0.05 between day 1 and day 3 predicting the development of ECM with 97% sensitivity. The Random Forest model predicted the day of ECM onset with high precision (mean absolute error: 0.43, R²: 0.64). Conclusion: Parasitemia dynamics can effectively identify mice at high risk of ECM, enabling more accurate modeling of early pathological processes and improving the consistency of experimental analyses. Full article

(This article belongs to the Special Issue Parasitic Diseases in the Contemporary World)

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14 pages, 1293 KiB

Open AccessArticle

Comprehensive Survey of PCV2 and PCV3 in Domestic Pigs and Wild Boars Across Portugal: Prevalence, Geographical Distribution and Genetic Diversity

by Bernardo Almeida, Margarida D. Duarte, Ana Duarte, Teresa Fagulha, Fernanda Ramos, Tiago Luís, Inês Caetano, Sílvia C. Barros, Fábio Abade dos Santos and Ana Margarida Henriques

Pathogens 2025, 14(7), 675; https://doi.org/10.3390/pathogens14070675 - 9 Jul 2025

Abstract

Porcine circoviruses are significant pathogens that affect swine populations worldwide, with implications for animal health and productivity. While PCV2 is well-documented, particularly due to widespread vaccination programs, PCV3 is less understood, and its epidemiological impact is still under investigation. This study screened for [...] Read more.

Porcine circoviruses are significant pathogens that affect swine populations worldwide, with implications for animal health and productivity. While PCV2 is well-documented, particularly due to widespread vaccination programs, PCV3 is less understood, and its epidemiological impact is still under investigation. This study screened for PCV2 and PCV3 in pigs and wild boars across Portugal to assess their prevalence. Also, nucleotide sequence determination was performed to evaluate the genetic diversity of these viruses. Stool samples from 160 pigs belonging to different groups (quarantine, nursery, fattening and adult pigs), as well as organ samples from 120 hunted wild boars, were analyzed. Samples were collected from twelve of the eighteen mainland Portuguese districts with positive cases being detected in nine of them. Pigs had a lower prevalence of PCV2 (1.9%) than PCV3 (11.2%), but the opposite was true in wild boars (76.7% for PCV2 and 55.0% for PCV3). The lower PCV2 prevalence in pigs can be attributed to the PCV2 vaccination program implemented. Additionally, these viruses were significantly more prevalent in wild boars (90.8% were infected with at least one of the viruses) than in domestic pigs (only 12.5%). This significant difference highlights the impact of the controlled environment in pig farms on disease prevention in contrast to the higher exposure risks faced by wild boars in their natural habitat. Compared to a previous study from 2023, we observed a slight decrease in the percentage of positive cases for both PCV2 and PCV3. Phylogenetic analysis of sequences obtained by Sanger sequencing allowed us to conclude that the samples from domestic pigs belong to the PCV2a and PCV3c clades, in contrast to the PCV2-positive cases detected in domestic pigs in 2023 that were classified in the PCV2d genotype. Conversely, samples from wild boars belong to the PCV2d and PCV3a clades. These results reveal genotype differences between wild and domestic pigs and shifts from 2023 to 2024. Our findings provide some information about the circulation of these viruses and emphasize the importance of vaccination and continued monitoring for a deeper understanding of their epidemiology to mitigate potential risks to swine health and production. Full article

(This article belongs to the Special Issue Current Challenges in Veterinary Virology)

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12 pages, 524 KiB

Open AccessArticle

Dientamoeba fragilis in Ulcerative Colitis: Analysis of Clinical Findings and Biochemical Parameters

by Ismail Taskiran, Erdogan Malatyali, Ibrahim Yildiz, Levent Durmus Guler, Evren Tileklioglu, Hatice Ertabaklar and Sema Ertug

Pathogens 2025, 14(7), 674; https://doi.org/10.3390/pathogens14070674 - 9 Jul 2025

Abstract

Although Dientamoeba fragilis is a common protozoan in humans, its pathogenicity and clinical significance in human diseases remain poorly understood. This study aimed to determine the frequency of D. fragilis in adult ulcerative colitis patients and to assess its relationship with clinical findings, [...] Read more.

Although Dientamoeba fragilis is a common protozoan in humans, its pathogenicity and clinical significance in human diseases remain poorly understood. This study aimed to determine the frequency of D. fragilis in adult ulcerative colitis patients and to assess its relationship with clinical findings, disease characteristics, and biochemical parameters. Patient data were analysed in a prospective, single-centre, cross-sectional design. Faecal samples were consecutively collected from June to December 2024 and screened for D. fragilis positivity using polymerase chain reaction. Of the 110 patients, 33 (30%) were in the active stage of the disease, while 77 (70%) were in remission. The overall frequency of D. fragilis was 10.9% (n = 12), with all isolates classified as genotype 1 according to SSU rRNA sequence analysis. Other protozoa identified were Blastocystis sp. (n = 5, 4.5%), Entamoeba coli (n = 1, 0.9%), and Iodamoeba bütschlii (n = 1, 0.9%). Two patients were co-infected with D. fragilis and Blastocystis. No significant associations were found between D. fragilis positivity and the disease stage, gastrointestinal symptoms, treatment response, or biochemical findings. In conclusion, despite the relatively small sample size, these findings highlight a limited clinical role of D. fragilis in adult ulcerative colitis patients. Full article

(This article belongs to the Section Parasitic Pathogens)

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20 pages, 1212 KiB

Open AccessSystematic Review

Undetectable = Untransmittable: A Cross-Population Systematic Review and Meta-Analysis on Awareness and Acceptance

by Nikolaos Georgiadis, Andreas Katsimpris, Perry N. Halkitis, Evridiki Kaba, Georgina Tzanakaki, Tonia Vassilakou, Apostolos Beloukas and Theodoros N. Sergentanis

Pathogens 2025, 14(7), 673; https://doi.org/10.3390/pathogens14070673 - 8 Jul 2025

Abstract

The Undetectable = Untransmittable (U=U) message is a cornerstone of HIV-related public health communication, yet global levels of awareness and acceptance remain unclear across key populations. This study aimed to assess the global prevalence of awareness and acceptance of the U=U message among [...] Read more.

