Pathogens

17 pages, 2502 KiB

Open AccessArticle

The Translationally Controlled Tumor Protein (TCTP), a Novel Antigen of Babesia bovis, Participates in the Establishment of Acute Infection and Contains Neutralizing B-Cell Epitopes

by Chyntia Pérez-Almeida, Diego Josimar Hernández-Silva, Edwin Esaú Hernández-Arvizu, Masahito Asada, Shin-ichiro Kawazu, Massaro W. Ueti, José Guadalupe Gomez-Soto, Urso Martín Dávila-Montero, Carlos A. Vega y Murguía and Juan Mosqueda

Pathogens 2025, 14(5), 502; https://doi.org/10.3390/pathogens14050502 - 20 May 2025

Abstract

Babesia bovis is a protozoan parasite that causes babesiosis in cattle. It has been hypothesized that in apicomplexan parasites, translationally controlled tumor protein (TCTP) interferes with the host immune response by inhibiting B cell proliferation. The aim of this study was the characterization [...] Read more.

Babesia bovis is a protozoan parasite that causes babesiosis in cattle. It has been hypothesized that in apicomplexan parasites, translationally controlled tumor protein (TCTP) interferes with the host immune response by inhibiting B cell proliferation. The aim of this study was the characterization of B. bovis TCTP (BboTCTP) and the evaluation of its expression, immunogenicity and role in infection. The tctp gene was identified and sequenced from B. bovis isolates and revealed a high conservation. Expression was confirmed in intraerythrocytic stages by Western blot and confocal microscopy. Synthetic peptides containing predicted B cell epitopes were used to immunize cattle, followed by a challenge with a virulent B. bovis strain. Immunized animals showed milder clinical signs and faster recovery compared to controls. Sera from non-immunized animals exhibited lower total IgG levels after challenge (p < 0.05), while sera from immunized animals induced significant in vitro invasion inhibition (32–33%). These results suggest that BboTCTP is immunogenic and may play a role in modulating the host immune response. The results provide novel insights into B. bovis biology and support BboTCTP as a promising candidate for further evaluation as a vaccine antigen. Future studies should explore its immunomodulatory mechanisms and potential use in combined vaccine formulations. Full article

(This article belongs to the Section Parasitic Pathogens)

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11 pages, 746 KiB

Open AccessArticle

Identification of Virulence Genes and Antibiotic Resistance in Extraintestinal Pathogenic Escherichia coli Isolated from Broiler Carcasses Using MALDI-TOF MS

by Jia-Tong Han, Yu-Xuan Tang, Si-Yi Wu, Yi-Ran Chen, Zhan-Peng Zou, Hang Zeng and Zhongjia Yu

Pathogens 2025, 14(5), 501; https://doi.org/10.3390/pathogens14050501 - 20 May 2025

Abstract

Escherichia coli contamination in poultry is a significant concern due to its potential to cause foodborne illness. The presence of extraintestinal pathogenic E. coli (ExPEC) strains in chicken carcasses can lead to severe human infections. This study investigates the prevalence, virulence, and antibiotic [...] Read more.

Escherichia coli contamination in poultry is a significant concern due to its potential to cause foodborne illness. The presence of extraintestinal pathogenic E. coli (ExPEC) strains in chicken carcasses can lead to severe human infections. This study investigates the prevalence, virulence, and antibiotic resistance of E. coli isolates from chicken carcasses processed in both wet market and industrial environments, with a focus on the detection capabilities of MALDI-TOF MS. A total of 119 E. coli isolates were obtained. Only a small proportion (5/119) carried enteropathogenic virulence genes. In contrast, 71.42% (85/119) of the isolates harbored multiple extraintestinal virulence genes. Among these, iucC and sitA, which are associated with systemic infections, were present in 68.24% (58/85) and 43.53% (37/85) of the isolates, respectively. Furthermore, 47.06% (56/119) of the isolates carrying at least two extraintestinal virulence genes were classified as ExPEC. Additionally, 94.6% (54/56) of ExPEC isolates were multidrug resistant (MDR), showing resistance to over three antibiotic classes, raising concerns about the spread of antibiotic resistance. MALDI-TOF MS profiling revealed significant heterogeneity among the ExPEC isolates, with no distinct clustering patterns based on processing environment or sampling site. These findings underscore the public health risks posed by ExPEC in poultry and emphasize the need for improved surveillance, stringent hygiene practices, and responsible antibiotic use in poultry production. Full article

(This article belongs to the Special Issue Foodborne Pathogens: The Antimicrobial Resistance from Farm to Fork)

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15 pages, 1729 KiB

Open AccessArticle

Red Fox (Vulpes vulpes) and Wolf (Canis lupus) as a Reservoir of Cryptosporidium spp. and Giardia intestinalis in Poland

by Dorota Dwużnik-Szarek, Ewa Julia Mierzejewska, Korneliusz Kurek, Małgorzata Krokowska-Paluszak, Patrycja Opalińska, Łukasz Stańczak, Grzegorz Górecki and Anna Bajer

Pathogens 2025, 14(5), 500; https://doi.org/10.3390/pathogens14050500 - 20 May 2025

Abstract

Infections with zoonotic pathogens have received increasing attention in recent years, as reflected in the literature of both veterinary and human medicine. Cryptosporidium and Giardia are recognised as the principal causes of waterborne outbreaks worldwide, but there is still limited data on the [...] Read more.

