Pathogens

10 pages, 1807 KiB

Open AccessArticle

Method to Generate Chlorine Dioxide Gas In Situ for Sterilization of Automated Incubators

by Cédric Schicklin, Georg Rauter, Philippe Claude Cattin, Manuela Eugster and Olivier Braissant

Pathogens 2024, 13(11), 1024; https://doi.org/10.3390/pathogens13111024 - 20 Nov 2024

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Pharmaceutical preclinical tests using cell cultures are nowadays commonly automated. Incubator microbial contaminations impact such tests. Chlorine dioxide (ClO₂) is widely used in aqueous solutions. However, a gaseous form, such as chlorine dioxide gas (gClO₂), can effectively access unreachable [...] Read more.

Pharmaceutical preclinical tests using cell cultures are nowadays commonly automated. Incubator microbial contaminations impact such tests. Chlorine dioxide (ClO₂) is widely used in aqueous solutions. However, a gaseous form, such as chlorine dioxide gas (gClO₂), can effectively access unreachable spaces, such as closed cell culture incubators. Steam sterilization requires a temperature rise to at least 121 °C, thus limiting the possibility of automation elements for sensors and actuators. gClO₂ sterilization is an ambient-temperature sterilization method. This article aims to demonstrate that gClO₂ generated from solid powder tablets is efficient for sterilizing incubators and can be automated. We selected (i) Bacillus subtilis strain, (ii) Saccharomyces cerevisiae, and (iii) T7 phages as representatives for (i) bacteria, (ii) fungi, and (iii) viruses for each domain to evaluate the sterilization efficiency. This study demonstrated that gClO₂ can be generated inside the incubator from a solid powder tablet without specific equipment and can effectively fight biological proxies in 15 min. After 30 sterilization cycles, the actuators and sensors mounted inside the incubator were still operating. Our proposed sterilization method seems to be generally applicable for automated in situ sterilization of incubators and medical robots. Full article

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10 pages, 8510 KiB

Open AccessCase Report

First Detection of West Nile Virus by Nasopharyngeal Swab, Followed by Phylogenetic Analysis

by Carlo Zuddas, Sergio Piras, Stefano Cappai, Federica Loi, Giulia Murgia, Giantonella Puggioni, Giovanni Savini, Federica Monaco, Andrea Polci, Fabrizia Valleriani, Giorgia Amatori, Valentina Curini, Maurilia Marcacci, Germano Orrù, Antonio Ledda, Elena Poma, Riccardo Cappai and Ferdinando Coghe

Pathogens 2024, 13(11), 1023; https://doi.org/10.3390/pathogens13111023 - 20 Nov 2024

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Abstract

West Nile Virus, an arthropod-borne RNA virus, may result in severe neurological disease. West Nile neuroinvasive disease is characterized by meningitis, encephalitis, and possible acute flaccid paralysis. Here, we report a case of neuroinvasive WNV in a 65-year-old woman hospitalized for hyperpyrexia, chills, [...] Read more.

West Nile Virus, an arthropod-borne RNA virus, may result in severe neurological disease. West Nile neuroinvasive disease is characterized by meningitis, encephalitis, and possible acute flaccid paralysis. Here, we report a case of neuroinvasive WNV in a 65-year-old woman hospitalized for hyperpyrexia, chills, intense asthenia, and continuous vomiting. Within days, her clinical condition worsened with the onset of severe neurological symptoms, leading to her death within 10 days despite supportive therapies being administered. The diagnosis of West Nile disease was made through nucleic acid amplification testing (NAAT) on blood and cerebrospinal fluid. However, in the final stages of the illness, cerebrospinal fluid collection was not possible due to the patient’s critical condition, and a nasopharyngeal swab was used instead. The nasopharyngeal swab facilitated the collection of a sample, which was subsequently analyzed for the presence of the virus and allowed for sequencing, showing that it was a strain that had been circulating in Sardinia for some time and had demonstrated its pathogenicity by causing the death of a hawk in 2021. This case report highlights the rapid progression and severity of WNV infection, particularly in vulnerable individuals, and suggests the potential utility of nasopharyngeal swabs as a less invasive option for sample collection. It also underscores the potential for the zoonotic transmission of the virus from birds to humans through vectors, emphasizing the importance of monitoring and controlling WNV outbreaks, especially in regions where such circulation is observed. Full article

(This article belongs to the Special Issue Emerging Zoonoses)

