Pathogens

20 pages, 2181 KiB

Open AccessArticle

Genetic Identification of Brazilian Mammalian Hosts of Trypanosoma cruzi: Improving Blood Meal Source Discrimination in Vector-Borne Transmission

by Quezia Moura Oliveira, Thaíla Santos Pessanha and Alena Mayo Iñiguez

Pathogens 2025, 14(6), 579; https://doi.org/10.3390/pathogens14060579 - 10 Jun 2025

Abstract

The detection of food sources of blood-sucking vectors is essential for a better understanding of the hosts, reservoirs, and other fauna that participate in the transmission web of hemoparasites. The molecular identification of triatomine blood meal sources (BMSs) has been shown to be [...] Read more.

The detection of food sources of blood-sucking vectors is essential for a better understanding of the hosts, reservoirs, and other fauna that participate in the transmission web of hemoparasites. The molecular identification of triatomine blood meal sources (BMSs) has been shown to be highly sensitive and taxonomically specific when compared to the immunological method. The application of molecular cloning makes it possible to identify multiple BMS species and/or different individuals/haplotypes of the same vertebrate species in a single triatomine specimen. In Brazil, the molecular detection of BMSs is incipient, with insufficient genetic information on the species of animals involved in the transmission of Trypanosoma cruzi. In this work, we evaluated the sensitivity and specificity of a molecular approach using molecular cloning for the detection of multiple Brazilian mammalian species. The DNA was extracted from blood clots of 13 species of canids, bats, xenarthral, marsupials, and rodents. Serial proportions were used to formulate mixtures combining taxonomically close (belonging to the same family or order) and taxonomically distant (different families) species. The results showed that GenBank lacks reference sequences for some native species tested, such as the sylvatic rodent, Necromys lasiurus, and the wild canid, Lycalopex gymnocercus, for cytb and 12S rDNA, and the rodent Oecomys cleberi for 12S rDNA. The study also demonstrated that it is possible to detect multiple different species, even for those that are taxonomically close. This approach was proven to be efficient for the detection of species in equal and even in disparate unequal proportions, which could represent complementary information about the diversity of potential hosts of T. cruzi. The detection of multiple BMS species in mixed samples provides a more comprehensive and accurate landscape of T. cruzi transmission in nature. Full article

(This article belongs to the Special Issue Chagas Disease: Celebrating the 115th Anniversary of the Discovery of Trypanosoma cruzi)

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Graphical abstract

13 pages, 602 KiB

Open AccessSystematic Review

An Umbrella Review of E-Cigarettes’ Impact on Oral Microbiota and Biofilm Buildup

by Beatriz Panariello, Fabrízio Dias Panariello, Ashminie Misir and Eliane Porto Barboza

Pathogens 2025, 14(6), 578; https://doi.org/10.3390/pathogens14060578 - 10 Jun 2025

Abstract

E-cigarettes, a form of electronic nicotine delivery system (ENDS), have gained significant popularity, particularly among adolescents who often view vaping as a “cool” lifestyle choice. This growing trend has spurred extensive research on the effects of ENDS on both oral and systemic health. [...] Read more.

E-cigarettes, a form of electronic nicotine delivery system (ENDS), have gained significant popularity, particularly among adolescents who often view vaping as a “cool” lifestyle choice. This growing trend has spurred extensive research on the effects of ENDS on both oral and systemic health. By synthesizing data from systematic reviews and meta-analyses, this umbrella review offers a comprehensive evaluation of the impact of e-cigarettes on oral biofilm accumulation and microbiota composition. A systematic search was conducted up to 12 March 2025, across PubMed/MEDLINE, Google Scholar, Cochrane Library, and Scopus for studies published between 2015 and 2025. Ten studies met the eligibility criteria. The quality of the selected papers, as assessed using the AMSTAR 2 tool, ranged from moderate to high. The findings of this review suggest that e-cigarette use may contribute to dysbiosis in the oral microbiota and foster biofilm accumulation, thereby increasing the risk of oral diseases such as periodontitis, peri-implantitis, oral candidiasis, and caries. The findings also highlight the need for further research into the long-term effects of e-cigarette use on oral health. This review is registered with PROSPERO (CRD420251025639). Full article

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18 pages, 4191 KiB

Open AccessBrief Report

Investigation of the Pathogenesis of Lumpy Skin Disease Virus in Indigenous Cattle in Kazakhstan

by Lespek Kutumbetov, Ainur Ragatova, Moldir Azanbekova, Balzhan Myrzakhmetova, Nurbek Aldayarov, Kuandyk Zhugunissov, Yergali Abduraimov, Raikhan Nissanova, Asylay Sarzhigitova, Nazerke Kemalova and Arman Issimov

Pathogens 2025, 14(6), 577; https://doi.org/10.3390/pathogens14060577 - 10 Jun 2025

Abstract

This study investigates the virulence properties and pathogenetic characteristics of the Kazakhstani strain of LSDV (LSDV KZ-Kostanay-2018) in indigenous cattle under controlled conditions. Twelve non-breed cattle were inoculated intradermally and monitored for clinical, pathological, and immunological responses. Clinical signs, including fever, skin nodules, [...] Read more.

This study investigates the virulence properties and pathogenetic characteristics of the Kazakhstani strain of LSDV (LSDV KZ-Kostanay-2018) in indigenous cattle under controlled conditions. Twelve non-breed cattle were inoculated intradermally and monitored for clinical, pathological, and immunological responses. Clinical signs, including fever, skin nodules, and lymphadenopathy, emerged as early as day 5 post-infection (pi), with peak severity observed between days 11 and 14. Rapid seroconversion was observed, with 100% of animals showing virus-neutralizing antibodies by day 13. Pathological findings revealed extensive necrosis, thrombosis, and edema, with pronounced damage in the spleen, lungs, and lymph nodes. Histological analyses identified widespread destructive changes in the dermis and systemic tissues, consistent with highly aggressive disease progression. Viral genome and replication were confirmed in blood, skin nodules, and lymph nodes, with peak viral loads between days 11 and 14 pi. These results align with findings in Russian cattle infected with the Saratov/2017 strain but demonstrate more rapid symptom onset and severe pathology, suggesting strain-specific virulence. These findings contribute to a deeper understanding of LSDV pathogenesis and underscore the importance of regional adaptations in disease management. Full article

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

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16 pages, 339 KiB

Open AccessArticle

Humoral and T-Cell-Mediated Immunity Against Phlebotomus perniciosus Salivary Proteins in Dogs from a Leishmaniosis-Endemic Area

by Núria Balsells-Aguilar, Maria Magdalena Alcover, Marta Baxarias, Alejandra Álvarez-Fernández, Lourdes Alarcón, Petra Sumova, Petr Volf and Laia Solano-Gallego

Pathogens 2025, 14(6), 576; https://doi.org/10.3390/pathogens14060576 - 10 Jun 2025

Abstract

Compounds in sand fly saliva elicit specific immune responses that may play a role in the establishment of canine Leishmania infection. Although canine antibodies to anti-sand fly saliva antigens have been extensively studied, little is known about cellular immune responses against Phlebotomus perniciosus [...] Read more.

