Pathogens

16 pages, 1017 KiB

Open AccessArticle

Assessment of Antibiotic Resistance Among Isolates of Klebsiella spp. and Raoultella spp. in Wildlife and Their Environment from Portugal: A Positive Epidemiologic Outcome

by Carolina Sabença, Rani de la Rivière, Paulo Barros, João Alexandre Cabral, Roberto Sargo, Luís Sousa, Maria de Lurdes Enes Dapkevicius, Filipe Silva, Filipa Lopes, Ana Carolina Abrantes, Madalena Vieira-Pinto, Manuela Caniça, Gilberto Igrejas, Carmen Torres and Patrícia Poeta

Pathogens 2025, 14(1), 99; https://doi.org/10.3390/pathogens14010099 - 20 Jan 2025

Viewed by 577

Abstract

One of the significant challenges facing modern medicine is the rising rate of antibiotic resistance, which impacts public health, animal health, and environmental preservation. Evaluating antibiotic resistance in wildlife and their environments is crucial, as it offers essential insights into the dynamics of [...] Read more.

One of the significant challenges facing modern medicine is the rising rate of antibiotic resistance, which impacts public health, animal health, and environmental preservation. Evaluating antibiotic resistance in wildlife and their environments is crucial, as it offers essential insights into the dynamics of resistance patterns and promotes strategies for monitoring, prevention, and intervention. Klebsiella and Raoultella genera isolates were recovered from fecal samples of wild animals and environmental samples using media without antibiotic supplementation. Antibiograms were performed for 15 antibiotics to determine the phenotypic resistance profile in these isolates. Extended-spectrum β-lactamase (ESBL) production was tested by the double-disc synergy test, and one ESBL-producing K. pneumoniae isolate was screened by PCR and whole-genome sequencing. Biofilm production was analyzed using the microtiter plate method. A total of 23 Klebsiella spp. and 3 Raoultella spp. isolates were obtained from 312 fecal samples from wild animals, 9 Klebsiella spp. and 4 Raoultella spp. isolates were obtained from 18 river and stream water samples, and 4 Klebsiella spp. and 3 Raoultella spp. isolates from 48 soil samples. Regarding antibiotic resistance, only one isolate of K. pneumoniae from soil samples was an ESBL-producer and showed resistance to six antibiotics. This isolate harbored multiple β-lactams genes (bla_CTX-M-15, bla_TEM-1, bla_SHV-28, and bla_OXA-1), as well as genes of resistance to quinolones, sulfonamides, tetracycline, aminoglycosides, and chloramphenicol, and belonged to the lineage ST307. Most of the Klebsiella spp. and Raoultella spp. isolates were biofilm producers (except for one Klebsiella isolate), and 45.6% were weak biofilm producers, with the remaining being moderate to strong biofilm producers. We can conclude that antibiotic resistance is not widespread in these environment-associated isolates, which is a positive epidemiological outcome. However, identifying a single ESBL-K. pneumoniae isolate should serve as a warning of potential hotspots of resistance emergence. Full article

(This article belongs to the Collection New Insights into Bacterial Pathogenesis)

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25 pages, 393 KiB

Open AccessReview

The Body of Chagas Disease Vectors

by Jean-Pierre Dujardin

Pathogens 2025, 14(1), 98; https://doi.org/10.3390/pathogens14010098 - 20 Jan 2025

Viewed by 548

Abstract

Morphometry is an effort to describe or measure the morphology of the body, or parts of it. It also provides quantitative data on the interactions of living organisms with their environment, external or internal. As a discipline, morphometrics has undergone significant developments in [...] Read more.

Morphometry is an effort to describe or measure the morphology of the body, or parts of it. It also provides quantitative data on the interactions of living organisms with their environment, external or internal. As a discipline, morphometrics has undergone significant developments in the last decade, making its implementation more visual and less laborious. Chagas disease vectors, often referred to by the common name of “kissing bugs”, belong to the subfamily Triatominae. Due to their apparent morphological plasticity, they have been the subject of numerous morphometric studies. Most of these have been applied taking into account the particularities of this group of vectors, such as domesticity (synanthropy), food preferences, dispersal ability, insecticide resistance, as well as some taxonomic issues. This brief review over nearly three decades is organized here according to the body organs considered by the authors. Full article

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

20 pages, 7537 KiB

Open AccessArticle

Diversity and Distribution of Phytophthora Species Along an Elevation Gradient in Natural and Semi-Natural Forest Ecosystems in Portugal

by Carlo Bregant, Eduardo Batista, Sandra Hilário, Benedetto Teodoro Linaldeddu and Artur Alves

Pathogens 2025, 14(1), 103; https://doi.org/10.3390/pathogens14010103 - 20 Jan 2025

Viewed by 612

Abstract

Globally, forests are constantly threatened by a plethora of disturbances of natural and anthropogenic origin, such as climate change, forest fires, urbanization, and pollution. Besides the most common stressors, during the last few years, Portuguese forests have been impacted by severe decline phenomena [...] Read more.

Globally, forests are constantly threatened by a plethora of disturbances of natural and anthropogenic origin, such as climate change, forest fires, urbanization, and pollution. Besides the most common stressors, during the last few years, Portuguese forests have been impacted by severe decline phenomena caused by invasive pathogens, many of which belong to the genus Phytophthora. The genus Phytophthora includes a large number of species that are invading forest ecosystems worldwide, chiefly as a consequence of global trade and human activities. This paper reports the results of a survey of Phytophthora diversity in natural and semi-natural forest ecosystems in Portugal along an elevation gradient. Isolations performed from 138 symptomatic plant tissues and rhizosphere samples collected from 26 plant species yielded a total of 19 Phytophthora species belonging to 6 phylogenetic clades, including P. cinnamomi (36 isolates), P. multivora (20), P. plurivora (9), P. cactorum (8), P. lacustris (8), P. pseudocryptogea (8), P. amnicola (6), P. hedraiandra (6), P. pseudosyringae (5), P. thermophila (5), P. bilorbang (4), P. inundata (4), P. asparagi (3), P. citricola (3), P. gonapodyides (3), P. rosacearum (3), P. chlamydospora (2), P. pachypleura (2), and P. syringae (1). Overall, the data obtained highlight the widespread occurrence of P. cinnamomi in natural ecosystems from sea level to mountain habitats. The results of the pathogenicity tests carried out on 2-year-old chestnut plants confirmed the key role of P. cinnamomi in the recrudescence of chestnut ink disease and the additional risk posed by P. pachypleura, P. plurivora, and P. multivora to Portuguese chestnut forests. Finally, three species, P. citricola, P. hedraiandra, and P. pachypleura, are reported for the first time in the natural ecosystems of Portugal. Full article

(This article belongs to the Special Issue Microbial Pathogenesis and Emerging Infections)

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9 pages, 4872 KiB

Open AccessCommunication

Complete Genome Sequence Analysis of the First Imported Mpox Virus Clade Ib Variant in China

by Yin Song, Yong Yan, Jingyu Xu, Shencong Lv, Ganglin Ren, Yamei Zhou, Wanchen Song, Rui Ge, Peihua Xu, Guoying Zhu and Zhongwen Chen

Pathogens 2025, 14(1), 102; https://doi.org/10.3390/pathogens14010102 - 20 Jan 2025

Viewed by 751

Abstract

Mpox, a zoonotic disease caused by the mpox virus (MPXV), has seen a significant shift in its epidemiological status since 2022, evolving from an initial local outbreak to a global epidemic. This recent outbreak of MPXV mainly emerged in several European and American [...] Read more.

