Pathogens

15 pages, 2127 KiB

Open AccessArticle

The Incidence and Trends of Yellow Fever from 1990 to 2021 in Major Endemic Regions: A Systematic Analysis Based on the 2021 Global Burden of Disease Study

by Xinwei Wang, Bin Li, Baoren He, Xipeng Yan, Linbin Huang, Jinlian Li, Rongji Lai, Mingshuang Lai, He Xie, Qiuhong Mo and Limin Chen

Pathogens 2025, 14(6), 594; https://doi.org/10.3390/pathogens14060594 - 16 Jun 2025

Abstract

As a re-emerging disease, the worldwide burden and trends of yellow fever (YF) remain inadequately quantified. This study aims to assess the incidence of YF both globally and in major endemic regions from 1990 to 2021. Utilizing data from the Global Burden of [...] Read more.

As a re-emerging disease, the worldwide burden and trends of yellow fever (YF) remain inadequately quantified. This study aims to assess the incidence of YF both globally and in major endemic regions from 1990 to 2021. Utilizing data from the Global Burden of Disease (GBD) database, we evaluated the burden of YF. We employed an age–period–cohort model to assess the influence of age, period, and cohort on the incidence of YF from 1992 to 2021. A secondary data analysis based on GBD database showed the following: in 2021, there were 86,509 incident cases of YF. Between 1990 and 2021, the number of incident cases decreased by 74.7%, while the age-standardized incidence rate (ASIR) declined at an EAPC of −4.76% (95% confidence interval: −5.10 to −4.42). In 2021, the highest ASIRs of YF were observed in Western Sub-Saharan Africa, Central Sub-Saharan Africa, and Eastern Sub-Saharan Africa. The analysis of age effects indicates that children aged 5–10 years old exhibit the highest incidence rate. Both period and cohort effects demonstrated a decline in morbidity risk. The decomposition analysis identified epidemiological changes as the primary factor contributing to the global reduction in the YF burden. Despite considerable reduction in incidence, YF remains a significant public health threat in Sub-Saharan Africa. Full article

(This article belongs to the Special Issue Arboviruses Worldwide: Epidemiology, Pathogenesis, and Global Health Challenges)

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14 pages, 1855 KiB

Open AccessArticle

Oxidative Stress and Apoptotic Markers in Goats Naturally Infected with Mycobacterium avium subsp. paratuberculosis

by Merve Ozturk, Muhammet Bahaeddin Dortbudak, Bayram Bekmez, Lucia Biagini, Nuri Altuğ, Giacomo Rossi, Yasin Ozturk and Alessandro Di Cerbo

Pathogens 2025, 14(6), 593; https://doi.org/10.3390/pathogens14060593 - 16 Jun 2025

Abstract

Paratuberculosis, caused by Mycobacterium avium subspecies paratuberculosis (MAP), is a chronic granulomatous enteritis with significant implications for ruminant health, economic productivity, and potential zoonotic risk. This study investigated the expression of biomarkers of oxidative stress and apoptosis in goats naturally infected with MAP, [...] Read more.

Paratuberculosis, caused by Mycobacterium avium subspecies paratuberculosis (MAP), is a chronic granulomatous enteritis with significant implications for ruminant health, economic productivity, and potential zoonotic risk. This study investigated the expression of biomarkers of oxidative stress and apoptosis in goats naturally infected with MAP, focusing on three biological matrices: serum, intestinal mucosa, and mesenteric lymph nodes. Twenty MAP-positive goats and ten healthy controls were included. Serum and tissue levels of malondialdehyde (MDA), glutathione S-transferase (GST), glutathione peroxidase (GPX), superoxide dismutase (SOD), glutathione reductase (GSR), and caspase-3 were quantitatively assessed using ELISA tests. Gross and histopathological analyses confirmed MAP infection. Infected animals showed significantly elevated serum levels of MDA and caspase-3 (p < 0.001), along with decreased antioxidant enzyme activities (GSR, GST, GPX, SOD). Tissue analysis revealed increased MDA and caspase-3 levels, particularly in the intestinal mucosa compared to mesenteric lymph nodes, suggesting localized oxidative damage and apoptosis. Conversely, antioxidant enzyme activity was higher in mesenteric lymph nodes, indicating a compensatory response and a pronounced involvement of the intestinal tract. These findings demonstrate that MAP infection induces marked oxidative stress and apoptotic processes, especially in the intestinal mucosa. The imbalance between pro-oxidant and antioxidant systems may play a key role in the pathogenesis and chronic progression of the disease. Caspase-3 and MDA, in particular, have been identified as promising diagnostic or prognostic biomarkers for MAP infection. This study highlights the importance of developing improved diagnostic tools and therapeutic strategies targeting oxidative stress pathways in paratuberculosis. Full article

