Pathogens

16 pages, 3894 KB

Open AccessArticle

Trends in Antibiotic Resistance of Escherichia coli Strains Isolated from Clinical Samples (2019–2023): A Hospital-Based Retrospective Analysis

by Claudia Daniela Goleanu (Vasiloiu), Corneliu Ovidiu Vrancianu, Daria Adelina Goleanu, Monica Marilena Tantu and Ortansa Csutak

Pathogens 2025, 14(9), 927; https://doi.org/10.3390/pathogens14090927 (registering DOI) - 13 Sep 2025

Abstract

Background: Antimicrobial resistance (AMR) is a major public health concern. Urinary tract infections (UTIs) account for up to 85–90% of community-acquired cases. The COVID-19 pandemic disrupted healthcare access and may have influenced resistance patterns. In this context, we retrospectively evaluated the antibiotic resistance [...] Read more.

Background: Antimicrobial resistance (AMR) is a major public health concern. Urinary tract infections (UTIs) account for up to 85–90% of community-acquired cases. The COVID-19 pandemic disrupted healthcare access and may have influenced resistance patterns. In this context, we retrospectively evaluated the antibiotic resistance dynamics of various bacterial strains isolated between 2019 and 2023 in a hospital unit; Methods: A total of 8217 clinical specimens (urine, wound secretions, sputum, pharyngeal exudate, nasal exudate, tracheal secretions, vaginal and cervical secretions, puncture fluids, purulent secretions, blood, ear secretions, eye secretions) were processed using standard microbiological techniques. Pathogen identification and susceptibility testing were performed with the VITEK 2 Compact system, following CLSI guidelines. Results: Following the analysis of 8217 clinical samples collected over a five-year period (2019–2023), a total of 2900 microorganisms were isolated and identified. Among these, the most frequently encountered were E. coli strains, with 1204 isolates. Urine cultures represented 71.3% of all processed samples. Out of these 5860 urine cultures, 1530 (26%) were positive. The resistance of E. coli strains to ampicillin (48–55.2%), trimethoprim/sulfamethoxazole (22.9–34%), and ciprofloxacin (21.4–31.5%) remained high throughout the period. ESBL-producing strains peaked at 17.6% in 2020, with multidrug resistance rates ranging from 14% to 22.4%. Conclusions: E. coli strains displayed persistently high resistance to ampicillin, trimethoprim/sulfamethoxazole, and ciprofloxacin, with peaks in ESBL production and multidrug resistance during the COVID-19 pandemic. These trends underscore the importance of continuous surveillance and antibiotic stewardship, with direct implications for empirical UTI therapy and broader strategies to mitigate the public health impact of antimicrobial resistance. Full article

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16 pages, 2348 KB

Open AccessArticle

Repellent Activity of the Botanical Compounds Thymol, Carvacrol, Nootkatone, and Eugenol Against Amblyomma sculptum Nymphs

by Mayara Macêdo Barrozo, Emilly Faria Santos, Haile Dean Figueiredo Chagas, Rafael Assunção Carvalho, Isabela Santos Silva, Ariel de Souza Oliveira, Laura Cristina Ferreira Faria, Ana Lúcia Coutinho Teixeira, Viviane Zeringota, Hermes Ribeiro Luz, Lorena Lopes Ferreira and Caio Monteiro

Pathogens 2025, 14(9), 926; https://doi.org/10.3390/pathogens14090926 (registering DOI) - 13 Sep 2025

Abstract

This study evaluated the repellent activity of the botanical compounds thymol, carvacrol, nootkatone, and eugenol (5%), as well as the combination of 2.5% nootkatone + 2.5% eugenol, against Amblyomma sculptum nymphs under laboratory and field conditions. In contact bioassays, carvacrol and nootkatone showed [...] Read more.

This study evaluated the repellent activity of the botanical compounds thymol, carvacrol, nootkatone, and eugenol (5%), as well as the combination of 2.5% nootkatone + 2.5% eugenol, against Amblyomma sculptum nymphs under laboratory and field conditions. In contact bioassays, carvacrol and nootkatone showed the highest mean repellency rates (94.5% and 93.7%), followed by thymol and eugenol (90.2% and 87.2%). The combination (nootkatone + eugenol) resulted in 92.8% repellency, with 100% efficacy in some evaluation periods. The repellency of 7% DEET (positive control) was 82.2%. Nootkatone 5% and the combination (nootkatone + eugenol) were also tested in a Y-tube olfactometer, showing repellency rates of 86.1% and 72.2%, respectively, both higher than 7% DEET (69.4%). In field trials, volunteers wore treated socks and walked through an area naturally infested with nymphs. A significant reduction (p < 0.05) in tick counts was observed in the treated groups (about five unfed nymphs) compared to the control (about 40 unfed nymphs). Cumulative efficacy exceeded 85% in both treatments. In the cost simulation, the estimated cost of a 100 mL formulation containing 5% nootkatone was USD 50.8, while the combination (nootkatone + eugenol) presented a cost USD 28.6, representing a 44% reduction with no loss of efficacy in field conditions. These results indicate that all compounds tested showed repellent activity in the laboratory. Nootkatone has high repellent activity, and its combination with eugenol is a promising and more economically viable alternative for tick repellency. Full article

(This article belongs to the Special Issue Vector Control and Parasitic Infection in Animals)

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9 pages, 681 KB

Open AccessArticle

In Vivo Efficacy of a Broad-Spectrum Antiviral Combination Against Yellow Fever in a Hamster Model

by Abbie E. Weight, Hunter Stanger, Robert J. Geraghty, Laurent F. Bonnac and Justin G. Julander

Pathogens 2025, 14(9), 925; https://doi.org/10.3390/pathogens14090925 (registering DOI) - 12 Sep 2025

Abstract

Yellow fever virus (YFV) recurrently causes severe outbreaks in tropical regions of South America and Africa and an average of 30 to 40 thousand deaths worldwide each year. An effective vaccine is available but the coverage of the population in countries at risk [...] Read more.

Yellow fever virus (YFV) recurrently causes severe outbreaks in tropical regions of South America and Africa and an average of 30 to 40 thousand deaths worldwide each year. An effective vaccine is available but the coverage of the population in countries at risk is not optimal. No antivirals are currently approved for YFV treatment. Herein, we describe the evaluation of 6-MMPr, a de-novo-purine-nucleotide biosynthesis inhibitor, as a potentiator for enhanced activity of the broad-spectrum antiviral drug favipiravir in a hamster model of yellow fever. Administration of 6-MMPr was well-tolerated and a combination of favipiravir and 6-MMPr did not cause overt toxicity as indicated by normal weight gain of treated hamsters. Treatment with a combination of a suboptimal dose of favipiravir with 6-MMPr was significantly more effective in improving survival, weight change and virus replication when compared with monotherapy. The initiation of treatment two days after virus challenge was also effective in improving survival when compared with monotherapy. Our results demonstrate the safety and efficacy of such a combination either as a preventive or delayed treatment. Full article

(This article belongs to the Section Viral Pathogens)

18 pages, 1049 KB

Open AccessArticle

Immune Sensitization to Mycobacterium tuberculosis Among Young Children with and Without Tuberculosis

by Jesús Gutierrez, LaShaunda L. Malone, Mitchka Mohammadi, John Mukisa, Michael Atuhairwe, Simon Peter G. Mwesigwa, Salome Athieno, Sharon Buwule, Faith Ameda, Sophie Nalukwago, Ezekiel Mupere, Catherine M. Stein and Christina L. Lancioni

Pathogens 2025, 14(9), 924; https://doi.org/10.3390/pathogens14090924 (registering DOI) - 12 Sep 2025

Abstract

Identification of young children with Mycobacterium tuberculosis (Mtb) infection is critical to curb pediatric morbidity and mortality. The optimal test to identify young children with Mtb infection remains controversial. Using a tuberculosis (TB) household contact (HHC) study design among 130 Ugandan children less [...] Read more.