Mpox, a zoonotic disease caused by the mpox virus (MPXV), has seen a significant shift in its epidemiological status since 2022, evolving from an initial local outbreak to a global epidemic. This recent outbreak of MPXV mainly emerged in several European and American countries and subsequently spread to over 100 countries and regions worldwide. The rapid evolution of MPXV, coupled with increased international interactions, has led to a gradual rise in mpox cases in certain regions of Asia, mostly involving MPXV clade II and its branch strains. In contrast, the more pathogenic and clinically severe MPXV clade Ib has been relatively rare, with no reports in China to date. Here, we analyzed the whole gene sequence of imported MPXV clade Ib variant from the first infection case detected in China. Through whole genome sequencing, we successfully obtained a full-length MPXV genome of 195,405 base pairs (bp). Phylogenetic analysis revealed that the genetic sequence of the MPXV in this case was predominantly clustered with MPXV clade Ib sequences previously reported from multiple African and European countries. Compared with the MPXV clade Ib reference strain DQ011155.1, there are 127 nucleotide alterations and 57 amino acid mutations in the MPXV genome of this case. Given that the MPXV clade Ib has started to appear in China, we must pay more attention to the prevention of and control measures for the spread of mpox. Full article

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16 pages, 2893 KiB

Open AccessArticle

Molecular Epidemiology, Drug-Resistant Variants, and Therapeutic Implications of Hepatitis B Virus and Hepatitis D Virus Prevalence in Nigeria: A National Study

by Oludare ‘Sunbo Adewuyi, Muhammad Shakir Balogun, Hirono Otomaru, Alash’le Abimiku, Anthony Agbakizu Ahumibe, Elsie Ilori, Que Anh Luong, Nwando Mba, James Christopher Avong, John Olaide, Oyeladun Okunromade, Adama Ahmad, Afolabi Akinpelu, Chinwe Lucia Ochu, Babatunde Olajumoke, Haruka Abe, Chikwe Ihekweazu, Adetifa Ifedayo, Michiko Toizumi, Hiroyuki Moriuchi, Katsunori Yanagihara, Jide Idris and Lay-Myint Yoshida Show full author list Hide full author list

Pathogens 2025, 14(1), 101; https://doi.org/10.3390/pathogens14010101 - 20 Jan 2025

Viewed by 1063

Abstract

Information on circulating HBV (sub-)genotype, variants, and hepatitis D virus (HDV) coinfection, which vary by geographical area, is crucial for the efficient control and management of HBV. We investigated the genomic characteristics of HBV (with a prevalence of 8.1%) and the prevalence of [...] Read more.

Information on circulating HBV (sub-)genotype, variants, and hepatitis D virus (HDV) coinfection, which vary by geographical area, is crucial for the efficient control and management of HBV. We investigated the genomic characteristics of HBV (with a prevalence of 8.1%) and the prevalence of HDV in Nigeria. We utilised 777 HBV-positive samples and epidemiological data from the two-stage sampled population-based, nationally representative Nigeria HIV/AIDS Indicator and Impact Survey conducted in 2018. We assessed 732 HBV DNA-extracted samples with detectable viral loads (VLs) for (sub-)genotypes and variants by whole-genome pre-amplification, nested PCR of the s-and pol-gene, and BigDye Terminator sequencing. We conducted HDV serology. In total, 19 out of the 36 + 1 states in Nigeria had a high prevalence of HBV (≥8%), with the highest prevalence (10.4%) in the north-central geopolitical zone. Up to 33.2% (95% CI 30.0–36.6) of the participants had detectable VLs of ≥300 copies/mL. The predominant circulating HBV genotype was E with 98.4% (95% CI 97.1–99.1), followed by A with 1.6% (95% CI 0.9–2.9). Drug-resistant associated variants and immune escape variants were detected in 9.3% and 0.4%, respectively. The seroprevalence of HDV was 7.34% (95% CI 5.5–9.2). Nigeria has subtype E as the major genotype with many variants. Full article

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

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6 pages, 2053 KiB

Open AccessCase Report

A Case of Dibothriocephalosis (Dibothriocephalus latus) from Iseo Lake (Northern Italy): An Update on a Persistent Sanitary Issue

by Vasco Menconi, Lisa Guardone, Elena Lazzaro, Romina Bottazzo, Valeria Besutti, Patrizia Danesi, Amedeo Manfrin, Andrea Basso, Giuseppe Arcangeli, Luana Cortinovis, Ewa Bilska-Zając and Giorgia Angeloni

Pathogens 2025, 14(1), 100; https://doi.org/10.3390/pathogens14010100 - 20 Jan 2025

Viewed by 597

Abstract

Dibothriocephalosis is a fish-borne parasitic zoonosis that is caused by tapeworms of the Dibothriocephalus (syn. Diphyllobothrium) genus. This paper describes a human case of dibothriocephalosis associated with the consumption of a presumably infected fish, prepared at a restaurant near Iseo Lake (northern [...] Read more.

Dibothriocephalosis is a fish-borne parasitic zoonosis that is caused by tapeworms of the Dibothriocephalus (syn. Diphyllobothrium) genus. This paper describes a human case of dibothriocephalosis associated with the consumption of a presumably infected fish, prepared at a restaurant near Iseo Lake (northern Italy). A month after, the patient found a segment of a worm in her stool. Molecular analysis identified it as Dibothriocephalus latus (syn. Diphyllobothrium latum). Several studies reported D. latus infections in patients from the subalpine regions of Switzerland, France, and Italy, but no cases have been reported from this area in the last 10 years. This report updates the epidemiology of dibothriocephalosis and emphasises the importance of collaboration among healthcare institutions for a prompt diagnosis and the need for food safety education for Food Business Operators and consumers to reduce transmission risks. Full article

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14 pages, 251 KiB

Open AccessArticle

Chlamydia trachomatis Serovars from the C-Complex and the B- and C-Related Complexes Are Significantly More Pathogenic than Those from the B-Complex in C3H/HeN but Not in BALB/c Mice

by Sukumar Pal, Jennifer R. Carmichael, Delia F. Tifrea, Olga Tatarenkova and Luis M. de la Maza

Pathogens 2025, 14(1), 97; https://doi.org/10.3390/pathogens14010097 - 19 Jan 2025

Viewed by 707

Abstract

Studies in humans indicate that certain Chlamydia trachomatis serovars are more pathogenic than others. Specifically, several studies concluded that serovars from the C-complex are more pathogenic than those from the B-complex, although there are reports that do not support this finding. To investigate [...] Read more.

Studies in humans indicate that certain Chlamydia trachomatis serovars are more pathogenic than others. Specifically, several studies concluded that serovars from the C-complex are more pathogenic than those from the B-complex, although there are reports that do not support this finding. To investigate these results in an animal model, the eight genitourinary C. trachomatis serovars were tested in two strains of mice: C3H/HeN and BALB/c. These two strains of mice were investigated because C3H/HeN is more susceptible to Chlamydia muridarum infections than BALB/c, indicative of differences in their immunogenetic background. Mice were infected transcervically with 10⁵ inclusion forming units of each of the C. trachomatis serovars, and vaginal cultures were collected. To determine the pathogenicity and its impact on fertility, at week seven post-infection, female mice were caged with male mice. In the C3H/HeN mice, significant differences in vaginal shedding and fertility were observed between serovars from the B-complex (D and E) and those from the C-complex (H, I, J) and B- and C-related complexes (G, F, and K). The animals infected with serovars F, G, H, I, J, and K shed less but had significantly more infertility than the mice infected with serovars D or E. The experiments in the BALB/c mice, however, did not show major differences in pathogenicity between the eight C. trachomatis serovars. These results support the findings in humans and emphasize the critical importance of the immunogenetic background of the host on the outcome of C. trachomatis infections. The data imply that management of C. trachomatis-infected patients may require a more personalized approach. Full article

(This article belongs to the Section Bacterial Pathogens)

12 pages, 949 KiB

Open AccessArticle

Seroprevalence of Borrelia, Anaplasma, Bartonella, Toxoplasma, Mycoplasma, Yersinia, and Chlamydia in Human Population from Eastern Poland

by Angelina Wójcik-Fatla, Anna Sawczyn-Domańska, Anna Kloc, Joanna Krzowska-Firych and Jacek Sroka

Pathogens 2025, 14(1), 96; https://doi.org/10.3390/pathogens14010096 - 18 Jan 2025

Viewed by 883

Abstract

The epidemiological situation related to infectious diseases is influenced by many factors. To monitor actual trends in selected zoonoses, a total of 473 serum samples from farmers, forestry workers, and veterinarians were collected for serological examination. Anti-Borrelia burgdorferi sensu lato (s.l.) antibodies [...] Read more.