(This article belongs to the Special Issue Biology of Mycobacterial Pathogens)

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16 pages, 876 KiB

Open AccessArticle

M72 Fusion Proteins in Nanocapsules Enhance BCG Efficacy Against Bovine Tuberculosis in a Mouse Model

by Federico Carlos Blanco, Renée Onnainty, María Rocío Marini, Laura Inés Klepp, Elizabeth Andrea García, Cristina Lourdes Vazquez, Ana Canal, Gladys Granero and Fabiana Bigi

Pathogens 2025, 14(6), 592; https://doi.org/10.3390/pathogens14060592 - 16 Jun 2025

Abstract

Mycobacterium bovis is the causative pathogen of bovine tuberculosis (bTB), a disease that affects cattle and other mammals, including humans. Currently, there is no efficient vaccine against bTB, underscoring the need for novel immunization strategies. The M72 fusion protein, composed of three polypeptides derived [...] Read more.

Mycobacterium bovis is the causative pathogen of bovine tuberculosis (bTB), a disease that affects cattle and other mammals, including humans. Currently, there is no efficient vaccine against bTB, underscoring the need for novel immunization strategies. The M72 fusion protein, composed of three polypeptides derived from Mycobacterium tuberculosis and M. bovis, has demonstrated protective efficacy against M. tuberculosis in clinical trials when combined with the AS01E adjuvant. Given the established efficacy of nanocapsule formulations as vaccine delivery systems, this study evaluated a novel immunization strategy combining BCG with either full-length M72 or a truncated M72 fused to a streptococcal albumin-binding domain (ABDsM72). Both antigens were encapsulated in chitosan/alginate nanocapsules and assessed in a murine M. bovis challenge model. Priming with BCG followed by an M72 boost significantly improved splenic protection compared to BCG alone, but it did not enhance pulmonary protection. Notably, boosting with ABDsM72 further increased the proportion of CD4+KLRG1-CXCR3+ T cells in the lungs of M. bovis-challenged mice, a key correlate of protective immunity. These findings demonstrate that chitosan/alginate-encapsulated antigens enhance BCG-induced immunity, supporting their potential as next-generation vaccine candidates for bTB control. Full article

(This article belongs to the Special Issue Mycobacterial Infection: Pathogenesis and Drug Development)

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20 pages, 4293 KiB

Open AccessArticle

Novel Antischistosomal Drug Targets: Identification of Alkaloid Inhibitors of SmTGR via Integrated In Silico Methods

by Valéria V. M. Paixão, Yria J. A. Santos, Adriana O. Fernandes, Elaine S. Conceição, Ricardo P. Rodrigues, Daniela A. Chagas-Paula, Silvio S. Dolabella and Tiago B. Oliveira

Pathogens 2025, 14(6), 591; https://doi.org/10.3390/pathogens14060591 - 15 Jun 2025

Abstract

Schistosomiasis mansoni is a neglected tropical disease caused by the parasite Schistosoma mansoni, affecting approximately 200 million people annually. Currently, treatment relies primarily on a single drug, praziquantel (PZQ), which shows limited efficacy against the parasite’s immature forms. As a result, Thioredoxin [...] Read more.

Schistosomiasis mansoni is a neglected tropical disease caused by the parasite Schistosoma mansoni, affecting approximately 200 million people annually. Currently, treatment relies primarily on a single drug, praziquantel (PZQ), which shows limited efficacy against the parasite’s immature forms. As a result, Thioredoxin Glutathione Reductase from S. mansoni (SmTGR) has emerged as a promising target for novel drug development. This study presents the development of integrated in silico methods to identify alkaloids from medicinal plants with potential activity against S. mansoni. Fourteen alkaloids were identified, with predicted activity ranging from 61.3 to 85.2%. Among these, lindoldhamine and daibucarboline A demonstrated, for the first time, potential SmTGR inhibition, with probabilities of 85.2% and 75.8%, respectively. These findings highlight the potential of these alkaloids as promising candidates for the development of new therapies against schistosomiasis. Full article

(This article belongs to the Section Parasitic Pathogens)