Infections with zoonotic pathogens have received increasing attention in recent years, as reflected in the literature of both veterinary and human medicine. Cryptosporidium and Giardia are recognised as the principal causes of waterborne outbreaks worldwide, but there is still limited data on the role of wild carnivores, such as red foxes and wolves, as reservoir hosts and in disseminating these pathogens in the environment. The aim of the current project was to analyse the prevalence and abundance of Cryptosporidium and Giardia infections in foxes from seven voivodeships and in wolves from the Warmia-Masuria Voivodeship in Poland and to conduct a phylogenetic analysis of the detected parasites. For the detection of both parasites, we used the commercial immunofluorescent assay MeriFluor Cryptosporidium/Giardia. For Cryptosporidium detection we also applied modified Ziehl–Neelsen (ZN) staining of faecal smears and, following PCR amplification, sequenced the 18S rDNA locus. For Giardia detection, we sequenced the glutamate dehydrogenase (gdh) gene. In total, 117 and 69 faecal samples obtained from red foxes and wolves, respectively, were screened for the presence of Cryptoporidium/Giardia. In red foxes, prevalence was 38.5% and 15.4% for Cryptosporidium spp. and G. intestinalis, respectively. In wolves, the prevalence of Cryptosporidium spp. was 14.5%, and only one sample was Giardia-positive. Cryptosporidium canis, Cryptosporidium sp. vole genotype, C. baileyi and Cryptosporidium sp. were identified in red foxes, while C. canis and Cryptosporidium sp. were detected in wolves. Our results indicate that red foxes and grey wolves act as reservoir hosts of Cryptosporidium spp. and G. intestinalis in natural areas in Poland. Full article

(This article belongs to the Section Parasitic Pathogens)

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12 pages, 975 KiB

Open AccessArticle

High Prevalence and Genetic Heterogeneity of Anaplasma marginale in Smallholder Bovine Populations of Pakistan, and Its Implications

by Abdul Ghafar, Waseem Shaukat, Muhammad Waqas, Charles G. Gauci, Robin B. Gasser and Abdul Jabbar

Pathogens 2025, 14(5), 499; https://doi.org/10.3390/pathogens14050499 - 20 May 2025

Abstract

Bovine anaplasmosis, caused by Anaplasma marginale, is a major tick-borne disease in tropical and subtropical regions of the world, leading to significant production losses. Prolonged convalescence periods are common and surviving animals often become subclinical carriers. This study aimed to detect and [...] Read more.

Bovine anaplasmosis, caused by Anaplasma marginale, is a major tick-borne disease in tropical and subtropical regions of the world, leading to significant production losses. Prolonged convalescence periods are common and surviving animals often become subclinical carriers. This study aimed to detect and characterise A. marginale in bovines in smallholder dairy farms across diverse climatic zones of Pakistan using molecular methods. In total, 321 blood DNA samples from apparently healthy cattle (n = 174) and buffaloes (n = 147) from six districts in Pakistan were tested for A. marginale using a nested PCR assay, targeting part of the major surface protein B gene (msp1β) as a genetic marker, followed by agarose gel electrophoresis and selective sequencing of amplicons from test-positive samples. Of the 321 DNA samples tested, 135 (42.1%) were test-positive for A. marginale. Prevalence was significantly higher in cattle (64.4%; 112/174) than in buffaloes (15.6%; 23/147), and female bovines (43.5%; 108/248) were more frequently infected than males (37%; 27/73). Phylogenetic analysis of the msp1β sequence data (n = 42) revealed that A. marginale from Pakistan clustered with those from Brazil, Thailand, South Africa, and the USA. This study represents the first comprehensive investigation of A. marginale from bovines from diverse agroecological zones of Pakistan and will further stimulate population genetic studies of A. marginale and investigations into the economic impact of subclinical infections in bovines in smallholder farming systems. Full article

(This article belongs to the Special Issue Tick-Borne Pathogens and Their Impact on Human and Animal Health)

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15 pages, 894 KiB

Open AccessArticle

Improvement of Cryopreservation and Production of Attenuated Babesia Parasites to Prevent Bovine Babesiosis

by Carmen Rojas-Martínez, José J. Lira-Amaya, Massaro W. Ueti, Roberto O. Castañeda-Arriola, Julio V. Figueroa Millán and Jesús A. Álvarez Martínez

Pathogens 2025, 14(5), 498; https://doi.org/10.3390/pathogens14050498 - 20 May 2025

Abstract

This study evaluated the effects of various concentrations of PVP-40 on the in vitro cryopreservation and recovery of Babesia bovis and Babesia bigemina. We also assessed a reduced dose of attenuated Babesia strains to determine its efficacy in preventing clinical disease. A [...] Read more.

