Pathogens

22 pages, 11866 KB

Open AccessArticle

Study on the Mechanism of RuHaoDaShi Granules in Treating H1N1 Viral Pneumonia Based on Network Pharmacology and Experimental Validation

by Aixin Chen, Tianhang Chen, Yu He, Jiehong Yang and Haitong Wan

Pathogens 2025, 14(8), 834; https://doi.org/10.3390/pathogens14080834 - 21 Aug 2025

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Abstract

Objective: This study aims to investigate the pharmacodynamic effects and underlying mechanisms of the Chinese herbal formula RuHaoDaShi (RHDS) granules against the influenza virus in experimental models. Methods: This study aims to employ network pharmacology to identify the active components of RHDS and [...] Read more.

Objective: This study aims to investigate the pharmacodynamic effects and underlying mechanisms of the Chinese herbal formula RuHaoDaShi (RHDS) granules against the influenza virus in experimental models. Methods: This study aims to employ network pharmacology to identify the active components of RHDS and its potential targets and mechanisms of action against H1N1. The molecular docking approach validated the interactions between the core targets and the RHDS compounds. In vitro, the antiviral activity of RHDS was assessed by therapeutic, prophylactic, and premixed administration to H1N1-infected A549 cells. An in vivo experiment was conducted using a mouse H1N1 pneumonia model. The model was treated with a dose of 1.04, 2.08, and 4.16 g/kg of RHDS, administered via gavage daily. The study’s objective was to evaluate the antiviral activity and mechanism of action of RHDS in mice. Mice were evaluated on day 6 by assessing survival, viral load (RT-qPCR), lung pathology (HE staining), inflammatory cytokines (ELISA, immunohistochemistry), and ferroptosis markers (WB, qPCR). Results: Network pharmacology identified 77 biologically active RHDS compounds (e.g., quercetin and kaempferol) and 32 core targets common to RHDS, H1N1, and ferroptosis. Molecular docking was used to verify a high affinity for binding between the core targets HIF-1α, MAPK3, and key RHDS compounds. In vitro studies demonstrated that RHDS exhibited protective properties against H1N1-infected cells, with the therapeutic delivery method proving the most efficacious. In vivo studies have shown that RHDS reduces mortality, lung index, and viral load in mice while attenuating histopathological damage. The study demonstrated a reduction in the release of inflammatory cytokines, including IL-6, IFN-γ, and IL-17A, and decreased expression levels of MPO and F4/80 proteins in lung tissue. Mechanistically, the administration of RHDS resulted in the up-regulation of the expression levels of GPX4, SLC7A11, and Nrf2 proteins while concomitantly inhibiting the expression of HIF-1α, COX2, and ACSL4. These findings confirm the modulatory effect of RHDS on the GPX4/SLC7A11/Nrf2 pathway. Conclusions: RHDS demonstrated a protective effect against H1N1-induced cytopathy in vitro and was effective in attenuating H1N1-induced pneumonia in murine models. The study suggests that RHDS has antiviral potential to treat H1N1 viral pneumonia by modulating inflammatory cytokines and the GPX4/SLC7A11/Nrf2 pathway. Full article

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10 pages, 851 KB

Open AccessArticle

Introduction and Spatial–Temporal Distribution of Oropouche Virus in Rio de Janeiro State, Brazil

by Fábio Burack da Costa, Andrea Cony Cavalcanti, Rafael Santos Erbisti, Vanessa Zaquieu Dias, Cristiane Gomes de Castro Moreira, Mateus Marques Grifo, Maria Carmelita dos Santos Vaz, Adriana Cardoso Camargo, Leandro Magalhães de Souza, Flávia Barreto dos Santos, Mário Sérgio Ribeiro, Viviana Malirat, Nildimar Alves Honório and Renata Campos Azevedo

Pathogens 2025, 14(8), 833; https://doi.org/10.3390/pathogens14080833 - 21 Aug 2025

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Abstract

The Oropouche virus (OROV) has been circulating in the Amazon region since the 1960s, with a progressive increase in outbreaks and human cases reported in Brazil and neighboring countries. In the Rio de Janeiro state, there has been a significant rise in suspected [...] Read more.

The Oropouche virus (OROV) has been circulating in the Amazon region since the 1960s, with a progressive increase in outbreaks and human cases reported in Brazil and neighboring countries. In the Rio de Janeiro state, there has been a significant rise in suspected cases of arboviruses, with only 30% of laboratory tests confirming infections with dengue, Zika, or chikungunya viruses. The investigation of OROV virus circulation in the Rio de Janeiro state was initiated and confirmed in April 2024. Our study aimed to retrospectively investigate OROV infections in dengue-suspected cases with inconclusive diagnosis in order to better understand the temporal and geographic introduction of OROV in the Rio de Janeiro state. Municipalities from Rio de Janeiro with arbovirus-like fever cases but a low percentage of dengue-positive RT-PCR test confirmations were identified in the laboratory database. Samples were selected for testing OROV infections using real-time RT-PCR as recommended by the Brazilian Ministry of Health. Municipalities in the Middle Paraíba region of the state showed 93% negative tests results for dengue, Zika, and chikungunya starting in September 2023. A total of 118 positive cases of Oropouche were recorded in the state of Rio de Janeiro between March and July 2024. Moreover, by genome sequencing of eight strains, it was shown that OROV circulating in Rio de Janeiro belongs to recently emergent M₁L₂S₂ lineage. Our findings retrospectively revealed a concentration of cases in the Middle Paraíba region and an outbreak in the rural village of Cacaria, located in the municipality of Piraí. According to our data, this region is the first area with sustained transmission in the Rio de Janeiro state. Full article

(This article belongs to the Special Issue Emerging Arboviruses: Epidemiology, Control, and Future Directions)

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19 pages, 1910 KB

Open AccessReview

Hansen’s Disease in Ecuador: Current Status, Knowledge Gaps, and Research Priorities: A Literature Review

by Manuel Calvopiña, Juan S. Izquierdo-Condoy, Esteban Ortiz-Prado, Jorge Vasconez-Gonzalez, Lorena Vaca and Elías Guamán

Pathogens 2025, 14(8), 832; https://doi.org/10.3390/pathogens14080832 - 21 Aug 2025

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Abstract

Background: Hansen’s disease or leprosy is one of the 21 neglected tropical diseases (NTDs). In Ecuador, leprosy is considered eliminated as a public health problem; however, new cases are reported annually. Additionally, Mycobacterium leprae infection was detected in nine-banded armadillos across the country, [...] Read more.

Background: Hansen’s disease or leprosy is one of the 21 neglected tropical diseases (NTDs). In Ecuador, leprosy is considered eliminated as a public health problem; however, new cases are reported annually. Additionally, Mycobacterium leprae infection was detected in nine-banded armadillos across the country, suggesting a potential zoonotic reservoir. This literature review aims to provide an updated overview of the epidemiological situation of leprosy in Ecuador, identify knowledge gaps, and outline research priorities to support the development of a comprehensive national strategy for achieving zero autochthonous cases. Methods: This article analyses the current situation of leprosy in Ecuador based on international and national publications. A retrospective literature search using five international, regional, and national publications on leprosy published between 1954 and 2024 (70 years) with no restriction on language or publication date, was performed. Findings: Our review identified 28 publications with the earliest article dating back to 1954. Of these, 14 were published in international journals, 15 (53.6%) were in Spanish. Four nationwide studies documented leprosy cases across Ecuador’s three continental regions (Coast, Andes, and Amazon) with a predominance in the tropical coast. No cases have been reported from the Galápagos Islands. From 1983, Ecuador started multi-drug therapy. Data from the Ministry of Public Health (MoH) system identified 1539 incident cases, showing a significant decline in new cases from 2000 to 2024, with no cases in children. New cases detection rate by 100,000 inhabitants was 0.51 in 2019 according to the World Health Organization (WHO). No study has genotyped the Mycobacterium spp. in human cases, other animal species, or environment. According to the MoH, multibacillary leprosy accounts for 78.95% of diagnosed cases, with confirmation based on Ziehl–Neelsen staining and histopathology. No survey has assessed disabilities, knowledge, attitudes, and practices (KAP) or stigma related to leprosy. Research is needed on transmission routes, Mycobacterium genotyping, genetic susceptibility, and antibiotic resistance. BCG vaccination coverage fell to 75.3% in 2021. Cases are currently diagnosed and treated on an outpatient basis in large hospitals. Conclusions: This comprehensive review highlights persistent gaps in leprosy research and critical information, despite seven decades of documented cases in Ecuador. The disease is still endemic across the country, particularly at subnational level in the subtropics and tropics of the Pacific coast and the Amazon. There is a need for nationwide epidemiological research on reservoirs and the environment applying the One Health concept. Increased laboratory facilities and readily available official data are required to improve our understanding of leprosy in Ecuador. Strengthening community-level efforts is essential for Ecuador to meet the targets of the “WHO’s Towards Zero Leprosy: Strategy 2021–2030.” Full article