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12 pages, 2254 KiB

Open AccessArticle

Latency-Associated Nuclear Antigen (LANA) Promotes Ferroptosis by Suppressing Nrf2/GPX4 and Upregulating MDM2

by Yuejia Cao, Shihan Shao, Yingying Zhang, Dandan Song, Fei Gui, Xinyi Chen, Yu Hong, Rong Chen, Yang Song, Dongmei Li, Xiaohua Tan and Chunhong Di

Pathogens 2025, 14(6), 590; https://doi.org/10.3390/pathogens14060590 - 15 Jun 2025

Abstract

Ferroptosis, an iron-dependent cell death driven by lipid peroxidation, is regulated by key mediators including glutathione peroxidase 4 (GPX4) and nuclear factor erythroid 2-related factor 2 (Nrf2). Kaposi’s sarcoma-associated herpesvirus (KSHV) encodes latency-associated nuclear antigen (LANA), a multifunctional protein critical for viral persistence. [...] Read more.

Ferroptosis, an iron-dependent cell death driven by lipid peroxidation, is regulated by key mediators including glutathione peroxidase 4 (GPX4) and nuclear factor erythroid 2-related factor 2 (Nrf2). Kaposi’s sarcoma-associated herpesvirus (KSHV) encodes latency-associated nuclear antigen (LANA), a multifunctional protein critical for viral persistence. Although studies reported that KSHV infection enhanced cellular resistance to ferroptosis, the specific role of LANA in this process remains unexplored. Here, we demonstrate that LANA unexpectedly promotes ferroptosis. In KSHV-positive iSLK.219 cells, LANA knockdown significantly attenuated RSL-3-induced ferroptosis, whereas LANA overexpression sensitized HeLa cells to ferroptotic death. Quantitative analysis revealed that LANA-depleted cells exhibited significantly elevated ROS accumulation (p < 0.01), whereas LANA-overexpressing cells maintained reduced ROS levels during challenge with the ferroptosis inducer RSl-3. Mechanistically, LANA suppressed glutathione peroxidase 4 (GPX4) expression, reduced nuclear factor erythroid 2-related factor 2 (Nrf2) expression and impaired its nuclear translocation, and upregulated mouse double minute 2 homolog (MDM2) expression. Pharmacological inhibition of Nrf2 (ML385) or MDM2 (nutlin3a) reversed the ferroptotic effects of LANA knockdown or overexpression, respectively. These findings reveal a pro-ferroptotic role of LANA via Nrf2/GPX4 suppression and MDM2 activation. Full article

(This article belongs to the Special Issue Herpesvirus Latency and Reactivation)

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29 pages, 2576 KiB

Open AccessReview

A Comprehensive Review of the Mechanisms of Human Q Fever: Pathogenesis and Pathophysiology

by José-Luis Pérez-Arellano, Jose Curbelo and Cristina Carranza-Rodriguez

Pathogens 2025, 14(6), 589; https://doi.org/10.3390/pathogens14060589 - 14 Jun 2025

Abstract

Coxiella burnetii infection has a worldwide distribution, although the incidence and clinical manifestations vary between and within countries. There are the following four basic forms: asymptomatic infection, acute Q fever, chronic Q fever, and post-Q fever fatigue syndrome. The aim of this review [...] Read more.

Coxiella burnetii infection has a worldwide distribution, although the incidence and clinical manifestations vary between and within countries. There are the following four basic forms: asymptomatic infection, acute Q fever, chronic Q fever, and post-Q fever fatigue syndrome. The aim of this review is to provide a comprehensive overview of the important aspects of its pathogenesis and pathophysiology. First, we provide a brief update of the taxonomic aspects, basic structures, and genotypes of C. burnetii necessary for the proper interpretation of the following sections. Routes of infection, different stages of pathogenesis (respiratory entry of C. burnetii; penetration into alveolar macrophages, life cycle, and effects; systemic dissemination), and innate, acquired humoral and cell-mediated immune responses in different forms of infection are described in detail. The pathophysiology and clinical manifestations of Q fever, such as the main mechanisms of injury, in isolation and in combination, are reviewed. The clinical and biological manifestations of the two main forms of Q fever (acute and chronic) are outlined, with a brief definition and mention of the mechanisms of post-Q fever fatigue syndrome. Full article

(This article belongs to the Section Bacterial Pathogens)

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19 pages, 266 KiB

Open AccessArticle

Characteristics of Neonates with Sepsis Associated with Antimicrobial Resistance and Mortality in a Tertiary Hospital in Mexico: A Retrospective Observational Study

by Uriel A. Angulo-Zamudio, Maria Luisa Velazquez-Meza, Jesus J. Martinez-Garcia, Nidia Leon-Sicairos, Jorge Velazquez-Roman, Hector Flores-Villaseñor, Claudia Leon-Sicairos, Francisco A. Martínez-Villa and Adrian Canizalez-Roman