Compounds in sand fly saliva elicit specific immune responses that may play a role in the establishment of canine Leishmania infection. Although canine antibodies to anti-sand fly saliva antigens have been extensively studied, little is known about cellular immune responses against Phlebotomus perniciosus salivary proteins. This study aimed to explore humoral and T-cell-mediated immunity against P. perniciosus salivary proteins in dogs (n = 85) from Mallorca (Spain), a leishmaniosis-endemic area, and find correlations with demographic (age, sex, and breed) and parasite-specific immunological parameters. Anti-sand fly saliva IgG was examined using a P. perniciosus whole salivary gland homogenate (SGH) ELISA and recombinant salivary protein rSP03B ELISA. Interferon gamma (IFN-γ) release whole blood assays with L. infantum soluble antigen (LSA), SGH, and rSP03B were also performed. Positive correlations were found between IgG levels in the SGH and rSP03B tests and between concentrations of SGH IFN-γ and rSP03B IFN-γ. While concentrations of SGH IFN-γ and rSP03B IFN-γ were low and produced only by a minority of dogs (less than 20%), high levels and frequencies of LSA IFN-γ as well as anti-saliva IgG for SGH and rSP03B were detected in a majority of dogs (61% and 75%, respectively). LSA IFN-γ levels were positively correlated with age and Leishmania-specific antibodies. In conclusion, dogs from a leishmaniosis-endemic area presented high humoral immunity against P. perniciosus salivary proteins, but their cellular immunity to these proteins was low and less frequent. Full article

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

10 pages, 214 KiB

Open AccessArticle

Comparative Evaluation of a Multistrain Indirect ELISA Targeting Anti- p26 and gp45 Antibodies for EIAV Detection

by Angela Ostuni, Raffaele Frontoso, Maria Antonietta Crudele, Lorella Barca, Mario Amati, Raffaele Boni, Jolanda De Vendel, Paolo Raimondi and Alfonso Bavoso

Pathogens 2025, 14(6), 575; https://doi.org/10.3390/pathogens14060575 - 8 Jun 2025

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Abstract

Equine Infectious Anemia Virus (EIAV), a lentivirus marked by considerable genetic variability, poses significant diagnostic challenges. Existing diagnostic tools encompass the Agar Gel Immunodiffusion Assay (AGID), enzyme-linked immunosorbent assay (ELISA), and Western blotting (WB). ELISA and AGID mainly utilize the p26 capsid protein, [...] Read more.

Equine Infectious Anemia Virus (EIAV), a lentivirus marked by considerable genetic variability, poses significant diagnostic challenges. Existing diagnostic tools encompass the Agar Gel Immunodiffusion Assay (AGID), enzyme-linked immunosorbent assay (ELISA), and Western blotting (WB). ELISA and AGID mainly utilize the p26 capsid protein, often sourced from the Wyoming reference strain. To broaden the range of viral proteins and strains employed in these immunoassays, we previously developed a novel p26/double-strain gp45 indirect ELISA. In this study, we evaluated the performance of this ELISA in comparison to two commercial EIAV ELISAs using Cohen’s Kappa test and Bayesian Latent Class Analysis (BLCA), a statistical method that estimates test performance without requiring a perfect reference standard. A comparison with the official classification of the sera by the Italian Veterinary Service was also performed. A total of 372 serum samples, including 96 that were positives by all three tests, were analyzed. Results from both Cohen’s Kappa test and BLCA, alongside comparison with official classifications, affirm the diagnostic reliability of the two commercial ELISAs and suggest that the novel ELISA, with its enhanced antigenic diversity, could offer an accurate and reliable diagnostic option for EIAV. This novel assay enhances existing commercial ELISAs and has the potential to strengthen routine diagnostic workflows. Full article

11 pages, 515 KiB

Open AccessArticle

Systemic Antifolate Chemotherapy Does Not Select for Fluconazole-Resistant Candida: A Multicenter Clinical Study

by Dawid Żyrek, Joanna Nowicka, Magdalena Pajączkowska, Mariola Paściak, Katarzyna Machnik, Tomasz Werner, Zygmunt Konieczny, Piotr Jędrzejczak, Dominika Raźniewska, Gabriela Fijałkowska, Michał Piątek, Barbara Radecka, Kinga Żyrek, Elżbieta Woźniak-Grygiel and Iwona Dzieńdziora-Urbińska

Pathogens 2025, 14(6), 574; https://doi.org/10.3390/pathogens14060574 - 7 Jun 2025

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Abstract

Previous studies have demonstrated that Candida spp. isolates exposed in vitro to the folic acid antagonist methotrexate may develop multidrug cross-resistance to azole antifungals. The aim of this study was to determine whether systemic therapy with antineoplastic antifolates—pemetrexed or methotrexate—constitutes a risk factor [...] Read more.

Previous studies have demonstrated that Candida spp. isolates exposed in vitro to the folic acid antagonist methotrexate may develop multidrug cross-resistance to azole antifungals. The aim of this study was to determine whether systemic therapy with antineoplastic antifolates—pemetrexed or methotrexate—constitutes a risk factor for colonization or infection with fluconazole-resistant yeasts. The study group comprised 44 cancer patients who received high-dose systemic antifolate therapy, while the control group consisted of 48 patients without prior exposure to either methotrexate or pemetrexed. Oral swabs and relevant clinical data were collected from all participants. In total, 109 fungal strains representing 13 species were isolated, identified, and subsequently tested for fluconazole susceptibility. Fluconazole-resistant isolates were identified in 4 out of 44 (9.1%) antifolate-treated patients and in 3 out of 48 (6.3%) control patients. Our findings suggest that, although this phenomenon occurs in vitro, systemic antineoplastic antifolate therapy does not induce azole resistance among endogenous yeast species in vivo. Full article

(This article belongs to the Special Issue New Insights into Fungal Pathogenesis, Mechanisms of Virulence and Resistance to Antifungals)

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29 pages, 2256 KiB

Open AccessArticle

Longitudinal Monitoring of Mono- and Coinfections Involving Primary Porcine Reproductive Viruses (PCV2, PPV1, and PRRSV) as Well as Emerging Viruses (PCV3, PCV4, and nPPVs) in Primiparous and Multiparous Sows and Their Litters

by Diana S. Vargas-Bermudez, Gina Polo, Jose Dario Mogollon and Jairo Jaime

Pathogens 2025, 14(6), 573; https://doi.org/10.3390/pathogens14060573 - 7 Jun 2025

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Abstract

Porcine reproductive failure (PRF) has multiple etiological origins, primarily involving the viruses PCV2, PPV1, and PRRSV. Some emerging viruses, such as PCV3, PCV4, and novel parvoviruses (nPPVs), have also been suggested as contributors. In this study, we longitudinally evaluated 40 healthy sows (20 [...] Read more.

Porcine reproductive failure (PRF) has multiple etiological origins, primarily involving the viruses PCV2, PPV1, and PRRSV. Some emerging viruses, such as PCV3, PCV4, and novel parvoviruses (nPPVs), have also been suggested as contributors. In this study, we longitudinally evaluated 40 healthy sows (20 gilts and 20 multiparous sows) over three phases: pregnancy (PP), farrowing (FP), and their litters during lactation (LP). We detected viruses through PCR and serology in mono- and coinfections. The results showed that primary viruses were present during all three phases, with PCV2 being the most frequently detected. PCV3 positivity was highest at the time of insemination, and PPV1 was found in all. Additionally, PPV1-positive fetuses and pre-suckling piglets were identified, indicating vertical transmission. PRRSV was also present in an unstable herd, with the PRRSV2 lineage A detected and evidence of vertical transmission. The majority of coinfections were either dual or triple. The most common coinfections in the PP and LP were PCV2/PPV1 and PCV2/PCV3, while in the FF, PCV2/PPV1 and PCV2/PRRSV predominated. Notably, coinfection PCV2/PPV1 impacted the replication of PCV2. In contrast, the likelihood of detecting PRRSV decreased in fetuses coinfected with PRRSV and either PCV2, PCV3, or PPV1. The detected viruses exhibited low viral loads, indicating subclinical infections. Therefore, we propose recognizing a subclinical presentation of PRF and establishing criteria to differentiate between this and symptomatic reproductive disease. Full article

(This article belongs to the Special Issue Swine Infectious Diseases: Diagnostics, Pathogenesis, and Control Strategies)

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13 pages, 2629 KiB

Open AccessArticle

Prevalence and Toxoplasma gondii Genotypes Circulating in Five Wild Corvid Species from Romania

by Călin Mircea Gherman, Anamaria Balea, Adriana Györke, Zsuzsa Kalmár, Angela Monica Ionică, Isabelle Villena, Furio Spano, Stéphane de Craeye and Vasile Cozma

Pathogens 2025, 14(6), 572; https://doi.org/10.3390/pathogens14060572 - 7 Jun 2025

Viewed by 96

Abstract

The apicomplexan parasite Toxoplasma gondii can potentially infect all warm-blooded animals, including birds, which, due to their high dispersal capabilities, are considered a significant candidate group of sentinel animals that reveal environmental contamination with this protozoan. In the present study, the serologic and [...] Read more.

