Pathogens

8 pages, 1562 KB

Open AccessBrief Report

Cannonballs in Trichomonas vaginalis Infection: Morphologic Evidence of Parasite-Associated Neutrophilic Aggregates

by Ruku Shinohara, Yukimi Misawa, Shuichi Mizuno, Saeka Honda, Koki Kikuchi, Rei Settsu, Yosuke Kato, Kaori Okayama, Mizue Oda and Mitsuaki Okodo

Pathogens 2026, 15(6), 588; https://doi.org/10.3390/pathogens15060588 (registering DOI) - 29 May 2026

Abstract

Cannonballs, compact aggregates of neutrophils observed in Papanicolaou (Pap) smears, are frequently associated with Trichomonas vaginalis (T. vaginalis) infection but are generally regarded as nonspecific inflammatory findings. To clarify their morphologic features, we analyzed cervicovaginal liquid-based cytology specimens from 29 cervicitis cases, including [...] Read more.

Cannonballs, compact aggregates of neutrophils observed in Papanicolaou (Pap) smears, are frequently associated with Trichomonas vaginalis (T. vaginalis) infection but are generally regarded as nonspecific inflammatory findings. To clarify their morphologic features, we analyzed cervicovaginal liquid-based cytology specimens from 29 cervicitis cases, including six positive for T. vaginalis. Cannonballs were evaluated using immunocytochemistry for T. vaginalis and cytokeratin, cell block analysis, and morphometric analysis, with negative cases as controls. All positive cases contained T. vaginalis-associated cannonballs, with a mean positivity rate of 58.5%. Parasites were intermingled with aggregated neutrophils, and cell block analysis demonstrated parasite-centered neutrophilic aggregates. Cytokeratin staining patterns differed morphologically between positive and negative cases. In addition, T. vaginalis-associated cannonballs were significantly smaller than adjacent squamous epithelial cells. These findings suggest that some cannonballs in trichomoniasis may represent parasite-associated neutrophilic structures and provide insight into host–parasite interactions in cervicovaginal inflammation. Full article

(This article belongs to the Special Issue Advancements in Host-Parasite Interactions)

15 pages, 1303 KB

Open AccessArticle

Underrecognized Tick-Borne Encephalitis in Serbia: Evidence from Patients with Suspected West Nile Virus Neuroinvasive Disease

by Dragana Mijatović, Ana Marija Radevska, Dejan Jakimovski, Lidija Popović-Dragonjić, Biljana Popovska Jovičić, Jagoda Gavrilović, Siniša Sević, Dajana Lendak, Irina Stojanac, Alejandro Cabezas-Cruz, Andreas Pilz, Tomás Cervantes Rincón, Jasmine Oberti-Cantergiani, Davide F. Robbiani and Pavle Banović

Pathogens 2026, 15(6), 587; https://doi.org/10.3390/pathogens15060587 (registering DOI) - 29 May 2026

Abstract

Tick-borne encephalitis (TBE) is an emerging vector-borne disease in Europe, but its epidemiology remains poorly defined in Serbia. In orthoflavivirus-endemic settings, diagnostic challenges may contribute to underrecognition of TBE, particularly among patients with suspected West Nile virus (WNV) infection. We conducted a multicenter [...] Read more.

Tick-borne encephalitis (TBE) is an emerging vector-borne disease in Europe, but its epidemiology remains poorly defined in Serbia. In orthoflavivirus-endemic settings, diagnostic challenges may contribute to underrecognition of TBE, particularly among patients with suspected West Nile virus (WNV) infection. We conducted a multicenter retrospective study including patients hospitalized between 2018 and 2023 with suspected WNV neuroinvasive disease or viral encephalitis of unknown etiology. Serum samples were tested for TBEV-neutralizing antibodies using a microneutralization assay. Among 79 patients, TBEV-neutralizing antibodies were detected in four (5.1%). Most reactive cases occurred in patients initially classified as having suspected WNV-associated meningoencephalitis, while TBE had not been onsidered in the differential diagnosis at admission. These findings suggest that TBE may be underrecognized in Serbia and highlight the importance of confirmatory testing in orthoflavivirus-endemic settings. Strengthening clinical awareness and surveillance will be essential to better define the burden of TBE and inform prevention strategies. Full article

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

32 pages, 8211 KB

Open AccessReview

Current Insights into the Epidemiology and Transmission Dynamics of African Swine Fever Virus and Future Control Perspectives

by Shanta Barua, Asep Gunawan, Autchara Kayan, Masa Tenaya, Mehmet Ulas Cinar, Made Kardena, Syeda Hasina Akter, Nurulfiza Mat Isa, Henry Annandale, Subir Sarker, David T. Williams, Sam Abraham and Jasim M. Uddin

Pathogens 2026, 15(6), 586; https://doi.org/10.3390/pathogens15060586 (registering DOI) - 29 May 2026

Abstract

African swine fever virus (ASFV) is an evolving threat to global swine health and food security, driven by its complex epidemiology, multi-host transmission cycles, and ongoing spread across countries. This review summarizes the global scenario and transmission pathways of ASFV, highlighting the outbreaks [...] Read more.