Pathogens 2025, 14(6), 588; https://doi.org/10.3390/pathogens14060588 - 14 Jun 2025

Abstract

The objective of this study was to determine the epidemiological, clinical, and laboratory characteristics of newborns with sepsis in northwestern Mexico, identify the microorganisms causing early- and late-onset sepsis, and assess antimicrobial resistance. Additionally, it sought to associate neonatal characteristics with antimicrobial resistance [...] Read more.

The objective of this study was to determine the epidemiological, clinical, and laboratory characteristics of newborns with sepsis in northwestern Mexico, identify the microorganisms causing early- and late-onset sepsis, and assess antimicrobial resistance. Additionally, it sought to associate neonatal characteristics with antimicrobial resistance or mortality. A retrospective study was conducted from August 2021 to April 2023, during which 8382 neonatal clinical records were analyzed to collect epidemiological, clinical, and laboratory characteristics, as well as microorganisms isolated from neonates and their antimicrobial resistance profiles. Of these, 314 neonates with sepsis were included. The incidence of neonatal sepsis was 4% (314/8382), and the mortality was 12.7% (40/314); late-onset sepsis (65.3%) was more frequent than early-onset sepsis (34.7%). Staphylococcus epidermidis was the most frequently isolated bacterium in neonates with sepsis (both early- and late-onset). Gram-positive bacteria, such as Staphylococcus hominis and Enterococcus faecium, were associated with early-onset sepsis, whereas fungi, particularly Candida albicans, were associated with late-onset sepsis. Of the microorganisms, 52.6% were multidrug resistant (MDR), 10.8% were extensively drug resistant (XDR), and 5.5% were pan-drug resistant (PDR). Low birth weight, prematurity, cesarean section, mechanical ventilation, tachycardia, and low hemoglobin and platelet levels, among others, were associated with XDR or MDR microorganisms. In contrast, low birth weight, mechanical ventilation, stroke, unexpected delivery, respiratory distress, tachycardia, convulsive crisis, high procalcitonin, urea, and AST/TGO levels, among others, were associated with mortality. The incidence, types of sepsis, antimicrobial resistance, and associations identified in this study will aid in diagnosing neonatal sepsis earlier and may reduce mortality in our region. Full article

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

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Graphical abstract

15 pages, 1423 KiB

Open AccessArticle

First Outbreak of Aeromoniasis, Caused by Aeromonas veronii, in Farmed European Seabass (Dicentrarchus labrax) in the Ionian Sea, Greece

by Eirini Lampou, Eleni Psychari, Konstantina Louka, Charalampos Kotzamanidis, Andigoni Malousi, Ioannis Petropoulos, Markos N. Kolygas, Dimitrios Doukas and Konstantina Bitchava

Pathogens 2025, 14(6), 587; https://doi.org/10.3390/pathogens14060587 - 14 Jun 2025

Abstract

This article documents the first outbreak of aeromoniasis caused by Aeromonas veronii in farmed European seabass (Dicentrarchus labrax) along the Greek Ionian Sea coast. In late spring 2024, commercially sized fish exhibited anorexia, hemorrhages, and ulcers on the skin, accompanied by [...] Read more.

This article documents the first outbreak of aeromoniasis caused by Aeromonas veronii in farmed European seabass (Dicentrarchus labrax) along the Greek Ionian Sea coast. In late spring 2024, commercially sized fish exhibited anorexia, hemorrhages, and ulcers on the skin, accompanied by elevated morbidity and mortality rates. The outbreak spread rapidly across local farms in Sagiada Bay, reaching its peak in late summer, and extending into the Astakos Gulf, southern in the Ionian Sea. The postmortem examination revealed hemorrhages, organomegaly, abscess formation, and granulomatous inflammation. Aeromonas veronii was isolated from all examined individuals in nutrient media and confirmed by biochemical and molecular methods. Whole genome sequencing and phylogenetic analysis demonstrated genetic homogeneity among two strains from two different areas along the Ionian Sea and a close evolutionary relationship with other Aeromonas veronii strains from the Aegean Sea. Although genetically similar, the isolates exhibited differences in phenotypic and biochemical characteristics, indicating regional variability. The present study provides an overview of the pathology, clinical characteristics and progression of aeromoniasis in Ionian Sea aquaculture, highlighting the need for continued monitoring, in-depth genomic and phenotypic assessment, and the design of region-specific preventive strategies, including autogenous vaccines, for effective disease management. Full article