Identification of young children with Mycobacterium tuberculosis (Mtb) infection is critical to curb pediatric morbidity and mortality. The optimal test to identify young children with Mtb infection remains controversial. Using a tuberculosis (TB) household contact (HHC) study design among 130 Ugandan children less than 5 years of age with Mtb exposure, this study was conducted to determine the following: (1) the prevalence of Mtb immune sensitization in young children heavily exposed to TB using both the tuberculin skin test (TST) and QuantiFERON Gold Plus (QFT-Plus) interferon gamma release assay, and to examine the concordance of these two tests; and (2) the diagnostic accuracy of TST and QFT-plus for confirmed and unconfirmed TB in young children. Prevalence of Mtb immune sensitization was determined using TST at both 5 mm and 10 mm thresholds for positivity; manufacturer’s thresholds were utilized to establish QFT-Plus positivity. Concordance analysis between TST and QFT-Plus results was performed, including correlation between QFT-Plus tube TB.1 and tube TB.2. The sensitivity and specificity of TST and QFT-Plus for confirmed and unconfirmed TB was determined, and a logistic regression model was utilized to estimate the odds of TB. A 5 mm TST threshold identified the most children with Mtb sensitization (49.2%) and had moderate agreement with QFT-Plus (Cohen’s Kappa 0.59). The odds of TB were two times higher among children with a positive TST using a 5 mm threshold. Concordance between 10 mm TST threshold and QFT-Plus was substantial (Cohen’s Kappa 0.65), with higher concordance observed among older children (2–5 years). The QFT-Plus tube TB.1 and tube TB.2 results were highly correlated. Positive TST using a 5 mm threshold demonstrated the highest sensitivity for TB (60%), whereas QFT-Plus testing demonstrated the highest specificity (72%). Overall, our findings support that among a population of young, BCG-vaccinated children with heavy household exposure to TB, the TST using a 5 mm threshold identified more children with evidence of Mtb immune sensitization, and children with TB disease, than the QFT-Plus. These findings are highly relevant for children who are TB HHCs in endemic settings, and most at risk for TB following an exposure. We recommend that TST testing continue to be performed to assess for Mtb sensitization in young, TB-exposed children in TB-endemic settings to both prioritize provision of preventive therapy and to aide in diagnosis of pediatric TB. Full article

(This article belongs to the Section Bacterial Pathogens)

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20 pages, 384 KB

Open AccessArticle

Phenotypic and Genotypic Characterization of Staphylococcus aureus Isolated from Patients with Chronic Furunculosis and Osteomyelitis from Northwestern Poland

by Aleksandra Wcisłek, Joanna Jursa-Kulesza, Helena Masiuk, Bartłomiej Grygorcewicz, Beata Hukowska-Szematowicz, Piotr Prowans, Paweł Ziętek and Danuta Kosik-Bogacka

Pathogens 2025, 14(9), 923; https://doi.org/10.3390/pathogens14090923 - 12 Sep 2025

Abstract

Staphylococcus aureus, a Gram-positive coccus commonly found in the environment, is indeed a common cause of both superficial and deep infections. The aim of the study was to determine the virulence factors of S. aureus characteristic of chronic infections, including chronic furunculosis [...] Read more.

Staphylococcus aureus, a Gram-positive coccus commonly found in the environment, is indeed a common cause of both superficial and deep infections. The aim of the study was to determine the virulence factors of S. aureus characteristic of chronic infections, including chronic furunculosis and chronic osteomyelitis. Phenotypic characteristics of the bacteria (ability to produce hemolysis, clumping factor, and coagulase; antibiotic susceptibility) and genotypic characteristics (presence of genes responsible for the production of enzymes and toxins; ability to form biofilm) were examined. The real-time PCR method was used to determine the presence of virulence genes. Biofilm production was confirmed using the crystal violet method. Antibiotic and chemotherapeutic susceptibility tests were performed using the disk diffusion method. In 90% of cases, S. aureus strains possessed the following virulence genes: clfA, clfB, spa, cna, eap, hlgA, hlgB, hlg, hld, bap, bbp, ebpS, fib, fnbA, fnbB, and pvl. A total of 82% of S. aureus strains showed susceptibility to methicillin (MSSA), whereas 12% of strains were susceptible to methicillin and simultaneously resistant to macrolides, lincosamides, and streptogramin B, including 10.5% with inducible resistance and 1.5% with constitutive resistance (MSSA/MLSB). In addition, 1.5% were methicillin-resistant S. aureus (MRSA) and susceptible to the remaining antimicrobial agents. The predominance of MSSA in the etiology of chronic furunculosis and chronic osteomyelitis was observed. It has been demonstrated that MSSA possesses a similar set of virulence genes to MRSA and that MSSA is responsible for most cases of chronic osteomyelitis and furunculosis. The findings indicate that S. aureus possesses numerous virulence factors that play a key role in the processes of adhesion to and proliferation within host cells. Full article

20 pages, 4315 KB

Open AccessArticle

Genomic Analysis of Lumpy Skin Disease Virus from Western and Central Africa Suggests a Distinct Sub-Lineage Within the 1.2 LSDV Cluster

by John Fadele, Olusola Ogunsanya, Oluwatobi Adedokun, Akeemat Ayinla, Mbitkebeyo Pami, Ayotunde Sijuwola, Femi Saibu, Harouna Soumare, Urbain Fanou, Corrie Brown, Bonto Faburay, Christian Happi and Anise Happi

Pathogens 2025, 14(9), 922; https://doi.org/10.3390/pathogens14090922 - 12 Sep 2025

Abstract

Lumpy Skin Disease Virus (LSDV) is a transboundary pathogen that affects cattle, causing significant economic losses, particularly in Africa and Asia. While the virus was originally endemic to sub-Saharan Africa, it has rapidly spread to Europe, the Middle East, and Asia, necessitating comprehensive [...] Read more.

Lumpy Skin Disease Virus (LSDV) is a transboundary pathogen that affects cattle, causing significant economic losses, particularly in Africa and Asia. While the virus was originally endemic to sub-Saharan Africa, it has rapidly spread to Europe, the Middle East, and Asia, necessitating comprehensive genomic surveillance. Despite LSDV’s African origins, genomic data from West and Central Africa remain scarce, limiting insights into regional viral evolution and vaccine compatibility. In this study, molecular detection of LSDV was carried out on cattle samples from Nigeria, Cameroon, and Benin. However, comparative genomic analysis was performed using two near-complete LSDV genomes obtained from Cameroon. Phylogenetic evaluation revealed that LSDV strains from Nigeria and Cameroon cluster within the classical 1.2 lineage. Furthermore, the two sequences from this study cluster with the only publicly available sequence from West and Central Africa, supporting earlier findings of the presence of a West/Central African sub-lineage. Functional genomic analysis identified mutations in genes encoding ankyrin repeat Kelch-like proteins, and envelope proteins involved in immune evasion and viral virulence, raising concerns about vaccine effectiveness. Furthermore, the detection of LSDV in flesh flies (Sarcophaga spp.) underlines their potential role in virus transmission. These findings highlight the importance of genomic monitoring and targeted surveillance. Full article

(This article belongs to the Special Issue Emergence and Re-Emergence of Animal Viral Diseases)

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8 pages, 668 KB

Open AccessArticle

Seroprevalence of Antibodies to Burkholderia pseudomallei in Mississippi Gulf Coast Residents, September 2023

by Katherine M. DeBord, Mindy G. Elrod, Claire Hartloge, William A. Meyer III, Brooke E. Swanson, Caroline A. Schrodt, Maria E. Negron and Zachary P. Weiner

Pathogens 2025, 14(9), 921; https://doi.org/10.3390/pathogens14090921 - 12 Sep 2025

Abstract

In 2022, Burkholderia pseudomallei was first identified in continental United States (U.S.) environmental samples from the Mississippi Gulf Coast following two autochthonous infections. To better understand the extent of exposure to this emerging bacterium, we tested a convenience sample of 825 residual sera [...] Read more.