The apicomplexan parasite Toxoplasma gondii can potentially infect all warm-blooded animals, including birds, which, due to their high dispersal capabilities, are considered a significant candidate group of sentinel animals that reveal environmental contamination with this protozoan. In the present study, the serologic and molecular prevalences of T. gondii infection were determined in 333 corvids from Romania. Paired meat juice (n = 333) and heart samples (n = 244) were collected and analyzed using the modified agglutination test for antibodies, polymerase chain reaction (PCR) for DNA, and SAG2 molecular marker sequencing for genotyping. The overall T. gondii antibodies prevalence was 19.5%, with 48.1% infected jackdaws, 72.8% rooks, 89.7% hooded crows, 77.5% magpies, and 42.9% jays. Of 244 heart samples analyzed with PCR amplification, only 3 (1.2%) resulted positive and were shown to belong to genotype III through the sequencing of the SAG2 amplicon. This is the first extensive study on T. gondii in crows from Romania. Full article

(This article belongs to the Section Parasitic Pathogens)

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13 pages, 504 KiB

Open AccessArticle

Type 2 Innate Lymphoid Cell (Ilc2)-Deficient Mice Are Transcriptionally Constrained During Nippostrongylus brasiliensis Infection

by Damarius S. Fleming, Fang Liu, Joseph F. Urban, Jr. and Robert W. Li

Pathogens 2025, 14(6), 571; https://doi.org/10.3390/pathogens14060571 - 7 Jun 2025

Viewed by 114

Abstract

Mouse models serve as a means of examining immune changes when genes of interest are knocked out (KO). One group of immune gene-producing cells that have been identified is type 2 innate lymphoid cells (Ilc2). These cells are involved in the production of [...] Read more.

Mouse models serve as a means of examining immune changes when genes of interest are knocked out (KO). One group of immune gene-producing cells that have been identified is type 2 innate lymphoid cells (Ilc2). These cells are involved in the production of Th2 equivalent immune responses and signal cytokine production during the resolution of Nippostrongylus brasiliensis parasite infection in mice lungs. However, many questions about Ilc2 activity in the gut remain. To study this, retinoic acid receptor (RAR)-related orphan receptor alpha (RORα)-deficient mice were infected with adult N. brasiliensis and arranged into four treatment groups. Ten days post-infection (dpi), mouse ileum tissue was extracted for RNA-Seq. The RORα-deficient mice showed little change in gene expression at 10 dpi (N = 51) when compared to the WT mice at 10 dpi (N = 915), displaying dysregulation within the mouse gut. Based on the results, the gene expression in the gut of Ilc2-deficient mice denoted that the inability to craft Ilc2 cells left the mice unable to mount classical helminth immune responses involving humoral, mast cell, and antibody Th2-driven reactions. Overall, the results showed the importance of Ilc2 in the gut during N. brasiliensis infections and the effect that the lack of these cells had on immunity. Full article

(This article belongs to the Special Issue Immunity and Immunoregulation in Helminth Infections)

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20 pages, 2304 KiB

Open AccessArticle

Genomic Insights into the Pathogenicity of Hypervirulent Aeromonas hydrophila Strain D4 Isolated from Diseased Blunt Snout Bream with the Epidemic Sequence Type 251 Clones

by Li Xu, Xingyu Kang, Zhicheng Wang, Zuyuan Xiao and Yi Luo

Pathogens 2025, 14(6), 570; https://doi.org/10.3390/pathogens14060570 - 6 Jun 2025

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Abstract

Aeromonas hydrophila ST251 is a crucial pathogen responsible for the outbreaks of Motile Aeromonas Septicemia (MAS) in global aquaculture. To elucidate the genetic basis underlying its hypervirulence, we investigated strain D4, an ST251 isolate recovered from diseased blunt snout bream. Phenotypic assays revealed [...] Read more.

Aeromonas hydrophila ST251 is a crucial pathogen responsible for the outbreaks of Motile Aeromonas Septicemia (MAS) in global aquaculture. To elucidate the genetic basis underlying its hypervirulence, we investigated strain D4, an ST251 isolate recovered from diseased blunt snout bream. Phenotypic assays revealed that, compared to the environmental strain ATCC 7966^T, D4 exhibited enhanced motility, hemolytic activity, and protease production. Average nucleotide identity (ANI) analysis demonstrated that D4 clustered within a distinct ST251 clade, with ANI values ≥ 99.74%. Comparative genomic analysis of D4, nine additional ST251 strains, and ATCC 7966^T identified multiple unique genomic islands in ST251 strains, including pathways for myo-inositol and L-fucose utilization and a pseudaminic acid biosynthesis gene cluster. These genetic elements are associated with nutrient acquisition and flagellar assembly, potentially enhancing colonization and environmental adaptability. In addition, distinct plasmids and prophages in ST251 strains may contribute to host adaptation and virulence by regulating stress responses and virulence-associated genes. These findings offer new insights into the molecular mechanisms driving the pathogenicity and adaptability of hypervirulent A. hydrophila ST251 strains. Full article

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12 pages, 828 KiB

Open AccessArticle

Molecular Prevalence of Leishmania infantum Infection from Oral Swabs Collected from Dogs in Region of Northwestern Spain

by Javier Merino-Goyenechea, Elora Valderas-García, Verónica Castilla Gómez de Agüero, Rafael Balaña-Fouce and María Martínez-Valladares

Pathogens 2025, 14(6), 569; https://doi.org/10.3390/pathogens14060569 - 6 Jun 2025

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Abstract

Leishmaniasis is a serious zoonotic parasitic disease caused by the protist Leishmania infantum and transmitted by phlebotomine sandflies in the countries of the Mediterranean basin. Dogs are the species most susceptible to the disease and serve as a reservoir for transmission to humans, [...] Read more.

Leishmaniasis is a serious zoonotic parasitic disease caused by the protist Leishmania infantum and transmitted by phlebotomine sandflies in the countries of the Mediterranean basin. Dogs are the species most susceptible to the disease and serve as a reservoir for transmission to humans, making the Iberian Peninsula an endemic region for this infection. Although the regions close to the Mediterranean coast are the most prevalent regions of leishmaniasis in Spain, climatic factors are favouring the expansion of the vectors to more northern latitudes, where the disease was hardly known decades ago. This paper presents a prevalence study of L. infantum infection in the province of Zamora (northwestern Spain) using a non-invasive sample from canine buccal swabs and an innovative qPCR method to determine the etiologic agent. The parasite load of 151 randomly selected dogs from different points of the province was analysed during the period 2021–2022, with an estimated prevalence of 30%. In addition, the most common clinical signs of leishmaniasis in the dogs are described, and intrinsic factors associated with the nature of the dogs—such as sex, size, age as well as other factors related to the habitat in which they live and their geographical location—which could favour the disease, are evaluated. Full article

(This article belongs to the Section Parasitic Pathogens)

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11 pages, 346 KiB

Open AccessArticle

The Effects of High-Dose Probiotic Supplementation on Immune Activation and Neurocognitive Disorders in People Living with HIV Undergoing Successful Antiretroviral Treatment: The Procog Study

by Matteo Vassallo, Margaux Zerlini, Roxane Fabre, Heloise Joly, Jacques Durant, Alain Makinson, Amandine Mauries, Jacqueline Capeau, Soraya Fellahi, Jean-Philippe Bastard, Christian Pradier and Christine Lebrun-Frenay

Pathogens 2025, 14(6), 568; https://doi.org/10.3390/pathogens14060568 - 6 Jun 2025

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Abstract

Background: The prevalence of HIV-associated neurocognitive disorders (HAND) remains high despite antiretroviral treatment (ART). Changes in gut microbiota and persistent immune activation have been suggested as possible causes, while the role of probiotic supplementation remains controversial. Methods: We included subjects with mild HAND [...] Read more.