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65 pages, 6478 KiB

Open AccessReview

The Challenge of Lyssavirus Infections in Domestic and Other Animals: A Mix of Virological Confusion, Consternation, Chagrin, and Curiosity

by Charles E. Rupprecht, Aniruddha V. Belsare, Florence Cliquet, Philip P. Mshelbwala, Janine F. R. Seetahal and Vaughn V. Wicker

Pathogens 2025, 14(6), 586; https://doi.org/10.3390/pathogens14060586 - 13 Jun 2025

Abstract

Lyssaviruses are RNA viruses in the Family Rhabdoviridae, Genus Lyssavirus. They represent the causative agents of acute, progressive encephalitis, known historically as rabies. Regardless of specific etiology, their collective viral morphology, biochemistry, pathobiology, associated clinical signs, diagnosis, epizootiology, and management are essentially [...] Read more.

Lyssaviruses are RNA viruses in the Family Rhabdoviridae, Genus Lyssavirus. They represent the causative agents of acute, progressive encephalitis, known historically as rabies. Regardless of specific etiology, their collective viral morphology, biochemistry, pathobiology, associated clinical signs, diagnosis, epizootiology, and management are essentially the same. Despite centuries of clinical recognition, these quintessential neurotropic agents remain significant pathogens today, with substantive consequences to agriculture, public health, and conservation biology. Notably, the singular morbidity caused by lyssaviruses is incurable and constitutes the highest case fatality of any viral disease. All warm-blooded vertebrates are believed to be susceptible. The dog is the only domestic animal that serves as a reservoir, vector, and victim. In contrast, felids are effective vectors, but not reservoirs. All other rabid domestic species, such as livestock, constitute spillover infections, as a bellwether to local lyssavirus activity. Frequently, professional confusion abounds among the veterinary community, because although the viral species Lyssavirus rabies is inarguably the best-known representative in the Genus, at least 20 other recognized or putative members of this monophyletic group are known. Frequently, this is simply overlooked. Moreover, often the ‘taxonomic etiology’ (i.e., ‘Lyssavirus x’) is mistakenly referenced in a biopolitcal context, instead of the obvious clinical illness (i.e., ‘rabies’). Global consternation persists, if localities believe they are ‘disease-free’, when documented lyssaviruses circulate or laboratory-based surveillance is inadequate to support such claims. Understandably, professional chagrin develops when individuals mistake the epidemiological terminology of control, prevention, elimination, etc. Management is not simple, given that the only licensed veterinary and human vaccines are against rabies virus, sensu lato. There are no adequate antiviral drugs for any lyssaviruses or cross-reactive biologics developed against more distantly related viral members. While representative taxa among the mammalian Orders Chiroptera, Carnivora, and Primates exemplify the major global reservoirs, which mammalian species are responsible for the perpetuation of other lyssaviruses remains a seemingly academic curiosity. This zoonosis is neglected. Clearly, with such underlying characteristics as a fundamental ‘disease of nature’, rabies, unlike smallpox and rinderpest, is not a candidate for eradication. With the worldwide zeal to drive human fatalities from canine rabies viruses to zero by the rapidly approaching year 2030, enhanced surveillance and greater introspection of the poorly appreciated burden posed by rabies virus and diverse other lyssaviruses may manifest as an epidemiological luxury to the overall global program of the future. Full article

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

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19 pages, 9515 KiB

Open AccessArticle

Survey of Piroplasmids in Wild Mammals, Unconventional Pets, and Ticks from Goiás State, Midwestern Brazil

by Raphaela Bueno Mendes Bittencourt, Ana Cláudia Calchi, Lucianne Cardoso Neves, Nicolas Jalowitzki de Lima, Gabriel Cândido dos Santos, Ennya Rafaella Neves Cardoso, Warley Vieira de Freitas Paula, Luciana Batalha de Miranda Araújo, Jessica Rocha Gonçalves, Elisângela de Albuquerque Sobreira, Luiz Alfredo Martins Lopes Baptista, Hermes Ribeiro Luz, Marcos Rogério André, Filipe Dantas-Torres and Felipe da Silva Krawczak

Pathogens 2025, 14(6), 585; https://doi.org/10.3390/pathogens14060585 - 12 Jun 2025

Abstract

Tick-borne piroplasmids are apicomplexan protozoa that infect a wide range of vertebrate hosts, with significant implications for animal and human health. This study investigated the occurrence and genetic diversity of piroplasmids in wild mammals, unconventional pets, and associated ticks in Goiás state, midwestern [...] Read more.