African swine fever virus (ASFV) is an evolving threat to global swine health and food security, driven by its complex epidemiology, multi-host transmission cycles, and ongoing spread across countries. This review summarizes the global scenario and transmission pathways of ASFV, highlighting the outbreaks associated with evolving risk patterns to support effective disease prevention and control. ASF has been reported in pig-producing regions across Africa, Europe, and, more recently, Asia, largely driven by the spread of genotype II strains. The virus is transmitted through direct contact with infected pigs or pig-products, indirectly via contaminated materials, and through soft ticks of the genus Ornithodoros spp., with epidemiological patterns varying according to wildlife reservoirs and regional factors. Control measures mainly rely on early detection, movement control, strict quarantine, robust biosecurity measures, and international trade regulations. Despite significant advances, persistent challenges, including the absence of a widely available commercial vaccine, long-term stability of the virus, human activities, and inconsistencies in global response capacities, continue to hinder disease eradication efforts. This review underscores the need for transnational strategies and policies that integrate economically sustainable disease management systems and reduce the long-term impact of ASFV. Full article

(This article belongs to the Section Viral Pathogens)

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14 pages, 3214 KB

Open AccessArticle

Campylobacter spp. in Poultry Slaughterhouses: Occurrence, Antimicrobial Resistance, and Virulence-Associated Genes

by Pietro Di Taranto, Fiorenza Petruzzi, Antonella Cristina Romano, Mariateresa Toce, Lucia Palazzo, Alessandra Alessiani, Loredana Capozzi, Stefano Castellana, Laura Del Sambro, Adelia Donatiello, Carmine Pedarra, Gilda Occhiochiuso, Giovanni Castelli, Alessandra Barlaam, Giovanni Normanno and Antonio Parisi

Pathogens 2026, 15(6), 585; https://doi.org/10.3390/pathogens15060585 (registering DOI) - 29 May 2026

Abstract

Poultry is the main reservoir of Campylobacter spp. and most human cases result from consuming undercooked poultry or handling raw meat. In 2022, a total of 55 samples, including neck skin, cecal contents, and processing waters, were collected at two poultry slaughterhouses in [...] Read more.

Poultry is the main reservoir of Campylobacter spp. and most human cases result from consuming undercooked poultry or handling raw meat. In 2022, a total of 55 samples, including neck skin, cecal contents, and processing waters, were collected at two poultry slaughterhouses in Italy and analysed according to ISO 10272-2:2017 at the Istituto Zooprofilattico Sperimentale della Puglia e della Basilicata laboratories. Overall, 51/55 (92.72%) samples tested positive for Campylobacter. Among the isolates, 64.71% were identified as C. coli, and 35.29% as C. jejuni. Phenotypic and genotypic analysis were performed to assess antimicrobial resistance and virulence characteristics. All C. jejuni isolates and 72.72% of C. coli showed resistance to fluoroquinolones. Resistances to tetracycline and carbapenem were observed in 60.78% and 45.09% of isolates, respectively. Genomic analysis confirmed the presence of the tet(O) gene, conferring tetracycline resistance. In addition, OXA-450 and OXA-466 genes, conferring beta-lactam resistance, were detected in 78.43% and 3.92% of isolates. Virulence-associated genes were detected. Specifically, the ciaB gene was found in 50/51 (98.04%) of isolates, whereas jlpA, cdtA, cdtB, and ctdC genes were exclusively identified in C. jejuni strains. The high prevalence of pathogenic and antimicrobial-resistant Campylobacter strains highlights the need for strengthened control measures along the poultry production chain. Full article

(This article belongs to the Special Issue Advances in Rapid Detection and Quantification of Campylobacter in Food Safety)

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20 pages, 1302 KB

Open AccessArticle

Genotype-Specific HPV mRNA Triage Improves Colposcopy Efficiency Compared with Cytology and ATHENA-Derived Triage: A Population-Based Study of HPV DNA-Positive Women

by Sveinung Wergeland Sørbye, Bente Marie Falang, Mona Antonsen and Elin Richardsen

Pathogens 2026, 15(6), 584; https://doi.org/10.3390/pathogens15060584 (registering DOI) - 28 May 2026

Abstract

Background: Effective triage of HPV DNA-positive women is needed to reduce unnecessary colposcopies while maintaining cervical cancer prevention. We evaluated genotype-specific 7-type HPV E6/E7 mRNA triage in a real-world screening cohort. Methods: In this population-based single-centre study at the University Hospital of North [...] Read more.

Background: Effective triage of HPV DNA-positive women is needed to reduce unnecessary colposcopies while maintaining cervical cancer prevention. We evaluated genotype-specific 7-type HPV E6/E7 mRNA triage in a real-world screening cohort. Methods: In this population-based single-centre study at the University Hospital of North Norway, 42,791 women underwent primary screening with the cobas HPV DNA assay during the period 2019–2024. Among 2370 HPV DNA-positive women, reflex cytology, 7-type HPV mRNA testing, and an ATHENA-derived triage strategy were compared using histologically confirmed CIN3+ through 31 December 2025 as the endpoint. Results: CIN3+ was detected in 60/2370 women (2.5%). Test positivity was 47.0% for cytology, 54.7% for ATHENA-derived triage, and 33.4% for HPV mRNA. Sensitivity was 78.3%, 86.7%, and 73.3%; specificity was 53.8%, 46.1%, and 67.7%; and PPV was 4.2%, 4.0%, and 5.6%, respectively. Colposcopies per CIN3+ detected were 23.7, 24.9, and 18.0. Conclusions: HPV mRNA triage improved referral precision and colposcopy efficiency, but with lower sensitivity than ATHENA-derived triage. These findings support 7-type HPV mRNA testing as a potentially useful molecular triage option where structured follow-up is feasible. Full article

