Pathogens

16 pages, 1564 KiB

Open AccessArticle

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

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

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

Abstract

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

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

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

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10 pages, 1764 KiB

Open AccessBrief Report

Cathelicidins Limit Intracellular Neospora caninum-Infection in Macrophages

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

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

Abstract

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

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

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

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14 pages, 3732 KiB

Open AccessArticle

Baicalein and Berberine Inhibit the Growth and Virulence of Clostridioides difficile

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

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

Abstract

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

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

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16 pages, 837 KiB

Open AccessArticle

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

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

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

Abstract

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

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

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

11 pages, 220 KiB

Open AccessArticle

Immune-Mediated Hemolytic Anemia in Cats with Feline Infectious Peritonitis

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

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

Abstract

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

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

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

10 pages, 1814 KiB

Open AccessArticle

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

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

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

Abstract

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

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

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

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15 pages, 1072 KiB

Open AccessArticle

Wastewater Surveillance for Group A Streptococcus pyogenes in a Small City

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

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

Abstract

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

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

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

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16 pages, 904 KiB

Open AccessReview

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

by Ben Enyetornye, Aurelle Yondo and Binu T. Velayudhan

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

Abstract

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

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

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

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11 pages, 1403 KiB

Open AccessArticle

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

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

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

Abstract

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

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

(This article belongs to the Section Bacterial Pathogens)

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18 pages, 9567 KiB

Open AccessArticle

Evaluating Entomopathogenic Nematodes as Biocontrol Agents Against Two Major Cockroach Species, Blattella germanica and Periplaneta americana, in Antalya, Türkiye

by Aysegul Cengiz, Burak Polat, Sevval Kahraman Kokten, Ummuhan Aslan Bıckı, Cansu Calıskan, Samed Koc, Emre Oz, Serap Kocaoglu-Cenkci, Ozge Tufan-Cetin and Huseyin Cetin

Pathogens 2025, 14(7), 655; https://doi.org/10.3390/pathogens14070655 - 1 Jul 2025

Abstract

Cockroaches, particularly the German cockroach (Blattella germanica Linnaeus, Blattodea: Ectobiidae) and the American cockroach (Periplaneta americana (Linnaeus), Blattodea: Blattidae), are major public health pests due to their ability to transmit pathogens and develop resistance to chemical insecticides, including synthetic pyrethroids, which [...] Read more.

Cockroaches, particularly the German cockroach (Blattella germanica Linnaeus, Blattodea: Ectobiidae) and the American cockroach (Periplaneta americana (Linnaeus), Blattodea: Blattidae), are major public health pests due to their ability to transmit pathogens and develop resistance to chemical insecticides, including synthetic pyrethroids, which are widely used worldwide. Given the increasing resistance, entomopathogenic nematodes (EPNs) have emerged as a potential biological control alternative. This study evaluates the efficacy of three EPN species, Steinernema carpocapsae (Weiser), S. feltiae (Filipjev), and Heterorhabditis bacteriophora Poinar, against B. germanica and P. americana collected from different regions of Antalya, Türkiye. Laboratory bioassays were conducted under controlled conditions, testing five EPN concentrations (100, 250, 500, 750, and 1000 IJs/mL). The results showed that S. carpocapsae was the most effective, causing mortality rates of 46.7% to 100% in adult German cockroaches and 20% to 66.7% in nymphs, while S. feltiae and H. bacteriophora exhibited lower efficacy. American cockroaches showed higher resistance, with S. carpocapsae achieving a maximum mortality of 33.3% at the highest concentration, whereas S. feltiae and H. bacteriophora had no significant lethal effect. These findings suggest that S. carpocapsae could be a promising biological control agent for B. germanica, particularly in pyrethroid-resistant populations. Full article

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Graphical abstract

34 pages, 981 KiB

Open AccessReview

Applying CRISPR Technologies for the Treatment of Human Herpesvirus Infections: A Scoping Review

by Chloë Hanssens and Jolien Van Cleemput

Pathogens 2025, 14(7), 654; https://doi.org/10.3390/pathogens14070654 - 1 Jul 2025

Abstract

Background: Human herpesviruses are double-stranded DNA viruses of which eight types have been identified at present. Herpesvirus infection comprises an active lytic phase and a lifelong latency phase with the possibility of reactivation. These infections are highly prevalent worldwide and can lead to [...] Read more.