The Undetectable = Untransmittable (U=U) message is a cornerstone of HIV-related public health communication, yet global levels of awareness and acceptance remain unclear across key populations. This study aimed to assess the global prevalence of awareness and acceptance of the U=U message among men who have sex with men (MSM), people living with HIV (PLWH), healthcare professionals, and the general population. A systematic review and meta-analysis was conducted using data from PubMed, Embase, and Google Scholar without language restrictions through 31 October 2023. Eligible studies included prospective cohort studies, randomized clinical trials, and cross-sectional studies reporting numerical data on U=U awareness and acceptance. From 1171 screened records, 43 studies were included. Data were analyzed using a random effects model. The findings showed that U=U awareness was high among PLWH, moderate among MSM and healthcare professionals, and low in the general population. Complete acceptance of U=U was low in MSM and the general population, and moderate in PLWH and healthcare professionals. Any acceptance was moderate among MSM and the general population, and high among PLWH and healthcare professionals. These results highlight the need for targeted education strategies to enhance understanding and reduce HIV-related stigma, particularly in populations with lower awareness and acceptance. Full article

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8 pages, 834 KiB

Open AccessBrief Report

Evaluating the Antiviral Activity of Termin-8 and Finio Against a Surrogate ASFV-like Algal Virus

by Amanda Palowski, Francisco Domingues, Othmar Lopez, Nicole Holcombe, Gerald Shurson and Declan C. Schroeder

Pathogens 2025, 14(7), 672; https://doi.org/10.3390/pathogens14070672 - 8 Jul 2025

Abstract

The objective of this study was to evaluate the time-course of incubation for the potential preventative mitigation of megaviruses using Termin-8 (a formaldehyde-based product) and Finio (non-formaldehyde solution) from Anitox. Emiliania huxleyi virus (EhV), an algal surrogate for African swine fever virus (ASFV), [...] Read more.

The objective of this study was to evaluate the time-course of incubation for the potential preventative mitigation of megaviruses using Termin-8 (a formaldehyde-based product) and Finio (non-formaldehyde solution) from Anitox. Emiliania huxleyi virus (EhV), an algal surrogate for African swine fever virus (ASFV), was treated with the recommended concentrations of Termin-8 (0.1% to 0.3%) and Finio (0.05% to 0.2%), and both viability qPCR (V-qPCR) and standard PCR (S-qPCR) were used to quantify EhV concentrations at 1 h, 5 h, 24 h and day 7 post-inoculation. Overall, Finio, and to a lesser extent Termin-8, at their highest treatment concentrations, showed the greatest log reduction of 4.5 and 2 log₁₀ units, respectively, at 1 h post-inoculation. Although Termin-8 efficacy did not improve with time, due to its fixing of viral particles and rendering them non-infectious, treatment with Finio showed 100% viable viral inactivation (>5 log₁₀ reduction units) at the lowest concentration after 7 days of exposure. Our results demonstrate that both Termin-8 and Finio can be used as effective chemical mitigants against megaviruses such as EhV and ASFV and can be used as effective preventive or mitigation strategies to prevent the transmission of ASFV by reducing particle viability in contaminated feed, although additional research is warranted. Full article

(This article belongs to the Special Issue Emergence and Control of African Swine Fever: Second Edition)

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11 pages, 2204 KiB

Open AccessArticle

Investigation of Leishmania infantum Infection and Feeding Preferences of Lutzomyia longipalpis During Deltamethrin (4%) Dog Collar Intervention

by Gabriel F. F. Rodrigues, Keuryn A. M. Luz-Requena, Bruno S. Mathias, Tania M. T. Suto, Rosemari Suto, Luciana T. R. Rocha, Osias Rangel, Katia D. S. Bresciani, Susy M. P. Sampaio, Lilian A. C. Rodas and Karin Kirchgatter

Pathogens 2025, 14(7), 671; https://doi.org/10.3390/pathogens14070671 - 8 Jul 2025

Abstract

Leishmaniasis is a zoonotic disease caused by protozoa of the genus Leishmania, transmitted by phlebotomine sand flies. Understanding the feeding behavior and infection rates of these vectors is crucial for disease surveillance and control. We aimed to investigate the natural infection rate [...] Read more.