Background: The prevalence of HIV-associated neurocognitive disorders (HAND) remains high despite antiretroviral treatment (ART). Changes in gut microbiota and persistent immune activation have been suggested as possible causes, while the role of probiotic supplementation remains controversial. Methods: We included subjects with mild HAND and successful ART. They were randomized to receive either 6 months of high-dose probiotic supplementation or to continue with only ART. Immune activation markers and neuropsychological testing were performed at baseline and the end of follow-up. Neuropsychological testing assessed learning, episodic memory, attention/concentration, executive functions, language, information processing speed, and motor skills. Z- and T-scores were calculated for all domains but motor skills, allowing the measurement of the global deficit score (GDS). The trajectories of neuropsychological performances and immune activation markers were compared between groups. Results: From September 2020 to July 2021, 31 PWHs were included (median age 62, 73% men, CD4 744 cc/mm³), and 28 completed the 6-month follow-up. The characteristics of the subjects and their neuropsychological performance at baseline in the two groups were similar. At the end of follow-up, probiotics did not have any impact on immune activation markers, while they were associated with better improvement in GDS (T-score 0.0 in controls vs. −0.3 in probiotics, p = 0.048) and the attention/concentration test (Z-score 0.4 in controls vs. 1.2 in probiotics, p = 0.035). Conclusions: Oral supplementation with high-dose probiotics for 6 months did not affect systemic immune activation but was associated with improved neurocognitive performance, suggesting benefits from probiotic supplementation for mild HAND. Full article

(This article belongs to the Special Issue Viral Infections and Their Effects on the Central Nervous System (CNS))

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17 pages, 964 KiB

Open AccessArticle

Using Digital PCR to Unravel the Occurrence of Piroplasmids, Bartonella spp., and Borrelia spp. in Wild Animals from Brazil

by Ana Cláudia Calchi, Anna Claudia Baumel Mongruel, Fernanda Beatriz Pereira Cavalcanti, Lilliane Bartone, José Maurício Barbanti Duarte, Emília Patrícia Medici, Danilo Kluyber, Mayara G. Caiaffa, Mario Henrique Alves, Arnaud Leonard Jean Desbiez, Taciana Fernandes Souza Barbosa Coelho, Rosangela Zacarias Machado, Edward B. Breitschwerdt, Ricardo G. Maggi and Marcos Rogério André

Pathogens 2025, 14(6), 567; https://doi.org/10.3390/pathogens14060567 - 6 Jun 2025

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Abstract

Piroplasmids (Babesia spp., Rangelia spp., Theileria spp., Cytauxzoon spp.) are tick-borne apicomplexan protozoa that infect, depending on the species, erythrocytes and leucocytes in a wide variety of mammals and birds. The genera Bartonella and Borrelia include vector-borne bacteria that can infect and [...] Read more.

Piroplasmids (Babesia spp., Rangelia spp., Theileria spp., Cytauxzoon spp.) are tick-borne apicomplexan protozoa that infect, depending on the species, erythrocytes and leucocytes in a wide variety of mammals and birds. The genera Bartonella and Borrelia include vector-borne bacteria that can infect and cause disease in both animals and humans. Detection of hemotropic bacteria and piroplasmids in wild animals is often challenging due to low bacteremia or parasitemia. Digital (d)PCR has proven to be an effective modality for the detection and quantification of DNA of hemotropic pathogens with low parasitemia. This study compared dPCR results from 366 biological samples from seven different Brazilian wild animal groups (5 Xenarthra species, 5 deer species, 3 felid species, 1 canid species, 3 rodent species, 1 bat species, 1 tapir species, and 12 bird species) to two other molecular diagnostic techniques: quantitative real-time (qPCR) and nested (nPCR). For this study, DNA extracted from wild animal blood and spleen samples were subjected to a multiplex dPCR assay for piroplasmids, Bartonella spp., and Borrelia spp. For comparison, the same primers and probes for each agent were used in qPCR assays. Additionally, an nPCR based on the 18S rRNA gene for piroplasmids was performed. The proportions of positive results obtained using dPCR were 85.5% for piroplasmids, 33.6% for Bartonella spp., and 16.7% for Borrelia spp. For all tested agents, dPCR proved to be the technique with the highest sensitivity, making it a useful tool for screening vector-borne agents in biological samples from wild animals with low parasitemia. Full article

(This article belongs to the Section Parasitic Pathogens)

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12 pages, 27477 KiB

Open AccessArticle

The Effects of Low Concentrations of Nisin on Biofilm Formation by Staphylococcus aureus Isolated from Dairy Cattle

by Edyta Kaczorek-Łukowska, Paweł Foksiński, Joanna Małaczewska, Roman Wójcik and Natalia Szyryńska

Pathogens 2025, 14(6), 566; https://doi.org/10.3390/pathogens14060566 - 5 Jun 2025

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Abstract

Staphylococcus aureus is one of the aetiological agents of mastitis in dairy cattle. Their biofilms are relevant for human and veterinary medicine. It has been shown that some antibiotics at low concentrations can stimulate the production of biofilms, but there is little information [...] Read more.

Staphylococcus aureus is one of the aetiological agents of mastitis in dairy cattle. Their biofilms are relevant for human and veterinary medicine. It has been shown that some antibiotics at low concentrations can stimulate the production of biofilms, but there is little information on the effects of low concentrations of nisin, which is considered a therapeutic agent and has been added to food products for years as a biopreservative. In our study, we used Staphylococcus aureus strains (n = 28) isolated from dairy cattle. The MIC of nisin were determined using the broth microdilution method. Based on the minimum inhibitory concentration (MIC) results, the following concentrations were selected for further analyses: nisin at 39, 19, 9 IU/mL; nisin in combination with tetracycline at 39 IU/mL + 0.06 μg/mL, 18 IU/mL + 0.06 μg/mL, and 9 IU/mL + 0.06 μg/mL; and tetracycline alone at 0.06 μg/mL. The biofilm-forming capacity was determined via crystal violet staining in 96-well plates, icaD gene expression was determined using the 2−ΔΔCt method, and microscopic evaluation was carried out using scanning electron microscopy. Results: The MICs were 156 IU/mL (46%) and 312 IU/mL (43%) for most strains. Due to large statistical deviations, there were no statistically significant changes in the biofilm-forming capacity or icaD gene expression despite a visible increasing trend. Despite the absence of statistically significant differences, it was observed that for all concentrations analysed biofilm formation was noticeably greater for both nisin alone and for tetracycline and its mixtures than for untreated cells. Conclusions: In our opinion, the effects of nisin, especially at low concentrations, on biofilm structure show a certain worrying trend that may pose a future threat. Full article

(This article belongs to the Section Bacterial Pathogens)

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22 pages, 3276 KiB

Open AccessArticle

Modulation of TvRAD51 Recombinase in Trichomonas vaginalis by Zinc and Cadmium as a Potential Mechanism for Genotoxic Stress Response

by Jonathan Puente-Rivera, José Jesús Flores-Vega, Marcos Morales-Reyna, Elisa Elvira Figueroa-Angulo, Yussel Pérez-Navarro, Alfonso Salgado-Aguayo, Ángeles Carlos-Reyes and Maria Elizbeth Alvarez-Sánchez

Pathogens 2025, 14(6), 565; https://doi.org/10.3390/pathogens14060565 - 5 Jun 2025

Viewed by 147

Abstract

Trichomonas vaginalis, the protozoan responsible for trichomoniasis, encounters fluctuating levels of metal cations in the male urogenital tract, notably zinc (Zn²⁺) and cadmium (Cd²⁺), which may induce genotoxic stress. While zinc is a key physiological component of the [...] Read more.