The epidemiological situation related to infectious diseases is influenced by many factors. To monitor actual trends in selected zoonoses, a total of 473 serum samples from farmers, forestry workers, and veterinarians were collected for serological examination. Anti-Borrelia burgdorferi sensu lato (s.l.) antibodies were tested with ELISA and Western blot (WB) tests; the detection of anti-Toxoplasma gondii antibodies was performed using an enzyme linked fluorescence assay (ELFA). Antibodies to bartonellosis, anaplasmosis, and chlamydiosis were determined by indirect immunofluorescent test (IFA), whereas antibodies to yersiniosis and mycoplasmosis were confirmed in the ELISA test. Positive or borderline results of antibodies against B. burgdorferi s.l. in the ELISA test were detected in 33.8% of the study population. The borderline or positive ELISA test results for at least one antibody class were confirmed by WB in 58.7% of cases. The IgG antibodies against Anaplasma phagocytophilum, Toxoplasma gondii, and Mycoplasma pneumoniae were detected in 9.6%, 51.7%, and 63.6% of samples, respectively. Antibodies against Yersinia spp., Bartonella henselae, and Chlamydia pneumoniae were found to vary between 43 and 47%. Full article

(This article belongs to the Special Issue Exploring the Biodiversity of Parasites in Humans, Wild and Domestic Animals)

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18 pages, 4648 KiB

Open AccessArticle

Potential Strategies Applied by Metschnikowia bicuspidata to Survive the Immunity of Its Crustacean Hosts

by Ji Zhang, Bingyu Li, Bingnan Zuo and Xiaodong Li

Pathogens 2025, 14(1), 95; https://doi.org/10.3390/pathogens14010095 - 18 Jan 2025

Viewed by 634

Abstract

Metschnikowia bicuspidata is the specific pathogen for “milky disease” in the Chinese mitten crab (Eriocheir sinensis), accounting for huge losses to the industry. And yet, there is no precise study describing the pathogenesis of M. bicuspidata, largely hindering the development [...] Read more.

Metschnikowia bicuspidata is the specific pathogen for “milky disease” in the Chinese mitten crab (Eriocheir sinensis), accounting for huge losses to the industry. And yet, there is no precise study describing the pathogenesis of M. bicuspidata, largely hindering the development of novel control methods against its causing diseases. Here, we compared the transcriptomes of M. bicuspidata cells collected from a control group (cultured without E. sinensis hemocytes) and a treatment group (cultured with E. sinensis hemocytes), using RNA sequencing. Through comprehensively analyzing the differentially expressed genes (DEGs), both the most regulated ones and the ones involved in crucial enriched KEGG pathways, we found that certain processes might be required for M. bicuspidata’s survival under hemocyte stress. Key genes involved in oxidative phosphorylation, fatty acid metabolism, upper glycolysis, and gluconeogenesis were upregulated, and those for β-glucan unmasking, autophagy, and cell polarity were downregulated, in the treatment group. Our results suggest that M. bicuspidata colonizes and therefore establishes an infection in E. sinensis via enhancing aerobic respiration, glucose-6-phosphate accumulation, and cell-wall masking. In addition, we applied multiple means to evaluate a series of candidate reference genes and found that PMA1 in combination with ACT1 is the most suitable choice for accurate normalization in quantitative real-time PCR (qRT-PCR) assays. Thus, we used this combination as the reference and performed qRT-PCR verification of several DEGs. It is shown that the expression trends of these tested DEGs in qRT-PCR assays are the same as those in RNA-Seq assays. This study not only provides insights into strategies facilitating M. bicuspidata’s survival within E. sinensis, initially elucidating the pathogenesis of this yeast, but also recommends a useful molecular tool regarding qRT-PCR assays in this pathogen. Full article

(This article belongs to the Section Fungal Pathogens)

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22 pages, 3050 KiB

Open AccessReview

The Role of Pathogens in Bumblebee Decline: A Review

by Huanhuan Chen, Nawaz Haider Bashir, Qiang Li, Chao Liu, Muhammad Naeem, Haohan Wang, Wenrong Gao, Richard T. Corlett, Cong Liu and Mayra C. Vidal

Pathogens 2025, 14(1), 94; https://doi.org/10.3390/pathogens14010094 - 18 Jan 2025

Viewed by 780

Abstract

Bumblebees, the most important wild pollinators in both agricultural and natural ecosystems, are declining worldwide. The global decline of bumblebees may threaten biodiversity, pollination services, and, ultimately, agricultural productivity. Several factors, including pesticide usage, climate change, habitat loss, and species invasion, have been [...] Read more.

Bumblebees, the most important wild pollinators in both agricultural and natural ecosystems, are declining worldwide. The global decline of bumblebees may threaten biodiversity, pollination services, and, ultimately, agricultural productivity. Several factors, including pesticide usage, climate change, habitat loss, and species invasion, have been documented in the decline of bumblebee species, but recent studies have revealed the dominating role of pathogens and parasites over any of these causes. Unfortunately, there is a lack of a full understanding of the role of pathogens and parasites in the decline of bumblebee species. The current study provides a comprehensive review of how pathogens and parasites contribute to the decline of bumblebee species. The study also explores the prevalence of each pathogen and parasite within bumblebee populations. Furthermore, we address the synergistic effects of pathogens and other stressors, such as pesticides, climatic effects, and habitat loss, on bumblebee populations. To summarize, we propose possible conservation and management strategies to preserve the critical role of bumblebees in pollination services and thus to support ecosystem and agricultural health. Full article

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12 pages, 2329 KiB

Open AccessArticle

Epidemiological Analysis of Fungal Infection Disease in Pediatric Population: Focus on Hospitalization from 2007 to 2022 in Veneto Region in Italy

by Lorenzo Chiusaroli, Claudia Cozzolino, Silvia Cocchio, Mario Saia, Carlo Giaquinto, Daniele Donà and Vincenzo Baldo

Pathogens 2025, 14(1), 93; https://doi.org/10.3390/pathogens14010093 - 18 Jan 2025

Viewed by 543

Abstract

Fungal infections (FIs) are widespread globally, affecting both immunocompromised and immunocompetent children, with varying clinical implications based on age and comorbidities. In immunocompromised children, particularly those with hematologic oncological conditions, FI leads to substantially longer hospital stays and increased in-hospital mortality, with reported [...] Read more.

Fungal infections (FIs) are widespread globally, affecting both immunocompromised and immunocompetent children, with varying clinical implications based on age and comorbidities. In immunocompromised children, particularly those with hematologic oncological conditions, FI leads to substantially longer hospital stays and increased in-hospital mortality, with reported rates ranging from 15% to 20%. Our study aims to analyze the epidemiological trends of fungal infections in the pediatric population within a specific region of Italy. We extracted ICD-9 codes related to fungal infections from hospital discharge records (HDRs) in the pediatric population of Veneto, located in the north-east of Italy, between 2007 and 2022. We included all children admitted to the hospital with a primary or secondary diagnosis during admission for other reasons. Data were stratified based on age, year, ward of admission, and type of diagnosis. Patients older than eighteen and HDRs related to a second admission within thirty days from the previous admission were excluded. A total of 1433 diagnoses were analyzed during the period, with 241 (16.8%) as main diagnoses and 1192 (83.2%) as secondary diagnoses. The overall hospitalization rate was 1084 cases/100,000 (1.69 cases/100,000 as primary diagnosis and 8.95 cases/100,000 as secondary). The hospitalization rate stratified for age was 11,055 cases/100,000 among infants younger than 1 year, 8.48 cases/100,000 among those aged 1-4 years, and 4.4 cases/100,000 among children older than 5. The more frequent infection was Candida spp. (62.8%), followed by Aspergillus spp. (14.6%) and skin mycosis (9.5%). Overall, the pediatric in-hospital case fatality rate due to FI was 2.09%. Our study elucidated the overall experience of fungal infections in the pediatric population of the Veneto region in Italy. Specifically, we underscored a relatively stable hospitalization rate for fungal diseases and a noteworthy mortality rate. Full article

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

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13 pages, 2180 KiB

Open AccessArticle

Towards Detecting Associations of Canine Astrovirus and Caliciviruses with Health and Living Characteristics of Dogs in Greece

by Efthymia Stamelou, Konstantinos Papageorgiou, Dimitrios Papadopoulos, Georgios Delis, Dimitrios Chatzopoulos, Zoi Athanasakopoulou, Efstratios Moschidis, Evanthia Petridou and Spyridon K. Kritas

Pathogens 2025, 14(1), 92; https://doi.org/10.3390/pathogens14010092 - 18 Jan 2025

Viewed by 502

Abstract

Astroviruses and caliciviruses are important causative agents of gastroenteritis in humans worldwide. They have been detected in a variety of animal species, including dogs, but their role in the induction of disease in animals remains uncertain. In a molecular study that was conducted [...] Read more.