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11 pages, 525 KB

Open AccessArticle

Susceptibility of Neisseria gonorrhoeae to Zoliflodacin and Quinolones in Hyogo Prefecture, Japan

by Takashi Yurube, Katsumi Shigemura, Yurino Kobayashi, Taishi Maeda, Nami Nishimura, Ayaka Yamada, Kotoko Kotani, Saki Horii, Hiroyuki Yoshida and Kayo Osawa

Pathogens 2025, 14(8), 831; https://doi.org/10.3390/pathogens14080831 - 21 Aug 2025

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Abstract

The DNA synthesis inhibitor zoliflodacin (ZFD) is expected to be effective against strains resistant to therapeutic agents for Neisseria gonorrhoeae infection. In addition to ZFD, we investigated the susceptibility of N. gonorrhoeae strains to ceftriaxone (CTRX), ciprofloxacin (CPFX), garenoxacin (GRNX), and sitafloxacin (STFX). [...] Read more.

The DNA synthesis inhibitor zoliflodacin (ZFD) is expected to be effective against strains resistant to therapeutic agents for Neisseria gonorrhoeae infection. In addition to ZFD, we investigated the susceptibility of N. gonorrhoeae strains to ceftriaxone (CTRX), ciprofloxacin (CPFX), garenoxacin (GRNX), and sitafloxacin (STFX). Minimum inhibitory concentration values for ZFD and four other drugs were determined for 147 strains of N. gonorrhoeae isolated at medical institutions in Hyogo Prefecture, Japan, from 2015 to 2022. Amino acid alterations in gyrA, gyrB, parC, and parE were examined by polymerase chain reaction and sequencing analysis. Sequence type (ST) was determined for epidemiological analysis, and N. gonorrhoeae strains were classified. The non-susceptibility rate was not observed in CTRX. The lowest non-susceptibility rate was observed in ZFD (39.5%) compared to CPFX (80.3%), GRNX (83.7%), and STFX (65.3%) (all p < 0.0001). The most common amino acid alterations in gyrA and parC had non-susceptibility rates exceeding 80% to quinolones except ZFD, suggesting that these alterations may have influenced the resistance trend. STs were different between isolates in 2015 and those in 2020 and later. ZFD showed potent antimicrobial activity against N. gonorrhoeae strains that are highly resistant to quinolones. It may become a new option in the treatment of gonococcal infections. Full article

(This article belongs to the Special Issue Cutting-Edge Research on Pathogenic Neisseria)

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17 pages, 1414 KB

Open AccessSystematic Review

Mechanistic Models of Virus–Bacteria Co-Infections in Humans: A Systematic Review of Methods and Assumptions

by Mani Dhakal, Brajendra K. Singh and Rajeev K. Azad

Pathogens 2025, 14(8), 830; https://doi.org/10.3390/pathogens14080830 - 21 Aug 2025

Viewed by 454

Abstract

Background: Viral–bacterial co-infections can amplify disease severity through complex biological mechanisms. Mathematical models are critical tools for understanding these threats, but it is unclear how well they capture the underlying biology. This systematic review addresses a central question: to what extent does the [...] Read more.

Background: Viral–bacterial co-infections can amplify disease severity through complex biological mechanisms. Mathematical models are critical tools for understanding these threats, but it is unclear how well they capture the underlying biology. This systematic review addresses a central question: to what extent does the current generation of models mechanistically represent co-infections, or do the mathematical assumptions underlying these models adequately represent the known biological mechanisms? Methods: Following PRISMA guidelines, we systematically reviewed the literature on mechanistic models of human virus–bacteria co-infections. A systematic search of articles on the scientific literature repositories PubMed, Scopus, and Dimensions was conducted and data on study objectives, model structure, assumptions about biological interactions (e.g., susceptibility, mortality), control measures (if evaluated), and the empirical sources used for key parameters were extracted. Results: We identified 72 studies for inclusion in this analysis. The reviewed models are consistently built on the established premise that co-infection alters disease severity and host susceptibility. However, we found they incorporate these dynamics primarily through high-level mathematical shortcuts, such as applying static “multiplicative factors” to transmission or progression rates. Our quantitative analysis also revealed questionable approaches; for example, 79% (57) of these studies relied on non-empirical sources (assumed or borrowed values) for parameter values including interaction parameters (e.g., increased susceptibility to a secondary pathogen following primary infection, or elevated mortality rates in co-infected individuals). Conclusions: An apparently unjustified practice exists in co-infection modeling, where complex biological processes are simplified to fixed numerical assumptions, often without empirical support. This practice limits the predictive reliability of current models. We identify an urgent need for data-driven parameterization and interdisciplinary collaboration to bridge the gap between biological complexity and modeling practice, thereby enhancing the public health relevance of co-infection modeling. Full article

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10 pages, 602 KB

Open AccessSystematic Review

Infective Endocarditis by Fusobacterium Species—A Systematic Review

by Petros Ioannou, Eleni Mourati, Styliani Thalasseli Kazali, Chrysoula Bali, Stella Baliou and George Samonis

Pathogens 2025, 14(8), 829; https://doi.org/10.3390/pathogens14080829 - 21 Aug 2025

Viewed by 524

Abstract

Background: Fusobacterium species are anaerobic Gram-negative bacilli that are part of the normal oropharyngeal and gastrointestinal flora. Although rare, they can cause infective endocarditis (IE), a severe condition with high morbidity. The clinical characteristics, treatment strategies, and outcomes of IE caused by Fusobacterium [...] Read more.

Background: Fusobacterium species are anaerobic Gram-negative bacilli that are part of the normal oropharyngeal and gastrointestinal flora. Although rare, they can cause infective endocarditis (IE), a severe condition with high morbidity. The clinical characteristics, treatment strategies, and outcomes of IE caused by Fusobacterium spp. remain incompletely defined. This systematic review aimed to synthesize available data on Fusobacterium IE and compare its features with IE caused by other pathogens. Methods: We conducted a systematic literature search in PubMed, Scopus, and the Cochrane Library up to 27 February 2025, using the terms “Fusobacterium” and “endocarditis.” Eligible studies included case reports or series describing adult or pediatric patients with Fusobacterium-associated IE. Data were extracted on demographics, risk factors, clinical features, microbiology, treatment, and outcomes. Results: A total of 21 studies (all case reports) involving 21 patients were included. The median age was 48 years, and 85.7% were male. Poor dental hygiene or recent dental work was the most common predisposing factor (47.6%). The mitral valve was most frequently affected (44.4%). Fever and sepsis were reported in nearly all cases, and embolic phenomena occurred in 81%. The most commonly isolated species were F. necrophorum (47.6%) and F. nucleatum (42.9%). Treatment commonly included metronidazole (61.9%), while surgical management was required in 23.8%. All-cause and infection-attributable mortality were both 9.5%. Conclusions: Fusobacterium IE predominantly affects younger males and is often linked to oral sources. This disease is associated with a high risk of systemic complications but seems to have a lower mortality compared to IE from other pathogens, including other anaerobic bacteria. Early diagnosis and appropriate antimicrobial treatment are of utmost importance for optimal outcomes. Further research is required to guide evidence-based management of this rare but serious infection. Full article

(This article belongs to the Special Issue Recent Research on Bloodstream Infections)

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16 pages, 297 KB

Open AccessReview

Host Immune Response in Chronic Hepatitis Delta: Implications for Pathogenesis and Therapy

by Arshi Khanam, Abutaleb Ameer, Poonam Mathur, Cihan Yurdaydin and Shyam Kottilil

Pathogens 2025, 14(8), 828; https://doi.org/10.3390/pathogens14080828 - 21 Aug 2025

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Abstract

Chronic hepatitis delta (CHD) represents the most severe form of viral hepatitis due to rapid disease progression towards liver cancer, leading to high morbidity and mortality. Hepatitis delta virus (HDV) can only infect individuals who are infected with hepatitis B. So far, there [...] Read more.