Trichomonas vaginalis, the protozoan responsible for trichomoniasis, encounters fluctuating levels of metal cations in the male urogenital tract, notably zinc (Zn²⁺) and cadmium (Cd²⁺), which may induce genotoxic stress. While zinc is a key physiological component of the male reproductive tract, both Zn²⁺ and Cd²⁺ can become genotoxic at elevated concentrations. However, their effect on DNA repair mechanisms in T. vaginalis remains poorly understood. This study characterizes, for the first time, the expression and modulation of the recombinase TvRAD51, a homologous recombination (HR) key enzyme, in response to UV irradiation and sublethal concentrations of Zn²⁺ (1.6 mM) and Cd²⁺ (0.1 mM). In silico analyses confirmed the presence and conserved structure of the tvrad51 gene and its interaction with HR-related proteins, such as TvBLM and TvBRCA2. Quantitative RT-PCR, Western blot, and immunofluorescence assays revealed that TvRAD51 is upregulated at both transcript and protein levels following UV- and cation-induced DNA damage, with distinct temporal expression patterns for Zn²⁺ and Cd²⁺ exposure. Notably, TvRAD51 showed nuclear localization at early time points post-exposure, suggesting active participation in DNA repair processes. These findings demonstrate that TvRAD51 is a central component of the genotoxic stress response in T. vaginalis, potentially contributing to parasite survival and adaptation in hostile environments through homologous recombination repair pathways. Full article

(This article belongs to the Special Issue Virulence Mechanisms Used by Pathogens Related to Human, Animal, and Environmental Health)

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9 pages, 973 KiB

Open AccessArticle

Detection and Characterization of Paslahepevirus balayani (Hepatitis E Virus) in Dairy Products from Hebei Province, China

by Xinyue Hu, Jinfeng Wang, Yinuo Wang, Wanzhe Yuan, Jianchang Wang and Xiangdong Xu

Pathogens 2025, 14(6), 564; https://doi.org/10.3390/pathogens14060564 - 5 Jun 2025

Viewed by 222

Abstract

Paslahepevirus balayani (hepatitis E virus), a zoonotic pathogen transmitted primarily via the fecal–oral route, has undergone shifting transmission dynamics in China, with foodborne and zoonotic routes becoming increasingly significant. To assess the potential risk of HEV transmission through dairy products, this study investigated [...] Read more.

Paslahepevirus balayani (hepatitis E virus), a zoonotic pathogen transmitted primarily via the fecal–oral route, has undergone shifting transmission dynamics in China, with foodborne and zoonotic routes becoming increasingly significant. To assess the potential risk of HEV transmission through dairy products, this study investigated HEV RNA presence in raw milk from cows, sheep, and goats in Hebei Province, China. From March 2024 to April 2025, we collected 102 cow milk, 18 sheep milk, and 59 goat milk samples, analyzing them using RT-qPCR, with positive samples confirmed by RT-Nested PCR and partial ORF2 sequencing. While no HEV RNA was detected in cow milk, 3/18 (16.67%) sheep milk and 1/59 (1.69%) goat milk samples tested positive. Phylogenetic analysis of two sheep-derived and one goat-derived HEV isolate showed 99.43–100% nucleotide identity to local swine HEV strains (HEV/HB-SJZ158/CHN/2021 and HEV/HB-CD28/CHN/2021), all clustering within genotype 4d, the dominant subtype in the region. This study provided the first evidence of HEV RNA in sheep and goat milk in Hebei Province, suggesting possible cross-species transmission from pigs to ruminants. These findings highlighted the need for further research on HEV transmission risks through dairy products and emphasize the importance of monitoring zoonotic HEV strains in food safety assessments. Full article

(This article belongs to the Special Issue Molecular Mechanisms and Ecological Interventions in Zoonotic Pathogens)

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12 pages, 622 KiB

Open AccessReview

Is African Swine Fever Driven by Flying Hematophagous Insects?

by Marek Walczak, Maciej Frant, Krzesimir Szymankiewicz, Małgorzata Juszkiewicz, Katarzyna Podgórska and Marcin Smreczak

Pathogens 2025, 14(6), 563; https://doi.org/10.3390/pathogens14060563 - 5 Jun 2025

Viewed by 184

Abstract

African swine fever (ASF) has become one of the most economically important diseases affecting swine and has a significant negative impact on the global pork production sector. In Europe, the main reservoir of the disease is the wild boar population, which poses a [...] Read more.

African swine fever (ASF) has become one of the most economically important diseases affecting swine and has a significant negative impact on the global pork production sector. In Europe, the main reservoir of the disease is the wild boar population, which poses a risk of transmitting the disease to pig farms. To date, no safe and effective vaccine is available on the market. Therefore, biosecurity measures and early recognition of the disease play a key role in preventing and combating ASF. In recent years, numerous insights into the nature of the virus have emerged; however, several knowledge gaps still need to be addressed. One of these gaps is an accurate understanding of all possible pathways through which the virus can reach a pig farm. Interrupting these pathways would significantly reduce the risk of disease outbreaks. Despite a general understanding of disease transmission, ASF can still affect farms with well-established high biosecurity measures. This article highlights the potential for mechanical transmission of ASF by flying hematophagous insects, considering several factors, including current knowledge of the putative role of insects in ASF transmission, insects’ abilities to transmit the virus, ASFV properties, the uncertainties regarding the effectiveness of indirect transmission, and the seasonality of disease outbreaks on domestic pig farms. Full article

(This article belongs to the Special Issue Laboratory Diagnostics in Epidemiological Diseases Caused by Animal Pathogens)

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11 pages, 1115 KiB

Open AccessArticle

Monitoring Multiple Sexually Transmitted Pathogens Through Wastewater Surveillance

by Balghsim Alshehri, Olivia N. Birch and Justin C. Greaves

Pathogens 2025, 14(6), 562; https://doi.org/10.3390/pathogens14060562 - 5 Jun 2025

Viewed by 165

Abstract

Wastewater-based epidemiology (WBE) offers a promising tool for sexually transmitted infection (STI) surveillance, especially in settings where underdiagnosis or social stigma complicates conventional reporting. To assess its utility, we conducted a year-long study examining six STIs, Chlamydia trachomatis, Treponema pallidum, Neisseria [...] Read more.

Wastewater-based epidemiology (WBE) offers a promising tool for sexually transmitted infection (STI) surveillance, especially in settings where underdiagnosis or social stigma complicates conventional reporting. To assess its utility, we conducted a year-long study examining six STIs, Chlamydia trachomatis, Treponema pallidum, Neisseria gonorrhoeae, human immunodeficiency virus (HIV), hepatitis C virus (HCV), and herpes simplex virus (HSV), in weekly composite samples from the primary influent of a small-sized Midwestern wastewater treatment plant. Pathogen detection and quantification were performed via digital PCR. Among the tested targets, Gonorrhea, HIV, HCV, and HSV were detected at the highest frequencies, often in 40–50% of the samples, while Chlamydia and Syphilis appeared less frequently. Despite the variability in detection patterns, this study demonstrates that even infrequent signals can reveal community-level shedding of poorly reported or asymptomatic infections. Although month-to-month wastewater data were not strongly correlated with corresponding clinical records, which could potentially reflect delayed healthcare seeking and pathogen-specific shedding dynamics, the overall findings underscore WBE’s ability to complement existing surveillance by capturing infections outside traditional healthcare channels. These results not only advance our understanding of STI prevalence and population shedding but also highlight the practical benefits of WBE as an early warning and targeted intervention tool. Full article

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

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14 pages, 290 KiB

Open AccessArticle

Tick Species Identification and Zoonotic Bacteria Detection from Healthcare-Extracted Specimens from Humans in the Basque Country, Northern Iberian Peninsula

by Patirke Ibarrondo-Mendiola, Patricia Vázquez, Miriam Alkorta, Cristina Zugazaga, Ana L. García-Pérez, Jesús F. Barandika and Aitor Cevidanes

Pathogens 2025, 14(6), 561; https://doi.org/10.3390/pathogens14060561 - 4 Jun 2025

Viewed by 181

Abstract

Hard ticks are vectors of a wide range of pathogens, with tick-borne bacteria being among the most frequently detected. This study presents a first approach to the identification of human-biting ticks in the Basque Country (northern Iberian Peninsula), where previous research has mainly [...] Read more.