In 2022, Burkholderia pseudomallei was first identified in continental United States (U.S.) environmental samples from the Mississippi Gulf Coast following two autochthonous infections. To better understand the extent of exposure to this emerging bacterium, we tested a convenience sample of 825 residual sera samples (550 from the Mississippi Gulf Coast, 275 from the northern U.S.) from a commercial diagnostic laboratory for the presence of antibodies to B. pseudomallei, using an indirect hemagglutination assay. We estimated seroprevalence of antibodies to B. pseudomallei in Mississippi Gulf Coast residents and in controls from northern regions of the U.S. where B. pseudomallei is less likely to persist in the environment. At a titer cut-off of ≥1:40, we observed a similar seropositivity between Mississippi Gulf Coast residents (14%, 95% CI: 11%, 17%) and controls (17%, 95% CI: 13%, 18%). Similarities in seropositivity suggest environmental exposure to B. pseudomallei in the Mississippi Gulf Coast may be limited; however, a lack of accompanying illness and exposure information limits our ability to conclusively interpret these findings. These estimates can serve as a baseline of seropositivity in the U.S. for future studies and to track the spread of B. pseudomallei in the U.S. over time. Full article

(This article belongs to the Special Issue Updates on Human Melioidosis)

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16 pages, 1226 KB

Open AccessArticle

Infectious Disease Awareness Among Future Health Professionals: A Comparison of Knowledge, Attitudes, and Practices Between Nursing Students in Japan and Laos

by Hansani Madushika Abeywickrama, Tiengkham Pongvongsa, Marcello Otake Sato, Rie Takeuchi, Yoichi Abiko, Tippayarat Yoonuan, Jun Kobayashi and Megumi Sato

Pathogens 2025, 14(9), 920; https://doi.org/10.3390/pathogens14090920 - 11 Sep 2025

Abstract

This study assessed and compared the knowledge, attitudes, and practices of nursing students in Japan and Laos, two countries with distinct epidemiological and socioeconomic contexts, regarding neglected tropical diseases (NTDs) and zoonoses. A cross-sectional survey was conducted using a self-administered questionnaire among 190 [...] Read more.

This study assessed and compared the knowledge, attitudes, and practices of nursing students in Japan and Laos, two countries with distinct epidemiological and socioeconomic contexts, regarding neglected tropical diseases (NTDs) and zoonoses. A cross-sectional survey was conducted using a self-administered questionnaire among 190 nursing students from Japan and 254 students from Laos. Descriptive statistics, chi-square test, and Fisher’s exact test were used to analyze intergroup differences. Most of the study participants were female and in their first or second year of their nursing program. Knowledge gaps were identified in both groups. Japanese students showed a higher perceived susceptibility to infections and greater adherence to hygiene practices. Laotian students reported higher exposure to endemic diseases and more frequent contact with livestock. Attitudes toward public health education were generally positive, although Japanese students expressed a greater willingness to engage in future educational roles. However, gaps between attitude and actual practice were apparent in both groups, including inconsistent use of soap and insect repellents. The findings reveal a gap in knowledge of NTDs and zoonoses and a deficit in translating knowledge and attitudes into preventive behaviors. Findings highlight the need for tailored educational strategies considering behavioral and cultural factors to strengthen the nurse’s role in infectious disease prevention. Full article

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

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14 pages, 618 KB

Open AccessReview

Rabies Surveillance in Mainland Tanzania: A Scoping Review of Animal Rabies Occurrences (1993–2023)

by Emmanuel Kulwa Bunuma, Julius Keyyu, Joseph Maziku, Stella Bitanyi, Robert Fyumagwa, Katendi Changula, Benjamin Mubemba, Edgar Simulundu, Simbarashe Chitanga, Daniel L. Horton, Abel Bulamu Ekiri, Hirofumi Sawa and Walter Muleya

Pathogens 2025, 14(9), 919; https://doi.org/10.3390/pathogens14090919 - 11 Sep 2025

Abstract

Animal rabies remains underreported in low-income countries, hindering effective control. This scoping review aimed to map reported animal rabies cases, identify key reservoir species, and assess gaps in surveillance coverage in mainland Tanzania from 1993 to 2023. Specifically, it addressed the distribution of [...] Read more.

Animal rabies remains underreported in low-income countries, hindering effective control. This scoping review aimed to map reported animal rabies cases, identify key reservoir species, and assess gaps in surveillance coverage in mainland Tanzania from 1993 to 2023. Specifically, it addressed the distribution of cases, species involved, and the extent of surveillance coverage during this period. Literature searches in PubMed, Google Scholar, and Science Direct were screened using Rayyan. Twenty articles published between 1993 and 2023 reported 7319 animal rabies cases across the Northern Zone (NZ), Southeastern Zone (SEZ), and Coastal Zone (CZ). In the NZ, domestic dogs accounted for most cases (5387), followed by jackals (225), cats (77), livestock (311), and various wildlife species including African wild dogs, bat-eared foxes, lions, cheetahs, and striped hyenas. Additionally, 102 cases involved unidentified animals. In SEZ, domestic dogs (588) were the primary source, followed by jackals (262), hyenas (8), cats (10), honey badgers (5), and leopards (2). In CZ, domestic dogs accounted for 94 cases. The findings confirm domestic dogs as the main rabies reservoir, highlighting the need for strengthened surveillance and control. The role of wildlife in rabies maintenance and spillover remains poorly understood and warrants further investigation, especially in enzootic hotspots. Full article

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17 pages, 1749 KB

Open AccessArticle

β-Giardin as an Immunomagnetic Enrichment Target for Multi-Host Detection of Giardia Duodenalis Cysts

by Hongyu Wang, Heng Yang, Chaofan Li, Mengge Chen, Xiaocen Wang, Xu Zhang, Pengtao Gong, Nan Zhang, Xichen Zhang, Jianhua Li and Xin Li

Pathogens 2025, 14(9), 918; https://doi.org/10.3390/pathogens14090918 - 11 Sep 2025

Abstract

Giardiasis is a globally prevalent waterborne zoonosis. Rapid enrichment and detection technologies for this disease are essential. Cyst outer wall proteins are ideal targets for the enrichment and detection of cysts in the environment, but there are few available targets with suboptimal effectiveness. [...] Read more.

Giardiasis is a globally prevalent waterborne zoonosis. Rapid enrichment and detection technologies for this disease are essential. Cyst outer wall proteins are ideal targets for the enrichment and detection of cysts in the environment, but there are few available targets with suboptimal effectiveness. In this study, Giardia duodenalis (G. duodenalis) cysts were purified, and outer wall proteins were biotinylated, followed by streptavidin magnetic bead purification and mass spectrometry. Sixty-three novel cyst wall proteins were identified, and their functions were annotated through Gene Ontology (GO) and KEGG analyses. The β-giardin and α-1 giardin were among the newly identified and predicted to be located on the outer wall of G. duodenalis cysts. For the characterization of these two targets, we applied sequence analysis, prokaryotic expression, preparation of polyclonal antibodies, and determination of subcellular localization. Finally, based on β-giardin immunomagnetic beads were prepared using the polyclonal antibodies and tested for their enrichment efficiency. Immunomagnetic beads targeting β-giardin achieved 65% cyst enrichment efficiency in fecal samples, comparable to conventional methods. Clinical evaluation across 163 multi-host fecal samples (ferrets, Siberian tigers, red-crowned cranes) demonstrated concordance with nested PCR, successfully enriching cysts from PCR-positive specimens. The immunomagnetic beads method targeting β-giardin demonstrated effective G. duodenalis cyst enrichment in multi-host fecal samples. These results provide a proteomic framework for the cyst wall proteins of G. duodenalis, expanding the detection targets for G. duodenalis cysts. It also establishes a theoretical foundation for subsequent research on the composition and function of G. duodenalis cysts. Full article

(This article belongs to the Special Issue Advances in Animal Parasitic Diseases)

14 pages, 1440 KB

Open AccessArticle

Association Between Pre-Existing Conditions and COVID-19 Hospitalization, Intensive Care Services, and Mortality: A Cross-Sectional Analysis of an International Global Health Data Repository

by Basant M. S. Elsayed, Lina Altarawneh, Habib Hassan Farooqui, Muhammad Naseem Khan, Giridhara Rathnaiah Babu, Suhail A. R. Doi and Tawanda Chivese

Pathogens 2025, 14(9), 917; https://doi.org/10.3390/pathogens14090917 - 11 Sep 2025

Abstract

Background: The use of globally shared individual-level data in answering epidemiological questions during health emergencies of international concern is still debatable. In this study, we investigated the association between pre-existing conditions and clinical outcomes of COVID-19 using data from a global data sharing [...] Read more.