Background: Human herpesviruses are double-stranded DNA viruses of which eight types have been identified at present. Herpesvirus infection comprises an active lytic phase and a lifelong latency phase with the possibility of reactivation. These infections are highly prevalent worldwide and can lead to a broad spectrum of clinical manifestations, ranging from mild symptoms to severe disease, particularly in immunocompromised individuals. Clustered regularly interspaced palindromic repeats (CRISPR)-based therapy is an interesting alternative to current antiviral drugs, which fail to cure latent infections and are increasingly challenged by viral resistance. Objective: This scoping review aimed to summarize the current state of CRISPR-based antiviral strategies against herpesvirus infections, highlighting the underlying mechanisms, study design and outcomes, and challenges for clinical implementation. Design: A literature search was conducted in the databases PubMed and Web of Science, using both a general and an individual approach for each herpesvirus. Results: This scoping review identified five main mechanisms of CRISPR-based antiviral therapy against herpesvirus infections in vitro and/or in vivo. First, CRISPR systems can inhibit the active lytic replication cycle upon targeting viral lytic genes or host genes. Second, CRISPR technologies can remove latent viral genomes from infected cells by targeting viral genes essential for latency maintenance or destabilizing the viral genome. Third, reactivation of multiple latent herpesvirus infections can be inhibited by CRISPR-Cas-mediated editing of lytic viral genes, preventing a flare-up of clinical symptoms and reducing the risk of viral transmission. Fourth, CRISPR systems can purposefully induce viral reactivation to enhance recognition by the host immune system or improve the efficacy of existing antiviral therapies. Fifth, CRISPR technology can be applied to develop or enhance the efficiency of cellular immunotherapy. Conclusions: Multiple studies demonstrate the potential of CRISPR-based antiviral strategies to target herpesvirus infections through various mechanisms in vitro and in vivo. However, aspects regarding the delivery and biosafety of CRISPR systems, along with the time window for treatment, require further investigation before broad clinical implementation can be realized. Full article

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

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2 pages, 322 KiB

Open AccessCorrection

Correction: Ortega-Carballo et al. Effect of Stevioside (Stevia rebaudiana) on Entamoeba histolytica Trophozoites. Pathogens 2024, 13, 373

by Karla Jocelyn Ortega-Carballo, Karla Montserrat Gil-Becerril, Karla Berenice Acosta-Virgen, Sael Casas-Grajales, Pablo Muriel and Víctor Tsutsumi

Pathogens 2025, 14(7), 653; https://doi.org/10.3390/pathogens14070653 - 1 Jul 2025

Abstract

In the original publication [...] Full article

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10 pages, 6593 KiB

Open AccessArticle

First Molecular Evidence of Ixodiphagus hookeri (Hymenoptera: Encyrtidae) in Ixodes ricinus and Haemaphysalis concinna (Acari: Ixodida) Ticks from Inland and Coastal Areas of the Balkan Peninsula

by Veronika Blažeková, Michal Stanko, Dana Zubriková, Lucia Vargová, Klaudia Mária Švirlochová and Bronislava Víchová

Pathogens 2025, 14(7), 652; https://doi.org/10.3390/pathogens14070652 - 1 Jul 2025

Abstract

Ixodiphagus hookeri (Howard, 1907) (Hymenoptera: Encyrtidae), a hyperparasitic wasp that parasitizes hard ticks, has been documented in various parts of Europe; however, data on its presence in southeastern regions has been lacking. This study provides the first molecular evidence of I. hookeri in [...] Read more.