Hard ticks are vectors of a wide range of pathogens, with tick-borne bacteria being among the most frequently detected. This study presents a first approach to the identification of human-biting ticks in the Basque Country (northern Iberian Peninsula), where previous research has mainly focused on ticks from vegetation and domestic and wild animals. The aim of this study was (i) to identify tick species collected in hospitals and health centres and (ii) to determine the presence and identify of pathogenic bacteria that they may carry using PCR, RLB and sequencing methods. A total of 181 ticks were collected and grouped in pools of one, two or three specimens, resulting in 157 samples. Morphological and molecular identification of collected ticks revealed that most specimens belonged to Ixodes ricinus (88.5%). Additionally, Rhipicephalus bursa, Haemaphysalis punctata, Dermacentor reticulatus and Dermacentor marginatus were also identified. A total of 25 samples (15.9%) tested positive for DNA from the targeted pathogens. The most prevalent vector-borne bacteria were Borrelia spp. (6.4%), followed by Rickettsia spp. (5.1%), Anaplasma phagocytophilum (2.5%) and Coxiella spp. (1.9%). Species identified included B. afzelii, B. garinii, B. miyamotoi, B. valaisiana, B. burgdorferi s. s., R. monacensis, “Candidatus R. rioja”, R. helvetica and R. massiliae. This study highlights the usefulness of combining molecular diagnostics with passive surveillance of human-attached ticks as an effective tool for regional monitoring of ticks and tick-borne pathogens. Full article

(This article belongs to the Special Issue Ticks and Tick-Borne Pathogens in a Changing World)

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14 pages, 1735 KiB

Open AccessArticle

Effect of Grape Pomace Inclusion in the Diet of Ewes Naturally Infected with Gastrointestinal Nematodes During Lactation

by Mateus Oliveira Mena, Gustavo Gabriel de Oliveira Trevise, César Cristiano Bassetto, Willinton Hernan Pinchao Pinchao, Helder Louvandini, Ricardo Velludo Gomes de Soutello, Ana Cláudia Alexandre Albuquerque and Alessandro Francisco Talamini do Amarante

Pathogens 2025, 14(6), 560; https://doi.org/10.3390/pathogens14060560 - 4 Jun 2025

Viewed by 247

Abstract

This study evaluated the feasibility of including grape pomace in the diet of Santa Inês ewes during lactation, aiming to reduce the effects of gastrointestinal nematode (GIN) infections and improve productive performance. Grape pomace, which contains phenolic compounds and tannins, was provided as [...] Read more.

This study evaluated the feasibility of including grape pomace in the diet of Santa Inês ewes during lactation, aiming to reduce the effects of gastrointestinal nematode (GIN) infections and improve productive performance. Grape pomace, which contains phenolic compounds and tannins, was provided as a dietary supplement, replacing 20% of the concentrate over 28 days of lactation, starting on day 7 after lambing. A total of 18 ewes were used, divided into two groups: supplemented (n = 8) and control (n = 10). The supplemented group showed significant reduction in fecal egg count on day 14 of lactation (p < 0.05) in comparison with the control group. Lambs from the supplemented group had higher weaning weights (p < 0.05). Additionally, the supplemented group showed higher values for packed cell volume and circulating eosinophils, indicating greater resilience to infections. Analysis of anti-L3 IgG against Haemonchus contortus revealed no significant differences between the groups. It is concluded that grape pomace supplementation demonstrated potential to improve hematological and productive parameters in lactating ewes, with more evident effects during early lactation. Although the results suggest a possible complementary role in controlling gastrointestinal nematodes, future studies are needed to confirm and optimize this nutritional strategy. Full article

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

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19 pages, 7482 KiB

Open AccessArticle

Tuberculosis and Impact of COVID-19 on Spread of Epidemics in Kazakhstan

by Zhandarbek Bekshin, Albert Askarov, Yergali Abduraimov, Aralbek Rsaliyev, Gulmira Bissenova, Nurgul Amirkhanova, Zhadyrassyn Nurbekova and Aliya Temirbekova

Pathogens 2025, 14(6), 559; https://doi.org/10.3390/pathogens14060559 - 4 Jun 2025

Viewed by 264

Abstract

This study examines the epidemiological situation of tuberculosis (TB) in the regions of the Republic of Kazakhstan over the past seven years (2018–2024), which cover the before-, during- and after-COVID-19 periods, with a focus on the risks of its emergence and spread. The [...] Read more.

This study examines the epidemiological situation of tuberculosis (TB) in the regions of the Republic of Kazakhstan over the past seven years (2018–2024), which cover the before-, during- and after-COVID-19 periods, with a focus on the risks of its emergence and spread. The analysis revealed that while TB incidence is declining, mortality remains high in the before- and during-COVID-19 periods, indicating a general decline in population health. The concentration of TB incidence in relation to geographic location was mainly in the northern, western and southern regions. Before COVID-19, TB incidence reached 48.2 cases and mortality reached a maximum of 2.4 cases per 100,000 people. In 2024, the incidence and mortality of tuberculosis significantly decreased to 33.5 (30.5%) and 1.0 (58.3%), respectively, reflecting an improvement in health indicators in the post-pandemic period. In the after-COVID-19 period, in regions with high unemployment, the incidence was higher than in the before- and during-COVID-19 periods. Nevertheless, it is important that the trend in tuberculosis incidence shows positive improvement after the COVID-19 period. In addition, a comparative analysis of tuberculosis incidence trends in different age groups and social factor groups shows that the adult population remains the most vulnerable category among the general population. The above-listed factors, as well as our analysis of tuberculosis incidence, shows that TB incidence does not always correlate with the level of vaccination in different regions of Kazakhstan, indicating a multifactorial influence on the tuberculosis epidemic. Full article

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

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17 pages, 1726 KiB

Open AccessArticle

Detection of Human Papillomavirus in Urogenital Swabs from Male Patients in Northern Serbia

by Gordana Kovačević, Vladimir Vuković, Nataša Nikolić, Branka Bašica, Jelena Radovanov, Aleksandra Čolović Popadić, Milica Pejaković Budinski, Tihomir Dugandžija, Zoran Golušin and Aleksandra Patić

Pathogens 2025, 14(6), 558; https://doi.org/10.3390/pathogens14060558 - 3 Jun 2025

Viewed by 244

Abstract

Human papillomavirus (HPV) is one of the most common sexually transmitted infections, affecting both men and women. However, in Serbia, the previous epidemiological and clinical research on HPV has primarily focused on women due to its established role in cervical cancer, while the [...] Read more.

Human papillomavirus (HPV) is one of the most common sexually transmitted infections, affecting both men and women. However, in Serbia, the previous epidemiological and clinical research on HPV has primarily focused on women due to its established role in cervical cancer, while the data on HPV prevalence in men remain scarce. This study analyzed 634 samples from men (mean age: 30 years, SD = 9.98; range: 18–79) from between 2012 and 2024. Overall, 30.76% of the tested men were HPV–positive, with the highest percentage of cases being observed in those aged 25–29 years (32.82%). The most common genotypes were HPV 16 and HPV 31 (22.05% each), followed by HPV6 (20.51%), HPV 56, and HPV 52 (9.23% each). The prevalence of HPV was the highest in the samples from men with genital warts (40.77%). Among the available vaccines, the nonavalent Gardasil 9 provides the broadest protection, covering genotypes found in 82% of the HPV–positive cases identified in this study. Our findings underscore the importance of comprehensive HPV prevention and control measures for the male population, contributing to ongoing research efforts aimed at reducing the burden of HPV–associated diseases in our region. Increasing the vaccination coverage among Serbian men could substantially reduce the overall burden of HPV–related diseases in both sexes. Full article

(This article belongs to the Special Issue Advances in HPV Diagnostic Testing—Improving Access and Accuracy)

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31 pages, 783 KiB

Open AccessSystematic Review

Global Epidemiology and Antimicrobial Resistance of Metallo-β-Lactamase (MBL)-Producing Acinetobacter Clinical Isolates: A Systematic Review

by Matthew E. Falagas, Dimitrios S. Kontogiannis, Maria Zidrou, Charalampos Filippou and Giannoula S. Tansarli

Pathogens 2025, 14(6), 557; https://doi.org/10.3390/pathogens14060557 - 3 Jun 2025

Viewed by 167

Abstract

This systematic review assessed the global epidemiology of metallo-β-lactamase (MBL)-producing Acinetobacter clinical isolates and the associated antimicrobial resistance. A total of 475 relevant articles from the Cochrane Library, Google Scholar, PubMed, Scopus, and Web of Science were identified and screened as potentially eligible [...] Read more.