Background: The use of globally shared individual-level data in answering epidemiological questions during health emergencies of international concern is still debatable. In this study, we investigated the association between pre-existing conditions and clinical outcomes of COVID-19 using data from a global data sharing repository. Methods: We used data of all cases recorded in the Global Health Data repository up to the 10th of March 2021 to carry out a cross-sectional analysis of associations between cardiovascular diseases (CVD), hypertension, diabetes, obesity, lung diseases, and kidney disease and hospitalization, ICU admission, and mortality due to COVID-19. The Global Health repository reported data from 137 countries, but only Brazil, Mexico, and Cuba reported more than 10 COVID-19 cases in participants with preexisting conditions. We used multivariable logistic regression to compute adjusted odds ratios (aOR) of the three outcomes for each pre-existing condition in ten-year age groups from 0 to 9 years and up to 110–120 years. Findings: As of March 10, the Global Health repository contained 25,900,000 records of confirmed cases of COVID-19, of which 2,900,000 cases from Brazil, Mexico, and Cuba had recorded data on pre-existing conditions. The overall aOR of ICU admission for each pre-existing condition were; CVD (aOR 2.1, 95%CI 1.8–2.4), hypertension (aOR 1.3, 95%CI 1.2–1.4), diabetes (OR 1.7, 95%CI 1.5–1.8), obesity (OR 2.2, 95%%CI 2.1–2.4), kidney disease (OR 1.4, 95%CI 1.2–1.7) and lung disease (aOR 1.1, 95%CI 0.9–1.3). Overall aORs of mortality for each pre-existing condition were: CVD (aOR 1.7, 95%CI 1.6–1.7), hypertension (aOR 1.3, 95%CI 1.3–1.4), diabetes (aOR 2.0, 95%CI 1.9–2.0), obesity (aOR 1.9, 95%CI 1.8–2.0), kidney disease (aOR 2.7, 95%CI 2.6–2.9), and lung disease (aOR 1.6, 95%CI 1.5–1.7). The odds of each adverse outcome were considerably larger in children and young adults with these preexisting conditions than for adults, especially for kidney disease, CVD, and diabetes. Conclusion: This analysis of a global health repository confirms associations between pre-existing diseases and clinical outcomes of COVID-19, and the odds of these outcomes were especially elevated in children and young adults with these preexisting conditions. This study shows that global data sharing can unlock answers to many epidemiological questions efficiently especially during the early stages of global health emergencies. Full article

(This article belongs to the Special Issue Leveraging Technological Advancement for Pandemic Preparedness)

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17 pages, 3128 KB

Open AccessArticle

Prevalence of Cytomegalovirus Antibodies in Croatian Childbearing-Aged and Pregnant Women: A Ten-Year Retrospective Study (2015–2024)

by Tatjana Vilibić-Čavlek, Klara Barbić, Tadej Ježek, Dan Navolan, Ana Sanković, Mario Sviben, Sara Glavaš, Daniel Mureșan, Laurentiu Pirtea and Maja Bogdanić

Pathogens 2025, 14(9), 916; https://doi.org/10.3390/pathogens14090916 - 11 Sep 2025

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Abstract

Due to possible congenital infections, cytomegalovirus (CMV) remains a significant public health concern in childbearing-aged and pregnant women. We analyzed the spatial, temporal, and age-related trends in CMV seroepidemiology in Croatia over 10 years. A total of 2838 childbearing-aged and pregnant women, aged [...] Read more.

Due to possible congenital infections, cytomegalovirus (CMV) remains a significant public health concern in childbearing-aged and pregnant women. We analyzed the spatial, temporal, and age-related trends in CMV seroepidemiology in Croatia over 10 years. A total of 2838 childbearing-aged and pregnant women, aged 16–45 years, tested between 2015 and 2024 were included in the study. CMV IgM/IgG antibodies were detected using a commercial ELISA. IgM/IgG-positive samples were tested for IgG avidity. CMV IgG antibodies were detected in 2006 (70.6%) of participants. No significant differences were observed between 2015–2019 and 2020–2024 (72.0% vs. 69.8%), while yearly differences were of borderline significance, ranging from 62.4 to 77.3%. The overall seropositivity increased progressively with age from 49.6% in the 16–20 age group to 77.5% in the 36–40 age group. Significant regional differences in IgG seroprevalence were observed: 68.6% in the City of Zagreb/Northern Croatia, 78.5% in Pannonian Croatia, and 71.9% in Adriatic Croatia, while differences between settlement types were not significant. IgG seroprevalence was higher in women with an unfavorable obstetric history (85.3%) than in non-pregnant women and those with a normal pregnancy (70.6% and 66.5%, respectively). IgM antibodies were detected in 278 (9.8%) of participants. Acute infections were more common in younger participants, with rates decreasing from 13.6% in the youngest age group to 6.7% in the oldest. Logistic regression showed that age was a significant predictor of both IgG and IgM positivity. Region and obstetric history were significant predictors of IgG seropositivity, while settlement was a significant predictor of IgM seropositivity. Full article

(This article belongs to the Section Viral Pathogens)

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14 pages, 1144 KB

Open AccessArticle

Sociability Linked to Reproductive Status Affects Intestinal Parasite Infections in the Red-Billed Chough

by Guillermo Blanco, Maria M. Garijo-Toledo, José Luis González del Barrio, Óscar Frías, Raymundo Montoya Ayala and Iñigo Palacios-Martínez

Pathogens 2025, 14(9), 915; https://doi.org/10.3390/pathogens14090915 - 11 Sep 2025

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Abstract

Social behaviour plays a crucial role in the dynamics of parasitic infections in wild bird populations. The red-billed chough (Pyrrhocorax pyrrhocorax), a corvid undergoing notable population declines, shows contrasting social structures linked to reproductive status: non-breeding individuals aggregate in communal roosts [...] Read more.

Social behaviour plays a crucial role in the dynamics of parasitic infections in wild bird populations. The red-billed chough (Pyrrhocorax pyrrhocorax), a corvid undergoing notable population declines, shows contrasting social structures linked to reproductive status: non-breeding individuals aggregate in communal roosts during winter, whereas breeding pairs often maintain territorial pair-bonds and roost at nesting sites. This study tested whether differences in sociality (communal roosting vs. territorial pairs) affect intestinal parasite infections. Fresh faecal samples were collected during winter in central Spain and analysed using flotation and McMaster techniques to detect and quantify coccidian oocysts and helminth eggs. The results revealed a relatively high positive rate of Isospora (36.2%, n = 116) and a low positive rate of helminths (9.5%, n = 116) among communally roosting non-breeders, while no parasites were detected in samples from territorial pairs. One communal roost in the Southern Plateau showed higher coccidian positive rate, possibly influenced by structural features that facilitate faecal contact. Although sample size for pairs was limited, the absence of parasites in this group suggests reduced infection risk, likely reflecting superior condition and immune defences rather than differences in exposure alone. These findings highlight the value of non-invasive parasite monitoring as an early-warning tool in wildlife health assessments and stress the importance of considering social behaviour and environmental heterogeneity into conservation strategies for threatened species. Full article

(This article belongs to the Special Issue Parasitic Helminths and Control Strategies)

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20 pages, 4590 KB

Open AccessArticle

Immunization with mRNA-LNPs Encoding Ornithodoros Argasid Tick Antigens Induces Humoral Immune Responses and Tick Resistance

by Ana Oleaga, Ana Laura Cano-Argüelles, María González-Sánchez, Rocío Vizcaíno-Marín and Ricardo Pérez-Sánchez

Pathogens 2025, 14(9), 914; https://doi.org/10.3390/pathogens14090914 - 11 Sep 2025

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Abstract

Argasid ticks Ornithodoros erraticus and Ornithodoros moubata are major vectors of zoonotic pathogens, including the African swine fever virus and relapsing fever Borrelia spp., and their control is essential to reduce disease transmission. In this study, we evaluated the immunogenicity and protective efficacy [...] Read more.