Chronic hepatitis delta (CHD) represents the most severe form of viral hepatitis due to rapid disease progression towards liver cancer, leading to high morbidity and mortality. Hepatitis delta virus (HDV) can only infect individuals who are infected with hepatitis B. So far, there is no cure or vaccine for HDV. Existing treatment options, including pegylated interferon-α and hepatocyte entry inhibitors, offer limited efficacy. Emerging therapeutic strategies are focused on targeting various steps of the HDV life cycle or enhancing the host immune response to promote viral elimination. A defective antiviral immune response is increasingly recognized as a culprit for HDV persistence; however, the precise immunological mechanism associated with disease progression and pathogenesis has not been well defined. This review provides an update on the current understanding of host immune response in CHD, highlighting its role in both disease pathogenesis and viral clearance. A deeper understanding of these immune correlates may lead the way to novel treatment strategies, including immunotherapies targeting host immune response that can be used in combination with other antiviral therapies to achieve more effective and durable treatment outcomes. Full article

(This article belongs to the Special Issue Viral Hepatitis and Therapeutic Strategies)

10 pages, 545 KB

Open AccessArticle

Anti-SARS-CoV-2 Antibodies in Urine of Individuals Vaccinated with Janssen AD26.COV2.S COVID-19 Vaccine

by Marina F. N. Melo, Rômulo C. D. Lira, Raquel S. B. Câmara, Isabela A. G. Pereira, Fernanda F. Ramos, Carolina S. F. Costa, Laura F. Amorim, Quezia D. Teixeira, Flávio G. da Fonseca, Vandack Nobre, Flavia G. F. Ferreira, Jorge Pinto, Eduardo A. F. Coelho, Fernanda Ludolf and Júlia F. M. Caporali

Pathogens 2025, 14(8), 827; https://doi.org/10.3390/pathogens14080827 - 21 Aug 2025

Viewed by 430

Abstract

Urine-based immunoassay is a non-invasive method with demonstrated utility in detecting anti-SARS-CoV-2 antibodies in unvaccinated patients with COVID-19. To evaluate urine’s potential for serological surveys in a real-world setting, SARS-CoV-2 serology was performed on urine samples from vaccinated individuals, both with and without [...] Read more.

Urine-based immunoassay is a non-invasive method with demonstrated utility in detecting anti-SARS-CoV-2 antibodies in unvaccinated patients with COVID-19. To evaluate urine’s potential for serological surveys in a real-world setting, SARS-CoV-2 serology was performed on urine samples from vaccinated individuals, both with and without prior confirmed COVID-19. (1) Methods: An in-house indirect ELISA was used to measure antibodies against recombinant spike (S) and nucleocapsid (N) proteins of SARS-CoV-2 in urine and paired serum from 149 individuals vaccinated with Janssen AD26.COV2.S, an S protein-based COVID-19 vaccine. (2) Results: Anti-S and anti-N levels were higher in the urine and serum of participants with confirmed prior COVID-19 compared to those without prior infection. Urinary anti-S effectively distinguished vaccinated individuals with (AUC = 0.96) and without (AUC = 0.88) prior infection from negative controls (non-vaccinated, non-previously infected individuals) (p < 0.0001). Among vaccinated participants, urinary anti-S and anti-N identified prior infection, with AUC values of 0.73 (p < 0.0001) and 0.60 (p = 0.03), respectively, being recorded. (3) Conclusions: Findings indicate that urinary anti-SARS-CoV-2 antibodies reflect AD26.COV2.S vaccination and previous COVID-19. To further advance the methodology, studies with larger sample sizes and a greater diversity of COVID-19 vaccines are required. Full article

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18 pages, 927 KB

Open AccessArticle

In Vitro Investigation of the Antimicrobial Properties of Gerês Propolis in Bacteria Isolated from Companion Animals and Safety Profile Characterization Using the Galleria mellonella Model

by Rafael Rodrigues, Rui Almeida, Soraia C. V. Rodrigues, Joana Castro, Ricardo Oliveira, Nuno Mendes, Carina Almeida, Sónia Silva, Daniela Araújo and Cristina Almeida-Aguiar

Pathogens 2025, 14(8), 826; https://doi.org/10.3390/pathogens14080826 - 21 Aug 2025

Viewed by 555

Abstract

Propolis, also known as bee glue, is a natural resinous mixture produced by Western honeybees and has long been recognized for its potential therapeutic properties. Recent research has focused on its diverse bioactivities, particularly its antimicrobial effects against a broad spectrum of microorganisms, [...] Read more.

Propolis, also known as bee glue, is a natural resinous mixture produced by Western honeybees and has long been recognized for its potential therapeutic properties. Recent research has focused on its diverse bioactivities, particularly its antimicrobial effects against a broad spectrum of microorganisms, including human but also animal pathogens. However, further investigation is required to fully elucidate the pharmacological properties and potential toxicity of propolis to support its broader application. This study investigated the antimicrobial efficacy and safety of an ethanol extract of Portuguese propolis collected from the Gerês region (G23.EE). The antimicrobial activity was assessed in vitro against both Gram-positive and Gram-negative bacteria associated with infections in companion animals, using the agar dilution method. To evaluate potential toxicity, the extract was administered via injection and topical application in an in vivo Galleria mellonella larval model, with health parameters monitored over a 96 hours period. The in vitro results showed that G23.EE was more effective against Gram-positive bacteria, including Staphylococcus spp. (e.g., S. felis, S. hominis, S. simulans, and S. pseudintermedius; MIC = 0.5 mg/mL) and Enterococcus faecium (MIC = 1.5 mg/mL), than against Gram-negative bacteria, such as Escherichia coli and Klebsiella oxytoca (MIC > 8.0 mg/mL). No significant adverse effects were observed in G. mellonella larvae following injection or brushing with propolis extract concentrations up to 8.0 mg/mL. Overall, these findings suggest that Portuguese propolis from Gêres is a promising, safe, and effective natural antimicrobial agent for targeting Gram-positive bacterial infections in companion animals. Full article

(This article belongs to the Special Issue Bacterial Pathogenesis and Antibiotic Resistance)

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5 pages, 149 KB

Open AccessEditorial

Pathogens of Wild Birds: Prevalence and Molecular and Morphological Characterization

by Alazne Díez-Fernández and Rafael Gutiérrez-López

Pathogens 2025, 14(8), 825; https://doi.org/10.3390/pathogens14080825 - 20 Aug 2025

Viewed by 369

Abstract

Wild birds are exposed to multiple infections by pathogenic microorganisms [...] Full article

(This article belongs to the Special Issue Pathogens of Wild Birds: Prevalence, Molecular and Morphological Characterization)

11 pages, 481 KB

Open AccessArticle

Human Tuberculosis in Migrant and Autocthonous Patients: A Ten-Year Single-Centre Experience

by Isabel García Soriano, Mónica Romero, Isabel Gascón, Verónica Solves, Reyes Pascual and Philip Erick Wikman-Jorgensen

Pathogens 2025, 14(8), 824; https://doi.org/10.3390/pathogens14080824 - 20 Aug 2025

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Abstract

In recent years, migratory movements have increased. This study aimed to compare tuberculosis cases in migrant and autochthonous patients. We conducted a retrospective analytical cohort study of patients diagnosed with tuberculosis in the Elda Health District (Alicante, Spain) between 2013 and 2023. Of [...] Read more.