This study evaluated the effects of various concentrations of PVP-40 on the in vitro cryopreservation and recovery of Babesia bovis and Babesia bigemina. We also assessed a reduced dose of attenuated Babesia strains to determine its efficacy in preventing clinical disease. A microaerophilic stationary phase blood culture system was used to recover Babesia parasites that were cryopreserved in solutions with various PVP-40 concentrations and Babesia parasites in 20% PVP-40 were used to vaccinate naïve cattle. The animals were vaccinated intramuscularly with frozen parasites cryopreserved in 20% PVP-40, with a dose of either 1 × 10⁸ or 1 × 10⁷ erythrocytes infected with both attenuated B. bigemina and B. bovis produced from blood cultures. The control group received uninfected erythrocytes. During the vaccination, clinical parameters such as rectal temperature and hematocrit levels were unaffected. The animals were relocated to a farm in a Babesia hyperendemic area to test the efficacy of these live vaccines in controlling disease onset. Some vaccinated animals showed mild disease. In the vaccinated groups, parasites were detected in blood smears for only one day during the challenge. In contrast, the control group experienced fever for three consecutive days, a decline in hematocrit levels, and significant health deterioration. In this group, parasites were detected in smears for four consecutive days. All the animals in the control group required treatment to manage their high parasitemia and prevent mortality. In this study, we demonstrated that increasing the concentration of PVP-40 to cryopreserve parasites improved the recovery and proliferation of Babesia spp. in blood culture, and we also showed that when animals were vaccinated with cryopreserved, in vitro cultured, attenuated Babesia parasites in 20% PVP-40, they were effectively protected from severe clinical babesiosis. Full article

(This article belongs to the Topic Advances in Infectious and Parasitic Diseases of Animals)

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Graphical abstract

1 pages, 122 KiB

Open AccessCorrection

Correction: Mo, J.; Mo, J. Infectious Laryngotracheitis Virus and Avian Metapneumovirus: A Comprehensive Review. Pathogens 2025, 14, 55

by Jongsuk Mo and Jongseo Mo

Pathogens 2025, 14(5), 497; https://doi.org/10.3390/pathogens14050497 - 19 May 2025

Abstract

There was an error in the original publication [...] Full article

29 pages, 730 KiB

Open AccessReview

Antimicrobial Resistant Staphylococcus spp., Escherichia coli, and Salmonella spp. in Food Handlers: A Global Review of Persistence, Transmission, and Mitigation Challenges

by Gustavo Guimarães Fernandes Viana, Marita Vedovelli Cardozo, Juliano Gonçalves Pereira and Gabriel Augusto Marques Rossi

Pathogens 2025, 14(5), 496; https://doi.org/10.3390/pathogens14050496 - 18 May 2025

Abstract

Antimicrobial resistance in foodborne pathogens represents a critical global health challenge, with food handlers serving as key contributors in their transmission. This comprehensive review synthesizes evidence on the prevalence, transmission dynamics, and antimicrobial resistance patterns of three major pathogens, Staphylococcus spp., Escherichia coli [...] Read more.

Antimicrobial resistance in foodborne pathogens represents a critical global health challenge, with food handlers serving as key contributors in their transmission. This comprehensive review synthesizes evidence on the prevalence, transmission dynamics, and antimicrobial resistance patterns of three major pathogens, Staphylococcus spp., Escherichia coli, and Salmonella spp., among food handlers worldwide. Analysis of studies across diverse geographical regions reveals considerable variation in colonization rates, with Staphylococcus spp. prevalence ranging from 19.5% to 95.0%, Escherichia coli from 2.8% to 89.3%, and Salmonella spp. from 0.07% to 9.1%. Resistance profiles demonstrate alarming trends, including widespread β-lactam resistance and emerging resistance to last-resort antibiotics like carbapenems. Particularly concerning is the high occurrence of multidrug resistant (MDR) strains and extended spectrum β-lactamase (ESBL) producers in low- and middle-income countries. This review identified inadequate handwashing, poor hygiene infrastructure, and asymptomatic carriage as critical factors facilitating the transmission of antimicrobial resistant strains. These findings underscore the urgent need for enhanced surveillance systems, targeted decolonization strategies, improved hygiene protocols, and food handler education to mitigate the spread of resistant pathogens through the food chain. Full article

(This article belongs to the Special Issue Foodborne Pathogens: The Antimicrobial Resistance from Farm to Fork)

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17 pages, 2051 KiB

Open AccessArticle

Repellent Activity of DEET Combined with Botanical Compounds Against Amblyomma sculptum Nymphs: Laboratory and Field Evaluations

by Mayara Macêdo Barrozo, Haile Dean Figueiredo Chagas, Gabrielly Bernardes Rodrigues Damaceno, Emilly Faria Santos, Rafael Assunção Carvalho, Isabela Santos Silva, Francisca Letícia Vale, Lainny Jordana Martins Pereira e Sousa, Hermes Ribeiro Luz, Lorena Lopes Ferreira and Caio Monteiro

Pathogens 2025, 14(5), 495; https://doi.org/10.3390/pathogens14050495 - 17 May 2025

Abstract

This study evaluated the repellent activity of DEET, eugenol, methyl eugenol, 1,8-cineole, and their combinations against Amblyomma sculptum nymphs through laboratory and field tests. Initially, the botanical compounds were tested individually in Petri dish assays to assess repellent activity over various intervals (1 [...] Read more.

This study evaluated the repellent activity of DEET, eugenol, methyl eugenol, 1,8-cineole, and their combinations against Amblyomma sculptum nymphs through laboratory and field tests. Initially, the botanical compounds were tested individually in Petri dish assays to assess repellent activity over various intervals (1 min to 168 h) at concentrations of 2%. Eugenol and methyl eugenol exhibited the highest average repellent percentages (60 to 90%), with significant effects (p < 0.05) across most evaluation periods, in contrast to 1,8-cineole. Therefore, eugenol and methyl eugenol were selected for combination testing with DEET. The combinations of 7% DEET + 1% eugenol and 7% DEET + 2% methyl eugenol showed the best results, with average repellent efficacy exceeding 85%. The first combination was selected for field evaluation. In this test, volunteers wore treated socks and walked for two hours in a naturally infested area. A significant reduction (p < 0.05) in the number of ticks recovered from the socks was observed in the 7% DEET and 7% DEET + 1% eugenol treatments, with efficacies of 82.5% and 88.5%, respectively, after 120 min. Isolated eugenol showed limited field efficacy, with significant repellent activity (p < 0.05) lasting only 20 min. The results highlight the potential of eugenol and methyl eugenol as repellent compounds, especially when combined with DEET. The field study confirmed the efficacy of the 7% DEET + 1% eugenol. Full article