(This article belongs to the Special Issue Epidemiology, Diagnostics, and Therapeutics of High-Risk Human Papillomavirus (HPV) Infections)

17 pages, 951 KB

Open AccessArticle

Age-Structured Clinical Background Is More Strongly Associated with C-Reactive Protein Levels than Individual Respiratory Viruses During Respiratory Virus Testing

by Sung Hun Jang, Bo Kyeung Jung, Jae-Sik Jeon, Jeong Su Han and Jae Kyung Kim

Pathogens 2026, 15(6), 583; https://doi.org/10.3390/pathogens15060583 (registering DOI) - 28 May 2026

Abstract

We aimed to characterize age-stratified C-reactive protein (CRP) patterns across respiratory virus infections, assess age-related CRP shifts in virus-not-detected-by-PCR episodes, and evaluate the independent associations of age and virus type with CRP levels. We retrospectively analyzed 19,002 test-level episodes with paired respiratory PCR [...] Read more.

We aimed to characterize age-stratified C-reactive protein (CRP) patterns across respiratory virus infections, assess age-related CRP shifts in virus-not-detected-by-PCR episodes, and evaluate the independent associations of age and virus type with CRP levels. We retrospectively analyzed 19,002 test-level episodes with paired respiratory PCR and serum CRP results from a single tertiary-care center between 2008 and 2024. Episodes were classified as virus-not-detected-by-PCR or virus-positive according to multiplex PCR results; the former was not considered a healthy control and may include off-panel infections, bacterial/mixed infections, false-negative results, or non-infectious inflammation. Descriptive analyses and multivariable linear regression of log-transformed CRP were used to assess adjusted associations. Median CRP increased with age in both virus-positive and virus-not-detected-by-PCR episodes, rising from 0.38 to 7.42 mg/dL across age groups in the latter. Age showed the strongest association with CRP. Adenovirus showed a positive adjusted association, whereas influenza A/B, respiratory syncytial virus A/B, and parainfluenza virus types 1 and 3 showed selective negative associations. Overall, CRP variation was more strongly associated with age-related clinical background than with virus type, although selective virus-associated differences were observed, supporting the interpretation of CRP as a non-specific, composite host-response indicator within broader clinical contexts. Full article

(This article belongs to the Special Issue Advances in the Epidemiology of Human Infectious Diseases)

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17 pages, 3885 KB

Open AccessArticle

Essential Oils of Thymus Species Against Phytophthora Species: A Structured Review and Novel In Vitro Evaluations

by Chiara Antonelli, Najwa Benfradj and Anna Maria Vettraino

Pathogens 2026, 15(6), 582; https://doi.org/10.3390/pathogens15060582 (registering DOI) - 28 May 2026

Abstract

Phytophthora species are among the most destructive plant pathogens worldwide, causing severe losses in agricultural, forest, and natural ecosystems. In recent years, the management of Phytophthora diseases has increasingly shifted toward eco-sustainable strategies, with growing interest in plant-derived extracts, particularly essential oils, as [...] Read more.

Phytophthora species are among the most destructive plant pathogens worldwide, causing severe losses in agricultural, forest, and natural ecosystems. In recent years, the management of Phytophthora diseases has increasingly shifted toward eco-sustainable strategies, with growing interest in plant-derived extracts, particularly essential oils, as low-risk alternatives to synthetic fungicides. In this study, a structured review was combined with new in vitro assays to assess the antifungal activity of essential oils from Thymus vulgaris (TV-EO) and T. serpyllum (TS-EO) against P. cinnamomi, P. drechsleri, P. cactorum, P. citrophthora, P. nicotianae, P. palmivora, and P. infestans. Literature searches were conducted in April 2025 using the Web of Science and Scopus databases, following PRISMA guidelines, with the search term “Thymus” or “Thyme” and “Phytophthora”. Twenty studies included in the review demonstrated that the activity of Thymus essential oils against Phytophthora species was highly variable and shaped by chemotype, Thymus species, pathogen, and experimental setup. Additional in vitro assays further confirmed a clear dose-dependent inhibitory effect for both TV-EO and TS-EO. TS-EO consistently exhibited stronger activity than TV-EO, likely reflecting its carvacrol-rich chemotype, while thymol-based TV-EO showed lower but still significant inhibition depending on the pathogen species. Overall, these results highlight the potential of Thymus essential oils as eco-friendly tools for the management of Phytophthora diseases. However, the strong dependence on chemotype, pathogen species, and assay conditions underscores the need for standardized testing, detailed chemical characterization, and in vivo validation. Full article

(This article belongs to the Section Fungal Pathogens)

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14 pages, 516 KB

Open AccessCase Report

Prosthetic Valve Endocarditis by Acinetobacter baumannii: Case Report and Systematic Descriptive Review

by Annabella Salvati, Loredana Alessio, Gloria Trombaccia, Giovanni Cimmino, Marisa De Feo, Fausto Ferraro, Stefania De Pascalis and Nicola Coppola

Pathogens 2026, 15(6), 581; https://doi.org/10.3390/pathogens15060581 (registering DOI) - 28 May 2026

Abstract

A 61-year-old woman developed prosthetic valve infective endocarditis after osteomyelitis caused by extensively drug-resistant (XDR) Acinetobacter baumannii. Moreover, a systematic descriptive review of published case reports was performed to describe the main features, treatment, and outcomes of this condition. Methods: Apart from [...] Read more.