Astroviruses and caliciviruses are important causative agents of gastroenteritis in humans worldwide. They have been detected in a variety of animal species, including dogs, but their role in the induction of disease in animals remains uncertain. In a molecular study that was conducted in Greece, including healthy and gastroenteritis-affected dogs of different ages, astrovirus (AstV) and sapovirus (SaV) were detected in 15% and 26% of the examined animals, respectively. A specialized questionnaire was filled out for each of the dogs participating in the study, including information about different characteristics and risk factors that could possibly affect their health status. This information was analyzed with the use of two innovative statistical methods, i.e., a Multiple Correspondence Analysis (MCA) and the Ascending Hierarchical Classification (AHC). Based on their results, it was possible to define various groups of dogs based on their characteristics. AstV seems to occur more often in low-health-status dogs, usually mongrels, living in rural areas, showing vomit, diarrhea, and diet changes. Dogs of this group usually live with other pets in the same household and have frequent contact with stray animals. The presence of SaV does not seem to be associated with any of the examined factors. Full article

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22 pages, 2499 KiB

Open AccessReview

The Importance of Lung Innate Immunity During Health and Disease

by Gusty Rizky Teguh Ryanto, Ratoe Suraya and Tatsuya Nagano

Pathogens 2025, 14(1), 91; https://doi.org/10.3390/pathogens14010091 - 17 Jan 2025

Viewed by 494

Abstract

The lung is a vital organ for the body as the main source of oxygen input. Importantly, it is also an internal organ that has direct contact with the outside world. Innate immunity is a vital protective system in various organs, whereas, in [...] Read more.

The lung is a vital organ for the body as the main source of oxygen input. Importantly, it is also an internal organ that has direct contact with the outside world. Innate immunity is a vital protective system in various organs, whereas, in the case of the lung, it helps maintain a healthy, functioning cellular and molecular environment and prevents any overt damage caused by pathogens or other inflammatory processes. Disturbances in lung innate immunity properties and processes, whether over-responsiveness of the process triggered by innate immunity or lack of responses due to dysfunctions in the immune cells that make up the innate immunity system of the lung, could be correlated to various pathological conditions. In this review, we discuss globally how the components of lung innate immunity are important not only for maintaining lung homeostasis but also during the pathophysiology of notable lung diseases beyond acute pulmonary infections, including chronic obstructive pulmonary disease (COPD), asthma, and pulmonary fibrosis. Full article

(This article belongs to the Special Issue Interactions between Pathogens and the Human Immune System)

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12 pages, 1120 KiB

Open AccessCommunication

Genomic and Drug Resistance Profile of Bovine Multidrug-Resistant Escherichia coli Isolated in Kazakhstan

by Pavel G. Alexyuk, Andrey P. Bogoyavlenskiy, Yergali S. Moldakhanov, Kuralay S. Akanova, Adolat N. Manakbayeva, Timur Kerimov, Vladimir E. Berezin and Madina S. Alexyuk

Pathogens 2025, 14(1), 90; https://doi.org/10.3390/pathogens14010090 - 17 Jan 2025

Viewed by 482

Abstract

While studying the prevalence and profile of antibiotic resistance among E. coli isolated from the feces of calves with signs of colibacillosis, a strain with a wide spectrum of drug resistance was isolated. Whole-genome sequencing, followed by bioinformatic processing and the annotation of [...] Read more.

While studying the prevalence and profile of antibiotic resistance among E. coli isolated from the feces of calves with signs of colibacillosis, a strain with a wide spectrum of drug resistance was isolated. Whole-genome sequencing, followed by bioinformatic processing and the annotation of genes of this strain, showed that the genome has a total length of 4,803,482 bp and contains 4986 genes, including 122 RNA genes. A total of 31% of the genes are functionally significant and represent 26 functional groups. Additionally, 55 antibiotic resistance genes were revealed. A phenotypic drug-resistance study was performed according to CASFM and CLSI guidelines, which showed that the investigated strain was resistant to eight antibacterial drugs of different classes, including colistin. This is the first report on the AMR profile of an E. coli isolate obtained from a sick calf with evidence of escherichiosis in Kazakhstan. The provided information on the genome will be valuable in studying the evolution and development of antibiotic-resistant forms of E. coli and increase our knowledge of pathogenicity. It may also be a source for future comparative studies of the virulence and drug resistance of E. coli isolates. Full article

(This article belongs to the Section Bacterial Pathogens)

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15 pages, 2891 KiB

Open AccessArticle

Maternal Infections, Antibiotics, Steroid Use, and Diabetes Mellitus Increase Risk of Early-Onset Sepsis in Preterm Neonates: A Nationwide Population-Based Study

by Hao-Yuan Lee, Yu-Lung Hsu, Wen-Yuan Lee, Kuang-Hua Huang, Ming-Luen Tsai, Chyi-Liang Chen, Yu-Chia Chang and Hung-Chih Lin

Pathogens 2025, 14(1), 89; https://doi.org/10.3390/pathogens14010089 - 17 Jan 2025

Viewed by 507

Abstract

The global evolution of pathogens causing early-onset sepsis (EOS), a critical condition in preterm infants, necessitates a re-evaluation of risk factors to develop updated prevention and treatment strategies. This nationwide case–control study in Taiwan analyzed data from the National Health Insurance Research Database, [...] Read more.

The global evolution of pathogens causing early-onset sepsis (EOS), a critical condition in preterm infants, necessitates a re-evaluation of risk factors to develop updated prevention and treatment strategies. This nationwide case–control study in Taiwan analyzed data from the National Health Insurance Research Database, Birth Reporting Database, and Maternal and Child Health Database from 2010 to 2019. The study included 176,681 mother–child pairs with preterm births. We identified 2942 clinical EOS cases from 5535 diagnosed sepsis cases, excluding unlikely cases. A control group of 14,710 preterm neonates without EOS was selected at a 1:5 ratio. Clinical EOS increased since 2017. Adjusted logistic regression identified significant EOS risk factors in preterm infants, including maternal fever, chorioamnionitis, maternal diabetes mellitus, maternal antibiotic usage, very preterm birth, birth weight (all with p < 0.001), maternal pneumonia (p = 0.002), and maternal CS (p = 0.004). Effective treatment of maternal conditions like diabetes, fever, and infections is essential to prevent EOS in preterm infants. Key measures include reducing unnecessary antibiotics or steroids, minimizing unnecessary cesarean sections, avoiding premature or prolonged rupture of membranes (PPROM), and increasing gestational age and neonatal birth weight. High-risk preterm neonates should be closely monitored for EOS and considered for antibiotics when warranted. Full article

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9 pages, 225 KiB

Open AccessCommunication

Pradofloxacin Minimum Inhibitory Concentration Profiling of Streptococcus suis Isolates: Insights into Antimicrobial Susceptibility in Swine

by Jessica Risser, Ronald Tessman, Don Bade, Orhan Sahin, Maria J. Clavijo, Saumya Dhup and Patrick Hoffmann

Pathogens 2025, 14(1), 88; https://doi.org/10.3390/pathogens14010088 - 17 Jan 2025

Viewed by 472

Abstract

This study evaluated the minimum inhibitory concentration (MIC) of pradofloxacin against various swine respiratory pathogens, including Bordetella bronchiseptica, Glaesserella parasuis, Mycoplasma hyopneumoniae, Pasteurella multocida, and Streptococcus suis (S. suis), associated with disease in swine. This research was [...] Read more.