This systematic review assessed the global epidemiology of metallo-β-lactamase (MBL)-producing Acinetobacter clinical isolates and the associated antimicrobial resistance. A total of 475 relevant articles from the Cochrane Library, Google Scholar, PubMed, Scopus, and Web of Science were identified and screened as potentially eligible articles. Data from 85 articles were extracted for the analysis. Most reports on MBL-producing Acinetobacter clinical isolates originated from Asia [68/85 (80%) studies] and Africa [14/85 (16.5%) studies]. There were also scarce reports from Europe and America. The bla_VIM (in 31 studies), bla_IMP (in 29 studies), and bla_NDM (in 21 studies) genes were the most commonly identified genes. In 22 out of 28 (78.6%) studies with comparable data, the proportions of MBL-producing pathogens detected using phenotypic methods were numerically higher than those using genotypic methods. MBL-producing Acinetobacter isolates showed high resistance (up to 100%) to several antibiotic classes, including carbapenems, cephalosporins, fluoroquinolones, and monobactams. However, they showed low resistance to colistin [ranging from 0% (in six studies) to 14.3% (in one study)] and to tigecycline [0% (in three studies)]. No risk of bias assessment was conducted. The findings emphasize the global spread of MBL-producing Acinetobacter and the need for enhanced antimicrobial stewardship, infection control measures, and surveillance. Full article

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17 pages, 1593 KiB

Open AccessArticle

Multiple Mechanisms of HIV-1 Resistance to PGT135 in a Chinese Subtype B’ Slow Progressor

by Yuanyuan Hu, Shasha Sun, Ying Liu, Li Ren, Xintao Hu, Yuhua Ruan, Liying Ma, Hao Liang, Yiming Shao, Kunxue Hong, Sen Zou and Yanling Hao

Pathogens 2025, 14(6), 556; https://doi.org/10.3390/pathogens14060556 - 3 Jun 2025

Viewed by 195

Abstract

We investigated HIV-1 immune evasion mechanisms in a slow progressor (CBJC515) by constructing pseudoviruses expressing autologous Env proteins. Intriguingly, all pseudoviruses exhibited resistance to the broadly neutralizing antibody (bNAb) PGT135. Using site-directed mutagenesis and chimeric Env construction, we identified distinct escape mechanisms: early [...] Read more.

We investigated HIV-1 immune evasion mechanisms in a slow progressor (CBJC515) by constructing pseudoviruses expressing autologous Env proteins. Intriguingly, all pseudoviruses exhibited resistance to the broadly neutralizing antibody (bNAb) PGT135. Using site-directed mutagenesis and chimeric Env construction, we identified distinct escape mechanisms: early 2005 strains lost the N332 glycan site, while 2006/2008 strains retained key epitopes but developed resistance through structural modifications in the V1/V4/C2 regions or acquired novel N-glycosylation sites (N398/N611). These findings provide insights into how HIV-1 can escape from N332-directed bNAb responses without altering the epitope itself. Furthermore, chimeric experiments also elucidated regional co-evolution and functional maintenance: the V1V2 region broadly interfered with envelope protein function, while the V3 region may exhibit compensatory activity, restoring functionality and mitigating deleterious polymorphisms in other regions to keep Env antigenic diversity. These results offer valuable mechanistic clues that may inform the development of next-generation HIV-1 vaccines. Full article

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20 pages, 387 KiB

Open AccessReview

Digestive Manifestations of Post-COVID-19: A Focus on Therapeutic Strategies

by Cristina Stasi and Massimo Bellini

Pathogens 2025, 14(6), 555; https://doi.org/10.3390/pathogens14060555 - 3 Jun 2025

Viewed by 734

Abstract

Post-COVID-19 is a chronic infection-related syndrome, including exacerbations of pre-existing or newly diagnosed conditions that have been established after the acute phase of COVID-19 and have demonstrated a wide range of systemic effects beyond the lungs. SARS-CoV-2 attaches to its receptor, angiotensin-converting enzyme [...] Read more.

Post-COVID-19 is a chronic infection-related syndrome, including exacerbations of pre-existing or newly diagnosed conditions that have been established after the acute phase of COVID-19 and have demonstrated a wide range of systemic effects beyond the lungs. SARS-CoV-2 attaches to its receptor, angiotensin-converting enzyme 2 (ACE-2). Transmembrane serine protease 2 (TMPRSS2) facilitates viral entry and spread. ACE-2 receptors are detectable in several tissues, including the respiratory mucosa, digestive tract, heart, kidney, and brain. Several investigations have demonstrated an increase in digestive manifestations post-acute COVID-19, likely related to an alteration in the intestinal microbiota following infection. These changes can lead to a loss of species diversity, resulting in an overgrowth of opportunistic pathogens and deprivation of commensal bacteria. In this context, post-infection irritable bowel syndrome shows an increased incidence compared to controls. Growing evidence also suggests the enduring presence of SARS-CoV-2 in the gut tissue. Studies are ongoing to investigate antiviral agents that counteract prolonged COVID-19 symptoms. Therefore, the objectives of this review were to summarize the digestive manifestations, focusing on irritable bowel syndrome and therapeutic strategies. This review gives an overview of studies published in English in the last two years on the PubMed database. Full article

(This article belongs to the Special Issue Antiviral Strategies Against Human Respiratory Viruses)

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15 pages, 1350 KiB

Open AccessReview

Advancements in Pathogen Detection: Argonaute-Based Nucleic Acid Detection Technology

by Meng Hong, Guodi Wu, Yanli Ren, Shanshan Wu, Haihong Zhu and Zhi Chen

Pathogens 2025, 14(6), 554; https://doi.org/10.3390/pathogens14060554 - 2 Jun 2025

Viewed by 304

Abstract

In recent years, global public health security has encountered significant challenges, with infectious diseases accounting for approximately 25% of global mortality annually. The worldwide pandemic instigated by the novel coronavirus, alongside the persistent threats posed by Ebola, influenza, and multidrug-resistant bacteria, has severely [...] Read more.

In recent years, global public health security has encountered significant challenges, with infectious diseases accounting for approximately 25% of global mortality annually. The worldwide pandemic instigated by the novel coronavirus, alongside the persistent threats posed by Ebola, influenza, and multidrug-resistant bacteria, has severely compromised human health, economic development, and social stability. Within this context, the development of rapid and precise pathogen detection technologies has emerged as a critical frontline defense for epidemic prevention and control, serving as a pivotal component in the implementation of the “early detection, early isolation, and early treatment” strategy. The Argonaute (Ago) protein, recognized as a programmable and target-specific activated nuclease, has demonstrated substantial potential in the realm of nucleic acid detection due to its distinctive biological properties, garnering considerable attention. In this study, we delineate the structural characteristics of Ago proteins and elucidate the mechanism underlying their nuclease activity. Furthermore, we review the principles of nucleic acid detection based on Argonaute and provide a comprehensive analysis of recent advancements in related detection systems. Additionally, we compare the advantages of detection based on Argonaute with other detection methodologies. Through a comprehensive analysis, we aim to provide a robust theoretical foundation and an advanced technical reference for the development of new-generation nucleic acid detection platforms with high sensitivity and high specificity. Full article

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18 pages, 326 KiB

Open AccessReview

Beyond Mosquitoes: A Review of Pediatric Vector-Borne Diseases Excluding Malaria and Arboviral Infections

by Giulia Carbone, Amina De Bona, Dragos Septelici, Alessandro Cipri, Andrea Nobilio and Susanna Esposito

Pathogens 2025, 14(6), 553; https://doi.org/10.3390/pathogens14060553 - 2 Jun 2025

Viewed by 375

Abstract

Vector-borne diseases (VBDs) significantly impact global child health, with mosquito-transmitted infections like malaria and arboviruses accounting for a substantial portion of this burden. However, other arthropod-borne diseases—transmitted by vectors such as ticks, fleas, sand flies, lice, and triatomine bugs—also pose serious health risks [...] Read more.