Leishmaniasis is a zoonotic disease caused by protozoa of the genus Leishmania, transmitted by phlebotomine sand flies. Understanding the feeding behavior and infection rates of these vectors is crucial for disease surveillance and control. We aimed to investigate the natural infection rate of Leishmania spp. in phlebotomines and analyze their blood-feeding patterns in one of the priority areas of the state of São Paulo for the implementation of insecticide-impregnated dog collars. Sand flies were collected from urban and peri-urban areas between 2022 and 2024 using CDC light traps, manual aspiration, and Shannon traps. PCR was used to detect Leishmania DNA (SSU rDNA gene), and blood meal sources (COI gene). A total of 414 sand flies were collected, with 222 engorged females analyzed for blood meals and 192 specimens tested for Leishmania spp. infection. The predominant blood source was humans (67%), followed by chickens (64.1%), and dogs (18.9%), considering that 45.1% of the samples presented mixed blood meals. Leishmania infantum was found in 1% of the samples. These findings highlight the feeding plasticity of sand flies and their potential role in disease transmission, reinforcing the need for continuous epidemiological surveillance and vector control strategies, particularly the implementation of insecticide-impregnated dog collars. Full article

(This article belongs to the Special Issue Leishmaniasis: Current Status and Future Perspectives)

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10 pages, 219 KiB

Open AccessArticle

The Quantitative Detection of Urogenital Mycoplasmas in Men with Urolithiasis

by Dominika Smolec, Małgorzata Aptekorz, Łukasz Filipczyk, Zygmunt Gofron, Jacek Zostawa, Robert Smolec, Tomasz J. Wąsik and Alicja Ekiel

Pathogens 2025, 14(7), 670; https://doi.org/10.3390/pathogens14070670 - 8 Jul 2025

Abstract

Urease-positive urogenital mycoplasmas are considered to be responsible for the formation of urinary stones. They are usually a part of the normal flora in the human urogenital tract, causing asymptomatic infections. However, many symptomatic infections with these bacteria have been reported. M. genitalium [...] Read more.

Urease-positive urogenital mycoplasmas are considered to be responsible for the formation of urinary stones. They are usually a part of the normal flora in the human urogenital tract, causing asymptomatic infections. However, many symptomatic infections with these bacteria have been reported. M. genitalium is recognized as a cause of male urethritis and other common genitourinary diseases. The role of other urogenital mycoplasmas is still unclear. The aim of this study was to estimate the quantitative prevalence of Ureaplasma spp., M. genitalium and M. hominis in men with urolithiasis using quantitative real-time PCR (qPCR). The study group comprised 100 men with urolithiasis. A total of 60 men were included in the control group. Urogenital mycoplasma DNA in urine samples was detected significantly more often among men with urolithiasis than in healthy subjects—43.0% vs. 26.6%, p = 0.0382, respectively. The majority of positive results (38/43) concerned U. parvum species, the frequency of which was higher in the study group (38.0% (38/100)) than in the control group (23.3% (14/60)), p = 0.0552. The median concentration of U. urealyticum DNA was higher in the study group compared with the control, p = 0.5714. However, further studies are needed to confirm the usefulness of quantitative studies in determining the role of urogenital mycoplasmas in pathology. Full article

(This article belongs to the Section Bacterial Pathogens)

1 pages, 148 KiB

Open AccessCorrection

Correction: Embo-Ibouanga et al. Peptide-Alkoxyamine Drugs: An Innovative Approach to Fight Schistosomiasis: “Digging Their Graves with Their Forks”. Pathogens 2024, 13, 482

by Ange W. Embo-Ibouanga, Michel Nguyen, Jean-Patrick Joly, Mathilde Coustets, Jean-Michel Augereau, Lucie Paloque, Nicolas Vanthuyne, Raphaël Bikanga, Anne Robert, Françoise Benoit-Vical, Gérard Audran, Philippe Mellet, Jérôme Boissier and Sylvain R. A. Marque

Pathogens 2025, 14(7), 669; https://doi.org/10.3390/pathogens14070669 - 8 Jul 2025

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Abstract

There was an error in the original publication [...] Full article

15 pages, 1885 KiB

Open AccessReview

Molecular Features of HPV-Independent Cervical Cancers

by Luca Giannella, Camilla Grelloni, Leonardo Natalini, Gianmarco Sartini, Mila Bordini, Giovanni Delli Carpini, Jacopo Di Giuseppe, Erica Dugo, Francesco Piva and Andrea Ciavattini

Pathogens 2025, 14(7), 668; https://doi.org/10.3390/pathogens14070668 - 8 Jul 2025

Viewed by 47

Abstract

HPV-independent cervical cancers represent a small proportion of these types of cancers, predominantly glandular lesions. It should be noted that some cases may depend on diagnostic problems that lead to false negative cases. However, the most recent classifications distinguish cervical tumors into HPV-associated [...] Read more.

HPV-independent cervical cancers represent a small proportion of these types of cancers, predominantly glandular lesions. It should be noted that some cases may depend on diagnostic problems that lead to false negative cases. However, the most recent classifications distinguish cervical tumors into HPV-associated and HPV-independent cancers. HPV-negative cervical carcinomas (5–11% of all cases) mainly include rare adenocarcinomas (gastric, mesonephric, clear, serous, and endometrioid) and present distinct clinical and molecular features. These tumors usually affect older women and are diagnosed at more advanced stages than HPV-positive tumors, with an overall worse prognosis. This concerning and notably worse prognosis highlights the need for further research and understanding. Unlike HPV-positive carcinomas (which depend on the viral oncogenes E6/E7), HPV-independent tumors accumulate genomic mutations that activate oncogenes and inactivate suppressor genes. Therefore, a comprehensive overview of these aspects can be the key to a better understanding and developing personalized treatments. In the present review, the main mutated genes, the signaling pathways involved, the differences from HPV-positive tumors, the distinctive immunohistochemical markers, and the diagnostic and therapeutic implications are explored in depth. Full article

(This article belongs to the Special Issue Oncogenic Viruses: Advances in Molecular Diagnosis, Prevention Strategies and Therapy)

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10 pages, 1229 KiB

Open AccessArticle

Copper-Induced Stimulation of Ectophosphatase Activity of Candida albicans

by Anita Leocadio Freitas-Mesquita, Fabiano Ferreira Esteves and José Roberto Meyer-Fernandes

Pathogens 2025, 14(7), 667; https://doi.org/10.3390/pathogens14070667 - 8 Jul 2025

Viewed by 26

Abstract

Candida albicans is an opportunistic fungal pathogen that can cause superficial and life-threatening infections, particularly in immunocompromised individuals. Its ability to adhere to host cells is critical for colonization and infection. In this context, investigating ectophosphatases is particularly relevant, as these enzymes have [...] Read more.