Ixodiphagus hookeri (Howard, 1907) (Hymenoptera: Encyrtidae), a hyperparasitic wasp that parasitizes hard ticks, has been documented in various parts of Europe; however, data on its presence in southeastern regions has been lacking. This study provides the first molecular evidence of I. hookeri in ticks from the coastal areas of the Balkan Peninsula, specifically Croatia and Bulgaria. A total of 1043 questing ticks were collected between 2011 and 2013 across 15 locations. Molecular screening revealed I. hookeri DNA in Ixodes ricinus (Linnaeus, 1758) (Acari: Ixodidae) nymphs from inland Croatia (overall prevalence: 18.72%) and in Haemaphysalis concinna (Koch, 1844) (Acari: Ixodidae) nymphs and larvae from coastal Bulgaria (prevalence: 17.2%). All I. hookeri-positive samples were co-infected with Wolbachia spp. (Rickettsiales: Anaplasmataceae). This detection marks the southernmost record of I. hookeri in Central Europe, expanding its known range to the Balkan Peninsula and supporting its relevance as a potential natural enemy in integrated tick management strategies. Full article

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

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11 pages, 1363 KiB

Open AccessCase Report

Molecular and Microscopic Challenges in Detecting Plasmodium cynomolgi Co-Infections with Plasmodium vivax: A Case Report

by Mohd Adilin Yaacob, Raden Shamilah Radin Hisam, Nor Parina Ismail, Noor Azian Md Yusuf, Jose Miguel Rubio Muñoz, Suhana Hashim and Tam Jenn Zhueng

Pathogens 2025, 14(7), 651; https://doi.org/10.3390/pathogens14070651 - 30 Jun 2025

Abstract

The risk of non-human primate (NHP) malaria transmission to humans is increasing, with Plasmodium knowlesi and Plasmodium cynomolgi emerging as significant zoonotic threats, particularly in Malaysia. While P. knowlesi is well-documented, P. cynomolgi infections in humans remain underreported, largely due to diagnostic challenges. [...] Read more.

The risk of non-human primate (NHP) malaria transmission to humans is increasing, with Plasmodium knowlesi and Plasmodium cynomolgi emerging as significant zoonotic threats, particularly in Malaysia. While P. knowlesi is well-documented, P. cynomolgi infections in humans remain underreported, largely due to diagnostic challenges. Routine microscopy and standard molecular diagnostic tools often misdiagnose P. cynomolgi infections as P. vivax due to morphological similarities and genetic homology. We report a new case of a human P. cynomolgi infection misdiagnosed as Plasmodium vivax in a 32-year-old male with no prior malaria history or travel to endemic countries. The initial diagnoses made by the microscopy and qPCR conducted by the Kota Bharu Public Health Laboratory in Kelantan identified the infection as P. vivax. However, cross-examination by the Institute for Medical Research (IMR) revealed the presence of mixed-species infection, prompting further analysis. The real-time PCR and sequencing performed at MAPELAB, Spain, confirmed the co-infection of P. vivax and P. cynomolgi. This case highlights the diagnostic limitations in detecting P. cynomolgi, which shares high genetic similarity with P. vivax, leading to potential cross-reactivity and diagnostic inaccuracies. As P. cynomolgi emerges as the second zoonotic malaria species after P. knowlesi capable of infecting humans in Southeast Asia, improved diagnostic methods are urgently needed. Enhanced molecular diagnostics and comprehensive epidemiological studies are essential to elucidate transmission dynamics, assess public health implications, and inform effective malaria control strategies. Full article

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

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15 pages, 9323 KiB

Open AccessArticle

Tissue Distribution and Abundance of the Parasitic Dinoflagellate Hematodinium perezi in Naturally Infected Portunus trituberculatus

by Ju Zhang, Meng Li, Qian Huang, Lijun Hu, Qi Xue, Jiayi Wang and Caiwen Li

Pathogens 2025, 14(7), 650; https://doi.org/10.3390/pathogens14070650 - 30 Jun 2025

Abstract

The parasitic dinoflagellate Hematodinium is an infectious pathogen that causes severe enzootic in numerous economically important marine crustaceans worldwide. Previous research has focused on investigating the identification and life stages of Hematodinium parasites, while the parasite abundance and tissue proliferation process of Hematodinium [...] Read more.