A 61-year-old woman developed prosthetic valve infective endocarditis after osteomyelitis caused by extensively drug-resistant (XDR) Acinetobacter baumannii. Moreover, a systematic descriptive review of published case reports was performed to describe the main features, treatment, and outcomes of this condition. Methods: Apart from the description of our case, a PubMed and Embase literature review was conducted up to January 2026 using the terms “A. baumannii” AND (“infective endocarditis” OR “endocarditis” OR “valvular infection”). We included clinical cases of IE caused by A. baumannii published as full-text articles in English. Results: After orthopedic osteosynthesis surgery following a femur fracture, our patient developed osteomyelitis by XDR A. baumannii and was treated for a short period of time. Later, prosthetic aortic valve endocarditis was diagnosed. Despite treatment with cefiderocol and eravacycline, she died. An additional 18 clinical cases of IE by A. baumannii were identified from the literature, bringing the total to 19 cases. IE affected prosthetic valves in nine cases, native valves in nine and involved a right atrial mass in one. Twelve cases were caused by MDR or XDR A. baumannii. Mortality occurred more frequently in cases not treated with surgery (9/13, 69%) compared to those treated with surgery (1/6, 16.7%). However, given the very small sample size, these data should be interpreted with caution. Conclusion: This case, together with previously reported observations, highlights the severity of EI by A. baumannii and the need of multidisciplinary management. Full article

(This article belongs to the Special Issue Challenges of Multidrug-Resistant Bacteria and Drug Development of Antibacterials)

10 pages, 3748 KB

Open AccessArticle

Comparison of Immunoprotective Efficacy of Six Antigenic Proteins of Pasteurella multocida Serotype a in KM Mice (Mus musculus)

by Wenjing Zhang, Yiming Guo, Lijun Guan, Lifang Si and Zhanqin Zhao

Pathogens 2026, 15(6), 580; https://doi.org/10.3390/pathogens15060580 - 28 May 2026

Abstract

Pasteurella multocida serotype A (P. multocida) is frequently associated with severe respiratory disease in swine (Sus scrofa), highlighting the need for effective preventive strategies. To identify protective antigens suitable for a subunit vaccine targeting porcine P. multocida infection, six [...] Read more.

Pasteurella multocida serotype A (P. multocida) is frequently associated with severe respiratory disease in swine (Sus scrofa), highlighting the need for effective preventive strategies. To identify protective antigens suitable for a subunit vaccine targeting porcine P. multocida infection, six recombinant proteins (rAspA, rLolA, rOmpP6, rOppA, rRps6, rSmpA) were expressed in a prokaryotic system, and their efficacy was evaluated in a Mus musculus (Kunming) mouse model. All proteins were purified using His-tag affinity chromatography, and SDS-PAGE analysis confirmed expression with bands at the expected molecular weights (61, 26, 21, 63, 19, and 17 kDa). Each protein, formulated with ISA 201 adjuvant, was administered to mice in two immunizations. Indirect ELISA of sera collected at multiple time points demonstrated that all vaccines induced high antigen-specific IgG levels. rOppA, rLolA, rOmpP6, and rRps6 were expressed in soluble form, whereas rAspA and rSmpA formed inclusion bodies. In a lethal challenge model, rLolA and rRps6 conferred the highest protection (60% each), followed by rAspA and rOmpP6 (30%), rOppA (20%), and rSmpA (10%). Under the conditions tested, the highest protection observed was 60%, and none of the six antigens achieved complete protection against homologous A7 challenge in mice. This first head-to-head comparison under identical conditions provides a reference framework for future antigen screening studies. Full article

(This article belongs to the Section Bacterial Pathogens)

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14 pages, 1278 KB

Open AccessArticle

Epidemiology and Clinical Impact of Pediatric Viral Gastroenteritis Before and After Rotavirus Vaccination in Sicily

by Floriana Bonura, Arcangelo Pepe, Dario Genovese, Emanuele Amodio, Chiara Filizzolo, Fabio Campisi, Mariangela Pizzo, Emilia Palazzotto, Simona De Grazia and Giovanni M. Giammanco

Pathogens 2026, 15(6), 579; https://doi.org/10.3390/pathogens15060579 - 28 May 2026

Abstract

Acute gastroenteritis (AGE) remains a leading cause of pediatric mortality and morbidity, with rotavirus as the leading cause of severe disease. Post-vaccine surveillance is essential to monitor circulating pathogens and assess vaccination impact. Sicily was the first Italian region to implement universal rotavirus [...] Read more.