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

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14 pages, 1712 KiB

Open AccessArticle

Bacterial Acute Otitis Media Complicated with Otorrhea in a Children’s Hospital in the Era of Pneumococcal Conjugate Vaccines

by Irene Tzovara, Anastasios Doudoulakakis, Georgios Kalogeras, Emmanouil Koutouzis, Charilaos Dellis, Sophia Pasparakis, Marietta Charakida, Evangelia Lebessi, Elisavet Bozavoutoglou, Michael Tsakanikos, Vassiliki Syriopoulou and Maria Tsolia

Pathogens 2025, 14(5), 494; https://doi.org/10.3390/pathogens14050494 - 17 May 2025

Abstract

Acute otitis media (AOM) is a common disease among children and can be complicated by otorrhea (AOMO). In 2010, the 13-valent Pneumococcal Conjugate Vaccine (PCV13) replaced the 7-valent vaccine (PCV7) in Greece. We aimed to describe the microbiological profile of bacterial ΑOMO among [...] Read more.

Acute otitis media (AOM) is a common disease among children and can be complicated by otorrhea (AOMO). In 2010, the 13-valent Pneumococcal Conjugate Vaccine (PCV13) replaced the 7-valent vaccine (PCV7) in Greece. We aimed to describe the microbiological profile of bacterial ΑOMO among children younger than 16 years across the two PCV periods in a tertiary children’s hospital. Middle ear fluid cultures from 2418 children with AOMO were collected from 2007 to 2022. Otopathogens were isolated and tested for antimicrobial susceptibility. Data were compared between the PCV7- (2007–2011) and PCV13-period (2012–2019). The most common otopathogen over the 16-year period was S. pyogenes (35.4%), followed by H. influenzae (33.8%), S. pneumoniae (26.6%), and M. catarrhalis (4.1%). Pneumococcal resistance to cefotaxime and clindamycin significantly increased from 2% to 4.5% (p = 0.019) and 16.1% to 22.8% (p = 0.039), respectively. Resistance of H. influenzae to ampicillin increased from 6.3% to 13.9% (p < 0.001). A significant reduction in cotrimoxazole-resistant S. pneumoniae from 31% to 22.4% (p = 0.012), and in clindamycin-resistant and erythromycin-resistant S pyogenes, from 17.4% to 9.3% and 21.4% to 10.8%, respectively (p ≤ 0.001), was observed. During 2013–2022, 38 S. pneumoniae serotypes were identified among 250 isolates. Serotype 3 (27.2%) and 19A (13.2%) prevailed, followed by 19F (7%). The most common causes after the shift to PCV13 are S. pyogenes and H. influenzae. However, S. pneumoniae remains an important otopathogen with significant antimicrobial resistance. Serotype 3 was mostly detected, followed by 19A. Full article

(This article belongs to the Special Issue Acute Respiratory Infections: Diagnosis, Treatment, Complications and Prevention)

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17 pages, 630 KiB

Open AccessArticle

Acute Respiratory and Influenza Viruses Circulating in Kazakhstan During 2018–2024

by Tatyana Glebova, Nailya Klivleyeva, Assem Baimukhametova, Galina Lukmanova, Nurbol Saktaganov, Nuray Ongarbayeva, Baiken Baimakhanova, Gulmira Kassymova, Madisha Sagatova, Almagul Rachimbayeva, Nazgul Zhanuzakova, Tatyana Naidenova, Nigina Rakhmonova and Richard Webby

Pathogens 2025, 14(5), 493; https://doi.org/10.3390/pathogens14050493 - 16 May 2025

Abstract

Respiratory tract infections cause serious morbidity and mortality and are a major public health problem. The objective of our study was detection of the prevalence of viral respiratory diseases in the territory of Kazakhstan during the epidemic period of 2018–2024. The presence of [...] Read more.

Respiratory tract infections cause serious morbidity and mortality and are a major public health problem. The objective of our study was detection of the prevalence of viral respiratory diseases in the territory of Kazakhstan during the epidemic period of 2018–2024. The presence of respiratory viruses in nasopharyngeal swabs was analyzed using real-time polymerase chain reaction. The level of specific antibodies in the blood serum was determined by hemagglutination inhibition assay and enzyme-linked immunosorbent assay. In rtRT-PCR, patients were diagnosed with non-influenza viral respiratory tract infections as well as influenza viruses A(H1N1), A(H3N2), and B. Antibodies were detected against A(H1N1)pdm09, influenza A(H3N2), and influenza B viruses and with simultaneous detection of both viruses. The circulation of influenza A(H3N2) viruses belonging to the 3C.2a1b.2a.2a.3a.1 clade was confirmed by whole-genome sequencing. According to the results, in the period 2018–2024, the spread of influenza A and B viruses and non-influenza respiratory tract infections was observed. The data of this study confirm the role of known causative agents of epidemic infection and indicate the need to continue monitoring their spread in Kazakhstan, which may add to the general quality of the health system. Full article

(This article belongs to the Section Viral Pathogens)

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16 pages, 295 KiB

Open AccessReview

Methods of Controlling Microbial Contamination of Food

by Renata Urban-Chmiel, Jacek Osek and Kinga Wieczorek

Pathogens 2025, 14(5), 492; https://doi.org/10.3390/pathogens14050492 - 16 May 2025

Abstract

The rapid growth of world population and increase in living standards have led to an increase in the demand for high-quality, safe food. The Food and Agriculture Organization of the United Nations (FAO) estimates that by 2050 the demand for food will increase [...] Read more.