The parasitic dinoflagellate Hematodinium is an infectious pathogen that causes severe enzootic in numerous economically important marine crustaceans worldwide. Previous research has focused on investigating the identification and life stages of Hematodinium parasites, while the parasite abundance and tissue proliferation process of Hematodinium in naturally infected crustacean hosts need to be further studied. In the present study, the tissue tropisms and intensity of H. perezi were investigated in the naturally infected Chinese swimming crabs Portunus trituberculatus by both the qualitative (hemolymph assay, histology) and quantitative analysis (cell count, quantitative PCR). The results showed that in P. trituberculatus with infection level I (4 ± 2 parasites in 200× microscopic field), filamentous trophonts were observed in the hemolymph and stomach tissues, with the average parasite number and ITS 1 copy number of H. perezi quantitatively detected in hemolymph (1.0 × 10² parasites/mL) and stomach tissues (1.7 × 10³ cells/g), respectively. H. perezi trophonts were observed in the hemolymph (4.3 × 10⁴ parasites/mL) and exhibited broad distribution in multiple tissues with its highest abundance of H. perezi in pereiopod muscles (1.1 × 10⁴ cells/g) followed by that in stomach (4.8 × 10³ cells/g) in P. trituberculatus with infection level II (80 ± 10 parasites in 200× microscopic field). In P. trituberculatus with infection level III (200 ± 35 parasites in 200× microscopic field), a high abundance of H. perezi sporoblasts was found in the hemolymph (3.1 × 10⁷ parasites/mL) and all of the other examined tissues, with its highest abundance detected in pereiopod muscles (3.5 × 10⁴ cells/g). In addition, the number of host’s hemocytes was significantly decreased during the Hematodinium infection. This study provides a comprehensive quantitative characterization of the tissue distribution and abundance of H. perezi in its natural crab host which will contribute to better understanding of the crustacean host–Hematodinium interactions. Full article

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21 pages, 253 KiB

Open AccessArticle

Anthelmintic Resistance in Livestock Farming: Challenges and Perceptions of Farmers and Veterinarians

by Naida Kapo, Adis Softić, Teufik Goletić, Šejla Goletić, Aleksandar Cvetkovikj and Jasmin Omeragić

Pathogens 2025, 14(7), 649; https://doi.org/10.3390/pathogens14070649 - 30 Jun 2025

Abstract

Anthelmintic resistance in livestock is a growing concern worldwide, with significant implications for animal health and agricultural productivity. This study explores the perceptions of veterinarians and farmers in Bosnia and Herzegovina regarding the factors contributing to anthelmintic resistance in Haemonchus contortus nematodes. Data [...] Read more.

Anthelmintic resistance in livestock is a growing concern worldwide, with significant implications for animal health and agricultural productivity. This study explores the perceptions of veterinarians and farmers in Bosnia and Herzegovina regarding the factors contributing to anthelmintic resistance in Haemonchus contortus nematodes. Data were collected through structured questionnaires completed by 106 veterinarians and 188 farmers in 2022 and 2023. The analysis focused on self-reported therapeutic practices, farm management and environmental variables. Logistic regression, including Firth’s penalized approach, was used to assess associations between these perceived factors and the reported occurrence of resistance. Notably, combination anthelmintic treatments were perceived as a significant risk factor (OR > 49.3), while higher altitude was seen as potentially protective (OR = 0.10). Routine prophylactic deworming was associated with an increased likelihood of perceived resistance (OR = 173.7), whereas staying informed about newly registered products was perceived as protective (OR = 0.34). Although the findings are based on the self-reported perceptions and practices of veterinarians and farmers, they align with globally recognized trends and offer the first structured insights into factors perceived to contribute to anthelmintic resistance in Bosnia and Herzegovina. This study underscores the importance of awareness and responsible anthelmintic use and the need for improved diagnostics and ongoing education to combat anthelmintic resistance. Full article