Acute gastroenteritis (AGE) remains a leading cause of pediatric mortality and morbidity, with rotavirus as the leading cause of severe disease. Post-vaccine surveillance is essential to monitor circulating pathogens and assess vaccination impact. Sicily was the first Italian region to implement universal rotavirus vaccination in 2012. We retrospectively studied 693 children hospitalized for suspected viral AGE at the Children’s Hospital of Palermo (March 2017–February 2020), testing stool samples for viral and bacterial enteric pathogens. Rotavirus remained the most common agent (13.3%), followed by norovirus (12.1%), adenovirus (11.3%), Salmonella spp. (4.6%) and astrovirus (3.2%). The study population was categorized as rotavirus-associated AGE (RV-AGE) or other-cause AGE (O-AGE). Epidemiological, clinical and virological features were compared with the pre-vaccine period (2011–2012). At least one pathogen was detected in 47.5% of samples. RV-AGE cases were older than those with O-AGE (median 32.6 vs. 30.5 months; p < 0.01) and had greater clinical severity, with higher frequency of vomiting, fever and dehydration. Rotavirus infection was significantly associated with unvaccinated status. Compared with the pre-vaccine era, rotavirus prevalence declined (32.6% vs. 13.3%), seasonal patterns were attenuated and genotype distribution shifted toward G2P[4], G9P[8] and equine-like G3P[8] strains. Despite the decline in RV-AGE following vaccine introduction, rotavirus remains a relevant cause of pediatric AGE, underscoring the need for high vaccination coverage and continued surveillance. Full article

(This article belongs to the Section Viral Pathogens)

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16 pages, 491 KB

Open AccessReview

Research Progress on Macrococcus: From Basic Biology to Clinical Antimicrobial Resistance Challenges

by Chenyu Zhan, Mingyu Zhang, Guijuan Hao, Yue Zhang and Fangkun Wang

Pathogens 2026, 15(6), 578; https://doi.org/10.3390/pathogens15060578 - 27 May 2026

Abstract

Macrococcus is a genus of Gram-positive cocci in the Staphylococcaceae family and a close phylogenetic relative of Staphylococcus. It is not a significant human pathogen but is known to widely colonize different environments, including animal skin and food products. Phylogenetically, Macrococcus is [...] Read more.

Macrococcus is a genus of Gram-positive cocci in the Staphylococcaceae family and a close phylogenetic relative of Staphylococcus. It is not a significant human pathogen but is known to widely colonize different environments, including animal skin and food products. Phylogenetically, Macrococcus is distinct from yet closely related to Staphylococcus, particularly the sciuri group. The species is effectively identified through such molecular markers as hsp60 and 16S rDNA. A key biochemical feature is an identified FAD-dependent oleate hydratase in Macrococcus equipercicus (M. equipercicus). Critically, Macrococcus carries various mobile antibiotic-resistance genes, especially against β-lactams (e.g., mecB, mecD) and macrolides (e.g., mef(F), msr(G)); these genes are located on plasmids, SCCmec-like elements, or resistance islands (e.g., McRI_mecD), which facilitates their horizontal transfer. Surveillance confirms the widespread presence of methicillin-resistant Macrococcus, often with a multidrug-resistant phenotype, in food animals and their products. Although its own pathogenicity is low, Macrococcus acts as a reservoir and transmission platform for resistance genes: through horizontal gene transfer, it can potentially confer resistance to pathogenic Staphylococcus, thereby posing a threat to animal and public health. This review summarizes the basic biological characteristics and drug resistance-related research progress of the genus Macrococcus; it aims to provide a reference for subsequent studies as well as to establish technical support and a theoretical basis for the epidemiological investigation, drug-resistant strain identification, and clinical drug-resistance risk prevention and control of Macrococcus. Full article

(This article belongs to the Section Bacterial Pathogens)

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16 pages, 1857 KB

Open AccessArticle

Species Diversity and Molecular Screening of Pyrethroid Resistance Mediated by the Voltage-Gated Sodium Channel in Ixodid Ticks from Puducherry, India

by Kaja Hussain Jasmiminal, Elango Ayyanar, Chakravarthi Madda, Vignesh Kumar Ramakrishnan, Hari Kishan Raju Konuganti and Manju Rahi

Pathogens 2026, 15(6), 577; https://doi.org/10.3390/pathogens15060577 - 27 May 2026

Abstract

Ticks are major ectoparasites of livestock in India, causing substantial economic losses and transmitting a wide range of pathogens. Control strategies rely heavily on synthetic pyrethroid acaricides; however, resistance associated with mutations in the voltage-gated sodium channel (VGSC) gene has been widely reported. [...] Read more.