Argasid ticks Ornithodoros erraticus and Ornithodoros moubata are major vectors of zoonotic pathogens, including the African swine fever virus and relapsing fever Borrelia spp., and their control is essential to reduce disease transmission. In this study, we evaluated the immunogenicity and protective efficacy of four Ornithodoros tick antigens formulated as mRNA–lipid nanoparticles (mRNA-LNPs): OeSOD, OeTSP1, OmPLA2, and Om86. Rabbits were immunised with three doses of each mRNA-LNP construct, and immune responses and tick biological parameters were assessed following infestation with both tick species. All mRNA-LNP constructs induced antigen-specific IgG responses that recognised native proteins in tick saliva and midgut extracts. Vaccination resulted in significant reductions in female oviposition and fertility, which correlated with antibody levels, and yielded protective efficacies of 21.9–41.6% against O. moubata and 23.1–41.6% against O. erraticus. Notably, the mRNA-LNPs of OeSOD and OeTSP1 outperformed their recombinant counterparts against O. moubata, and Om86 mRNA-LNP conferred markedly improved protection against both O. moubata and O. erraticus. These findings highlight the potential of mRNA-LNP vaccines to induce effective anti-argasid tick immunity and provide a promising platform for the development of sustainable strategies to control argasid ticks and associated pathogens. Full article

(This article belongs to the Section Ticks)

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14 pages, 1481 KB

Open AccessArticle

E2 Tyrosine 102 Regulates MmuPV1 Pathogenesis In Vivo

by Jessica Gonzalez, Marsha DeSmet, Kennedy Stoll, Leny Jose, Neil Christensen and Elliot J. Androphy

Pathogens 2025, 14(9), 913; https://doi.org/10.3390/pathogens14090913 - 11 Sep 2025

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Abstract

The papillomavirus (PV) life cycle is strictly controlled and can be divided into the following three distinct stages: initial infection, maintenance, and amplification. The papillomavirus E2 gene encodes a multifunctional protein responsible for regulating transcription and replication by recruiting viral and host factors [...] Read more.

The papillomavirus (PV) life cycle is strictly controlled and can be divided into the following three distinct stages: initial infection, maintenance, and amplification. The papillomavirus E2 gene encodes a multifunctional protein responsible for regulating transcription and replication by recruiting viral and host factors to the viral DNA genome. Our lab has previously reported that tyrosine 102 may impact bovine (BPV) and human (HPV) viral replication in cell culture systems. This tyrosine is conserved in the E2 protein of the murine papillomavirus MmuPV1. To investigate how this amino acid impacts the MmuPV1 lifecycle in vivo, we generated potential phosphorylation mimetic (Y102E) and phosphorylation deficient (Y102F) mutants in the E2 open reading frame. The Y102F mutant protein supported both transcriptional activation and transient replication, while Y102E was defective. However, Y102E was capable of associating with E1 and the Brd4 C-terminal motif. When these E2-mutated MmuPV1 genomes were introduced into the skin of immunocompromised mice, only Y102F was capable of inducing papilloma development and production of infectious progeny virus. These findings demonstrate that while highly conserved, tyrosine at this position is not required by the virus. These data suggest that the chemical nature of the amino acid at this position can influence E2 activity and viral replication. Full article

(This article belongs to the Section Viral Pathogens)

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25 pages, 1146 KB

Open AccessReview

Emerging Challenges in Salmonella Control: The Need for Innovative and Sustainable Disinfection Strategies in Poultry Farming

by Nicla Gentile, Laura Lorenzo-Rebenaque, Ana Marco-Fuertes, Laura Montoro-Dasi and Clara Marin

Pathogens 2025, 14(9), 912; https://doi.org/10.3390/pathogens14090912 - 10 Sep 2025

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Abstract

Salmonella is one of the primary causes of foodborne infections worldwide and is often linked to the consumption of poultry products. Despite the implementation of numerous control programmes, the persistence of Salmonella in poultry environments remains a challenge, exacerbated by the emergence of [...] Read more.

Salmonella is one of the primary causes of foodborne infections worldwide and is often linked to the consumption of poultry products. Despite the implementation of numerous control programmes, the persistence of Salmonella in poultry environments remains a challenge, exacerbated by the emergence of strains resistant to traditional disinfectants. This review examines the key factors associated with the limitations of disinfection and the new strategies employed in poultry production, underscoring the need for more sustainable and effective alternative solutions. Various chemical (nanoparticles), physical (ultraviolet light, heat, pressurised steam, infrared radiation) and biological (bacteriophages, essential oils, and positive biofilm) treatments are examined. Of the various alternatives assessed, some have shown promising antimicrobial activity against Salmonella in vitro and under experimental conditions. However, their application in real-field settings is still limited, and few studies evaluate their effectiveness on a commercial scale. The review emphasises the importance of integrating these alternatives within broader biosecurity programmes, supported by clear regulations to minimise the risk of transmission. In conclusion, the adoption of innovative and sustainable approaches, combined with strengthened biosecurity measures, represents a key strategy to reduce Salmonella contamination in poultry farms, protect public health and promote responsible production systems. Full article

(This article belongs to the Special Issue Salmonella: A Global Health Threat and Food Safety Challenge)

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20 pages, 677 KB

Open AccessReview

Balancing Host Defense and Viral Tolerance for the Development of Next-Generation Broad-Spectrum Antiviral Agents

by Xiujuan Zhao, Ruikun Du and Lijun Rong

Pathogens 2025, 14(9), 911; https://doi.org/10.3390/pathogens14090911 - 10 Sep 2025

Viewed by 208

Abstract

The human immune system has evolved to cope with various virus infections using two major strategies: host defense and viral tolerance. The host defense relies on the innate and adaptive immune responses to control virus replication, while the viral tolerance allows sufficient virus [...] Read more.

The human immune system has evolved to cope with various virus infections using two major strategies: host defense and viral tolerance. The host defense relies on the innate and adaptive immune responses to control virus replication, while the viral tolerance allows sufficient virus replication in the host with minimal or no clinical signs. Generally, a balanced host defense and viral tolerance can guarantee health from infections, while disturbed immune homeostasis usually results in diseases. It is desirable to develop drugs/therapeutics to enhance the invulnerability of host immune equilibrium to combat viral infections. In this review, we summarize the advanced understanding on mechanisms of both the host defense against and tolerance to virus infections, and therapeutic agents/approaches that work by modulating the balance between host defense and tolerance. Full article

(This article belongs to the Section Viral Pathogens)

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12 pages, 4204 KB

Open AccessArticle

Establishment of a Pseudovirus-Based Golden Hamster Model for the Attachment and Entry Stages of Hendra Virus Infection and Evaluation of Protective Immunity

by Tao Li, Binfan Liao, Danfeng Li, Jie Zhang, Chunhui Zhao, Yunfei Pei, Liping Chen, Meng Wang, Yawen Liu, Xi Wu, Weijin Huang and Jianhui Nie

Pathogens 2025, 14(9), 910; https://doi.org/10.3390/pathogens14090910 - 10 Sep 2025

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Abstract

Objective: Establish an in vivo evaluation model focused on the attachment and entry stages of Hendra virus infection for protective immunity assessment. Methods: A golden hamster infection model based on recombinant Hendra-F/G pseudovirus was developed, and a luciferase luminescence assay was used to [...] Read more.