(This article belongs to the Special Issue Pathogenesis, Epidemiology, and Drug Resistance in Nematode Parasites)

25 pages, 1347 KiB

Open AccessArticle

Antibiotic Resistance, Virulence Genes, and Molecular Diversity of Clinical Klebsiella pneumoniae Isolates from Patients of District Hospital in Central Poland

by Barbara Kot, Małgorzata Witeska, Piotr Szweda, Małgorzata Piechota, Elżbieta Kondera, Elżbieta Horoszewicz, Izabela Balak, Ahmer Bin Hafeez and Alicja Synowiec

Pathogens 2025, 14(7), 648; https://doi.org/10.3390/pathogens14070648 - 30 Jun 2025

Abstract

In hospital environments, pathogenic bacteria spread easily and acquire virulence and antibiotic resistance genes. The aim of the study was an evaluation of the genetic diversity of 109 K. pneumoniae isolates recovered from patients of a district hospital in central Poland. The frequencies [...] Read more.

In hospital environments, pathogenic bacteria spread easily and acquire virulence and antibiotic resistance genes. The aim of the study was an evaluation of the genetic diversity of 109 K. pneumoniae isolates recovered from patients of a district hospital in central Poland. The frequencies of genes coding for β-lactamases, efflux pumps, and virulence factors were determined. Genotyping of the isolates was performed with ERIC (Enterobacterial Repetitive Intergenic Consensus) and REP (Repetitive Element Sequence Based) PCR techniques, with 21 and 19 genotypes being identified, respectively. The bla_SHV-1 (92.7%), bla_CTX-M group 1 (83.5%), bla_TEM-1 (28.4%), bla_NDM-1 (16.5%), bla_VEB-1 (11.0%), bla_CTX-M group 9 (3.7%), bla_KPC (1.8%), bla_IMP, bla_OXA-48, bla_CTX-M group 2, bla_CTX-M groups 8, and 25/26 (0% each) and efflux pumps: AcrAB (100%), tolC (93.6%), and mdtk (60.5%), and virulence genes coding: urease subunit ureA (94.5%) endotoxins wabG (92.7%) and uge (64.2%), and siderophore iucB (3.7%) were detected. The bla_SHV-1, bla_CTX-M group 1, mdtk, tolC, AcrAB (16.5%); bla_SHV-1, bla_CTX-M group 1, tolC, AcrAB (15.6%), and bla_SHV-1, bla_CTX-M group 1, bla_NDM-1, mdtk, tolC, AcrAB (11.9%) were the most common resistance patterns. The distribution of resistance and virulence genes varied more between hospital wards than between different clinical materials. Hospital’s antibiotic-resistant and virulent K. pneumoniae, able to spread among humans, animals, and in the environment, pose a significant threat to public health. Full article

(This article belongs to the Section Bacterial Pathogens)

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20 pages, 5311 KiB

Open AccessArticle

Projected Distribution and Dispersal Patterns of Potential Distribution Fasciola hepatica and Its Key Intermediate Host Radix spp. in Qinghai-Tibet Plateau, China, Under Plateau Climatic Conditions

by Luyao Xu, Yunhai Guo, Zengkui Li, Mingjia Guo, Ming Kang, Daoxin Liu, Limin Yang, Zhongqiu Li, Panpan Wang, Wenhui Luo and Ying Li

Pathogens 2025, 14(7), 647; https://doi.org/10.3390/pathogens14070647 - 30 Jun 2025

Abstract

(1) Background: As a prominent zoonotic parasitic disease, fascioliasis threatens the sustainable development of animal husbandry and public health. Current research focuses mainly on individual species (parasite or intermediate host), neglecting systematic evaluation of the transmission chain and exposure risks to animal husbandry. [...] Read more.