Tick-borne piroplasmids are apicomplexan protozoa that infect a wide range of vertebrate hosts, with significant implications for animal and human health. This study investigated the occurrence and genetic diversity of piroplasmids in wild mammals, unconventional pets, and associated ticks in Goiás state, midwestern Brazil. Between April 2023 and January 2024, 105 blood samples, 22 tissue samples, and 300 ticks were collected from 21 mammalian species housed in wildlife screening centers, zoos, and veterinary clinics. Molecular screening targeting the 18S rRNA gene of piroplasmids detected a 25.7% (27/105) overall positivity, with gray brockets (Subulo gouazoubira) and South American tapirs (Tapirus terrestris) showing the highest infection rates. Three tick samples tested positive, including two Amblyomma sculptum nymphs and a male of Amblyomma dubitatum collected from a tapir and capybara (Hydrochoerus hydrochaeris). Cytauxzoon brasiliensis was reported, for the first time, in cougars (Puma concolor) from Goiás state, midwestern Brazil, indicating the role of this feline as a host of this parasite. Babesia goianiaensis was confirmed in a capybara, and Theileria terrestris in tapirs. Phylogenetic analyses clustered gray brockets-associated Theileria sequences with Theileria sp. previously detected in Neotropical deer from Brazil and Theileria cervi. While the phylogenetic analysis of amino acid sequences of the cytochrome c oxidase subunit III separated Theileria genotypes detected in S. gouazoubira from T. cervi, hsp70-based phylogenetic inferences clustered the genotypes detected in Tapirus terrestris with Theileria terrestris, suggesting host-specific evolutionary lineages. These findings contribute to the understanding of Piroplasmida diversity and circulation in South American wild mammals, emphasizing the need for enhanced molecular surveillance to elucidate transmission dynamics, assess potential health risks, and contribute to the establishment of wildlife conservation and One Health strategies. Full article

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16 pages, 1357 KiB

Open AccessArticle

EmsB Microsatellite Analysis of Echinococcus multilocularis Specimens Isolated from Belgian Patients with Alveolar Echinococcosis and from Animal Hosts

by Sabrina Egrek, Jenny Knapp, Rosalie Sacheli, Khalid El Moussaoui, Philippe Léonard, Eva Larranaga Lapique, Laurence Millon, Sara Engelskirchen, Olivier Detry, Annick Linden and Marie-Pierre Hayette

Pathogens 2025, 14(6), 584; https://doi.org/10.3390/pathogens14060584 - 12 Jun 2025

Abstract

Alveolar echinococcosis (AE), caused by Echinococcus multilocularis (E. multilocularis), is a severe parasitic zoonosis that is potentially fatal for humans. The parasite is primarily transmitted by wildlife, with red foxes acting as definitive hosts and rodents as intermediate hosts, while humans [...] Read more.

Alveolar echinococcosis (AE), caused by Echinococcus multilocularis (E. multilocularis), is a severe parasitic zoonosis that is potentially fatal for humans. The parasite is primarily transmitted by wildlife, with red foxes acting as definitive hosts and rodents as intermediate hosts, while humans can become accidental but dead-end hosts. The aim of this study is to use EmsB typing on E. multilocularis isolates from human AE cases and local animals such as foxes and rodents. In this study, retrospective EmsB typing was performed on 39 samples, including 11 tissue samples from 10 patients, 18 fecal swabs from foxes, and 10 tissue samples from rodents. A dendrogram was created to determine the EmsB profiles present. The results showed that all the rodent samples were associated with the EmsB P1 profile (10/10), while the human and fox samples shared the EmsB profile P1 (5/11 humans and 8/18 foxes), a profile near P4 (2/11 humans and 3 foxes), and a profile near P8 (1/11 humans and 1/18 foxes). The study demonstrates that the same EmsB profiles circulate among humans and animals, confirming that wildlife reservoirs play a key role in transmission. Full article

(This article belongs to the Section Parasitic Pathogens)

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7 pages, 315 KiB

Open AccessBrief Report

Implications of Intravenous and Inhaled Amikacin Breakpoint Reporting for Mycobacterium avium Complex Pulmonary Isolates

by Christian M. Gill, Robin Chamberland and Getahun Abate

Pathogens 2025, 14(6), 583; https://doi.org/10.3390/pathogens14060583 - 12 Jun 2025

Abstract

The treatment of Mycobacterium avium complex (MAC) remains a clinical challenge as multidrug regimens are needed and may be limited by treatment-related toxicity. The Clinical and Laboratory Standards Institute (CLSI) endorses breakpoints for several agents used for MAC infection treatment. Amikacin has distinct [...] Read more.