Objective: Establish an in vivo evaluation model focused on the attachment and entry stages of Hendra virus infection for protective immunity assessment. Methods: A golden hamster infection model based on recombinant Hendra-F/G pseudovirus was developed, and a luciferase luminescence assay was used to assess the optimal pseudoviral challenge in terms of route of infection, dose and detection time. The biodistribution of the pseudovirus in infected organs was evaluated using the IVIS spectral CT system. The protective effect of antibody prophylaxis was evaluated by measuring the luminescence intensity of pseudoviruses. Results: Intraperitoneal injection was identified as the optimal route of infection, and the optimal time of detection was 6 h post-challenge. Our model simulates the infection of the brain and lungs by live viruses, with the strongest infection occurring in the abdomen, especially in the intestinal organs. The dose of pseudovirus was linearly correlated with luminescence intensity. The infection model was able to differentiate the protective effect of monoclonal antibodies, with complete protection in the high-dose group. Conclusions: The recombinant Hendra-F/G pseudovirus hamster model allows the effective evaluation of prophylactic monoclonal antibodies, providing a crucial tool for studying Hendra virus infection and control strategies. Full article

(This article belongs to the Special Issue Pseudovirus-Based Strategies for Understanding and Controlling Viral Infections)

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15 pages, 2208 KB

Open AccessArticle

Effect of Surface Modification of a Dental Composite on the Adhesion of Streptococcus mitis, Streptococcus mutans, and Candida albicans: An In Vitro Study

by Santiago Arango-Santander, Claudia María Bedoya-Correa, Camila Soto, Santiago Bustamante and John Franco

Pathogens 2025, 14(9), 909; https://doi.org/10.3390/pathogens14090909 - 10 Sep 2025

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Abstract

Adhesion of different microorganisms to the surface of dental materials has generated significant interest since one of the most important requirements of biomaterials to be considered successful is their ability to withstand the damage caused by microorganisms that may lead to failure and [...] Read more.

Adhesion of different microorganisms to the surface of dental materials has generated significant interest since one of the most important requirements of biomaterials to be considered successful is their ability to withstand the damage caused by microorganisms that may lead to failure and the onset of different pathologies, such as caries. In vitro testing has demonstrated that surface modification is an alternative approach to reduce the adhesion of microorganisms to surfaces. The objective of this work was to assess the adhesion of Streptococcus mitis, Streptococcus mutans, and Candida albicans to a dental composite surface modified following a biomimetic approach and coated with salivary proteins. Soft lithography was used to copy the topography from the Crocosmia aurea leaf and then transfer it to the surface of dental composite discs that were coated with saliva proteins. Surfaces were characterized by contact angle and atomic force microscopy. S. mitis, S. mutans, and C. albicans were used to assess bacterial and fungal adhesion in monoculture and co-culture. The topographic modification of the surface of a dental composite reduced the adhesion of assessed microorganisms and the adhesion of these species in monoculture and co-culture on saliva-coated surfaces was higher than on topographically modified surfaces. Full article

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20 pages, 1324 KB

Open AccessReview

Mycobacterium marinum Immune Evasion in Zebrafish

by Priyank Kumar, Joshua Cameron, Beatrice Saviola and Vishwanath Venketaraman

Pathogens 2025, 14(9), 908; https://doi.org/10.3390/pathogens14090908 - 10 Sep 2025

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Abstract

Fish mycobacteriosis, a chronic progressive disease caused by nontuberculous mycobacteria (NTM), affects marine, brackish, and freshwater fish. Mycobacterium marinum (M. marinum), the most important of the NTM, infects fresh and marine water fish causing necrotizing granulomas and associated morbidity and mortality. [...] Read more.

Fish mycobacteriosis, a chronic progressive disease caused by nontuberculous mycobacteria (NTM), affects marine, brackish, and freshwater fish. Mycobacterium marinum (M. marinum), the most important of the NTM, infects fresh and marine water fish causing necrotizing granulomas and associated morbidity and mortality. M. marinum causes disease in zebrafish in a dose-dependent fashion. The M. marinum-induced disease in the zebrafish is associated with the development of necrotizing granulomas with abundant bacteria in the necrotic areas. Acute infection with high infectious doses of M. marinum infection in zebrafish was characterized by uncontrolled replication of the pathogen and death of all fish within 16 days, while chronic infections were marked by the formation of granulomas in different organs and longer survival in the range of 4–8 weeks. This review therefore synthesizes recent advances in our understanding of M. marinum’s infection of zebrafish, molecular pathogenesis, virulence mechanisms, and immune evasion strategies in zebrafish, while also highlighting the host immune effector responses and the virulence mechanisms of M. marinum. Full article

(This article belongs to the Special Issue Infectious Diseases in Aquatic Animals)

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13 pages, 239 KB

Open AccessArticle

First Signs at Admission: Prognostic Value of Initial Proinflammatory Markers for Sepsis and Mortality in ICU Patients

by Fatma Aktaş, Yasemin Zer, Gülseren Elay and Mehmet Erinmez

Pathogens 2025, 14(9), 907; https://doi.org/10.3390/pathogens14090907 - 10 Sep 2025

Viewed by 354

Abstract

Sepsis is a life-threatening condition and due to its non-specific symptoms, diagnosing sepsis and determining its severity remains difficult. Delays in recognizing sepsis can significantly increase mortality despite advances in antimicrobial therapy and resuscitation procedures. Biomarkers can help detect the presence and severity [...] Read more.

Sepsis is a life-threatening condition and due to its non-specific symptoms, diagnosing sepsis and determining its severity remains difficult. Delays in recognizing sepsis can significantly increase mortality despite advances in antimicrobial therapy and resuscitation procedures. Biomarkers can help detect the presence and severity of sepsis, distinguish between types of infections, and evaluate treatment response. This prospective study aims to determine whether biomarker levels measured at the time of intensive care unit (ICU) admission can assist in the early prediction, diagnosis, and prognosis of sepsis and bacteremia. Blood samples were collected from 132 ICU patients upon admission and analyzed for CRP, IL-6, PCT, SAA, and endotoxin levels. Patients were monitored for sepsis development, blood culture results, and mortality. IL-6 levels demonstrated a significant association with prognosis and identified as an independent risk factor. CRP and PCT levels exhibit a significant effect on the development of sepsis in both univariate and multivariate analyses. Also, our study demonstrated that the presence of bacteremia in the initial blood sample taken from intensive care patients holds significant diagnostic and prognostic value even without waiting for species-level identification when combined with markers such as PCT, CRP, and IL-6. Full article

10 pages, 1384 KB

Open AccessArticle

Forest Density and Invasive Carnivores Are Related to Trichinella Infection in Wild Boars in Poland

by Jakub Kubacki, Daniel Klich, Aneta Bełcik, Weronika Korpysa-Dzirba, Tomasz Cencek, Jacek Karamon, Jacek Sroka, Małgorzata Samorek-Pieróg, Michał Gondek and Ewa Bilska-Zając

Pathogens 2025, 14(9), 906; https://doi.org/10.3390/pathogens14090906 - 9 Sep 2025

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Abstract

The purpose of this study was to investigate and update the spatial distribution of Trichinella spp. in wild boars tested between 2015 and 2022 and to test the correlation of the population density of chosen animals (wild boars, red foxes (Vulpes vulpes [...] Read more.