The rapid growth of world population and increase in living standards have led to an increase in the demand for high-quality, safe food. The Food and Agriculture Organization of the United Nations (FAO) estimates that by 2050 the demand for food will increase by 60%, and production of animal protein will increase by 1.7% a year, with meat production to increase by nearly 70%, dairy products by 55%, and aquaculture by as much as 90%. Microbial contamination of food is a significant problem for the accessibility of safe food which does not pose a threat to the life and health of consumers. Campylobacter, Salmonella, and Yersinia are responsible for thousands of food-borne infections in humans. Currently, numerous programs are being developed to combat pathogenic bacteria in the food supply chain, especially at the primary production stage. These approaches include physical, chemical, biological, and other strategies and methods used to inhibit the bacterial growth of bacteria or completely eliminate the pathogens from the food chain. Therefore, an extremely important goal is to provide safe food and control its quality by eliminating pathogenic and spoilage microorganisms. However, the use of chemicals in food preservation has negative effects for both the consumption values of food and the natural environment. Therefore, it seems absolutely necessary to implement measures utilizing the most environmentally friendly and effective techniques for controlling microbial contamination in food. There is a great need to develop ecological methods in food production which guarantee adequate safety. One of these methods is the use of bacteriophages (bacterial viruses) naturally occurring in the environment. Given the above, the aim of this study was to present the most natural, ecological, and alternative methods of food preservation with regard to the most common foodborne zoonotic bacteria. We also present methods for reducing the occurrence of microbial contamination in food, thus to produce maximally safe food for consumers. Full article

(This article belongs to the Special Issue Epidemiology and Molecular Pathogenesis of Antimicrobial Resistance and Virulence for Foodborne Pathogens: 2nd Edition)

17 pages, 1417 KiB

Open AccessArticle

Validation of a High-Throughput Microfluidic Real-Time PCR for the Detection of Vector-Borne Agents in Wild Birds from the Brazilian Pantanal

by Amir Salvador Alabí Córdova, João Batista Pinho, Amanda Garcia Pereira, Clémence Galon, Tiago Valadares Ferreira, Lorena Freitas das Neves, Gabrielly de Oliveira Lopes, Rosangela Zacarias Machado, Sara Moutailler and Marcos Rogério André

Pathogens 2025, 14(5), 491; https://doi.org/10.3390/pathogens14050491 - 16 May 2025

Abstract

Despite numerous studies on haemosporidians in wild birds from Brazil, the presence of other vector-borne agents (VBA) such as Anaplasma spp., Bartonella spp., and Onchocercidae filariids in avian hosts remains largely unknown. The low occurrence of these VBAs might be due to the [...] Read more.

Despite numerous studies on haemosporidians in wild birds from Brazil, the presence of other vector-borne agents (VBA) such as Anaplasma spp., Bartonella spp., and Onchocercidae filariids in avian hosts remains largely unknown. The low occurrence of these VBAs might be due to the low sensitivity of traditional molecular techniques. The microfluidic real-time PCR assay, known for its high sensitivity, has emerged as a promising method to detect and study the occurrence and diversity of VBAs in both arthropod vectors and vertebrate hosts. To validate previously and standardize newly designed microfluidic real-time PCR protocols, selected positive avian blood DNA samples for Anaplasma spp., Bartonella spp., haemosporidians, and filariids were used. The molecular occurrence rates for the selected VBAs were 18.2% for Anaplasma spp., 0.36% for Bartonella spp., 6.2% for Plasmodium spp., 4.7% for Haemoproteus spp., and 6.5% for Onchocercidae filariids. The Plasmodium spp. cytB sequence detected in a Volatinia jacarina clustered with Plasmodium tejerai, whereas the Haemoproteus spp. cytB sequence detected in a Columbina squamata clustered with Haemoproteus columbae. While Onchocercidae filariid cox-1 sequences were detected in specimens of Ramphocelus carbo, Turdus amaurocalinus and Synallaxis albilora grouped with Aproctella spp., one sequence detected in R. carbo was ancestral to the clade comprising Splendidofilaria spp. and Eufilaria spp. High-throughput microfluidic real-time PCR assay can be used for screening VBAs in avian hosts from South America, but new primers/probe sets should be designed for VBA genotypes present in Brazil. Full article

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

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15 pages, 5060 KiB

Open AccessArticle

Evaluation of Fusarium Head Blight Resistance Through a Genome-Wide Association Study in CIMMYT and South Asian Wheat Germplasm

by Rupsanatan Mandal, Xinyao He, Gyanendra Pratap Singh, Muhammad Rezaul Kabir, Arun Kumar Joshi and Pawan Kumar Singh

Pathogens 2025, 14(5), 490; https://doi.org/10.3390/pathogens14050490 - 16 May 2025

Abstract

Fusarium head blight (FHB) is an important disease throughout the world due to its strong association with yield reduction, quality deterioration, and mycotoxin contamination in wheat. The use of FHB-resistant genotypes in wheat production can significantly reduce damage. The current study screened a [...] Read more.