Candida albicans is an opportunistic fungal pathogen that can cause superficial and life-threatening infections, particularly in immunocompromised individuals. Its ability to adhere to host cells is critical for colonization and infection. In this context, investigating ectophosphatases is particularly relevant, as these enzymes have been associated with fungal adhesion to host cells. This study aimed to investigate the nature of copper-induced stimulation of ectophosphatase activity in C. albicans. Ectophosphatase activity was measured using p-nitrophenyl phosphate as substrate. Micromolar concentrations of CuCl₂ markedly stimulated ectophosphatase activity, and its response to reducing agents and metal chelators suggested that this modulation does not involve redox reactions. The significant differences between the biochemical properties of basal (Cu²⁺-independent) and Cu²⁺-dependent ectophosphatase activities suggest the presence of at least two distinct ectophosphatases in C. albicans. Cu²⁺-independent ectophosphatase activity presented an acidic profile and was insensitive to Mg²⁺, whereas Cu²⁺-dependent ectophosphatase activity exhibited an alkaline profile and was also stimulated by Mg²⁺. Both activities were negatively modulated by classical phosphatase inhibitors, but Cu²⁺-dependent ectophosphatase had lower sensitivity compared to the basal activity. These findings highlight the role of copper as a modulator of C. albicans ectophosphatase activity and suggest potential implications for fungal adaptation during infection. Full article

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9 pages, 789 KiB

Open AccessArticle

Pharmacokinetics of Molnupiravir in Cats with Naturally Occurring Feline Infectious Peritonitis

by Petra Černá, Luke Wittenburg, Jennifer Hawley, McKenna Willis, Britta Siegenthaler and Michael R. Lappin

Pathogens 2025, 14(7), 666; https://doi.org/10.3390/pathogens14070666 - 7 Jul 2025

Viewed by 140

Abstract

Antiviral drugs like EIDD-2801 (molnupiravir; MPV) have been successfully used in the treatment of feline infectious peritonitis (FIP). The previous study of the pharmacokinetics of MPV in healthy cats showed promise for its use and safety. The objective was to determine the pharmacokinetics [...] Read more.

Antiviral drugs like EIDD-2801 (molnupiravir; MPV) have been successfully used in the treatment of feline infectious peritonitis (FIP). The previous study of the pharmacokinetics of MPV in healthy cats showed promise for its use and safety. The objective was to determine the pharmacokinetics of molnupiravir in cats with naturally occurring FIP by measuring MPV and EIDD-193 (β-D-N4-hydroxycytidine; NHC) serum levels. Blood was collected from seven cats diagnosed with naturally occurring FIP treated at 1, 2, 4, 6 and 12 h post oral MPV administration and at 12 h post pill administration 7 days later. Serum concentrations of MPV and NHC were determined using a previously published high-performance liquid chromatography–tandem mass spectrometry (HPLC-MS/MS) method. The mean dose of MPV was 15.44 mg/kg (SD ± 1.82). The mean peak serum concentration of MPV (C_max) after a single PO dose of MPV was 38 ng/mL (SD ± 5). The mean peak serum concentration of NHC (C_max) after a single PO dose of MVP was 1551 ng/mL (SD ± 720). the time to reach NHC C_max (T_max) was 2.6 h (SD ± 1.4), and the NHC elimination half-life was 1.6 h (SD ± 1.1). Minimal drug accumulation was seen in trough concentrations following twice-daily dosing for 7 days. The low MPV levels may be explained by fast conversion to its active metabolite NHC. The mean NHC concentrations at all time points were at least 4 times the reported in vitro IC₅₀ for feline coronavirus strains and twice-daily dosing for seven days did not lead to drug accumulation within the serum. The results support the use of MPV in the treatment of FIP, and if therapeutic drug monitoring is to be performed, NHC should be measured. Full article

(This article belongs to the Special Issue Feline Coronavirus Infections)

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9 pages, 589 KiB

Open AccessArticle

Occurrence and Molecular Characterization of Cryptosporidium spp. in Swine Farms in Northeastern Spain

by Laura Garza-Moreno, Celia León and Joaquín Quílez

Pathogens 2025, 14(7), 665; https://doi.org/10.3390/pathogens14070665 - 5 Jul 2025

Viewed by 119

Abstract

Cryptosporidium spp. are protozoan parasites that cause cryptosporidiosis, an enteric disease that can affect a wide range of vertebrate hosts. Pigs play a potential role in the transmission of Cryptosporidium spp. to humans, although infections are most often subclinical. This study aimed to [...] Read more.