Vector-borne diseases (VBDs) significantly impact global child health, with mosquito-transmitted infections like malaria and arboviruses accounting for a substantial portion of this burden. However, other arthropod-borne diseases—transmitted by vectors such as ticks, fleas, sand flies, lice, and triatomine bugs—also pose serious health risks to children worldwide. This review specifically excludes mosquito-borne diseases to concentrate on these less-discussed yet clinically important pediatric VBDs. We examine their clinical manifestations, diagnostic challenges, and treatment options, highlighting the unique vulnerabilities of children, including immature immune systems, behavioral factors, and communication barriers that can delay diagnosis. Additionally, we explore how environmental and anthropogenic factors, such as climate change and urbanization, are expanding the geographic range of these vectors, leading to the emergence of diseases like Lyme disease and leishmaniasis in new regions. By focusing on non-mosquito VBDs, this review aims to raise awareness and inform healthcare providers and public health practitioners about the comprehensive landscape of pediatric vector-borne diseases. Full article

(This article belongs to the Special Issue Emerging and Re-emerging Infections in Pediatrics)

15 pages, 1418 KiB

Open AccessArticle

Detection and Inhibition of Clostridium perfringens by Cocktail of Star Anise and Thymus Extracts in Chicken Meat Products

by Gamal M. Hamad, Shenoda Gaber Monir Henry, Gamal E. A. El-Rokh, Nadia H. A. Ramadan, Hany S. Abdel Raoof, Ahmed M. Sulaiman, Ahmed M. El-Mesallamy, Samy E. Elshaer, Sara M. Gaber, Ibrahim M. Rabah, Ahmed R. Mahmoud, Mahmoud S. A. Salama, Taha Mehany and Hesham E. A. Abdelfttah

Pathogens 2025, 14(6), 552; https://doi.org/10.3390/pathogens14060552 - 1 Jun 2025

Viewed by 357

Abstract

Background: The presence of C. perfringens in chicken meat products leads to significant economic losses for the industry and is associated with human food poisoning. It inspires creative answers for novel substances and techniques, such natural antibacterials, for improved prospects in the future. [...] Read more.

Background: The presence of C. perfringens in chicken meat products leads to significant economic losses for the industry and is associated with human food poisoning. It inspires creative answers for novel substances and techniques, such natural antibacterials, for improved prospects in the future. The objective of this study was to examine the antibacterial properties of thymus and star anise extracts for the suppression of Clostridium perfringens in products made from chicken meat. Methods: Thymus and star anise extracts were phytochemically analyzed using the Folin–Ciocalteu spectrophotometric method, High-Performance Liquid Chromatography (HPLC) to determine the phenolic compounds, DPPH to determine the antioxidant activity, and the agar disk diffusion assay to determine the antibacterial effect of the star anise. Following treatment with the mixture extract, an experimental application was conducted on chicken burgers. C. perfringens was also found in poultry samples. Lastly, the sensory evaluation of the chicken burger was detected. Results: The findings showed that Clostridium perfringens was present in a large number of chicken meat products, with a significant concentration in chicken thighs (84%), as well as in the mixture extract has the largest concentration of phytochemical components (TPC:123.88 ± 7.42 mg GAE/g, TFC: 69.04 ± 6.37 mg quercetin/g) with strong antibacterial action (the minimum inhibitory concentration (MIC) of the mixed extract was 3.12 mg/mL, with an inhibition zone of 13.06 ± 0.91 mm). Also, the addition of mix. extract to chicken burgers extends their shelf life, particularly when the mix content is high. Additionally, the mix. extract shows non-significant effect (p < 0.05) on the chicken burger’s sensory qualities. Conclusions: Finally, we can make use of the mix. extract of thymus and star anise as a natural preservative in chicken meat products especially the chicken burger. Full article

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12 pages, 766 KiB

Open AccessArticle

by Raffaele Vitale, Marco Denina, Laura Badiali, Matteo Sandei, Giulia Mazzetti, Carlotta Canavese, Aba Tocchet, Antonia Versace and Silvia Garazzino

Pathogens 2025, 14(6), 551; https://doi.org/10.3390/pathogens14060551 - 1 Jun 2025

Viewed by 247

Abstract

Introduction: While influenza-associated encephalopathy (IAE) in children remains a serious concern, recent evidence suggests a shift in its epidemiology, with a possible decline in incidence and severity over time. Methods: This retrospective review includes patients aged 0–18 admitted to a tertiary hospital in [...] Read more.

Introduction: While influenza-associated encephalopathy (IAE) in children remains a serious concern, recent evidence suggests a shift in its epidemiology, with a possible decline in incidence and severity over time. Methods: This retrospective review includes patients aged 0–18 admitted to a tertiary hospital in Northern Italy between November 2023 and February 2025. Inclusion criteria were a positive influenza test, influenza-like symptoms, and neurological involvement. Findings are interpreted alongside current literature. Results: Twenty-five unvaccinated children met criteria for IAE (11 in 2023/24; 14 in 2024/25). Neurological comorbidities were present in 40%. All patients had pathological EEGs. Lumbar puncture was performed in 40%, with abnormalities in 33%. Brain imaging was conducted in 56% of cases, revealing findings in 15%. All received oseltamivir; 60% were also treated with dexamethasone. Severe complications occurred in 16%, while 12% had persistent symptoms or required therapy at discharge. After adjusting for seasonal peak timing, no significant inter-seasonal difference was observed. Discussion: Although IAE continues to present serious risks, recent trends suggest a changing burden. The lack of vaccination among cases underscores the need for prevention. Study limitations include its single-center design and the absence of long-term follow-up. Broader prospective studies are needed. Full article

(This article belongs to the Special Issue Influenza A Virus: Updates and Insights)

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15 pages, 928 KiB

Open AccessArticle

Neutrophil-to-Lymphocyte Ratio (NLR) and Platelet-to-Lymphocyte Ratio (PLR) as Prognostic Markers of COVID-19 Disease Irrespective of Immunosuppression Status: A Case-Control Retrospective Single-Center Study

by Amalia Papanikolopoulou, Vasiliki Rapti, Polyxeni Alexiou, Charalampos M. Charalampous, Maria Effrosyni Livanou, Vissaria Sakka, Konstantinos N. Syrigos and Garyfallia Poulakou

Pathogens 2025, 14(6), 550; https://doi.org/10.3390/pathogens14060550 - 1 Jun 2025

Viewed by 293

Abstract

Neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR) have been studied as predictors for severe COVID-19 outcomes. The aim of the present study is to identify prognostic cut-off values of NLR and PLR for intubation and death in hospitalized COVID-19 patients, with or without [...] Read more.

Neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR) have been studied as predictors for severe COVID-19 outcomes. The aim of the present study is to identify prognostic cut-off values of NLR and PLR for intubation and death in hospitalized COVID-19 patients, with or without immunosuppression. From June 2021 to December 2022, we retrospectively analyzed 393 consecutively admitted COVID-19 patients, who were divided in two cohorts according to immunosuppression status (hematological malignancy and/or autoimmune condition vs. non-immunocompromised), using a propensity score-matching in 1:2 ratio. Higher NLR and PLR values were observed on days 1 and 4 for severe COVID-19, irrespective of immunosuppression status. NLR on day 1 >5.06 and day 4 >6.40 (p < 0.001), as well as PLR on day 1 >262.2 and day 4 >217.3 (p = 0.003), were associated with a greater probability for intubation. Similarly, a higher probability for death was found in the subset of patients with NLR on day 1 >4.82 (p < 0.001) and day 4 >6.41 (p < 0.001) and PLR on day 1 >229 (p = 0.009) and day 4 >205.4 (p = 0.003). Both PLR and NLR exhibited consistently higher negative predictive values (NPVs) (>93%) compared to positive predictive values (PPVs) for intubation and death. NLR and PLR displayed strong prognostic potential in hospitalized COVID-19 patients regarding intubation and death, irrespective of immunosuppression status, thus the surveillance of these biomarkers may help clinicians identify high-risk COVID-19 patients at an early stage. Full article

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Pathogens, Volume 14, Issue 6 (June 2025) – 69 articles

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