Fusarium head blight (FHB) is an important disease throughout the world due to its strong association with yield reduction, quality deterioration, and mycotoxin contamination in wheat. The use of FHB-resistant genotypes in wheat production can significantly reduce damage. The current study screened a panel of bread wheat from CIMMYT and South Asian countries for FHB resistance to identify promising genotypes useful for wheat breeding and to map the associated genomic regions and linked molecular markers through a genome-wide association study (GWAS). Spray-inoculated field experiments were conducted at CIMMYT, Mexico, over three years, and a wide range of phenotypic variations was observed. Four lines, CIM-39, CIM-29, CIM-9, and CIM-3, exhibited consistent resistance across experiments, with FHB indices ranging from 6.5 to 8.1. Genotyping was conducted using the Illumina Infinium 15 K Bead Chip, and 11,184 high-quality SNP markers were obtained and used for GWAS. Nineteen significant marker-trait associations (MTAs) were detected, among which MTAs at Ra_c58315_265 on 1A and Tdurum_contig102328_129 and Ku_c20136_198 on 7B showed reproducible results, with phenotypic effects on FHB resistance of 6.05%, 3.54%, and 3.92%, respectively. Several genes associated with disease resistance were found near the significant SNPs. The identified resistant genotypes and markers may be useful in future marker-assisted breeding in wheat. Full article

(This article belongs to the Special Issue Current Research on Fusarium: 2nd Edition)

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12 pages, 1897 KiB

Open AccessArticle

Molecular Detection of Encephalitozoon cuniculi in Migratory Waterfowl of the Genus Anser (Anseriformes: Anatidae) in Poland

by Piotr Solarczyk, Agnieszka Perec-Matysiak, Agnieszka Wojtkowiak-Giera and Mike Heddergott

Pathogens 2025, 14(5), 489; https://doi.org/10.3390/pathogens14050489 - 16 May 2025

Abstract

Microsporidiosis is a zoonotic disease that derives from disparate sources. Most of the microsporidial agents are host-specific but some are capable of interspecies transmission, causing disease in various animals including humans. Human microsporidiosis may be caused by 17 species, with Encephalitozoon cuniculi, [...] Read more.

Microsporidiosis is a zoonotic disease that derives from disparate sources. Most of the microsporidial agents are host-specific but some are capable of interspecies transmission, causing disease in various animals including humans. Human microsporidiosis may be caused by 17 species, with Encephalitozoon cuniculi, E. intestinalis and E. hellem mostly being responsible for human infections worldwide. Wildlife and migratory waterfowl can serve as reservoirs of these human-infectious agents and play a significant role in disseminating these pathogens into the environment. The aim of the study was to detect E. cuniculi, E. intestinalis and E. hellem in wild, migratory greater white-fronted geese (Anser albifrons) and other Anatidae members in feacal samples obtained in north-western Poland, using a molecular method. We collected 189 fecal droppings from Anatidae species (75 samples from greater white-fronted geese and 114 from other Anser spp.) during autumn migration. New species specific primers for PCR amplification were used to amplify a fragment of the small subunit ribosomal (SSU) rRNA of E. cuniculi, E. intestinalis and E. hellem. All fecal droppings were negative for E. intestinalis and E. hellem whereas E cuniculi was detected in 6 of 189 fecal samples (3.2%; 95% CI: 1.3–6.3%). In total, 1 of 75 tested fecal samples of greater white-fronted geese was positive (1.3%; 95% CI: 0.08–5.7%) while 5 of 114 (4.4%; 95% CI: 1.6–9.1%) tested fecal samples without exact species affiliation (only Anser sp.) were also positive. The phylogenetic analysis placed the sequences obtained from the birds’ droppings in the clade E. cuniculi from various rodents, wild carnivores and humans. Our results provide the first description of the occurrence and genotyping of the microsporidian E. cuniculi in greater white-fronted geese and in other members of the Anserinae Subfamily. Our findings support the results of other authors that E. cuniculi may originate from diverse sources, including common waterfowl. Our results are important in a One Health context, as wild migrating waterfowl may disseminate this zoonotic agent in remote regions through their migratory behaviour. These species should be considered significant sources of zoonotic pathogens, potentially hazardous to domestic and farmed animals as well as humans. Full article

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

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13 pages, 2372 KiB

Open AccessCase Report

Reactivation of Latent Tuberculosis Following COVID-19 and Epstein-Barr Virus Coinfection: A Case Report

by Iryna Halabitska, Pavlo Petakh, Valentyn Oksenych and Oleksandr Kamyshnyi

Pathogens 2025, 14(5), 488; https://doi.org/10.3390/pathogens14050488 - 16 May 2025

Abstract

Background: This case is unique in demonstrating the reactivation of latent tuberculosis (TB) following co-infection with SARS-CoV-2 and Epstein–Barr virus (EBV) in an otherwise healthy young adult. It highlights a rare clinical scenario in which viral immune dysregulation likely facilitated TB progression. To [...] Read more.