Cryptosporidium spp. are protozoan parasites that cause cryptosporidiosis, an enteric disease that can affect a wide range of vertebrate hosts. Pigs play a potential role in the transmission of Cryptosporidium spp. to humans, although infections are most often subclinical. This study aimed to assess the occurrence and molecular characterization of Cryptosporidium spp. in swine farms located in Aragón, northeastern Spain. Fecal samples (n = 72) were collected from 10 breeding farms, encompassing various production stages (lactation, nursery, fattening, and/or wean-to-finish). Data regarding the type of production system (two- or three-stage), production stages, and farming facilities (the type of flooring and water source) associated with the parasite presence were also analyzed using a questionnaire. The results showed that Cryptosporidium spp. were more frequently detected in fecal samples originating from three-stage production systems (21.9%) compared to two-stage systems (12.5%). Samples from the fattening stage exhibited the highest positivity rate and estimated oocyst count (3.0 oocyst/microscopic field). Furthermore, the molecular characterization of Cryptosporidium spp. revealed the circulation of multiple species both among farms and within the same pig production flow, with Cryptosporidium scrofarum being the most prevalent species (7/72; 9.7%), followed by Cryptosporidium suis (1/72; 1.4%). These findings underscore the importance of the surveillance and molecular characterization of Cryptosporidium spp. for controlling infections in pigs, considering the potential for the zoonotic transmission of this parasite to humans. Full article

(This article belongs to the Special Issue Parasites and Zoonotic Diseases)

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16 pages, 1564 KiB

Open AccessArticle

Antiprotozoal Effects of Pediococcus acidilactici-Derived Postbiotic on Blastocystis Subtypes ST1/ST3

by Selahattin Aydemir, Yunus Emre Arvas, Mehmet Emin Aydemir, Fethi Barlık, Esra Gürbüz, Yener Yazgan and Abdurrahman Ekici

Pathogens 2025, 14(7), 664; https://doi.org/10.3390/pathogens14070664 - 5 Jul 2025

Viewed by 226

Abstract

Blastocystis, a common intestinal protozoan in humans, is associated with gastrointestinal disorders, irritable bowel syndrome, urticaria, and colorectal cancer. Its genetic diversity and potential for treatment resistance make it a focus of ongoing research. This study evaluated the in vitro antiprotozoal activity [...] Read more.

Blastocystis, a common intestinal protozoan in humans, is associated with gastrointestinal disorders, irritable bowel syndrome, urticaria, and colorectal cancer. Its genetic diversity and potential for treatment resistance make it a focus of ongoing research. This study evaluated the in vitro antiprotozoal activity of a postbiotic derived from Pediococcus acidilactici as a natural alternative treatment. P. acidilactici cultures were grown in MRS broth under anaerobic conditions, and the postbiotic was collected and characterized for pH, yield, organic acid composition, and phenolic compound content. Human isolates of Blastocystis subtypes ST1 and ST3 were cultured in Jones’ medium and exposed to varying postbiotic concentrations for 72 h. Viability was assessed microscopically. The cytotoxic effect of the postbiotic-derived P. acidilactici was evaluated by investigating its impact on the viability of HT-29 cells using the Cell Counting Kit 8. The postbiotic showed a 7% yield and a pH of 4.52 ± 0.11. It contained seven different organic acids, predominantly lactic acid, and eleven phenolic compounds, with naringin as the most abundant. At 4.38 mg/mL, the postbiotic achieved over 94% inhibition and 100% inhibition at 8.75 mg/mL and above. A pH analysis confirmed that the inhibition was independent of the culture medium acidity. Cell viability was not affected at the postbiotic concentration showing 100% antiprotozoal activity (8.75 mg/mL). These findings suggest that the P. acidilactici postbiotic is effective on a mixed culture of ST1 and ST3 subtypes and holds promise as a safe, natural antiprotozoal agent. Further in vivo studies are needed to confirm this. Full article

(This article belongs to the Special Issue Exploring Natural Products as Antiparasitic Agents: Efficacy Against Parasites of Veterinary and Public Health Significance)

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10 pages, 1764 KiB

Open AccessBrief Report

Cathelicidins Limit Intracellular Neospora caninum-Infection in Macrophages

by Franco Fiorani, Priyoshi Lahiri, Rodrigo Puentes, Peter John Bradley, Dadin Prando Moore and Eduardo Ruben Cobo

Pathogens 2025, 14(7), 663; https://doi.org/10.3390/pathogens14070663 - 5 Jul 2025

Viewed by 240

Abstract

Infections with the protozoan Neospora caninum cause abortion in cattle, likely due to the parasite’s replication and excessive inflammation in the placenta. Cathelicidins are host defense peptides known for their antimicrobial and immunomodulatory functions, but their role in N. caninum infections remains elusive. [...] Read more.

Infections with the protozoan Neospora caninum cause abortion in cattle, likely due to the parasite’s replication and excessive inflammation in the placenta. Cathelicidins are host defense peptides known for their antimicrobial and immunomodulatory functions, but their role in N. caninum infections remains elusive. Using bone marrow-derived macrophages (BMDMs) isolated from mice expressing (wild-type, Camp⁺^/+) and lacking (Camp⁻^/−) cathelicidins, we investigated the role of endogenous cathelicidin in infections with N. caninum. We show that Camp⁻^/− macrophages primed with lipopolysaccharide (LPS) had an increased number of intracellular N. caninum tachyzoites, and these macrophages released higher amounts of IL-1β and lactate dehydrogenase (LDH), a marker of cytotoxicity. These findings indicate that cathelicidins contribute to intracellular N. caninum control and inflammation by limiting the activation of the inflammasome, particularly under LPS-induced conditions. This insight reveals the immunomodulatory role of cathelicidins in controlling N. caninum-associated pathologies. Full article