This study evaluated the minimum inhibitory concentration (MIC) of pradofloxacin against various swine respiratory pathogens, including Bordetella bronchiseptica, Glaesserella parasuis, Mycoplasma hyopneumoniae, Pasteurella multocida, and Streptococcus suis (S. suis), associated with disease in swine. This research was conducted in two phases: the initial phase examined isolates from the lungs that could be either commensal or pathogenic, while the second phase focused on systemic S. suis strains that spread from the respiratory tract to the brain. The pradofloxacin MIC values of the second phase were within the MIC range of the initial phase, with MIC₅₀ and MIC₉₀ values highlighting its potential as an effective antimicrobial agent. Quality control data validated the reliability of our MIC findings, with all pradofloxacin MIC values for control organisms within approved ranges. These findings suggest that pradofloxacin has broad-spectrum activity against Gram-positive and Gram-negative bacteria and may serve as a reliable therapeutic option for managing S. suis and other swine respiratory infections. This study highlights pradofloxacin as an alternative antimicrobial therapy for swine respiratory diseases, offering a potential solution amidst rising concerns over antibiotic resistance. Full article

(This article belongs to the Special Issue Understanding the Pathogenesis of Streptococcus suis)

19 pages, 1145 KiB

Open AccessArticle

Inverted Patterns of Schistosomiasis and Fascioliasis and Risk Factors Among Humans and Livestock in Northern Tanzania

by Ephrasia A. Hugho, Yakob P. Nagagi, Lucille J. Lyaruu, Victor V. Mosha, Ndealilia Senyael, Magweiga M. Mwita, Ruth W. Mabahi, Violet M. Temba, Mapulish Hebel, Mohamed Nyati, Blandina T. Mmbaga, Theonest O. Ndyetabura and AbdulHamid S. Lukambagire

Pathogens 2025, 14(1), 87; https://doi.org/10.3390/pathogens14010087 - 17 Jan 2025

Viewed by 594

Abstract

Fascioliasis and schistosomiasis are parasitic trematodiases of public health and economic concern in humans and livestock. However, data on the distribution and risk factors for fascioliasis remain limited, while epidemiological gaps hinder schistosomiasis control in Tanzania. This One Health, cross-sectional study examined the [...] Read more.

Fascioliasis and schistosomiasis are parasitic trematodiases of public health and economic concern in humans and livestock. However, data on the distribution and risk factors for fascioliasis remain limited, while epidemiological gaps hinder schistosomiasis control in Tanzania. This One Health, cross-sectional study examined the prevalence and risk factors of schistomiasis and fascioliasis in northern Tanzania, involving 310 livestock and 317 human participants from Arusha, Kilimanjaro, and Manyara regions. Using standard parasitological methods, livestock fascioliasis prevalence was 21.3%, while schistosomiasis prevalence was 1.0%. Human fascioliasis prevalence was 1.9%, while schistosomiasis prevalence was 12.6%. Female animals, particularly cattle in Kilimanjaro and Manyara, had higher odds of fascioliasis. Human–animal contact through husbandry increased schistosomiasis risk (aOR = 4.21; 95% CI: 1.81–9.80), while the use of borehole-water was protective (aOR = 0.33; 95% CI: 0.11–0.97). Fascioliasis risk was higher among individuals aged 36–55 years (aOR = 7.66; 95% CI: 1.36–43.23), with cabbage consumption offering protection (aOR = 0.08; 95% CI: 0.01–0.89). The study revealed inverted prevalence patterns of fascioliasis and schistosomiasis in humans and livestock, driven by vector-dependent transmission dynamics. These findings emphasize the need for an integrated One Health approach to manage shared human and animal health risks in Tanzania. Full article

(This article belongs to the Special Issue Epidemiology, Diagnosis and Clinical Management of Human Parasitic Infections)

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21 pages, 830 KiB

Open AccessReview

Parasites in Sewage: Legal Requirements and Diagnostic Tools

by Oliwia Obuch-Woszczatyńska, Klaudia Bylińska, Małgorzata Krzyżowska, Karol Korzekwa and Piotr Bąska

Pathogens 2025, 14(1), 86; https://doi.org/10.3390/pathogens14010086 - 16 Jan 2025

Viewed by 567

Abstract

Despite the vast amount of water on Earth, only a small percent is suitable for consumption, and these resources are diminishing. Moreover, water resources are unevenly distributed, leading to significant disparities in access to drinking water between countries and populations. Increasing consumption and [...] Read more.

Despite the vast amount of water on Earth, only a small percent is suitable for consumption, and these resources are diminishing. Moreover, water resources are unevenly distributed, leading to significant disparities in access to drinking water between countries and populations. Increasing consumption and the expanding human population necessitate the development of novel wastewater treatment technologies and the use of water treatment byproducts in other areas, such as fertilisers. However, water treatment sludge often cannot be used to enhance crop production due to the presence of parasite eggs, particularly from roundworms (Ascaridae family), which are resistant to environmental factors and can pose a threat for several years. Legislation prohibits the use of sludge containing parasite eggs as fertiliser. In some cases, water may not contain parasite eggs but larvae, which require different detection methods. Additionally, the presence of eggs does not necessarily indicate danger since they may lose infectivity due to prolonged storage or exposure to chemical compounds in the sewage. This paper reviews European Union regulations on wastewater treatment, the selected parasitic diseases related to the presence of parasites in wastewater, the spectrum of detection methods, and highlights differences in viability and invasiveness, which is intended to draw attention to the need to determine both biological properties of parasites. Full article

(This article belongs to the Section Parasitic Pathogens)

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10 pages, 440 KiB

Open AccessArticle

Prevalence of Leptospira spp. in Lithuanian Wild Boars (Sus scrofa)

by Birutė Karvelienė, Inga Stadalienė, Jūratė Rudejevienė, Evelina Burbaitė, Dalia Juodžentė, Marius Masiulis, Jūratė Buitkuvienė, Jurgita Šakalienė and Gintaras Zamokas

Pathogens 2025, 14(1), 85; https://doi.org/10.3390/pathogens14010085 - 16 Jan 2025

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Abstract

Leptospira is a bacteria responsible for a widespread zoonosis that affects both humans and animals. Leptospirosis is a challenging pathology to diagnose and treat since its signs are unspecific and symptoms vary greatly. The disease seems to be highly prevalent in environments where [...] Read more.

Leptospira is a bacteria responsible for a widespread zoonosis that affects both humans and animals. Leptospirosis is a challenging pathology to diagnose and treat since its signs are unspecific and symptoms vary greatly. The disease seems to be highly prevalent in environments where reservoir animals such as rats and small mammals are common. Even though leptospirosis in humans in Lithuania is rare, it remains a disease of significance in Europe. Information on reservoir animals and prevalence of Leptospira in wild animals in Lithuania is lacking. The aim of this country-wide study was to evaluate the seroprevalence of Leptospira in wild boars in Lithuania. Hunted animals were collected from ten counties that represented the boar population of the country. The sera of 451 collected boars were evaluated for eight Leptospira serovars using the microscopic agglutination test. Seropositivity was observed in 102 (22.6%) boars. Overall, 194 positive reactions occurred. Boars older than 2 years were affected by more serovars and were more seropositive than younger boars (p < 0.05). The highest number of positive reactions was observed in Panevėžys (87.9%) and Vilnius (69.1%) counties. The results of this study might indicate that the wild boar is a reservoir animal of Leptospira and plays a role in its transmission in Lithuania. Full article

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

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17 pages, 10117 KiB

Open AccessArticle

A Conserved Di-Lysine Motif in the E2 Transactivation Domain Regulates MmuPV1 Replication and Disease Progression

by Jessica Gonzalez, Marsha DeSmet and Elliot J. Androphy

Pathogens 2025, 14(1), 84; https://doi.org/10.3390/pathogens14010084 - 16 Jan 2025

Viewed by 413

Abstract

The papillomavirus E2 protein regulates the transcription, replication, and segregation of viral episomes within the host cell. A multitude of post-translational modifications have been identified which control E2 functions. A highly conserved di-lysine motif within the transactivation domain (TAD) has been shown to [...] Read more.