The treatment of Mycobacterium avium complex (MAC) remains a clinical challenge as multidrug regimens are needed and may be limited by treatment-related toxicity. The Clinical and Laboratory Standards Institute (CLSI) endorses breakpoints for several agents used for MAC infection treatment. Amikacin has distinct breakpoints for intravenous (IV) therapy and inhaled therapy using amikacin liposome inhalation suspension (ALIS) for MAC pulmonary disease. The purpose of the present retrospective cohort study of MAC pulmonary isolates was to assess the number of amikacin non-susceptible isolates by the IV breakpoints that remain susceptible to the inhaled breakpoints. One isolate per patient per year was assessed and susceptibility was described for amikacin IV, amikacin inhaled, clarithromycin, moxifloxacin, and linezolid per the CLSI. Of the 218 isolates, 94% [204/218] tested as susceptible to amikacin per the IV breakpoints compared with 99.5% [217/218] to the inhaled breakpoints. Of the amikacin IV non-susceptible isolates, 93% [13/14] were susceptible by the inhaled breakpoints. For comparison, clarithromycin was the next most active agent followed by moxifloxacin and linezolid with 97% [211/218], 82% [178/218], and 66% [143/218] of isolates testing as susceptible to each, respectively. These data highlight the importance of laboratories to report both the IV and inhaled amikacin interpretive criteria so that clinicians do not disregard potential therapeutic options for the treatment of MAC pulmonary disease. Full article

(This article belongs to the Section Bacterial Pathogens)

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21 pages, 3837 KiB

Open AccessArticle

Evaluation of the In Vitro Activity of Bedaquiline, Delamanid, and Clofazimine Against Mycobacterium abscessus Complex and Their Antibiofilm Potential

by Katarzyna Kania, Katarzyna Wójcik, Alicja Skórkowska and Karolina Klesiewicz

Pathogens 2025, 14(6), 582; https://doi.org/10.3390/pathogens14060582 - 11 Jun 2025

Abstract

Mycobacterium abscessus complex (MABc) poses a major therapeutic challenge due to its intrinsic multidrug resistance and ability to form biofilms. This study evaluated the in vitro activity of three antimycobacterial agents—bedaquiline, delamanid, and clofazimine—on 20 clinical MABc isolates, including M. abscessus subsp. abscessus [...] Read more.

Mycobacterium abscessus complex (MABc) poses a major therapeutic challenge due to its intrinsic multidrug resistance and ability to form biofilms. This study evaluated the in vitro activity of three antimycobacterial agents—bedaquiline, delamanid, and clofazimine—on 20 clinical MABc isolates, including M. abscessus subsp. abscessus, massiliense, and bolletii, with a focus on biofilm-forming phenotypes. Biofilm analysis showed that the rough colony morphotypes were mostly weak biofilm formers, while the smooth and mixed morphotypes were predominantly moderate or strong biofilm formers. A statistically significant association was observed between the mixed colony morphology and strong biofilm formation (p = 0.032). Importantly, bedaquiline exhibited potent and consistent activity across all isolates, regardless of the biofilm-forming ability, with MIC values ranging from 0.125 to 1 µg/mL. In contrast, delamanid and clofazimine showed limited efficacy, with MIC values exceeding 16 µg/mL and 8 µg/mL, respectively. These findings strongly support the role of bedaquiline as a promising core agent for future combination therapies targeting drug-resistant MABc infections, including biofilm-associated infections. Our results, among the first from Poland, highlight the critical need for incorporating novel agents such as bedaquiline into therapeutic strategies against this difficult-to-treat pathogen. Full article

(This article belongs to the Special Issue Pathogenesis and Treatment of Non-Tuberculous Mycobacterial Infections)

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16 pages, 1065 KiB

Open AccessArticle

First Serologic Analysis of Antibodies Against African Swine Fever Virus Detected in Domestic Pig Farms in South Korea from 2019 to 2024

by Seong-Keun Hong, Mugyeom Moon, Ki-Hyun Cho, Hae-Eun Kang, Jong-Soo Lee and Yeon-Hee Kim

Pathogens 2025, 14(6), 581; https://doi.org/10.3390/pathogens14060581 - 11 Jun 2025

Abstract

Background: African swine fever (ASF) is a crucial socioeconomic setback to South Korea’s swine industry. This study aimed to determine seropositivity for ASF virus (ASFV) in pigs that appeared to be infected on farms with reported ASF outbreaks. Methods: A total of 2232 [...] Read more.