(This article belongs to the Special Issue Genetics and Molecular Evolution of Parasitic Protozoa)

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13 pages, 3732 KiB

Open AccessArticle

Baicalein and Berberine Inhibit the Growth and Virulence of Clostridioides difficile

by Xue Yang, Dongming Zheng, Jiangyan Yong, Yuchen Li, Yunzhi Sun, Fei Zhao, Daiyan Tang, Yi Xie and Dongming Bi

Pathogens 2025, 14(7), 662; https://doi.org/10.3390/pathogens14070662 - 4 Jul 2025

Viewed by 170

Abstract

Clostridioides difficile is a leading pathogen involved in healthcare-associated diarrhea. With its increasing incidence, mortality, and antibiotic resistance, there is an urgent need for novel therapeutic strategies to address the infection and prevent its recurrence. Gegen Qinlian Decoction (GQD) is a traditional Chinese [...] Read more.

Clostridioides difficile is a leading pathogen involved in healthcare-associated diarrhea. With its increasing incidence, mortality, and antibiotic resistance, there is an urgent need for novel therapeutic strategies to address the infection and prevent its recurrence. Gegen Qinlian Decoction (GQD) is a traditional Chinese medicine for the treatment of diarrhea, but its main active ingredient is not known. Therefore, in this study, we evaluated the biological activity of berberine (BER) and baicalein (BAI), key components of GQD, against C. difficile. Time–kill curves and scanning electron microscopy were employed to assess their effects on C. difficile growth, while Enzyme-Linked Immunosorbnent Assay (ELISA) and cytotoxicity assays were used to examine their impact on toxin production. We also employed Quantitative Reverse Transcription PCR (qRT-PCR) to examine how BER and BAI influenced the expression of toxin-associated genes. At sub-inhibitory concentrations, these compounds exerted antibacterial activity against C. difficile by disrupting the integrity of the cell membrane and cell wall. Furthermore, BER and BAI also suppressed toxin production, demonstrating effects comparable to those of vancomycin. This suppression likely resulted from their bactericidal activity and the inhibition of toxin gene expression. This study not only highlights the potential application of GQD in treating C. difficile infections but also offers promising options for developing drugs targeting the growth and virulence of this pathogen. C. difficile infection (CDI) is a leading cause of severe diarrhea, and its treatment remains challenging due to limited drug options and its high recurrence rate. BAI and BER, the main active components of the traditional Chinese medicinal formula GQD, inhibited the growth of C. difficile by disrupting its cellular structure and significantly reduced the production of toxins associated with disease severity. Furthermore, the effects of BAI and BER on C. difficile were comparable to those of conventional antibiotics, suggesting that these compounds could be potential alternative therapies for CDI. This study not only highlights the therapeutic potential of GQD in treating CDI but also provides a replicable research strategy for the development of novel anti-CDI agents. Full article

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16 pages, 1910 KiB

Open AccessArticle

Targeting the Risk of Diptera-Borne Zoonoses by a Sentinel Equestrian Centers Program

by Cristiana Cazapal-Monteiro, David Boso, Inês Abreu, Mercedes Camiña, Jaime Sanchís, Adolfo Paz-Silva, Luis Cardoso, Rita Sánchez-Andrade, María Sol Arias and José Ángel Hernández

Pathogens 2025, 14(7), 661; https://doi.org/10.3390/pathogens14070661 - 4 Jul 2025

Viewed by 112

Abstract

Diptera-borne diseases pose a major threat to global health, and their distribution is constantly changing due to climate change, globalization, and environmental changes. To improve the knowledge of dipteran species and their distribution in equine facilities, CDC-UV and oviposition traps were placed, and [...] Read more.

Diptera-borne diseases pose a major threat to global health, and their distribution is constantly changing due to climate change, globalization, and environmental changes. To improve the knowledge of dipteran species and their distribution in equine facilities, CDC-UV and oviposition traps were placed, and the dipping technique was performed in 16 equestrian centers of Northwest (NW) Spain (Galicia and Castilla y León Autonomous Communities) between July and November 2023. A questionnaire was distributed among the horse owners to obtain additional information. Four genera of culicids, Culex (51.8%), Culiseta (38.6%), Anopheles (8.4%), and Aedes/Ochlerotatus (1.2%) were identified in the equestrian centers. Culex pipiens s.l. was the most prevalent and well-distributed species (93.8% of the centers), whereas Anopheles maculipennis s.l. and An. claviger/petragnani, the anopheline species, were the most frequent (37.5% and 31.2%, respectively). The Culiseta genus was found in approximately 81.2% of the equine facilities. All genera were collected at medium and high altitudes and in Csb (warm-summer Mediterranean climate) areas. Equestrian centers from NW Spain albeit a variety of culicids with high vectorial capacity, together with an ideal environment for their breeding, the presence of vectors and hosts (humans and animals). This potential problem for global health enhances the need for entomological surveillance. Full article

(This article belongs to the Special Issue Pets, Wildlife and Parasites—2nd Edition)

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11 pages, 220 KiB

Open AccessArticle

Immune-Mediated Hemolytic Anemia in Cats with Feline Infectious Peritonitis

by Petra Černá, Marieke Knies, Marleen Assink, Samantha Evans, Séverine Tasker, Danièlle A. Gunn-Moore, Katrin Hartmann, Katharina Buchta, Samantha Taylor, Solène Meunier, Regina Hofmann-Lehmann, Nicole Jacque, Allison Koonce, Casandra Jacobs, Ashley Gillett and Michael R. Lappin

Pathogens 2025, 14(7), 660; https://doi.org/10.3390/pathogens14070660 - 4 Jul 2025

Viewed by 195

Abstract

Feline infectious peritonitis (FIP) is caused by mutated feline coronaviruses. Immune-mediated hemolytic anemia (IMHA) arises due to immune-mediated erythrocyte destruction and can be non-associative or associative with diseases such as FIP. Records of cats with FIP were reviewed to find those with associative [...] Read more.