In recent years, migratory movements have increased. This study aimed to compare tuberculosis cases in migrant and autochthonous patients. We conducted a retrospective analytical cohort study of patients diagnosed with tuberculosis in the Elda Health District (Alicante, Spain) between 2013 and 2023. Of the 98 patients analyzed, 28 (29.6%) were migrants, predominantly male (65%), with a mean age of 35.6 years. Pulmonary tuberculosis was present in 82% of patients in both groups, and nine cases of drug-resistant tuberculosis were identified. No significant differences were observed between groups in treatment cure rates, mortality, or hospitalization. Unfavourable outcomes—a composite endpoint comprising mortality, treatment failure, and loss to follow-up—were more frequent in males and in patients with elevated C-reactive protein (CRP) levels (p = 0.033) or a higher CRP/albumin ratio. Migrants accounted for a substantial proportion of total TB cases and tended to be younger, with fewer comorbidities and lower rates of substance use. They showed a non-significant trend toward higher loss to follow-up and drug resistance. Overall, unfavourable outcomes were associated with elevated CRP levels and the CRP/albumin ratio. Full article

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15 pages, 1806 KB

Open AccessArticle

Acute HSV-1 Ocular Infection Is Impaired in KLF15 Knockout Mice but Stress-Induced Reactivation from Latency Is Prolonged in Male KLF15 Knockout Mice

by Kelly S. Harrison and Clinton Jones

Pathogens 2025, 14(8), 823; https://doi.org/10.3390/pathogens14080823 - 20 Aug 2025

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Abstract

Acute human alpha-herpesvirus 1 (HSV-1) infection culminates in a latent infection of neurons in trigeminal ganglia (TG) and the central nervous system. Following infection of mucosal epithelial cells, certain neurons survive infection and life-long latency is established. Periodically, stressful stimuli trigger reactivation from [...] Read more.

Acute human alpha-herpesvirus 1 (HSV-1) infection culminates in a latent infection of neurons in trigeminal ganglia (TG) and the central nervous system. Following infection of mucosal epithelial cells, certain neurons survive infection and life-long latency is established. Periodically, stressful stimuli trigger reactivation from latency, which result in virus shedding, transmission to other people, and, occasionally, recurrent disease. The glucocorticoid receptor (GR) and Krüppel-like factor 15 (KLF15) comprise a feed-forward transcriptional loop that cooperatively transactivate key HSV-1 promoters that drive expression of infected cell protein 0 (ICP0), ICP4, and ICP27. Silencing KLF15 significantly reduces HSV-1 replication in cultured mouse neuroblastoma cells. Consequently, we hypothesized that KLF15 mediates certain aspects of reactivation from latency. To test this hypothesis, we compared HSV-1 replication in KLF15^−/− mice versus wild-type (wt) parental C57BL/6 mice. Virus shedding during acute infection was reduced in KLF15^−/− mice. Male KLF15^−/− mice shed higher titers of virus during late stages of reactivation from latency compared to KLF15^−/− females and wt mice regardless of sex. At 15 d after explant-induced reactivation, virus shedding was higher in male KLF15^−/− mice relative to wt mice and female KLF15^−/− mice. These studies confirm KLF15 expression enhances viral replication during acute infection and reactivation from latency. Full article

(This article belongs to the Section Viral Pathogens)

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11 pages, 377 KB

Open AccessArticle

Coronary Artery Disease in People Living with HIV May Reflect Their Sensitivity to Inflammation Associated with Cytomegalovirus

by Luna-faye Veld, Shelley Waters, Silvia Lee, Anna C. Hearps, Janine Trevillyan, Ari S. Mushin, Damien Foo, Jennifer Hoy and Patricia Price

Pathogens 2025, 14(8), 822; https://doi.org/10.3390/pathogens14080822 - 20 Aug 2025

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Abstract

Cytomegalovirus (CMV) is implicated in cardiovascular disease in healthy adults and after transplantation, but analyses in people living with HIV (PLWH) are difficult as almost all have CMV co-infections. Here, we address whether coronary artery disease (CAD) is associated with levels of CMV-reactive [...] Read more.

Cytomegalovirus (CMV) is implicated in cardiovascular disease in healthy adults and after transplantation, but analyses in people living with HIV (PLWH) are difficult as almost all have CMV co-infections. Here, we address whether coronary artery disease (CAD) is associated with levels of CMV-reactive antibodies or with sensitivity to inflammation associated with CMV. PLWH stable on antiretroviral therapy (ART) with a recent diagnosis of CAD were matched with PLWH without CAD. Plasma samples stored at the time of the CAD event and 6, 12, 24 or 36 months earlier (n = 34–55 per group) were used for analyses. Antibodies reactive with a lysate from CMV infected cells were quantitated using an in-house ELISA, and inflammatory biomarkers were assessed using commercial kits. Bivariate analyses demonstrated similar levels of CMV antibodies in PLWH with and without CAD at all time points (p > 0.5). However, in PLWH with CAD, levels of CMV antibody correlated directly with plasma sCD14, LBP, CXCL10 and/or IL-6 at the earlier points. These correlations were not impacted by detectable plasma HIV RNA. Our findings suggest that individual differences in sensitivity to the inflammatory effects of CMV impact upon the development of CAD. Full article

(This article belongs to the Special Issue Immune Pathways and Mechanisms Involved in Viral Infections)

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18 pages, 2226 KB

Open AccessArticle

The Clonal Spread and Persistence of Campylobacter in Danish Broiler Farms and Its Association with Human Infections

by Katrine Grimstrup Joensen, Gitte Sørensen, Pernille Gymoese, Louise Gade Dahl and Eva Møller Nielsen

Pathogens 2025, 14(8), 821; https://doi.org/10.3390/pathogens14080821 - 19 Aug 2025

Viewed by 447

Abstract

Campylobacter is the most common cause of bacterial foodborne illness in the EU, primarily linked to poultry. To better understand its transmission dynamics, we applied whole-genome sequencing (WGS) to Campylobacter isolates collected at slaughterhouses over a two-year period from broilers originating from 26 [...] Read more.

Campylobacter is the most common cause of bacterial foodborne illness in the EU, primarily linked to poultry. To better understand its transmission dynamics, we applied whole-genome sequencing (WGS) to Campylobacter isolates collected at slaughterhouses over a two-year period from broilers originating from 26 Danish farms. The samples included cloacal swabs and boot sock samples from broiler houses and surrounding farm environments. We identified 150 distinct cgMLST types among 883 isolates. While most cgMLST types were flock-specific, some persisted across production cycles or appeared at different farms, indicating entrenched contamination or potential common-source introductions. Notably, 39% of broiler-associated cgMLST types overlapped with human clinical isolates from the same period, with the strongest overlap among persistent and cross-farm types, particularly in conventional production systems. Our findings underscore the need for strengthened biosecurity, targeted surveillance of high-risk genotypes, and real-time WGS integration to mitigate the burden of human Campylobacteriosis. This study supports a One Health approach to managing zoonotic risk in poultry production. Full article

(This article belongs to the Special Issue Feature Papers on the Epidemiology of Infectious Diseases)

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18 pages, 4035 KB

Open AccessArticle

Antiviral Activity of Medicinal Plant Extracts Vitex negundo and Macaranga tanarius Against SARS-CoV-2

by Muhareva Raekiansyah, Mya Myat Ngwe Tun, Alexandra Ang, Alexandra Lee, Stephani Joy Macalino, Junie Billones, Yuki Takamatsu, Takeshi Urano, Lyre Anni E. Murao, Noel Quiming, Kouichi Morita and Maria Constancia Carrillo

Pathogens 2025, 14(8), 820; https://doi.org/10.3390/pathogens14080820 - 19 Aug 2025

Viewed by 718

Abstract

Natural products possess a wide range of biological and biochemical potentials, with plant-derived compounds being significant sources for discovering new drugs. In this study, extracts of Vitex negundo and Macaranga tanarius prepared with different solvents were tested for their antiviral activity against the [...] Read more.