Background: African swine fever (ASF) is a crucial socioeconomic setback to South Korea’s swine industry. This study aimed to determine seropositivity for ASF virus (ASFV) in pigs that appeared to be infected on farms with reported ASF outbreaks. Methods: A total of 2232 sera from ASF outbreaks (2019–2024) in South Korea were collected. Two enzyme-linked immunosorbent assay (ELISA) kits were used to detect ASFV antibodies, and an immunoperoxidase test (IPT) was used as a confirmatory test following the method recommended by the World Organisation for Animal Health in the Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. Also, spatial clustering was identified using the Density-Based Spatial Clustering of Applications with Noise (DBSCAN) model to understand ASF hotspots in the wild boar population and assess the spatial relationship between the hotspots and ASF antibody-positive domestic pig farms. Results: Antibodies were first detected in Hwacheon in 2020, but by 2024, only 1.43% of pigs had detectable antibodies against ASFV. Although this percentage is still low, the number of antibody-positive pigs is gradually increasing. Additionally, 32 positive samples were found from nine pig farms with outbreaks, and these samples were confirmed positive in both the two ELISA tests and the IPT. The highest seropositivity was recorded at the finishing stage of pig production. When compared to the confirmatory IPT, both blocking and competition ELISA demonstrated high diagnostic sensitivities. The statistical association between ASF antibody-positive farms and wild boars were analyzed using Fisher’s exact test, yielding a significant p-value of 0.007. This indicates a strong correlation, as eight out of nine ASF-seropositive farms were located within hotspots that were significantly associated. Conclusions: Our findings provide valuable insights into ASFV antibody detection in South Korea and demonstrate a statistical association between farms housing pigs with ASFV antibodies and hotspots of ASFV-infected wild boars. Confirmatory tests, such as the IPT, are needed. These insights will contribute to the improvement of surveillance and biosecurity measures for swine farms. Full article

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

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16 pages, 1381 KiB

Open AccessArticle

Characterization of Microbial Growth, Pathogen Presence, and Histamine Accumulation in Chilled Rainbow Trout and Mackerel Samples Collected from Romanian Markets

by Vida Silviu, Alexandra Tabaran, Oana Lucia Crişan Reget, Mihaela Niculina Duma, Luciana Cătălina Panait and Sorin Daniel Dan

Pathogens 2025, 14(6), 580; https://doi.org/10.3390/pathogens14060580 - 11 Jun 2025

Abstract

This study aimed to evaluate microbial growth, pathogen presence, and histamine production in rainbow trout and mackerel stored on flaked ice over their shelf life. A total of 72 fish samples (rainbow trout and mackerel) were analyzed across four storage intervals (day 1, [...] Read more.

This study aimed to evaluate microbial growth, pathogen presence, and histamine production in rainbow trout and mackerel stored on flaked ice over their shelf life. A total of 72 fish samples (rainbow trout and mackerel) were analyzed across four storage intervals (day 1, 3, 9, 12/11) on flaked ice. TVC increased from 2.59 to 5.04 log cfu/g in rainbow trout and from 3.18 to 4.88 log cfu/g in mackerel over the storage period. Significant increases were observed in Pseudomonas, Aeromonas, and Enterobacteriaceae populations, especially after the ninth day. Microbial identification revealed spoilage-associated bacteria, such as Pseudomonas fluorescens and Aeromonas salmonicida, as well as opportunistic pathogens, including Francisella tularensis, Yersinia spp., and Chromobacterium violaceum. Histamine levels rose with storage time but remained below toxic thresholds (<200 mg/kg), peaking at 1.56 mg/kg in trout and 1.87 mg/kg in mackerel. A strong positive correlation was found between TVC and histamine levels (Pearson’s r = 0.85 for trout, 0.82 for mackerel). Proper hygiene and storage are crucial, and consumption is recommended before day 9 of storage on flaked ice. Hygiene measures remain essential to minimize contamination risks and preserve product safety. Full article

(This article belongs to the Special Issue Globalization and Emerging Zoonotic Diseases: One Health Perspectives on Veterinary Public Health and Epidemiology)

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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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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)

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