Feline infectious peritonitis (FIP) is caused by mutated feline coronaviruses. Immune-mediated hemolytic anemia (IMHA) arises due to immune-mediated erythrocyte destruction and can be non-associative or associative with diseases such as FIP. Records of cats with FIP were reviewed to find those with associative IMHA based on exclusion of other causes of anemia and a positive saline agglutination test and/or Coombs test. The inclusion criteria were met for 45 cats (26 (58%) cats with effusive and 19 (42%) with non-effusive FIP). Median hematocrit was 18% (interquartile range [IQR] 13–20). Anemia was non-regenerative in 36 (80%) cats and regenerative in 5 (11%) cats; 4 (9%) cats had no reticulocyte count available. Concurrent thrombocytopenia was present in 18 (40%) cats. All 45 cats were treated with nucleoside analogs, and 44 (98%) cats with glucocorticoids; in 5 (11%) cats, glucocorticoids were added after starting antiviral treatment due to persistent anemia. Median follow-up was 72 days (IQR 14–246); at the time of last follow-up 33 (73%) cats had survived while 12 (27%) had died or were euthanized. Of the 33 surviving cats, 17 achieved remission of both FIP and IMHA. In three cats, FIP remission was achieved, but IMHA relapsed; in one of these, IMHA relapsed twice. FIP relapsed without IMHA in two cats, and both FIP and IMHA relapsed in one cat. In 9 cats the antiviral and glucocorticoid treatment is still ongoing at the time of the publication. Although FIP is likely an uncommon cause of associative IMHA, as more cats with FIP are treated with antiviral therapy, it is important to consider IMHA as a possible cause of anemia in cats with FIP. Full article

(This article belongs to the Special Issue Feline Coronavirus Infections)

10 pages, 1814 KiB

Open AccessArticle

First Molecular Identification of Cotylophoron cotylophorum in Ecuador and Its Phylogenetic Relationship with Fasciola hepatica

by Geanella Barragán-López, Fausto Bedoya-Páez, María Lugo-Almarza, Carolina Fonseca-Restrepo, Francisco Angulo-Cubillán, Edison J. Romero, Jacobus H. de Waard and Armando Reyna-Bello

Pathogens 2025, 14(7), 659; https://doi.org/10.3390/pathogens14070659 - 4 Jul 2025

Viewed by 268

Abstract

Trematode infections caused by Fasciolidae and Paramphistomidae remain widespread in livestock, resulting in substantial economic losses. The two distinct fluke families are difficult to distinguish morphologically, and molecular identification provides the most reliable means of accurate diagnosis. In Ecuador, however, molecular data on [...] Read more.

Trematode infections caused by Fasciolidae and Paramphistomidae remain widespread in livestock, resulting in substantial economic losses. The two distinct fluke families are difficult to distinguish morphologically, and molecular identification provides the most reliable means of accurate diagnosis. In Ecuador, however, molecular data on these parasites are scarce. In this study, we collected trematodes from cattle rumen and bile ducts, molecularly identified them, and assessed their phylogenetic relationship to Fasciola hepatica to determine their introduction pathways into South America. Genomic DNA was extracted, and PCR was used to amplify the ITS2 (~500 bp) and COXI (~266 bp) regions; all amplicons were Sanger-sequenced. Phylogenetic trees for both markers were constructed using a Maximum Likelihood approach with 1000 bootstrap replicates in CIPRES v3.3. The rumen fluke exhibited 99% ITS2 and COXI similarity to an Indian Cotylophoron cotylophorum strain, while the bile-duct fluke showed 99% ITS2 and 100% COXI similarity to F. hepatica isolates from Australia and Nigeria, respectively. Distinct single-nucleotide polymorphisms (SNPs) in the ITS2 chromatograms suggest a diploid genome structure in both trematode species. This is the first report of C. cotylophorum in Ecuador, and its presence may be linked to the late 19th-century introduction of Zebu cattle (Bos taurus indicus) from India. Full article

(This article belongs to the Special Issue Parasites and Parasitic Diseases in the Livestock Industry in Latin America)

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15 pages, 1072 KiB

Open AccessArticle

Wastewater Surveillance for Group A Streptococcus pyogenes in a Small City

by Olivia N. Birch, Frankie M. Garza and Justin C. Greaves

Pathogens 2025, 14(7), 658; https://doi.org/10.3390/pathogens14070658 - 3 Jul 2025

Viewed by 249

Abstract

Streptococcus pyogenes is a bacterial pathogen known to be the causative agent in many different illnesses, with Group A Streptococcus (GAS) pharyngitis (strep throat), being one of the more prevalent. The spread and severity of GAS pharyngitis can grow exponentially if individuals are [...] Read more.