The papillomavirus E2 protein regulates the transcription, replication, and segregation of viral episomes within the host cell. A multitude of post-translational modifications have been identified which control E2 functions. A highly conserved di-lysine motif within the transactivation domain (TAD) has been shown to regulate the normal functions of the E2 proteins of BPV-1, SfPV1, HPV-16, and HPV-31. This motif is similarly conserved in the E2 of the murine papillomavirus, MmuPV1. Using site-directed mutagenesis, we show that the first lysine (K) residue within the motif, K112, is absolutely required for E2-mediated transcription and transient replication in vitro. Furthermore, mutation of the second lysine residue, K113, to the potential acetyl-lysine mimic glutamine (Q) abrogated E2 transcription and decreased transient replication in vitro, while the acetylation defective arginine (R) mutant remained functional. Both K113 mutants were able to induce wart formation in vivo, though disease progression appeared to be delayed in the K113Q group. These findings suggest that acetylation of K113 may act as a mechanism for repressing MmuPV1 E2 activity. Full article

(This article belongs to the Section Viral Pathogens)

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17 pages, 1570 KiB

Open AccessArticle

Archival and Newly Isolated Historical Bacillus anthracis Strains Populate the Deeper Phylogeny of the A.Br.075(Sterne) Clade

by Markus Antwerpen, Peter Braun, Wolfgang Beyer, Dirk Aldenkortt, Michael Seidel and Gregor Grass

Pathogens 2025, 14(1), 83; https://doi.org/10.3390/pathogens14010083 - 16 Jan 2025

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Abstract

The anthrax pathogen Bacillus anthracis can remain dormant as spores in soil for many years. This applies to both natural foci and to sites of anthropogenic activity such as tanneries, abattoirs, or wool factories. The A.Br.075 (A-branch) clade (also known as A.Br.Sterne) is [...] Read more.

The anthrax pathogen Bacillus anthracis can remain dormant as spores in soil for many years. This applies to both natural foci and to sites of anthropogenic activity such as tanneries, abattoirs, or wool factories. The A.Br.075 (A-branch) clade (also known as A.Br.Sterne) is prominent not only because it comprises several outbreak strains but even more so because spore preparations of its namesake, the Sterne strain, are counted among the most utilized anthrax animal vaccines. In this study, we genome-sequenced and analyzed 56 additional B. anthracis isolates of the A.Br.075 clade. Four of these we recently retrieved from soil samples taken from a decades-long abandoned tannery. The other 52 strains originated from our archival collection from the 20th century. Notably, the extended phylogeny of the A.Br.075 clade indicated that many of the newly added chromosomes represent basal members, some of which are among the most basal strains from this lineage. Twelve new strains populate a very deep-branching lineage we have named A.Br.Ortho-Sterne (also known as A.Br.076). A further 11 isolates amend the clade named A.Br.Para-Sterne (A.Br.078). Finally, some of the terminal clusters of the clade named A.Br.Eu-Sterne appear to be replete with (near) identical isolates, possibly a result of widespread use of the Sterne vaccine and of its re-isolation from vaccination-related animal anthrax outbreaks. From the accrued new phylogenetic information, we designed and tested a variety of new SNP-PCR assays for rapid and facile genotyping of unassigned B. anthracis genomes. Lastly, the successful isolation of live B. anthracis from a long-abandoned tannery reemphasizes the need for continued risk awareness of such sites. Full article

(This article belongs to the Section Bacterial Pathogens)

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19 pages, 4179 KiB

Open AccessArticle

Pneumocystis Pneumonia Severity Is Associated with Taxonomic Shifts in the Respiratory Microbiota

by Valentina Del Prete, Antonia Piazzesi, Matteo Scanu, Francesca Toto, Stefania Pane, Federica Berrilli, Giovangiacinto Paterno, Lorenza Putignani and David di Cave

Pathogens 2025, 14(1), 82; https://doi.org/10.3390/pathogens14010082 - 16 Jan 2025

Viewed by 494

Abstract

Pneumonia caused by Pneumocystis jirovecii infection (PCP) is a potentially life-threatening illness, particularly affecting the immunocompromised. The past two decades have shown an increase in PCP incidence; however, the underlying factors that promote disease severity and fatality have yet to be fully elucidated. [...] Read more.

Pneumonia caused by Pneumocystis jirovecii infection (PCP) is a potentially life-threatening illness, particularly affecting the immunocompromised. The past two decades have shown an increase in PCP incidence; however, the underlying factors that promote disease severity and fatality have yet to be fully elucidated. Recent evidence suggests that the microbiota of the respiratory tract may play a role in stimulating or repressing pulmonary inflammation, as well as the progression of both bacterial and viral pneumonia. Here, we employed 16S rRNA metataxonomic sequencing to profile the respiratory microbiota of patients with mild-moderate and severe PCP. Our results show that the upper and lower airways of PCP patients have bacterial profiles which have been associated with a pro-inflammatory response. Furthermore, we find that severe PCP is associated with lower bacterial diversity and an increase in Prevotella and a decrease in Neisseria. Functionally, severe PCP was associated with a decrease in metabolic pathways of molecules with anti-inflammatory and antimicrobial properties. To our knowledge, this is the first study showing an association of PCP severity with shifts in the respiratory microbiome and may provide some insight into which patients are more susceptible to the more severe manifestations of the disease. Full article

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14 pages, 3950 KiB

Open AccessArticle

Is Intestinal Microbiota Fully Restored After Chickens Have Recovered from Coccidiosis?

by Jiaqing Guo, Zijun Zhao, Chace Broadwater, Isabel Tobin, Jing Liu, Melanie Whitmore and Guolong Zhang

Pathogens 2025, 14(1), 81; https://doi.org/10.3390/pathogens14010081 - 16 Jan 2025

Viewed by 604

Abstract

The intestinal microbiota is known to be altered by Eimeria-induced coccidiosis, but it remains unclear whether the microbiota is fully restored after recovery. To address this, 110 newly hatched Cobb male broiler chickens were challenged with 2 × 10⁴ sporulated oocysts [...] Read more.

The intestinal microbiota is known to be altered by Eimeria-induced coccidiosis, but it remains unclear whether the microbiota is fully restored after recovery. To address this, 110 newly hatched Cobb male broiler chickens were challenged with 2 × 10⁴ sporulated oocysts of Eimeria maxima (EM) strain M6 or mock-infected with saline on day 10. Body weight and feed intake were recorded. Additionally, 10 mock- and 12 EM-infected birds were randomly selected to assess the small intestinal lesion, fecal oocyst shedding, and ileal and cecal microbiota compositions using 16S rRNA gene sequencing at 3, 5, 7, 14, and 21 days post-infection (dpi). EM infection significantly decreased (p < 0.001) body weight by 5 dpi, persisting through 21 dpi. The infection also reduced (p < 0.05) weight gain, feed intake, and feed efficiency in the first week; however, these parameters became comparable in the second and third weeks. At 7 dpi, during the peak of infection, major lactic acid bacteria were enriched, while short-chain fatty acid-producing bacteria were mostly suppressed in both the ileum and cecum. Opportunistic pathogens such as Escherichia and Clostridium perfringens transiently bloomed at 7 dpi. By 14 dpi, differential bacterial enrichment subsided, and nearly all commensal bacteria returned to healthy levels by 21 dpi. Coupled with comparable growth performance between healthy and EM-recovered chickens, we conclude that the intestinal microbiota is largely restored to its healthy state after recovery. Understanding the microbiota’s responses to coccidiosis may inform probiotic-based mitigation strategies. Full article

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15 pages, 5931 KiB

Open AccessArticle

Survey and Identification of Fusarium Head Blight Pathogens of Wheat in the Western Cape Region of South Africa

by Ali Al-Hashimi, Augustine Innalegwu Daniel, Omolola Aina, Morné Du Plessis, Marshall Keyster and Ashwil Klein

Pathogens 2025, 14(1), 80; https://doi.org/10.3390/pathogens14010080 - 16 Jan 2025

Viewed by 613

Abstract

Fusarium head blight (FHB) is a major disease affecting wheat production worldwide, caused by multiple Fusarium species. In this study, seven Fusarium strains were isolated from wheat fields across the Western Cape region of South Africa and identified through phylogenetic analysis. The strains [...] Read more.