Natural products possess a wide range of biological and biochemical potentials, with plant-derived compounds being significant sources for discovering new drugs. In this study, extracts of Vitex negundo and Macaranga tanarius prepared with different solvents were tested for their antiviral activity against the original SARS-CoV-2 Wuhan strain and its variants using plaque assay, quantitative real time RT-PCR, and immunofluorescence assay (IFA). Our results showed that at their maximum non-toxic concentrations, Vitex-Dichloromethane (DCM) and Macaranga extracts significantly inhibited SARS-CoV-2 Wuhan strain growth in Vero E6 cells, showing a 5-log reduction in plaque assay and confirmed by IFA. Meanwhile, Vitex-Hexane showed moderate activity with a 2-log decrease. The inhibition was shown in a dose-dependent manner. The antiviral efficacy of these extracts was further demonstrated against various SARS-CoV-2 variants including Alpha, Beta, Delta, and Omicron. Both Vitex-DCM and Macaranga showed strong virucidal activity. In addition, Vitex-DCM and Macaranga inhibited the transcriptional activity of purified SARS-CoV-2 RdRp, indicating that RdRp inhibition may contribute to viral suppression as shown at the post-infection stage. Furthermore, combining Vitex-DCM or Macaranga with remdesivir showed a synergistic effect against SARS-CoV-2. These results suggest that Vitex negundo and Macaranga tanarius extracts are promising candidates for anti-SARS-CoV-2 treatments. Their synergy with remdesivir also underscores the potential of drug combinations in fighting SARS-CoV-2 and preventing the emergence of mutant variants. Full article

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10 pages, 278 KB

Open AccessCase Report

Paid Organ Donation: Case Report and Review of the Literature on Health Implications for Kidney Donors and Recipients

by Małgorzata Marchelek-Myśliwiec, Krzysztof Korzeniewski, Emilia Marchelek, Joanna Stępniewska and Danuta Kosik-Bogacka

Pathogens 2025, 14(8), 819; https://doi.org/10.3390/pathogens14080819 - 19 Aug 2025

Viewed by 447

Abstract

The shortage of organs for use in transplantation has contributed to the development of an international commercial market for organ transplantation. Unfortunately, transplant tourism (TT) is associated with risks for surgical complications, poor graft outcome, increased mortality, and infectious complications. TT increases the [...] Read more.

The shortage of organs for use in transplantation has contributed to the development of an international commercial market for organ transplantation. Unfortunately, transplant tourism (TT) is associated with risks for surgical complications, poor graft outcome, increased mortality, and infectious complications. TT increases the risk of several viral (HIV and hepatitis B and C viruses), bacterial (Mycobacterium tuberculosis, Pseudomonas sp., Enterococcus sp., Escherichia coli, and Acinetobacter sp.), fungal (Aspergillus sp., Zygomycetes, Ramichloridium sp., Scedosporium apiospermum, and Trichosporon sp.), and parasitic (Plasmodium sp., Trypanosoma cruzi and Strongyloides sp., and Microsporidia sp.) infections. This paper presents a case report of an anonymous patient who travelled to Pakistan and underwent a commercial kidney transplant. He developed infection from extended-spectrum β-lactamase-producing Escherichia coli (ESBL-EC). Moreover, we reviewed all published cases of bacterial, viral, and fungal infections in kidney transplant recipients who bought their organs abroad. Full article

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20 pages, 984 KB

Open AccessReview

Maternal HIV Infection and Antiretroviral Therapy in Pregnancy: Implications for Vertical Transmission, Fetal Safety, and Long-Term Infant Outcomes

by Tudor Fleșeriu, Lorena Elena Meliț, Cristina Oana Mărginean, Adrian Vlad Pop and Anca-Meda Văsieșiu

Pathogens 2025, 14(8), 818; https://doi.org/10.3390/pathogens14080818 - 19 Aug 2025

Viewed by 737

Abstract

HIV mother-to-child transmission (MTCT) continues to pose a significant public health challenge, especially in regions with limited resources, although the worldwide distribution of antiretroviral therapy (ART) has drastically lowered the risk of vertical transmission to even below 1% in some regions. There are [...] Read more.

HIV mother-to-child transmission (MTCT) continues to pose a significant public health challenge, especially in regions with limited resources, although the worldwide distribution of antiretroviral therapy (ART) has drastically lowered the risk of vertical transmission to even below 1% in some regions. There are still uncertainties regarding the safety of some ART regimens during pregnancy and their longer-term effects on infants who are perinatally exposed to HIV but remain uninfected. This review explores current evidence regarding the interplay between maternal HIV infection, ART during pregnancy, and both maternal and pediatric outcomes. Particular attention is given to the risk/benefit ratio surrounding different drug classes, with integrase inhibitors seeming promising choices in MTCT due to their rapid viral suppression and favorable safety profiles. Meanwhile, regimens containing protease inhibitors or nucleoside reverse transcriptase inhibitors have been linked to some adverse outcomes such as low birth weight, growth restriction, and potential mitochondrial or metabolic disturbances. Although ART remains central in preventing MTCT, a deeper understanding of its effects on fetal development and postnatal health is needed, and it should be thoroughly monitored through future research and longitudinal surveillance. Full article

(This article belongs to the Special Issue Understanding HIV Pathogenesis and Targeting for the Prevention and Cure)

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11 pages, 1097 KB

Open AccessArticle

Development of a Droplet Digital PCR Assay Targeting the Internal Transcribed Spacer Gene for Rapid Detection of Talaromyces marneffei in AIDS Patients

by Yingyin Yang, Qinzhi Zhang, Pengle Guo, Meijun Chen, Yeyang Zhang, Weiping Cai, Xiaoping Tang and Linghua Li

Pathogens 2025, 14(8), 817; https://doi.org/10.3390/pathogens14080817 - 18 Aug 2025

Viewed by 437

Abstract

Talaromycosis caused by Talaromyces marneffei is a life-threatening mycosis in patients with acquired immunodeficiency syndrome (AIDS). The gold-standard diagnostic method relies on time-consuming cultures, which delay treatment and increase mortality. In this study, we developed a rapid and sensitive droplet digital PCR (ddPCR) [...] Read more.

Talaromycosis caused by Talaromyces marneffei is a life-threatening mycosis in patients with acquired immunodeficiency syndrome (AIDS). The gold-standard diagnostic method relies on time-consuming cultures, which delay treatment and increase mortality. In this study, we developed a rapid and sensitive droplet digital PCR (ddPCR) assay targeting the internal transcribed spacer (ITS) gene for detecting T. marneffei and compared its performance with blood culture and quantitative PCR (qPCR) assays. The ddPCR assay had a detection limit of one copy/reaction, making it 10-fold more sensitive than qPCR. It demonstrated 100% specificity for T. marneffei, with no cross-reactivity to 15 other fungal pathogens, six bacterial pathogens, and plasma from 119 AIDS patients without talaromycosis. In 119 AIDS patients with talaromycosis, ddPCR exhibited better overall sensitivity (92.44%) than blood culture (86.55%) and qPCR (87.29%). The sensitivity of ddPCR was 97.8% (89/91) and 75% (21/28) in plasma collected before and after antifungal therapy, respectively. Moreover, fungal load measured by ddPCR negatively correlated with the time to blood culture positivity. Fungal loads in patients receiving antifungal therapy were significantly lower than those in untreated patients. These findings indicate that ddPCR facilitates rapid diagnosis of T. marneffei infection in AIDS patients and can assist clinicians in evaluating treatment efficacy by quantifying fungal load. Full article

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30 pages, 361 KB

Open AccessReview

Risk of Venous Thromboembolism in Infectious Diseases: A Literature Review

by Ilaria Pati, Francesca Masiello, Vanessa Piccinini, Lucia De Fulvio, Maria Simona Massari, Vincenzo De Angelis and Mario Cruciani

Pathogens 2025, 14(8), 816; https://doi.org/10.3390/pathogens14080816 - 18 Aug 2025

Viewed by 647

Abstract

Systemic or localized infections increase the risk of venous thromboembolism (VTE). All types of infection can elevate the risk of VTE thrombosis, although some appear to increase risk more than others. In the current narrative review, we seek to overview the available evidence [...] Read more.