(1) Background: As a prominent zoonotic parasitic disease, fascioliasis threatens the sustainable development of animal husbandry and public health. Current research focuses mainly on individual species (parasite or intermediate host), neglecting systematic evaluation of the transmission chain and exposure risks to animal husbandry. Thus, comprehensive studies are urgently needed, especially in the ecologically fragile alpine region of the Qinghai-Tibet Plateau; (2) Methods: Distribution data of Radix spp. and Fasciola hepatica in the Qinghai-Tibet Plateau and adjacent areas were gathered to establish a potential distribution model, which was overlaid on a map of livestock farming in the region; (3) Results: The key environmental factors influencing Radix spp. distribution were temperature seasonality (21.4%), elevation (16.4%), and mean temperature of the driest quarter (14.7%). For F. hepatica, the main factors were elevation (41.3%), human footprint index (30.5%), and Precipitation of the driest month (12.1%), with all AUC values exceeding 0.9. Both species exhibited extensive suitable habitats in Qinghai and Tibet, with higher F. hepatica transmission risk in Qinghai than Tibet; (4) Conclusions: The significant transmission risk and its impacts on the livestock industry in the Qinghai-Tibet Plateau highlight the need for proactive prevention and control measures. This study provides a scientific foundation for targeted alpine diseases control, establishes an interdisciplinary risk assessment framework, fills gaps in high-altitude eco-epidemiology, and offers insights for ecological conservation of the plateau. Full article

(This article belongs to the Section Parasitic Pathogens)

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17 pages, 2362 KiB

Open AccessArticle

Growth Inhibition and Additive Effect to Antimalarial Drugs of Brucea javanica Extracts on Asexual Blood-Stage Plasmodium falciparum

by Niwat Kangwanrangsan, Gamolthip Niramolyanun, Chonnipa Praikongkatham, Pathanin Chantree, Pongsakorn Martviset and Viriya Pankao

Pathogens 2025, 14(7), 646; https://doi.org/10.3390/pathogens14070646 - 30 Jun 2025

Abstract

Malaria is a parasitic infectious disease that is endemic in many tropical countries. Even though several effective antimalarial agents have been implemented, treatment failure still occurs, and malaria continues to cause neurological complications and death, particularly in severe or drug-resistant cases. Hence, novel [...] Read more.

Malaria is a parasitic infectious disease that is endemic in many tropical countries. Even though several effective antimalarial agents have been implemented, treatment failure still occurs, and malaria continues to cause neurological complications and death, particularly in severe or drug-resistant cases. Hence, novel therapeutic agents with distinct mechanisms of action, as well as alternative chemical compounds that can overcome resistance, are still needed to improve malaria therapy. This study aimed to investigate the antimalarial activities of Brucea javanica, a tropical plant extracts against Plasmodium falciparum, the major species associated with severe malaria. In this study, malaria parasites were treated with plant extracts using single and co-incubation methods, along with artesunate and chloroquine, and their inhibitory effect on parasite development was determined by microscopy. The results show that all tested doses of the extracts that effectively inhibited malaria parasites did not cause hemolysis of red blood cells (RBCs). The root extract (RE) and fruit extract (FE) inhibited parasite growth at IC₅₀ values of 0.41 ± 1.14 µg/mL and 0.26 ± 1.15 µg/mL, respectively. These plant extracts significantly interrupted malaria development at the ring stage, as presented by a reduction in the conversion rate to trophozoites and schizonts. The defective parasites treated with plant extracts were characterized by nuclear clumping, leading to pyknotic cell death. Moreover, RE and FW extracts elicited an additive effect with artesunate and chloroquine, significantly reducing IC₉₀ levels for the inhibition of parasite development. In conclusion, B. javanica extracts inhibited the asexual blood-stage development of malaria parasites. They distinctively show the additive effects of ATS and CRQ, elucidating their potential for further studies on novel formulas of antimalarial drug regimens. Full article

(This article belongs to the Section Parasitic Pathogens)

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