Fusarium head blight (FHB) is a major disease affecting wheat production worldwide, caused by multiple Fusarium species. In this study, seven Fusarium strains were isolated from wheat fields across the Western Cape region of South Africa and identified through phylogenetic analysis. The strains were classified into three species complexes: the Fusarium graminearum species complex (FGSC), Fusarium incarnatum-equiseti species complex (FIESC), and Fusarium tricinctum species complex (FTSC). Disease severity was highest in the South coast regions of Swellendam (42.73%) and Caledon (38.00%), with the dough stage of wheat development showing the highest disease rate (0.3 in Swellendam and Caledon). The phylogenetic analysis showed distinct clustering of these isolates with known species from the NCBI database, confirming their classification. F. ipomoeae was uniquely found in Swellendam and Caledon, while F. tricinctum occurred only in Klipheuwel and Caledon, highlighting geographical variation in species distribution. Mycotoxin profiling revealed that F. culmorum and F. pseudograminearum produced zearalenone, F. culmorum and F. tricinctum produced 15-acetyl-deoxynivalenol (15-ADON), while F. pseudograminearum produced nivalenol (NIV). These findings provide significant insights into the distribution of Fusarium species and their associated trichothecene chemotypes in the Western Cape, which is crucial for developing effective FHB management strategies and ensuring food security and safety. Full article

(This article belongs to the Section Fungal Pathogens)

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19 pages, 1944 KiB

Open AccessArticle

Antibiotic Susceptibility Patterns and Virulence Profiles of Classical and Hypervirulent Klebsiella pneumoniae Strains Isolated from Clinical Samples in Khyber Pakhtunkhwa, Pakistan

by Azra, Taj Ali Khan, Ihtisham Ul Haq, Woranich Hinthong, Susana Campino, Aisha Gohar, Noman Khan, Muhammad Kashif, Ihsan Ullah and Taane G. Clark

Pathogens 2025, 14(1), 79; https://doi.org/10.3390/pathogens14010079 - 15 Jan 2025

Viewed by 716

Abstract

The emergence of hypervirulent and carbapenem-resistant hypermucoviscous Klebsiella pneumoniae strains presents a significant public health challenge due to their increased virulence and resistance to multiple antibiotics. This study evaluates the antibiotic susceptibility patterns and virulence profiles of classical and hypervirulent K. pneumoniae strains [...] Read more.

The emergence of hypervirulent and carbapenem-resistant hypermucoviscous Klebsiella pneumoniae strains presents a significant public health challenge due to their increased virulence and resistance to multiple antibiotics. This study evaluates the antibiotic susceptibility patterns and virulence profiles of classical and hypervirulent K. pneumoniae strains isolated from various clinical samples. A total of 500 clinical samples were collected from patients at the Mardan Medical Complex and Ayub Medical Complex in KPK between July 2022 and June 2024. Among these, 64 K. pneumoniae strains were isolated and subsequently subjected to antimicrobial susceptibility testing (AST) and phenotypic virulence detection. Among the 64 isolates, 21 (32.8%) exhibited hypermucoviscosity, a characteristic associated with increased pathogenicity. Hemagglutination was observed in 35 (54.1%) of the isolates, indicating the presence of surface adhesins that facilitate bacterial adherence to host tissues. A high prevalence of biofilm formation was noted, with 54 (84%) isolates capable of forming biofilms, which are known to protect bacteria from antibiotics and the host immune response. Most isolates (59/64, 92.1%) were resistant against ampicillin, highlighting its limited efficacy against these strains. Conversely, the lowest resistance was observed for tigecycline, with only 15% (10/64) of the isolates showing resistance, indicating its potential utility as a treatment option. The study also found that 38 (59.3%) of the isolates were extended-spectrum beta-lactamase (ESBL) producers, 42 (65.6%) were multidrug-resistant (MDR), 32 (50%) were extensively drug-resistant (XDR), and 13 (20.3%) were resistant to carbapenems. The genetic study revealed biofilm producer and enhancer genes (mrkD, pgaABCD, fimH, treC, wzc, pilQ, and luxS) mainly in the hypervirulent strains. These hypervirulent strains also show a high number of resistance genes. The findings of this study underscore the critical need for the active surveillance of antimicrobial resistance and virulence determinants in K. pneumoniae. The coexistence of high levels of antibiotic resistance and virulence factors in these isolates poses a severe threat to public health, as it can lead to difficult-to-treat infections and increased morbidity and mortality. Full article

(This article belongs to the Special Issue Virulence and Resistance Mechanisms in Multidrug-Resistant Gram-Negative Bacteria)

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13 pages, 2083 KiB

Open AccessBrief Report

Use of Tick Cell Lines in Co-Infection Studies with a Preliminary Study of Co-Culture of Borrelia burgdorferi and Anaplasma phagocytophilum

by Violetta Zając, Lesley Bell-Sakyi and Angelina Wójcik-Fatla

Pathogens 2025, 14(1), 78; https://doi.org/10.3390/pathogens14010078 - 15 Jan 2025

Viewed by 521

Abstract

Ixodes ricinus is an important vector of infectious human and livestock diseases in Europe. Co-infections of pathogens in ticks and hosts have been reported. Tick cell lines offer a useful model system for study of co-infections. We present a review of the existing [...] Read more.

Ixodes ricinus is an important vector of infectious human and livestock diseases in Europe. Co-infections of pathogens in ticks and hosts have been reported. Tick cell lines offer a useful model system for study of co-infections. We present a review of the existing literature on co-infections in tick cell lines. Previous studies have demonstrated the usefulness of tick cell lines in studies on co-infection of different pathogens and their interaction with the tick microbiome. We also carried out a preliminary study to investigate the effects of co-culturing Borrelia burgdorferi and Anaplasma phagocytophilum on their growth and interactions with the Ixodes ricinus cell line IRE/CTVM19 over a 13-day period. Replication of both pathogens was quantified by real-time PCR. The presence of A. phagocytophilum appeared to have a slight inhibitory effect on the multiplication of B. burgdorferi, that were added subsequently. In contrast, the prior presence of B. burgdorferi appeared to have a stimulatory effect on A. phagocytophilum after 6 days in culture. We conclude that the IRE/CTVM19 tick cell line is suitable for simultaneous and continuous cultivation of both bacteria and can be applied in future research. Full article

(This article belongs to the Special Issue Exploring the Biodiversity of Parasites in Humans, Wild and Domestic Animals)

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13 pages, 932 KiB

Open AccessArticle

The Anthelmintic Activity of Nepeta racemosa Lam. Against Gastrointestinal Nematodes of Sheep: Rosmarinic Acid Quantification and In Silico Tubulin-Binding Studies

by Büşra Karpuz Ağören, Mahmut Sinan Erez, Esma Kozan, Aydın Dağyaran, Mevlüt Akdağ, Eduardo Sobarzo-Sánchez and Esra Küpeli Akkol

Pathogens 2025, 14(1), 77; https://doi.org/10.3390/pathogens14010077 - 15 Jan 2025

Viewed by 888

Abstract

Gastrointestinal nematodes (GINs) inflict significant economic losses on sheep and goat farming globally due to reduced productivity and the development of anthelmintic resistance. Sustainable control strategies are urgently needed including the exploration of medicinal plants as safer alternatives to chemical anthelmintics. This genus [...] Read more.