Ticks are major ectoparasites of livestock in India, causing substantial economic losses and transmitting a wide range of pathogens. Control strategies rely heavily on synthetic pyrethroid acaricides; however, resistance associated with mutations in the voltage-gated sodium channel (VGSC) gene has been widely reported. Information on tick species diversity and molecular resistance status in Puducherry remains limited. This study investigated tick diversity, host-associated infestation patterns, and the presence of key VGSC knockdown resistance (kdr) mutations (C190A and G215T). A cross-sectional survey was conducted across 15 villages in Puducherry, where ticks were collected from cattle, goats, and dogs and identified using standard morpho-taxonomic keys. A total of 3779 ticks representing eight ixodid species were identified, showing clear host-associated infestation patterns. Individual ticks were selected from stratified pools (based on village, host, tick species, and sex) and processed individually for genomic DNA extraction. A fragment of the VGSC gene was amplified by PCR and analysed using Sanger sequencing. Molecular analysis of 62 high-quality VGSC sequences representing all eight species across the surveyed villages revealed no mutations at the investigated loci (C190A and G215T) in the analysed samples. All sequences exhibited the wild-type genotype at these positions. These findings provide baseline molecular information on the screened VGSC loci in ixodid ticks from Puducherry and highlight the importance of continued surveillance, phenotypic validation, and integrated tick management strategies. Full article

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20 pages, 7628 KB

Open AccessArticle

Marine-Derived Sterols from Saccharina japonica: Potential Antibacterial Activity and Target Prediction Against Bacterial Pathogens Through Integrated In Vitro and In Silico Approaches

by Eun-Seop Lee, Chae-Min Kim, Si-Heon Song, Su-Bin Jeon, Byeong-Su Kang, Md Sekendar Ali and Eon-Bee Lee

Pathogens 2026, 15(6), 576; https://doi.org/10.3390/pathogens15060576 - 27 May 2026

Abstract

Marine natural products, including seaweeds, sponges, and marine microorganisms, have emerged as promising sources of bioactive compounds with diverse pharmacological properties. We investigated the antibacterial and antioxidant potential of ethanol extracts (30%, 60%, and 90%) from Saccharina japonica collected from two Korean coastal [...] Read more.

Marine natural products, including seaweeds, sponges, and marine microorganisms, have emerged as promising sources of bioactive compounds with diverse pharmacological properties. We investigated the antibacterial and antioxidant potential of ethanol extracts (30%, 60%, and 90%) from Saccharina japonica collected from two Korean coastal regions, Gijang and Wando, and evaluated their bioactive metabolites through integrated in vitro and in silico approaches. Among the extracts, the 60% ethanol fraction exhibited the highest total phenolic content and strongest 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging activity, indicating superior antioxidant capacity. Antibacterial assays revealed selective concentration-dependent inhibition against Staphylococcus aureus, while limited effects were observed against Escherichia coli. Kinetic analysis further demonstrated growth suppression of S. aureus at higher concentrations. Molecular docking was performed against multiple bacterial proteins, including DNA gyrase, topoisomerase IV, and tyrosyl-tRNA synthetase. Sterol compounds, particularly fucosterol and 24-methylene cholesterol, showed strong binding affinities across key targets, suggesting their potential role as multitarget antibacterial agents. ADMET predictions indicated favorable pharmacokinetic properties, although high lipophilicity and potential hERG II inhibition were noted. Overall, while the antibacterial effects observed were moderate, these findings suggest that marine-derived sterols from S. japonica may represent compounds of interest for further mechanistic investigation and optimization in complementary antibacterial strategies. Full article

(This article belongs to the Special Issue Targeted Antibacterial Strategies Against Pathogens in Fish and Farm Animals)

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19 pages, 11239 KB

Open AccessArticle

Blocking Host Factors IAP and DDX3 Activates HIV-1 Transcription and Increases Apoptosis Sensitivity of HIV-1 Infected Cells

by Jade Jansen, Shirley Man, Fenna Kootstra, Ad C. van Nuenen, Karel A. van Dort, Claudio Zamperini, Conraed Willem Houck, Neeltje A. Kootstra and Teunis B. H. Geijtenbeek

Pathogens 2026, 15(6), 575; https://doi.org/10.3390/pathogens15060575 - 27 May 2026

Abstract

Antiretroviral therapy (ART) effectively suppresses HIV-1 replication but does not purge the latent HIV-1 reservoir. Strategies aimed at HIV-1 latency reversal and subsequent elimination of infected cells are being explored. Targeting the inhibitor of apoptosis proteins (IAP) and DEAD-box polypeptide 3 (DDX3) RNA [...] Read more.

Antiretroviral therapy (ART) effectively suppresses HIV-1 replication but does not purge the latent HIV-1 reservoir. Strategies aimed at HIV-1 latency reversal and subsequent elimination of infected cells are being explored. Targeting the inhibitor of apoptosis proteins (IAP) and DEAD-box polypeptide 3 (DDX3) RNA helicase reduces the HIV-1 reservoir ex vivo. However, the mechanisms driving apoptosis of HIV-1 infected cells remain unclear. Here, we uncovered the mechanism regarding HIV-1 transcriptional activation and induction of apoptosis specific for HIV-1 infected cells using an acute in vitro infection model. Inhibition of IAP by second mitochondrial-derived activator of caspases mimetic (SMACm; AZD5582) resulted in activation of non-canonical NF-κB pathway (RelB/p52) that induced HIV-1 transcription, confirming previous reports, whereas inhibition of DDX3 sensitized HIV-1 infected cells for apoptosis (DDX3i; FH1321). Transcriptome analysis revealed that HIV-1 actively suppressed apoptosis-related genes in HIV-1 infected cells. SMACm treatment resulted in a broad induction of these genes irrespective of infection. Notably, DDX3 inhibition specifically restored the expression of the majority of HIV-1 suppressed genes, and when combined with SMACm, restored almost all HIV-1 downregulated genes, thereby rendering HIV-1 infected cells sensitive to apoptosis. Thus, our data strongly suggest that inhibition of host factors IAP and DDX3 not only induces activation of HIV-1 transcription but also restores HIV-1 suppressed apoptotic processes in infected cells. Full article