Systemic or localized infections increase the risk of venous thromboembolism (VTE). All types of infection can elevate the risk of VTE thrombosis, although some appear to increase risk more than others. In the current narrative review, we seek to overview the available evidence related to the epidemiology of VTE caused by infections. We focused on patients with infection in community setting or hospitalized, on patients with COVID-19, HIV infection, tuberculosis, HCV infection, and CMV infection, as well as on individuals with other types of infection that might increase the risk of VTE. Moreover, we tried to evaluate how the risk of VTE in person with different types of infections could be addressed in clinical practice with the use of anticoagulants. Extended VTE prophylaxis may not be warranted for all infections, but may be very helpful for some, such as those with intra-abdominal infection, systemic bloodstream infection, lower respiratory infection, and symptomatic urinary tract infection. Full article

21 pages, 4238 KB

Open AccessArticle

Relationship Between Cell Surface Viral Glycoprotein Expression and Resistance of Parainfluenza Virus Persistently Infected Cells to Complement-Mediated Lysis

by Nasser N. Yousef and Griffith D. Parks

Pathogens 2025, 14(8), 815; https://doi.org/10.3390/pathogens14080815 - 17 Aug 2025

Viewed by 387

Abstract

Persistent RNA virus infections (PI) are often characterized by extended viral shedding and maintained cycles of inflammation. The innate immune Complement (C′) pathways can recognize acute infected (AI) cells and result in their lysis, but the relative sensitivity of PI cells to C′-directed [...] Read more.

Persistent RNA virus infections (PI) are often characterized by extended viral shedding and maintained cycles of inflammation. The innate immune Complement (C′) pathways can recognize acute infected (AI) cells and result in their lysis, but the relative sensitivity of PI cells to C′-directed killing is incompletely understood. Here, we extended our previous studies on the interactions of C′ with parainfluenza virus AI and PI A549 cells to two additional respiratory tract cell lines. AI Hep2 and H1975 cells infected with Parainfluenza virus 5 (PIV5) were found to be highly sensitive to C′ lysis. By contrast, PIV5 PI cells were highly resistant to killing by C″. Surface deposition of membrane attack complex (MAC) and C3 was also greatly reduced on the surface of PI cells compared to AI cells. PI cells had lower levels of surface viral glycoprotein expression compared to AI cells. Treatment of AI cells with ribavirin (RBV) showed a dose-dependent decrease in both viral glycoprotein expression and sensitivity to C′-mediated lysis. When surface viral glycoprotein levels were reduced in AI cells to those in PI cells, AI cells became similarly resistant to C′. While sialic acid levels on PI cell surfaces matched that of naïve cells, enzymatic removal of this sialic acid did not increase sensitivity to C′-mediated lysis. Despite their varying profiles of C′ activation and deposition, these studies indicate downregulation of viral gene expression as a common mechanism of C′ resistance across various parainfluenza virus PI cell lines. Full article

(This article belongs to the Special Issue Virus–Host Cell Interactions and Research of New Antivirals)

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10 pages, 1184 KB

Open AccessArticle

Differential Exposure to Borrelia spp. and Spotted Fever Group Rickettsia in Serbia and North Macedonia: A Comparative Study

by Dejan Jakimovski, Sofija Mateska, Marija Najdovska, Angela Stamenkovska, Verica Pavleva, Mile Bosilkovski, Dragana Mijatović, Verica Simin, Ivana Bogdan, Jasmina Grujić, Milica Simeunović, Miodrag Vranješ, Eleftherios Meletis, Polychronis Kostoulas, Olympia Lioupi and Pavle Banović

Pathogens 2025, 14(8), 814; https://doi.org/10.3390/pathogens14080814 - 17 Aug 2025

Viewed by 466

Abstract

Several diseases caused by tick-borne pathogens, including Lyme borreliosis (LB) and spotted fever group rickettsioses, are endemic in the Balkan Peninsula, positioned between Central Europe and the Middle East. This cross-sectional study aimed to assess serological exposure to Borrelia spp. and spotted fever [...] Read more.

Several diseases caused by tick-borne pathogens, including Lyme borreliosis (LB) and spotted fever group rickettsioses, are endemic in the Balkan Peninsula, positioned between Central Europe and the Middle East. This cross-sectional study aimed to assess serological exposure to Borrelia spp. and spotted fever group Rickettsiae (SFGR) among individuals with recent tick bites and healthy controls in two Balkan countries—Serbia and North Macedonia. Serum samples from 223 participants were tested for anti-Borrelia and anti-SFGR IgG antibodies. SFGR exposure was significantly higher in tick-exposed individuals from Skopje (North Macedonia) compared to those from Novi Sad (Serbia) (30.9% vs. 8.0%; p = 0.003). In contrast, anti-Borrelia IgG was more frequently detected in Novi Sad, though differences did not reach statistical significance. The findings support a north-to-south gradient in Borrelia exposure and a reverse trend for SFGR, consistent with earlier studies and regional tick infection data. Given the high SFGR exposure and limited clinical reporting in North Macedonia, the results highlight the likelihood that tick-borne rickettsioses remain under-recognized. Additionally, Borrelia exposure in North Macedonia warrants further investigation. These findings emphasize the need for enhanced tick-borne disease surveillance, identification of endemic zones, and improved diagnostic and public health infrastructure in both countries. Full article

(This article belongs to the Section Ticks)

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17 pages, 1502 KB

Open AccessReview

Perspectives on the History and Epidemiology of the Varicella Virus Vaccine and Future Challenges

by Masayuki Nagasawa

Pathogens 2025, 14(8), 813; https://doi.org/10.3390/pathogens14080813 - 16 Aug 2025

Viewed by 537

Abstract

The varicella attenuated virus vaccine, developed in Japan in the 1970s, has dramatically reduced the number of pediatric chickenpox cases over the past 30 years due to its widespread use. However, a small number of cases of chickenpox, shingles, aseptic meningitis, and acute [...] Read more.

The varicella attenuated virus vaccine, developed in Japan in the 1970s, has dramatically reduced the number of pediatric chickenpox cases over the past 30 years due to its widespread use. However, a small number of cases of chickenpox, shingles, aseptic meningitis, and acute retinal necrosis caused by vaccine strains have been reported. There are also issues that need to be addressed, such as breakthrough infections and the persistence of the preventive effect of vaccination. In addition, there is the possibility of the emergence of revertants or mutations in the vaccine strain. In recent years, subunit vaccines have been developed, their immune-stimulating effects have been demonstrated, and they are being applied clinically. In addition, development of an mRNA varicella vaccine is underway. In this review, the history and impact of the varicella vaccine are overviewed, as well as its future challenges. Full article

(This article belongs to the Special Issue Emerging and Re-Emerging Human Infectious Diseases)

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14 pages, 2032 KB

Open AccessArticle

Pandemic Disruptions to Private Pathology Testing Uptake in Australia, 2019–2024

by Melanie Keech, Shane Kavanagh, John Crothers, Liliana Orellana and Catherine M. Bennett

Pathogens 2025, 14(8), 812; https://doi.org/10.3390/pathogens14080812 - 15 Aug 2025

Viewed by 380

Abstract

A new human pathogen triggering a pandemic can impact health directly through disease resulting from infection and indirectly through health system disruption. The COVID-19 pandemic is hypothesised to have impacted pathology testing by impacting healthcare and pathology operations and reducing healthcare attendance for [...] Read more.