Gastrointestinal nematodes (GINs) inflict significant economic losses on sheep and goat farming globally due to reduced productivity and the development of anthelmintic resistance. Sustainable control strategies are urgently needed including the exploration of medicinal plants as safer alternatives to chemical anthelmintics. This genus of plants is used for anti-inflammatory, antioxidant, and antimicrobial activities. In this study, we aimed to evaluate the anthelmintic activities of Nepeta racemosa Lam. MeOH extract, n-hexane, dichloromethane (DCM), ethyl acetate (EtOAc), n-buthanol (n-BuOH) and aqueous (H₂O) subextracts, and quantify rosmarinic acid in the active extract by the HPLC method, and perform in silico molecular docking studies of rosmarinic acid to examine its binding interactions with tubulin. The anthelmintic activity of the plant extracts on gastrointestinal nematode eggs and larvae (L3) of the sheep was assessed using in vitro test methods such as the egg hatch assay and larval motility assay, conducted over a 24 h period (1, 2, 3, 4, 6, 8, 24). All extracts exhibited 100% effectiveness in the egg hatch inhibition assay, regardless of concentration (50–1.5625 mg/mL). The EtOAc subextract shows the highest effectiveness at 79.66%, followed by the MeOH extract at 74.00%, water at 64.00%, n-hexane at 67.00%, and DCM at 61.00%, and the lowest effectiveness is observed with n-BuOH at 51.66% in the larval motility assay. The major compound of EtOAc extract, the most active extract of N. racemosa, was determined as rosmarinic acid and its amount in the extract was determined as 14.50 mg/100 mg dry extract. The amount of rosmarinic acid in the MeOH extract was found to be 0.21 mg/100 mg dry extract. n-Hexane, DCM, n-BuOH, and H₂O extracts’ rosmarinic acid content was lower than the LOQ value. As tubulin plays an important role in the mechanism of anthelmintics, the major compound of the most active extract (NR-EtOAc) rosmarinic acid was docked onto the colchicine-binding site of the tubulin (5OV7) protein. Rosmarinic acid showed a similar activity spectrum to the anthelmintic drug albendazole. The discovery of low-cost and low-toxicity anthelmintic compounds is very important. 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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15 pages, 5067 KiB

Open AccessArticle

Ex Vivo Propagation of Pinctada Birnavirus Using Mantle Tissue Fragment Culture: Application for Measuring Replication at Different Temperatures, TCID₅₀ Assay, and UV Sensitivity

by Tomomasa Matsuyama, Takashi Atsumi, Ikunari Kiryu, Kousuke Umeda and Natsuki Morimoto

Pathogens 2025, 14(1), 76; https://doi.org/10.3390/pathogens14010076 - 15 Jan 2025

Viewed by 396

Abstract

Pinctada birnavirus (PiBV) is the causative agent of summer atrophy in pearl oyster (Pinctada fucata (Gould)). The disease, which induces mass mortality in juveniles less than 1 year old and abnormalities in adults, was first reported in Japan in 2019. Research on [...] Read more.

Pinctada birnavirus (PiBV) is the causative agent of summer atrophy in pearl oyster (Pinctada fucata (Gould)). The disease, which induces mass mortality in juveniles less than 1 year old and abnormalities in adults, was first reported in Japan in 2019. Research on the disease has been hindered by the lack of cell lines capable of propagating PiBV. We established an ex vivo method for PiBV propagation using mantle tissue, the primary infection site of the virus. The method was used to investigate the proliferation characteristics of the virus at different culture temperatures and the sensitivity of the virus to UV radiation. The marginal zone of the mantle was found to be the most suitable for PiBV replication in terms of both viral yield and reproducibility. PiBV showed optimal propagation at an incubation temperature of 25 °C, with minimal to no increase at 15 °C or 32.5 °C. Using the tissue culture infectious dose 50 (TCID₅₀) measurement system developed in this study, we found that PiBV propagation was no longer detectable after UV irradiation at 6150 J/m² or higher. The tissue fragment culture method developed in this study is expected to facilitate both ex vivo experiments and PiBV research. Full article

(This article belongs to the Section Viral Pathogens)

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10 pages, 271 KiB

Open AccessCommunication

Screening Wild Birds for Tick-Borne Zoonotic Pathogens in Portugal

by Filipa Loureiro, João R. Mesquita, Luís Cardoso, Sérgio Santos-Silva, Guilherme Moreira, Jaqueline T. Bento, Vanessa Soeiro, Andreia Gonçalves, Filipe Silva, Patrícia F. Barradas, Ana C. Matos, Manuela Matos and Ana Cláudia Coelho

Pathogens 2025, 14(1), 75; https://doi.org/10.3390/pathogens14010075 - 15 Jan 2025

Viewed by 949

Abstract

Wild birds may be involved in the transmission of agents of infectious diseases, including zoonoses, a circumstance which raises a number of public and animal health issues. Migratory bird species play a significant role in the introduction of tick-borne pathogens to new geographic [...] Read more.

Wild birds may be involved in the transmission of agents of infectious diseases, including zoonoses, a circumstance which raises a number of public and animal health issues. Migratory bird species play a significant role in the introduction of tick-borne pathogens to new geographic areas, contributing to the dissemination of various etiological agents. This preliminary study aimed to assess the occurrence of four potentially zoonotic pathogens (Hepatozoon spp., Borrelia spp., Babesia spp. and Theileria spp.) in the wild birds of Portugal. Blood and tissue samples were taken from 103 birds admitted at wildlife rehabilitation centers. Through the use of conventional PCR, our findings indicate no evidence of the circulation of these pathogens among the studied bird populations in the region. In the One Health context, it is relevant to understand how faraway avian populations play a role in the epidemiology of infectious diseases. Further molecular studies are needed to deepen the knowledge of avian piroplasmosis, borreliosis and hepatozoonosis. Full article

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

22 pages, 27465 KiB

Open AccessArticle

Inferring Tripartite Associations of Vector-Borne Plant Pathogens Using a Next-Generation Sequencing Approach

by Ava M. Gabrys, Christopher H. Dietrich and Valeria Trivellone

Pathogens 2025, 14(1), 74; https://doi.org/10.3390/pathogens14010074 - 14 Jan 2025

Viewed by 704

Abstract

Phytoplasmas are a group of plant-pathogenic, cell-wall-less bacteria vectored primarily by leafhoppers (Hemiptera Cicadellidae), one of the most diverse families of insects. Despite the importance of documenting associations between phytoplasmas, their insect vectors, and plant hosts to prevent disease outbreaks, such knowledge is [...] Read more.

Phytoplasmas are a group of plant-pathogenic, cell-wall-less bacteria vectored primarily by leafhoppers (Hemiptera Cicadellidae), one of the most diverse families of insects. Despite the importance of documenting associations between phytoplasmas, their insect vectors, and plant hosts to prevent disease outbreaks, such knowledge is currently highly incomplete and largely neglects the diversity of the system in natural areas. Here, we used anchored hybrid enrichment (AHE) to recover the DNA of five plant genes (rbcL, matK, ITS1, ITS2, and trnH-psbA) in 58 phloem-feeding leafhoppers from around the world that had previously tested positive for phytoplasma infection. Using BLASTn and a strict filtering approach, we assigned taxonomic classifications to the plant sequences and tested for cophylogenetic signals between potential Deltocephalinae leafhopper vectors and their associated plants. We observed incongruence between plant and insect phylogenies. Many leafhopper species, including presumed grass specialists, fed on distantly related plant lineages; 66% of sampled leafhoppers fed on plants from at least two different orders. By disentangling phytoplasma–leafhopper–plant interactions, we identify locations at risk of phytoplasma disease outbreaks. Furthermore, the observed wide diet breadth raises questions about how phytoplasma infection may manipulate the feeding preference of their insect host and helps fill the gaps in understanding the ecology and diversification of the tripartite association. Full article

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Pathogens, Volume 14, Issue 1 (January 2025) – 103 articles

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