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22 pages, 928 KB

Open AccessSystematic Review

A Systematic Review of Gastrointestinal and Respiratory Pathogen Detection in Wastewater in Africa, with Focus on Rwanda: Implications for Early Warning and Public Health Surveillance

by Sylvie Bambara, Marie Claire Isingizwe, Taofeek Tope Adegboyega and Leon Mutesa

Pathogens 2026, 15(6), 574; https://doi.org/10.3390/pathogens15060574 - 27 May 2026

Abstract

In Africa, the disease burden of diarrheal and respiratory diseases is amplified by limited surveillance capacity, diagnostic limitations, and socioeconomic inequalities. In rapidly urbanizing settings such as Kigali (Rwanda), integrating wastewater-based epidemiology (WBE) into existing surveillance systems offers a promising strategy for generating [...] Read more.

In Africa, the disease burden of diarrheal and respiratory diseases is amplified by limited surveillance capacity, diagnostic limitations, and socioeconomic inequalities. In rapidly urbanizing settings such as Kigali (Rwanda), integrating wastewater-based epidemiology (WBE) into existing surveillance systems offers a promising strategy for generating real-time epidemiological intelligence, identifying community-level hotspots, and addressing gaps in traditional reporting systems. Gastrointestinal and respiratory infections remain major causes of morbidity and mortality globally, particularly in low- and middle-income countries (LMICs), where traditional clinical surveillance systems frequently underreport the true disease burden. This systematic review synthesizes current evidence on the detection of gastrointestinal and respiratory pathogens in wastewater and evaluates the utility of WBE for early warning and public health action. A narrative review approach was used to identify peer-reviewed literature, global health reports, and surveillance studies focusing on the wastewater detection of gastrointestinal and respiratory pathogens. Databases including PubMed, Scopus, and Google Scholar were searched for studies published between 2000 and 2026. The search yielded 1247 records, of which 312 duplicates were removed. After title/abstract screening, 228 full-text articles were retrieved and assessed for eligibility. After a detailed evaluation, 108 studies were excluded for the following reasons: absence of pathogen-specific wastewater data (n = 46), a focus on environmental monitoring without public health relevance (n = 25), insufficient methodological description (n = 21), or other eligibility limitations such as a lack of primary data (n = 16). WBE provides a non-invasive, cost-effective approach for monitoring symptomatic and asymptomatic infections. Challenges involve variability in sampling, environmental factors affecting viral decay, and differences in laboratory workflows. WBE is a powerful complement to traditional infectious disease surveillance, offering early warning capabilities, population-level coverage, and real-time insights into pathogen circulation. Integrating WBE into surveillance programs, especially in LMICs such as Rwanda, can significantly strengthen epidemic preparedness, guide resource allocation, and improve outbreak response. Sustained investment in laboratory capacity, standardized protocols, and multisector collaboration is essential to fully leverage WBE for public health protection. Full article

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11 pages, 239 KB

Open AccessArticle

Enumeration and Genomic Confirmation of Viable Shiga-Toxin-Producing Escherichia coli from Ground Beef by Integrating Serial Plating with Long-Read Sequencing

by Katrina L. Counihan and Shannon Tilman

Pathogens 2026, 15(6), 573; https://doi.org/10.3390/pathogens15060573 - 27 May 2026

Abstract

Consumption of food contaminated with Shiga toxin-producing Escherichia coli (STEC) causes thousands of illnesses in the United States annually. Long-read sequencing could reduce the time needed to test food for STEC, but sequencing is not quantitative and cannot differentiate between live and dead [...] Read more.

Consumption of food contaminated with Shiga toxin-producing Escherichia coli (STEC) causes thousands of illnesses in the United States annually. Long-read sequencing could reduce the time needed to test food for STEC, but sequencing is not quantitative and cannot differentiate between live and dead bacteria. Therefore, this study investigated combining serial plating with long-read sequencing to quantify only live STEC in a sample. Ground beef was inoculated with STEC and homogenized with a stomacher. The liquid was filtered to remove eukaryotic cells and then centrifuged to pellet the bacteria. Serial dilutions of the pellet were plated on selective agar, and single colonies were subsequently sequenced. Initial experiments revealed that processing samples at room temperature led to a 1 log increase in STEC levels from the initial inoculated concentration, confounding accurate enumeration. In subsequent experiments, samples and reagents were kept cold, and the amount of STEC recovered did not significantly differ from the amount inoculated. The DNA extracted from a single colony provided enough data to identify all virulence genes of interest multiple times. The amount of STEC in a sample could be quantified down to 1 cfu g⁻¹. Quantification of STEC in food samples using this method would improve risk assessment and guide mitigation efforts in industry. Full article

(This article belongs to the Section Bacterial Pathogens)

17 pages, 2959 KB

Open AccessArticle

Spatiotemporal Dynamics of the 2025–2026 Measles Outbreak in Mexico: Jalisco as the Emerging Epicenter and the Impact of Intensive Immunization Efforts

by Elena Sandoval-Pinto, Rosa Cremades, Héctor Raúl Pérez-Gómez, Roberto Carlos Rivera-Ávila, Cesar Augusto Domínguez Barbosa, Lakshmanane Premkumar, Erick Sierra-Díaz, Diana Emilia Martínez-Fernández and José Ángel Regla-Nava

Pathogens 2026, 15(6), 572; https://doi.org/10.3390/pathogens15060572 - 27 May 2026

Abstract

Measles represents a critical public health challenge due to its high transmissibility and the resurgence of outbreaks of the disease, linked to gaps in vaccination coverage. In this study, we analyze the epidemiological dynamics of the outbreak that began in Mexico in 2025 [...] Read more.