A new human pathogen triggering a pandemic can impact health directly through disease resulting from infection and indirectly through health system disruption. The COVID-19 pandemic is hypothesised to have impacted pathology testing by impacting healthcare and pathology operations and reducing healthcare attendance for fear of infection. The impacts of COVID-19 incidence and pandemic control measures on non-COVID pathology testing were assessed in four Australian states/territories using pathology data (histology, prostate-specific antigen, gynaecological cytology, complete blood count, haemoglobin A1c, and human immunodeficiency virus) from a large national private pathology provider (January 2019–December 2024). Weekly testing volumes from lockdown periods were compared to the equivalent weeks in 2019. All pathology tests demonstrated a substantial decline during the initial national lockdown in March 2020. Subsequent lockdowns were also associated with disruption. For example, complete blood count testing in Victoria was −22% in March 2020 and −5% in the second wave that year. Total annual testing volumes were lower for all tests in 2020 compared to 2019, excluding haemoglobin A1c, and reduced testing persisted through to 2024. The findings indicate substantial and sustained negative pandemic impacts on pathology testing. Reductions in pathology testing signal heightened risk of delayed disease diagnosis, disrupted chronic disease management, and poorer health outcomes. Full article

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6 pages, 168 KB

Open AccessCommunication

A Cell-Free DNA Plasma Next-Generation Sequencing Test—Is It Worth the Cost?

by Sean Jung, Francesca Torriani, Shira Abeles and Ahnika Kline

Pathogens 2025, 14(8), 811; https://doi.org/10.3390/pathogens14080811 - 15 Aug 2025

Viewed by 511

Abstract

Background: The Karius Test (KT), a microbial cell-free DNA next-generation sequencing assay, is increasingly utilized in challenging infectious syndromes. However, its real-world clinical utility and cost-effectiveness remain uncertain. Methods: We conducted a retrospective review of 88 KT results from adult patients at UC [...] Read more.

Background: The Karius Test (KT), a microbial cell-free DNA next-generation sequencing assay, is increasingly utilized in challenging infectious syndromes. However, its real-world clinical utility and cost-effectiveness remain uncertain. Methods: We conducted a retrospective review of 88 KT results from adult patients at UC San Diego Health between July 2017 and April 2024. Each case was evaluated for clinical impact using standardized criteria. We analyzed diagnostic yield, turnaround time, number needed to test (NNT), and institutional billing data for reimbursement and cost implications. Results: Of 88 unique tests, forty-nine (55.7%) identified at least one pathogen. Eleven (12.5%) had a positive clinical impact—eight provided the only microbiologic diagnosis, and three were faster than conventional methods. Vascular/graft infections showed the highest yield. Twenty-one tests had a neutral impact; fifty-six showed no clinical benefit. The Median turnaround time was 3 days. The NNT was 6.1 or 2.75 including neutral cases. Cost analysis revealed a substantial financial burden without transparent reimbursement mechanisms in inpatient settings. Conclusion: The KT demonstrates modest clinical utility with noteworthy benefits in select scenarios. Given its high cost and variable impact, we advocate for diagnostic stewardship led by infectious disease specialists to optimize test use and minimize unnecessary expense. Full article

10 pages, 2586 KB

Open AccessCase Report

Comprehensive Laboratory Analysis of a Scrub Typhus and H1N1 Influenza Co-Infection: A Case Report from Hainan, China

by Siqi Chen, Fahui Wang, Shannan Wu, Yuanze Chen, Yi Niu, Yijia Guo, Dachuan Lin, Xiuji Cui, Ruoyan Peng, Zhao Xu, Biao Wu, Min Liao, Yongguo Du, Liyuan Zhang and Feifei Yin

Pathogens 2025, 14(8), 810; https://doi.org/10.3390/pathogens14080810 - 15 Aug 2025

Viewed by 389

Abstract

Co-infection of Orientia tsutsugamushi and influenza A virus complicates diagnosis and treatment in endemic regions because of overlapping clinical features and potential synergistic inflammation. We describe a 68-year-old woman from Hainan, China, who presented with five days of high fever (39.2 °C), nonproductive [...] Read more.

Co-infection of Orientia tsutsugamushi and influenza A virus complicates diagnosis and treatment in endemic regions because of overlapping clinical features and potential synergistic inflammation. We describe a 68-year-old woman from Hainan, China, who presented with five days of high fever (39.2 °C), nonproductive cough, eschar formation, lymphadenopathy, cytopenias, elevated liver enzymes, and raised inflammatory markers. On the day of admission, influenza A was confirmed by rapid antigen test and Orientia tsutsugamushi IgM/IgG was detected via colloidal-gold immunochromatography, prompting concurrent oseltamivir and doxycycline therapy. Quantitative PCR on day 2 measured an Orientia tsutsugamushi load of 2.85 × 10⁴ copies/mL (Cq 28.86), and targeted next-generation sequencing on day 3 revealed a high H1N1pdm09 viral burden (>1 × 10⁶ copies/mL) with low-level human herpesvirus 1 co-detection. Nested PCR and Sanger sequencing assigned Orientia tsutsugamushi to the Karp_A lineage and influenza A to clade 6B.1A.5a.2a. The patient defervesced by hospital day 2, laboratory indices normalized by day 3, and radiographic abnormalities resolved by day 6. This first documented Orientia tsutsugamushi–influenza A co-infection in China highlights the value of integrating rapid serology, qPCR quantification, nested PCR genotyping, and tNGS for early, precise dual-pathogen identification. Systematic multi-pathogen screening during overlapping transmission seasons is recommended to guide timely combination therapy and enhance epidemiological surveillance. Full article

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13 pages, 3546 KB

Open AccessArticle

Ticks of the Genus Amblyomma and Lice of the Genus Eutrichophilus: Potential Vectors of Brazilian Porcupinepox Virus

by Nathana Beatriz Martins, Matias Pablo Juan Szabó, Julio Cesar de Souza Junior, Mario Henrique Alves, Marcio de Barros Bandarra, Paulo Eduardo Brandão and Aline Santana da Hora

Pathogens 2025, 14(8), 809; https://doi.org/10.3390/pathogens14080809 - 15 Aug 2025

Viewed by 466

Abstract

Brazilian porcupinepox virus (BPoPV) is a recently described pathogen associated with severe cutaneous and systemic disease in Coendou spp. porcupines, posing potential conservation and zoonotic risks. Given the solitary behavior of porcupines and the unclear mechanisms of BPoPV transmission, this study investigated the [...] Read more.

Brazilian porcupinepox virus (BPoPV) is a recently described pathogen associated with severe cutaneous and systemic disease in Coendou spp. porcupines, posing potential conservation and zoonotic risks. Given the solitary behavior of porcupines and the unclear mechanisms of BPoPV transmission, this study investigated the presence of BPoPV DNA in porcupines and their associated ectoparasites (ticks and lice). We analyzed ticks and lice collected from 17 porcupines (C. longicaudatus and C. spinosus), with or without clinical signs of BPoPV infection. Ectoparasites were identified morphologically, separated into distinct pools for ticks and lice by host, and screened by PCR. BPoPV DNA was detected in all symptomatic porcupines and their ectoparasites—including Amblyomma longirostre, A. sculptum ticks, and Eutrichophilus spp. lice—except for one tick pool. Notably, an asymptomatic, BPoPV-negative porcupine harbored A. longirostre ticks that tested positive for the virus. Sequencing confirmed 100% identity with the BPoPV reference strain. These findings suggest that Eutrichophilus lice, A. sculptum, and particularly A. longirostre ticks may play a role in BPoPV transmission. Further studies are needed to elucidate whether these ectoparasites act as biological or mechanical vectors and to assess the zoonotic potential of BPoPV in contact with humans and domestic animals. Full article

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

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11 pages, 1158 KB

Open AccessArticle

Can Oral Fluids Replace Nasal Swabs in Swine Influenza A Virus (swIAV) PCR Diagnostics?

by Aleksandra Woźniak, Piotr Cybulski, Pia Ryt-Hansen, Lars Erik Larsen, Kinga Biernacka, Dagmara Miłek and Tomasz Stadejek

Pathogens 2025, 14(8), 808; https://doi.org/10.3390/pathogens14080808 - 14 Aug 2025

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Abstract

The diagnosis of swine influenza A virus (swIAV) has to involve laboratory detection, as the clinical signs are not pathognomonic. Nasal swabs (NSs) have been the preferred sample material for swIAV PCR diagnostics, but oral fluid (OF) is a convenient alternative material. In [...] Read more.