Background: This case is unique in demonstrating the reactivation of latent tuberculosis (TB) following co-infection with SARS-CoV-2 and Epstein–Barr virus (EBV) in an otherwise healthy young adult. It highlights a rare clinical scenario in which viral immune dysregulation likely facilitated TB progression. To date, few reports have explored the complex interplay between COVID-19, EBV reactivation, and TB in a single patient, particularly with isolated extrapulmonary involvement. Case Presentation: A 24-year-old woman presented with persistent low-grade fever, fatigue, night sweats, unintentional weight loss, and progressive cervical and supraclavicular lymphadenopathy. These symptoms emerged shortly after a moderate COVID-19 infection. Laboratory studies revealed elevated inflammatory markers and pronounced lymphopenia. EBV reactivation was confirmed via serology and PCR. Despite antiviral therapy, symptoms persisted, and imaging revealed necrotic lymphadenopathy. Tuberculous lymphadenitis was diagnosed through fine-needle aspiration cytology and PCR detection of Mycobacterium tuberculosis. The patient was treated with a standard anti-tuberculosis regimen, resulting in clinical, radiological, and immunological improvement. Conclusions: This case underscores the importance of considering latent TB reactivation in patients with persistent lymphadenopathy and recent viral infections, particularly in regions with high TB prevalence. It also emphasizes the need for thorough immunological and microbiological assessment in complex post-viral syndromes. The main clinical takeaway is that COVID-19 and EBV co-infection may create a permissive environment for TB reactivation through immune system compromise. Full article

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

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13 pages, 230 KiB

Open AccessArticle

Genetic and Antigenic Diversity of Neisseria meningitidis Serogroup B Strains in Vietnam

by Trieu Phi Long, Vo Viet Cuong, Bui Thi Lan Anh, Trinh Van Toan, Vu Thi Loan, Pham Viet Hung, Le Thi Lan Anh, Nguyen Ngoc Tan, Luong Thi Mo, Le Van Khanh and Hoang Van Tong

Pathogens 2025, 14(5), 487; https://doi.org/10.3390/pathogens14050487 - 15 May 2025

Abstract

Background: Neisseria meningitidis (N. meningitidis) is a leading cause of acute meningitis and is classified into 13 serogroups, six of which are predominantly associated with invasive meningococcal disease. This study aimed to investigate the genotype, subgenotype, and antigenic profiles of N. [...] Read more.

Background: Neisseria meningitidis (N. meningitidis) is a leading cause of acute meningitis and is classified into 13 serogroups, six of which are predominantly associated with invasive meningococcal disease. This study aimed to investigate the genotype, subgenotype, and antigenic profiles of N. meningitidis serogroup B strains isolated in Vietnam. Methods: Genotyping was performed on 106 N. meningitidis strains isolated from clinical samples from Vietnamese patients and nasopharyngeal swabs of healthy adolescents between 2019 and 2024. The genetic profiles, including the porA, porB, fetA, fHbp, abcZ, adk, aroE, fumC, gdh, pdhC, and pgm genes, were analyzed using Sanger sequencing and bioinformatic methods. Results: We found that 84.9% of the strains carried VR3 families 36 or 35-1, with VR1, VR2, and VR3 families 22-25, 14, and 36 being the most prevalent. Among the 106 serogroup B isolates, 20 variants of the porB allele 3 were identified, with porB 3-1212 being the most frequent (30.2%). Dominant PorB variable loops included L1.6, L4.5, L5.7, L6.6, and L7.13. fHbp variant group 2 was predominant (104/106 strains), and 12 FetA allele variants were identified, with F1-7 being the most common (47.2%). Three clonal complexes were identified, and clonal complex ST-32 was the most predominant. Fifty-five strains (51.9%) belonged to sequence types that have not yet been assigned to any clonal complexes, and 15 strains (14.1%) with allelic profiles were not assigned to STs. The 3-253 and 3-1212 alleles of porB, the F1-7 variant of FetA, the ST-44 and ST-1576 sequence types, and the ST-41/44 complex were observed more frequently in patients compared to asymptomatic carriers, suggesting their association with more virulence. Conclusions: This study showed a high genetic and antigenic diversity of N. meningitidis serogroup B isolates in Vietnam, with VR3 family 36 most common and porB 3-1212 as the predominant allele. fHbp variant group 2 and FetA allele F1-7 were most frequent. ST-32 was the dominant clonal complex, though many strains remained unassigned, highlighting the need for ongoing molecular surveillance. Full article

13 pages, 1789 KiB

Open AccessArticle

Genetic Epidemiology and Resistance Investigations of Clinical Yeasts in Alexandria, Egypt

by Bram Spruijtenburg, Carolina Melchior do Prado, Mats van Kempen, Sherine M. Shawky, Jacques F. Meis, Vânia Aparecida Vicente, Flavio Queiroz-Telles, Theun de Groot, Mohammed A. El-Kholy and Eelco F. J. Meijer

Pathogens 2025, 14(5), 486; https://doi.org/10.3390/pathogens14050486 - 15 May 2025

Abstract

Yeast bloodstream infections lead to high mortality and morbidity and are mostly observed in immunocompromised patients. In Africa, only a few studies have characterized clinical yeasts. To increase insight into yeast resistance and transmission in Africa, we identified various yeasts from Alexandria, Egypt [...] Read more.