Measles represents a critical public health challenge due to its high transmissibility and the resurgence of outbreaks of the disease, linked to gaps in vaccination coverage. In this study, we analyze the epidemiological dynamics of the outbreak that began in Mexico in 2025 and focus on Jalisco as the epicenter during the first quarter of 2026. A descriptive, retrospective, ecological time-series analysis was conducted using official epidemiological surveillance data, correlating incidence with immunization strategies. The results reveal that 6681 cases were recorded in Jalisco, with the highest prevalence among adults aged 20 to 49 years (41.96%), which highlights the accumulated gaps in immunity coverage. The public health response, comprising the administration of three million doses, focusing on zero-dose children and vaccination blockades, contributed to a notable decrease in infections by epidemiological week 17 of 2026. It was concluded that the outbreak was controlled through intensive and coordinated intervention. However, the proximity of the 2026 International Federation of Association Football (FIFA) World Cup requires vaccination coverage to be maintained at over 95%, in addition to vigilance, to mitigate the risk of viral reintroduction and to safeguard global health security. Full article

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

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13 pages, 1243 KB

Open AccessCommunication

Isolation and Identification of Emerging Equine Encephalosis Virus Serotype 6 in Israel, 2023

by Natalia Golender, Zvia Mildenberg, Michel Bellaiche and Bernd Hoffmann

Pathogens 2026, 15(6), 571; https://doi.org/10.3390/pathogens15060571 - 27 May 2026

Abstract

Equine encephalosis (EE) is an arthropod-borne viral disease resembling a mild form of African horse sickness disease, which affects all species of equids. Previously, only EEV serotype 4 (EEV-4) caused sporadic outbreaks of the disease in Israel. In the summer of 2023, EE [...] Read more.

Equine encephalosis (EE) is an arthropod-borne viral disease resembling a mild form of African horse sickness disease, which affects all species of equids. Previously, only EEV serotype 4 (EEV-4) caused sporadic outbreaks of the disease in Israel. In the summer of 2023, EE was clinically diagnosed in horses and donkeys in Israel. For laboratory confirmation, tissue cultures were inoculated with whole-blood samples obtained from sick animals. Five EEV isolates were successfully recovered in tissue and confirmed by conventional RT-PCR and Sanger sequencing of several viral genes. BLAST analysis of genome segment 2 revealed that the isolates belonged to serotype 6. Full-genome sequencing of one representative strain and subsequent phylogenetic analysis demonstrated a close relationship to South African and Indian EEV strains, suggesting possible epidemiological links that warrant further investigation. This observation can indirectly point to a broader geographic circulation of EEV than was previously thought. Full article

(This article belongs to the Section Viral Pathogens)

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22 pages, 2264 KB

Open AccessReview

Advances in CRISPR-Cas for Diagnosis and Treatment of Klebsiella pneumoniae

by Changmei Feng and Jun Yin

Pathogens 2026, 15(6), 570; https://doi.org/10.3390/pathogens15060570 - 26 May 2026

Abstract

Carbapenem-resistant Klebsiella pneumoniae (CRKP) is a significant pathogen for both hospital-acquired and community-acquired infections, characterized by its strong epidemic potential and high mortality rate, posing a severe threat to global public health. CRKP spreads widely across the globe through the horizontal transfer of [...] Read more.

Carbapenem-resistant Klebsiella pneumoniae (CRKP) is a significant pathogen for both hospital-acquired and community-acquired infections, characterized by its strong epidemic potential and high mortality rate, posing a severe threat to global public health. CRKP spreads widely across the globe through the horizontal transfer of plasmid-mediated resistance genes such as *blaKPC*, *blaNDM*, and *blaOXA-48*. The clinical treatment options for this bacterium are limited, and its resistance has been increasing year by year, urgently necessitating the development of new antimicrobial drugs or alternative strategies. In recent years, the CRISPR-Cas system has shown great potential in the diagnosis and treatment of CRKP, including rapid detection and identification, gene editing, antimicrobial strategies, and resistance inhibition. For instance, CRISPR-Cas12a/13a can be used for the rapid detection and identification of CRKP, while CRISPR-Cas9/Cas3 can target resistance genes to reverse the resistance of strains. With the advancement of delivery and biotechnologies, the CRISPR-Cas system is expected to become an important tool against drug-resistant CRKP. This review focuses on the application of the CRISPR-Cas system in the detection and treatment of CRKP, analyzing its technical advantages, limitations, and future development directions. Full article

(This article belongs to the Section Bacterial Pathogens)

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