The diagnosis of swine influenza A virus (swIAV) has to involve laboratory detection, as the clinical signs are not pathognomonic. Nasal swabs (NSs) have been the preferred sample material for swIAV PCR diagnostics, but oral fluid (OF) is a convenient alternative material. In this study, NSs and OFs from 35 Polish swine herds were collected and tested with real-time RT-PCR in order to assess swIAV circulation patterns in Poland and improve protocols for efficient, non-invasive and cost-effective swIAV surveillance in pig farms. The study showed that the swIAV RNA was detected in 65.7% of the tested farms. In total, 21.2% of NS pools and 48.6% of OF samples were positive for swIAV. The Ct values in NS pools and OFs were similar (p > 0.05), but a significant reduction (p < 0.05) in swIAV prevalence in NSs was observed in nursery pigs from farms applying swIAV vaccinations. Successful subtyping was achieved more effectively with OFs compared to NSs, and the H1avN2 was most prevalent subtype detected. The results emphasized that OF can be useful for monitoring swIAV and subtyping. However, OFs cannot replace NSs, which were more useful in the assessment of the effect of swIAV vaccinations in nursery pigs. Full article

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

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17 pages, 2415 KB

Open AccessArticle

Dancing with the Dust Devil: Examining the Lung Mycobiome of Sonoran Desert Wild Mammals and the Effect of Coccidioides Presence

by Ana Fabio-Braga, Jaida Salois, Mitchell L. Bryant, Daniel R. Kollath and Bridget Barker

Pathogens 2025, 14(8), 807; https://doi.org/10.3390/pathogens14080807 - 14 Aug 2025

Viewed by 466

Abstract

Microbiome studies report a decrease in diversity associated with active infections. Under the endozoan hypothesis, Coccidioides can inhabit a host without causing disease. In this study, we describe and compare the lung mycobiome of Coccidioides-positive and -negative samples obtained from wildlife. If [...] Read more.

Microbiome studies report a decrease in diversity associated with active infections. Under the endozoan hypothesis, Coccidioides can inhabit a host without causing disease. In this study, we describe and compare the lung mycobiome of Coccidioides-positive and -negative samples obtained from wildlife. If Coccidioides is not causing infection, we predict there will be no differences in the mycobiome between positive and negative samples. Lung samples were obtained from mammals previously trapped in Tucson, Arizona, USA (n = 26), and Mesa, Arizona, USA (n = 14). Samples were screened for Coccidioides with CocciDx, and mycobiome was characterized through Illumina-based amplicon sequencing of the internal transcribed spacer 2 (ITS2). We compared alpha and beta diversity of the mycobiome to assess the effects of Coccidioides’ presence and host taxonomy. A greater number of reads were captured from Tucson samples (114,706.4 ± 57,945.8) than from Mesa (384.9 ± 953.5); however, Mesa (16.8 ± 8.8) and Tucson (12 ± 7.8) had a similar number of fungal genera per sample. CocciDx detected Coccidioides in more samples than the ITS2 amplicon sequencing. All samples from Mesa and five from Tucson tested positive for Coccidioides. Therefore, Mesa samples were excluded from statistical analysis. No difference in alpha and beta diversity was associated with Coccidioides presence, which is consistent with the endozoan hypothesis. Host taxonomy had a significant effect on beta diversity. This effect is likely driven by host behavioral and physiological differences. Full article

(This article belongs to the Special Issue Epidemiology and Molecular Detection of Emerging Fungal Pathogens)

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35 pages, 1649 KB

Open AccessReview

Candidemia: An Update on Epidemiology, Risk Factors, Diagnosis, Susceptibility, and Treatment

by Juan Pablo Cabrera-Guerrero, Eduardo García-Salazar, Graciela Hernandez Silva, Alberto Chinney Herrera, Erick Martínez-Herrera, Rodolfo Pinto-Almazán, María Guadalupe Frías-De-León and Carlos Alberto Castro-Fuentes

Pathogens 2025, 14(8), 806; https://doi.org/10.3390/pathogens14080806 - 14 Aug 2025

Viewed by 1011

Abstract

Candidemia is a highly prevalent invasive fungal infection caused primarily by C. albicans, C. parapsilosis, C. glabrata (currently Nakaseomyces glabratus), C. tropicalis, and C. krusei (currently Pichia kudriavzevii). Risk factors for the development of candidemia include steroid-induced immunosuppression [...] Read more.

Candidemia is a highly prevalent invasive fungal infection caused primarily by C. albicans, C. parapsilosis, C. glabrata (currently Nakaseomyces glabratus), C. tropicalis, and C. krusei (currently Pichia kudriavzevii). Risk factors for the development of candidemia include steroid-induced immunosuppression used in solid organ or hematopoietic transplantation, and neutropenia secondary to infectious or tumorous processes. Alterations in the gut microbiota in people living with HIV, caused by antiretroviral therapy, increase the possibility of colonization by C. albicans. Likewise, the presence of a central venous catheter, parenteral nutrition, and abdominal surgery stand out as the main risk factors for the development of candidemia. New diagnostic tools have been developed for the diagnosis of this mycosis that allow the identification of the main species, from improvements in conventional stains such as calcofluor white, which increases sensitivity, as well as technologies such as T2 Candida, MoiM assay, biomarker panel (1,3 β-D-glucan, C-reactive protein, presepsin, and procalcitonin), and, more recently, the development of biosensors for the identification of Candida spp. Regarding treatment, the use of micafungin and anidulafungin in patients with obesity defined by a BMI > 30 kg/m² has shown higher survival rates and therapeutic success. Meanwhile, newer antifungals such as rezafungin and fosmanogepix have demonstrated excellent results in the treatment of these patients. Therefore, this review aims to update the epidemiology and risk factors of candidemia, as well as analyze the diagnostic tools and treatments currently available. Full article

(This article belongs to the Special Issue An Update on Fungal Infections)

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10 pages, 351 KB

Open AccessArticle

Serotype Distribution of Aggregatibacter actinomycetemcomitans in Periodontitis Patients

by Nabil Khzam, Omar Kujan, Dorte Haubek and Leticia Algarves Miranda

Pathogens 2025, 14(8), 805; https://doi.org/10.3390/pathogens14080805 - 13 Aug 2025

Viewed by 404

Abstract

Aim: The aim of the current investigation was to detect serotypes of Aggregatibacter actinomycetemcomitans in a cohort of Western Australians diagnosed with periodontitis. Materials and Methods: A total of 64 subjects were selected. Intra-oral samples were taken from every subject in the present [...] Read more.

Aim: The aim of the current investigation was to detect serotypes of Aggregatibacter actinomycetemcomitans in a cohort of Western Australians diagnosed with periodontitis. Materials and Methods: A total of 64 subjects were selected. Intra-oral samples were taken from every subject in the present investigation. Periodontal, radiographical, and microbiological analyses were conducted. A polymerase chain reaction was employed to investigate the presence of Aggregatibacter actinomycetemcomitans serotypes. Results: Only twelve (18.75%) patients were tested positive for Aggregatibacter actinomycetemcomitans. The most dominant serotypes of Aggregatibacter actinomycetemcomitans in this group were serotype e (80.55%), followed by serotype c (52.77%). Both serotypes b and d were absent in the present investigation. Serotype e presented in isolation or combined with other serotypes. The other serotypes tend to be present alone, but when they were isolated together, they were always combined with serotype e. It seems that serotype e of Aggregatibacter actinomycetemcomitans is associated with those who live in rural areas (p = 0.003), and those with low education (p = 0.041), and severe forms of periodontitis in this cohort. Conclusions: In patients diagnosed with severe periodontitis, serotype e was dominant in this population. Serotypes b and d did not appear in the present study. Full article

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