The purpose of this study was to investigate and update the spatial distribution of Trichinella spp. in wild boars tested between 2015 and 2022 and to test the correlation of the population density of chosen animals (wild boars, red foxes (Vulpes vulpes), raccoon dogs (Nyctereutes procyonoides), and European badgers (Meles meles)) with the prevalence of Trichinella spp. in wild boars in Poland. In addition, to understand the distribution of infected animals, we sought to see if there were a correlation of Trichinella spp. infections in wild boars with land cover type. Among the wild carnivore species analyzed, only the population density of the raccoon dog (Nyctereutes procyonoides)—an invasive alien species—was significantly associated with infection rates in wild boars, particularly at the regional scale. As scavengers and competent reservoir hosts for all four European Trichinella species, raccoon dogs are likely to play a key role in the sylvatic transmission cycle. The positive rate of Trichinella spp. infection in wild boars during 2015–2022 was 0.22%, compared to 0.3% in 2009–2016. Moreover, forest density was positively correlated with infection rates, underlining the role of forest habitats in sustaining Trichinella transmission. Full article

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10 pages, 1699 KB

Open AccessArticle

Detection of Pseudorabies Virus in Hunting Dogs in Greece: The Role of Wild Boars in Virus Transmission

by Konstantinos Papageorgiou, Ilias Bouzalas, Kiriaki Giamoustari, Małgorzata Wróbel, Dimitrios Doukas, Aikaterini Stoikou, Zoi Athanasakopoulou, Dimitrios Chatzopoulos, Dimitrios Papadopoulos, Spyridon Pakos, Chrysanthi Karapetsiou, Charalambos Billinis, Evanthia Petridou and Spyridon K. Kritas

Pathogens 2025, 14(9), 905; https://doi.org/10.3390/pathogens14090905 - 9 Sep 2025

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Abstract

Aujeszky’s disease, or pseudorabies, is a viral infection caused by Suid herpesvirus 1 (pseudorabies virus), with swine as its natural host. Although eradicated in domestic pigs in many European countries, PRV remains endemic in wild boar populations, posing a risk to other species, [...] Read more.

Aujeszky’s disease, or pseudorabies, is a viral infection caused by Suid herpesvirus 1 (pseudorabies virus), with swine as its natural host. Although eradicated in domestic pigs in many European countries, PRV remains endemic in wild boar populations, posing a risk to other species, including carnivores. In this study, we report eight fatal cases of PRV infection in hunting dogs from Epirus and Thessaly, Greece, all of which followed direct contact with hunted wild boars. Postmortem brain samples tested positive for PRV via PCR targeting the glycoprotein C (gC) gene. Partial sequencing and phylogenetic analysis of the amplified gC fragments revealed genetic divergence among the examined isolates. The Epirus-derived strains formed a distinct cluster, closely related to previously reported Greek strains from the region of Central Macedonia as well as to the French strain FRA 527 and the German isolate GER614BW. In contrast, the two Thessaly sequenced isolates were phylogenetically distant from all other Greek strains, potentially representing an independently evolving lineage, and clustered more closely with the Kaplan strain. These findings underscore the persistent threat of PRV transmission from wild to domestic species and highlight the genetic heterogeneity of PRV strains circulating in Greece. Veterinary practitioners should consider PRV in the differential diagnosis of encephalitic symptoms in hunting dogs. Enhanced molecular surveillance and public awareness are critical to mitigating the risks posed by this emerging threat. Full article

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22 pages, 4172 KB

Open AccessArticle

Impact of Plasmodiophora brassicae on Canola Root and Rhizosphere Microbiomes and Its Implications for Clubroot Biocontrol

by Jorge Cordero-Elvia, Leonardo Galindo-González, Rudolph Fredua-Agyeman, Sheau-Fang Hwang and Stephen E. Strelkov

Pathogens 2025, 14(9), 904; https://doi.org/10.3390/pathogens14090904 - 9 Sep 2025

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Abstract

Clubroot, caused by the obligate parasite Plasmodiophora brassicae, is a soilborne disease affecting canola (Brassica napus) and other crucifers. Although planting resistant cultivars remains the primary strategy for managing clubroot, the emergence of resistance-breaking P. brassicae pathotypes continues to threaten [...] Read more.

Clubroot, caused by the obligate parasite Plasmodiophora brassicae, is a soilborne disease affecting canola (Brassica napus) and other crucifers. Although planting resistant cultivars remains the primary strategy for managing clubroot, the emergence of resistance-breaking P. brassicae pathotypes continues to threaten canola production. In this context, soil and root microorganisms may play a role in suppressing the disease. This study investigated the impact of P. brassicae infection on the microbial communities of soil, seeds, roots, and the rhizosphere in susceptible and resistant canola lines, with the aim of analyzing host–pathogen–microbiome interactions and identifying microbial taxa potentially associated with disease resistance. Our findings showed that resistant canola lines inoculated with P. brassicae (pathotype 3A) exhibited reduced disease severity compared to their susceptible counterparts. Diversity analyses of microbial communities revealed that clubroot-resistant canola lines tended to maintain more stable and diverse fungal communities, with a higher Shannon index than susceptible lines. Inoculation with P. brassicae induced more pronounced changes in the root microbiome than in the rhizosphere. Additionally, the seed microbiomes of resistant and susceptible lines displayed distinct bacterial and fungal profiles, suggesting that clubroot susceptibility may influence seed-associated microbial community composition. Differential abundance analysis of root and rhizosphere microbiomes indicated that certain microbial taxa, including bacterial genera such as Acidovorax, Bacillus, Cupriavidus, Cytophaga, Duganella, Flavobacterium, Fluviicola, Luteimonas, Methylotenera, Pedobacter, and Peredibacter, as well as fungal genera such as Aspergillus, Candida, Fusicolla, Paecilomyces, and Rhizophlyctis, may be recruited or enriched in resistant canola lines following P. brassicae inoculation, potentially contributing to reduced clubroot severity. Full article

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10 pages, 641 KB

Open AccessCase Report

Candidozyma auris Spondylodiscitis: A Case Report from Saudi Arabia and Literature Review

by Rakan Sambas, Khalid Bin Aziz, Faisal N. Alqahtani, Hussam Alhathlol, Marwan Alhumaidi, Abdulrahman Alsaedy and Thamer S. Alhowaish

Pathogens 2025, 14(9), 903; https://doi.org/10.3390/pathogens14090903 - 8 Sep 2025

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Abstract

Candidozyma auris (formerly known as Candida auris) is an emerging multidrug-resistant fungal pathogen that has become increasingly implicated in healthcare-associated infections; however, its involvement in spondylodiscitis is exceedingly rare. We report the case of a 65-year-old Saudi male with multiple comorbidities who [...] Read more.

Candidozyma auris (formerly known as Candida auris) is an emerging multidrug-resistant fungal pathogen that has become increasingly implicated in healthcare-associated infections; however, its involvement in spondylodiscitis is exceedingly rare. We report the case of a 65-year-old Saudi male with multiple comorbidities who presented with altered mental status and was subsequently diagnosed with Candidozyma auris spondylodiscitis and bilateral psoas abscesses. Despite broad-spectrum antifungal therapy and multidisciplinary management, the patient’s condition rapidly deteriorated. This case highlights the significant challenges in diagnosing and managing multidrug-resistant C. auris infections and underscores the need for early suspicion, effective source control, and novel antifungal strategies in high-risk populations. Full article

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13 pages, 1993 KB

Open AccessArticle

An Oral Salmonella-Based Vaccine Expressing Viral M43 Protein Elicits Effective Immunity Against Murine Cytomegalovirus in Mice

by Yujun Liu, Hao Gong, Jiaming Zhu and Fenyong Liu

Pathogens 2025, 14(9), 902; https://doi.org/10.3390/pathogens14090902 - 8 Sep 2025

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Abstract

Human cytomegalovirus (HCMV) is the leading viral cause of congenital infections and causes substantial morbidity and mortality in neonates and immunosuppressed people. Generating an anti-HCMV vaccine is required for preventing viral-associated diseases and infections. Oral vaccines based on attenuated Salmonella are an attractive [...] Read more.