Streptococcus pyogenes is a bacterial pathogen known to be the causative agent in many different illnesses, with Group A Streptococcus (GAS) pharyngitis (strep throat), being one of the more prevalent. The spread and severity of GAS pharyngitis can grow exponentially if individuals are not taking the proper precautions. Wastewater surveillance has been used to test for numerous different pathogens that humans spread throughout a community and in this study, we utilized wastewater surveillance to monitor GAS pharyngitis in a small city. Over a year, 57 wastewater influent samples were tested for S. pyogenes and three commonly tested respiratory viruses (Respiratory Syncytial Virus (RSV), SARS-CoV-2, Influenza A). Three microbial indicators and population normalizers (CrAssphage, Pepper mild mottle virus (PMMoV), and Mycobacterium) were tested as well to compare and contrast each indicator’s value and range over time. Wastewater data was then compared to publicly available search term data as clinical data was not readily available. There was a high correlation between the collected molecular data and the publicly available search term data for Streptococcus pyogenes. Additionally, this study provided more information about the seasonal trend of S. pyogenes throughout the year through molecular data and allowed for the ability to track peak infection months in this small city. Overall, these results highlight the substantial benefits of using wastewater surveillance for the monitoring of GAS pharyngitis. This study also provides helpful insights into future studies about the prevalence of respiratory bacteria and their seasonal trends in wastewater, allowing for public health systems to provide mitigation strategies. Full article

(This article belongs to the Special Issue Wastewater Surveillance and Public Health Strategies)

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16 pages, 904 KiB

Open AccessReview

Point-of-Care Diagnostic Testing for Emerging and Existing Poultry Viral Respiratory Pathogens Using Loop-Mediated Isothermal Amplification

by Ben Enyetornye, Aurelle Yondo and Binu T. Velayudhan

Pathogens 2025, 14(7), 657; https://doi.org/10.3390/pathogens14070657 - 2 Jul 2025

Viewed by 156

Abstract

Accurate, rapid and inexpensive diagnosis of poultry respiratory pathogens remains a challenge, especially in many developing countries. Meanwhile, poultry respiratory pathogens are a major threat to poultry production worldwide, accounting for billions of dollars in economic loss to the sector. Early and accurate [...] Read more.

Accurate, rapid and inexpensive diagnosis of poultry respiratory pathogens remains a challenge, especially in many developing countries. Meanwhile, poultry respiratory pathogens are a major threat to poultry production worldwide, accounting for billions of dollars in economic loss to the sector. Early and accurate diagnosis of these diseases is critical for economic poultry production. Molecular diagnostic methods, including PCR-based techniques, have been developed and used to fill this gap, but unfortunately, these techniques require skilled technicians, relatively costly equipment and reagents and can only be performed in a laboratory setting. This warrants the development of other diagnostic tools, which can be used in the field even by unskilled personnel. In this review, we discussed the genesis, challenges, advances and prospects of loop-mediated isothermal amplification (LAMP) for the detection of poultry respiratory pathogens at the flock side, especially in resource-constrained countries. We highlighted the application of LAMP in routine poultry disease surveillance and early outbreak detection, underscoring its value as a transformative diagnostic tool in poultry production. The development and use of a point-of-care (POC) LAMP assay that can be used to screen for these poultry respiratory pathogens simultaneously enhance disease surveillance and diagnosis. Full article

(This article belongs to the Special Issue Diagnostics of Emerging and Re-Emerging Pathogens)

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11 pages, 1403 KiB

Open AccessArticle

Clinical Features of Pulmonary Nocardiosis and Diagnostic Value of Metagenomic Next-Generation Sequencing: A Retrospective Study

by Yanbin Chen, Hailong Fu, Qiongfang Zhu, Yalu Ren, Jia Liu, Yining Wu and Jie Xu

Pathogens 2025, 14(7), 656; https://doi.org/10.3390/pathogens14070656 - 2 Jul 2025

Viewed by 259

Abstract

Pulmonary nocardiosis (PN) is a rare, opportunistic, and potentially life-threatening infection, especially in disseminated cases. This retrospective study aimed to characterize the clinical features of PN and assess the diagnostic utility of metagenomic next-generation sequencing (mNGS). We reviewed data from 19 patients diagnosed [...] Read more.

Pulmonary nocardiosis (PN) is a rare, opportunistic, and potentially life-threatening infection, especially in disseminated cases. This retrospective study aimed to characterize the clinical features of PN and assess the diagnostic utility of metagenomic next-generation sequencing (mNGS). We reviewed data from 19 patients diagnosed with PN between September 2019 and August 2022, including 3 with disseminated disease. Common symptoms included fever, cough, and sputum production, while chest imaging frequently revealed nodules, consolidations, exudates, cavities, and pleural effusions. The sensitivity of mNGS for detecting Nocardia was significantly higher than that of culture (100% vs. 36.84%, p < 0.001). mNGS successfully identified Nocardia species and co-infected pathogens. The most common species was Nocardia farcinica. Four PN cases were co-infected with Rhizomucor pusillus, Cryptococcus neoformans, Lichtheimia ramosa, and Aspergillus spp. Eighteen patients (94.7%) received trimethoprim-sulfamethoxazole (TMP-SMZ). Sixteen cases (84.2%) were improved or cured. Misdiagnosis is common due to the nonspecificity of clinical and imaging presentations of pulmonary nocardiosis. The timely combination of mNGS represents a promising approach to enhance the diagnosis of pulmonary nocardiosis and inform targeted antimicrobial therapy. TMP-SMZ is the first line of treatment. Full article

(This article belongs to the Section Bacterial Pathogens)

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Pathogens, Volume 14, Issue 7 (July 2025) – 78 articles

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