Yeast bloodstream infections lead to high mortality and morbidity and are mostly observed in immunocompromised patients. In Africa, only a few studies have characterized clinical yeasts. To increase insight into yeast resistance and transmission in Africa, we identified various yeasts from Alexandria, Egypt and performed antifungal susceptibility testing (AFST) and genotyping. A total of 1307 single isolates from unique patients, recovered from different anatomical sites including the bloodstream, retrieved from a reference laboratory in Alexandria, Egypt were studied. All isolates were identified with MALDI-TOF MS, while some were initially identified with a Vitek 2 Compact system. Short tandem repeat (STR) genotyping was performed for the most common species, and AFST was performed with microbroth dilution. Among bloodstream isolates (n = 71), C. albicans was the most common etiological agent, followed by C. tropicalis and C. parapsilosis. Comparison of yeast identification methods demonstrated that 22% of isolates were incorrectly identified with the Vitek 2 Compact system compared to MALDI-TOF MS. Multiple rare yeasts showed reduced antifungal susceptibility. STR genotyping demonstrated potential events of nosocomial transmission with N. glabratus and C. parapsilosis. Moreover, an azole-resistant C. tropicalis clade identified earlier in Alexandria was still present. To conclude, clinical yeasts in Alexandria, Egypt, are overall susceptible common species. Full article

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

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25 pages, 12805 KiB

Open AccessArticle

From the Gut to the Brain: Transcriptomic Insights into Neonatal Meningitis Escherichia coli Across Diverse Host Niches

by Lekshmi K. Edison and Subhashinie Kariyawasam

Pathogens 2025, 14(5), 485; https://doi.org/10.3390/pathogens14050485 - 15 May 2025

Abstract

Neonatal Meningitis-causing Escherichia coli (NMEC) is the leading cause of neonatal meningitis and exhibits remarkable adaptability to diverse host environments. Understanding its transcriptional responses across different host niches is crucial for deciphering pathogenesis and identifying potential therapeutic targets. We performed a comparative transcriptomic [...] Read more.

Neonatal Meningitis-causing Escherichia coli (NMEC) is the leading cause of neonatal meningitis and exhibits remarkable adaptability to diverse host environments. Understanding its transcriptional responses across different host niches is crucial for deciphering pathogenesis and identifying potential therapeutic targets. We performed a comparative transcriptomic analysis of NMEC RS218, the prototype strain of NMEC, under four distinct host-mimicking conditions: colonic fluid (CF), serum (S), human brain endothelial cells (HBECs), and cerebrospinal fluid (CSF). Differential gene expression analysis was conducted to assess metabolic shifts, virulence factor regulation, and niche-specific adaptation strategies, in which RS218 demonstrated niche-specific transcriptional reprogramming. In CF, genes associated with biofilm formation, motility, efflux pumps, and cell division regulation were upregulated, aiding gut colonization. The serum environment triggered the expression of siderophore-mediated iron acquisition, enterobactin biosynthesis, and heme utilization genes, facilitating immune evasion and bacterial persistence. In HBECs, NMEC upregulated genes linked to nucleoside metabolism, membrane remodeling, pilus organization, and blood–brain barrier (BBB) traversal. In CSF, genes related to oxidative stress resistance, chemotaxis, DNA repair, biofilm formation, and amino acid biosynthesis were enriched, reflecting NMEC’s adaptive mechanisms for survival under nutrient-depleted conditions. Energy-intensive pathways were consistently downregulated across all niches, highlighting the need for an energy conservation strategy. This study provides novel insights into NMEC’s adaptive strategies across different host environments, emphasizing its metabolic flexibility, virulence regulation, and immune evasion mechanisms, offering potential targets for therapeutic intervention. Full article

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27 pages, 3928 KiB

Open AccessArticle

3,3′-((3-Hydroxyphenyl)azanediyl)dipropionic Acid Derivatives as a Promising Scaffold Against Drug-Resistant Pathogens and Chemotherapy-Resistant Cancer

by Povilas Kavaliauskas, Waldo Acevedo, Eglė Mickevičiūtė, Ramunė Grigalevičiūtė, Birutė Grybaitė, Birutė Sapijanskaitė-Banevič, Guoda Pranaitytė, Vidmantas Petraitis, Rūta Petraitienė and Vytautas Mickevičius

Pathogens 2025, 14(5), 484; https://doi.org/10.3390/pathogens14050484 - 15 May 2025

Abstract

The synthesis and antimicrobial and anticancer activity of 3,3′-((3-hydroxyphenyl)azanediyl)dipropionic acid derivatives (2–25) against drug-resistant bacterial pathogens and FaDu head and neck cancer cells were investigated. The derivatives were synthesized through various methods, including esterification, hydrazinolysis, and condensation reactions. The [...] Read more.

The synthesis and antimicrobial and anticancer activity of 3,3′-((3-hydroxyphenyl)azanediyl)dipropionic acid derivatives (2–25) against drug-resistant bacterial pathogens and FaDu head and neck cancer cells were investigated. The derivatives were synthesized through various methods, including esterification, hydrazinolysis, and condensation reactions. The compounds demonstrated structure-dependent antimicrobial activity, predominantly targeting Gram-positive pathogens. Compounds containing 4-nitrophenyl, 1-naphthyl, and 5-nitro-2-thienyl groups exhibited enhanced activity against S. aureus and E. faecalis. Additionally, compounds 5, 6, and 25 showed antiproliferative activity in cisplatin-resistant FaDu cells at low micromolar concentrations. The in silico modeling revealed that compound 25 interacts with the HER-2 and c-MET proteins. These compounds also induced significant oxidative stress in FaDu cells and demonstrated low cytotoxic activity in non-cancerous HEK293 cells. These results highlight the potential of N-aryl-substituted β-amino acid derivatives as promising scaffolds for the further development of novel amino acid-based antimicrobial and anticancer agents targeting drug-resistant pathogens and cancers. Full article

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