Human cytomegalovirus (HCMV) is the leading viral cause of congenital infections and causes substantial morbidity and mortality in neonates and immunosuppressed people. Generating an anti-HCMV vaccine is required for preventing viral-associated diseases and infections. Oral vaccines based on attenuated Salmonella are an attractive solution, since these vaccines can be applied orally and easily for mass immunization. In this report, we constructed an attenuated Salmonella strain for the expression of the murine cytomegalovirus (MCMV) M43 protein and studied its ability as an oral vaccine candidate to stimulate antiviral immunity in mice. In orally immunized mice, the constructed vaccine, Sal-M43, elicited both serum IgG and mucosal IgA levels as well as T cell responses that were specific against the MCMV M43 protein. Moreover, the Sal-M43 immunization substantially inhibited the viral growth and infection in various organs and tissues and offered complete immune protection against both intraperitoneal and intranasal MCMV challenges. Thus, the Salmonella-based vaccine expressing the M43 antigen is effective in inducing anti-MCMV immunity. These findings also reveal the promise of developing oral anti-CMV vaccines based on attenuated Salmonella vectors expressing different viral antigens. Full article

(This article belongs to the Section Viral Pathogens)

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18 pages, 2429 KB

Open AccessArticle

Isolation, Molecular Characterization and In Vitro Propagation of an Anaplasma platys-Like Bacterium in Tick Cells

by Erica Rodrigues de Matos, Priscilla Nunes dos Santos, Erich Peter Zweygarth, Talys Henrique Assumpção Jardim, Huarrisson Azevedo Santos, Matheus Dias Cordeiro, Bruna de Azevedo Baêta, Lesley Bell-Sakyi, Adivaldo Henrique da Fonseca and Claudia Bezerra da Silva

Pathogens 2025, 14(9), 901; https://doi.org/10.3390/pathogens14090901 - 7 Sep 2025

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Abstract

The family Anaplasmataceae comprises etiological agents of infectious diseases of significant importance. This study aimed to achieve the in vitro isolation and propagation of an Anaplasma sp. using tick-derived cell lines. The study was realized in Seropédica municipality, Rio de Janeiro, Brazil. Blood [...] Read more.

The family Anaplasmataceae comprises etiological agents of infectious diseases of significant importance. This study aimed to achieve the in vitro isolation and propagation of an Anaplasma sp. using tick-derived cell lines. The study was realized in Seropédica municipality, Rio de Janeiro, Brazil. Blood smears from a naturally infected bovine revealed cytoplasmic inclusions in blood cells. To isolate and propagate the organism, IDE8 and ISE6 tick cell lines derived from Ixodes scapularis were used. Two methods of inoculum preparation were employed: Histopaque^® density gradient and platelet-rich plasma separation. Following infection, cells were maintained in L-15B medium without antibiotics at 34 °C, and infection was monitored weekly by Giemsa-stained cytocentrifuge smears. After achieving ≥ 70% infection, bacteria were subcultured and successfully cryopreserved and resuscitated. PCR amplification and sequencing of 16S rDNA, 23S rDNA, rpoB, and groEL genes were performed for molecular characterization. Phylogenetic analyses revealed that the isolated strain clustered within the A. platys-like clade. This study reports the successful in vitro isolation, propagation, and cryopreservation of the ‘A. platys-like strain Natal’ bacterium in tick cell lines and provides molecular evidence supporting its phylogenetic classification. These findings contribute to the understanding of genetic variability and host–cell interactions of Anaplasma spp., laying the groundwork for future research. Full article

(This article belongs to the Special Issue Vector Control and Parasitic Infection in Animals)

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16 pages, 329 KB

Open AccessArticle

Bayesian Assessment of True Prevalence of Paratuberculosis Infection in Dairy Herds and Their Parity Subgroups

by Katalin Veres, Zsolt Lang and László Ózsvári

Pathogens 2025, 14(9), 900; https://doi.org/10.3390/pathogens14090900 - 7 Sep 2025

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Abstract

Paratuberculosis is a widespread infectious disease in ruminants that leads to significant economic losses in livestock production. In this study, we developed a practical method for predicting the likelihood of the herd-level presence of the infection and estimating its prevalence in subgroups of [...] Read more.

Paratuberculosis is a widespread infectious disease in ruminants that leads to significant economic losses in livestock production. In this study, we developed a practical method for predicting the likelihood of the herd-level presence of the infection and estimating its prevalence in subgroups of a dairy herd—specifically, first-time calving cows (primiparous) and those that have calved more than once (multiparous). We fit a Bayesian hierarchical model to cow-level data, incorporating prior knowledge about regional prevalence of infection to improve the accuracy and reliability of the estimates. The model was tested using synthetic data representing six regional scenarios in four countries (Chile, Denmark, Italy, and Hungary). The likelihood that a herd is infected is evaluated using Bayes factors and posterior probability of infection. Both the Bayes factor and the posterior probability of infection classified the simulated herds in accordance with the proportions of infected herds. Summary measures obtained for within-herd true prevalence estimates demonstrated acceptable accuracy. The R and STAN codes of the model are available as an open-access tool. The model can be customized for any region using real local data and prior information. The relationship between true and apparent prevalence is linear and stable and therefore can be estimated well. We found that, in Hungary, the TP/AP ratios were 1.6 and 1.5 for primi- and multiparous cows, respectively. Full article

17 pages, 579 KB

Open AccessReview

Hepatitis Delta Virus Infection: An Overview

by Vitor Duque and Diana Duque

Pathogens 2025, 14(9), 899; https://doi.org/10.3390/pathogens14090899 - 6 Sep 2025

Viewed by 660

Abstract

The burden of hepatitis delta virus (HDV) infection is currently unknown and may affect 12 to 72 million people distributed across various hot spots in different regions of the globe. Screening for antibodies to HDV infection in patients positive for the hepatitis B [...] Read more.

The burden of hepatitis delta virus (HDV) infection is currently unknown and may affect 12 to 72 million people distributed across various hot spots in different regions of the globe. Screening for antibodies to HDV infection in patients positive for the hepatitis B surface antigen (HBsAg) is generally available in most parts of the world, but systematic testing for HDV is needed. Chronic HDV infection is associated with a higher risk of progression to cirrhosis, liver failure, and hepatocellular carcinoma compared to hepatitis B virus (HBV) mono-infection. Bulevirtide is the recently available treatment against hepatitis delta. The results of efficacy studies and new drugs (lonafarnib) are under discussion. New therapeutic strategies are in development, revealing a critical need for valid next-generation treatments to cure HDV. Full article

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

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17 pages, 1578 KB

Open AccessArticle

Independent Development of Resistance to Main Classes of Anthelmintics by Gastrointestinal Nematodes of Ruminants and Horses

by Jacques Cabaret

Pathogens 2025, 14(9), 898; https://doi.org/10.3390/pathogens14090898 - 5 Sep 2025

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Abstract

Resistance to anthelmintics in gastrointestinal nematodes (GINs) is highly prevalent, as these parasites have been treated with anthelmintics for decades in ruminants and horses. Anthelmintics belong to different classes, each with a different mode of action. The most used are benzimidazoles and macrocyclic [...] Read more.

Resistance to anthelmintics in gastrointestinal nematodes (GINs) is highly prevalent, as these parasites have been treated with anthelmintics for decades in ruminants and horses. Anthelmintics belong to different classes, each with a different mode of action. The most used are benzimidazoles and macrocyclic lactones and, to a lesser extent, levamisole and pyrantel in herbivores, as estimated from the literature. Combining these classes should be effective in controlling GIN. However, several farmers’ practices tend to promote GIN resistance. Therefore, it is unclear whether the use of anthelmintic associations is a sustainable solution for controlling resistance in natural conditions. It is not easy to establish the association of anthelmintic resistances on farms since rarely several anthelmintics and their combinations are used on a single farm. Composed probability calculations were employed when literature data indicated the presence of resistance (to benzimidazoles, levamisole, or macrocyclic lactones) in several ruminant GIN cases. The efficacy of different anthelmintics (benzimidazoles, pyrantel, or macrocyclic lactones) was evaluated in terms of the correlation between faecal nematode egg counts in horses in the available literature. No associations of anthelmintic resistance were found between the different classes of anthelmintics in either ruminants or horses. However, the association between anthelmintic resistance in GIN may appear in the long term. It is presumed that combining drugs may reduce the development of resistance and allow better control of infection on farms where resistance is already established to a low level. Full article

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