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13 pages, 263 KiB

Open AccessArticle

Loop-Mediated Isothermal Amplification (LAMP): An Innovative Approach for the Environmental Monitoring of SARS-CoV-2

by Simona Spiteri, Federica Marino, Luna Girolamini, Maria Rosaria Pascale, Carlo Derelitto, Laura Caligaris, Simone Paghera and Sandra Cristino

Pathogens 2024, 13(11), 1022; https://doi.org/10.3390/pathogens13111022 - 20 Nov 2024

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Abstract

The rapid and accurate detection of SARS-CoV-2 in environmental settings is crucial for effective public health management during the COVID-19 pandemic. This study compares the performance of the Reverse Transcription quantitative polymerase chain reaction (RT-qPCR) and the Reverse Transcription loop-mediated isothermal amplification (RT-LAMP) [...] Read more.

The rapid and accurate detection of SARS-CoV-2 in environmental settings is crucial for effective public health management during the COVID-19 pandemic. This study compares the performance of the Reverse Transcription quantitative polymerase chain reaction (RT-qPCR) and the Reverse Transcription loop-mediated isothermal amplification (RT-LAMP) for SARS-CoV-2 detection from 100 surface samples collected in healthcare environments. The reference method, RT-qPCR, identified a percentage of 25% of positive samples, while RT-LAMP detected a percentage of 27% of positive surfaces. Our findings reveal a sensitivity of 32% and specificity of 75% for RT-LAMP, with a positive predictive value of 30% and a negative predictive value of 77%. The overall accuracy and concordance with RT-qPCR was 64% for both methods. Despite its lower sensitivity compared to RT-qPCR, RT-LAMP had an advantage due to its rapid screening and environmental surveillance, which is particularly useful for confirming negative results. These results underscore the potential of RT-LAMP not only as a valuable method in the environmental monitoring of SARS-CoV-2 but also as a system to control the sanitation process in ordinary and emergency conditions, providing further optimization and validation for its reliability in routine surveillance and outbreak response efforts. Full article

14 pages, 1190 KiB

Open AccessArticle

Analysis of the Correlation Between Toxoplasma gondii Seropositivity and Alzheimer’s Disease

by Jianjun Wang, Ping Lin, Dan Li, Biyu Yang, Jiaqi Wang, Meng Feng and Xunjia Cheng

Pathogens 2024, 13(11), 1021; https://doi.org/10.3390/pathogens13111021 - 20 Nov 2024

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Abstract

Alzheimer’s disease (AD) is a multifactorial brain disorder and infectious diseases are considered as one of the predisposing factors for AD. Toxoplasma gondii, an obligate intracellular parasitic protozoan, is suspected of being associated with AD. Serum samples were collected from 109 AD [...] Read more.

Alzheimer’s disease (AD) is a multifactorial brain disorder and infectious diseases are considered as one of the predisposing factors for AD. Toxoplasma gondii, an obligate intracellular parasitic protozoan, is suspected of being associated with AD. Serum samples were collected from 109 AD patients and 114 age-matched healthy controls. ELISA was performed using recombinant T. gondii cyst wall protein 1 (CST1) to detect T. gondii antibodies. A parallel experiment was performed with Toxoplasma gondii tachyzoites lysate protein. To analyze whether factors associated with the onset of AD included chronic T. gondii infection, a multivariate logistic regression model was applied, further validating the correlation between chronic T. gondii infection and AD. AD patients exhibited significantly higher levels of Toxoplasma-specific antibodies in their serum compared to the control group, with statistically significant differences (p < 0.05). Multivariate logistic regression analysis revealed that Toxoplasma infection is a risk factor for AD (p < 0.01), and the CST1 antigen can significantly improve the model’s performance in predicting the occurrence of AD. The results indicate that chronic infection with Toxoplasma gondii could be one of the risk factors for the development of AD, potentially predisposing individuals with underlying health conditions to the disease. This further validates the correlation between Toxoplasma gondii and AD. Full article

(This article belongs to the Section Parasitic Pathogens)

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15 pages, 2923 KiB

Open AccessArticle

Coxsackievirus A6 U.K. Genetic and Clinical Epidemiology Pre- and Post-SARS-CoV-2 Emergence

by Alice M. Joyce, Jack D. Hill, Theocharis Tsoleridis, Stuart Astbury, Louise Berry, Hannah C. Howson-Wells, Nancy Allen, Ben Canning, Carl B. Jones, Gemma Clark, William L. Irving, Alexander W. Tarr and C. Patrick McClure

Pathogens 2024, 13(11), 1020; https://doi.org/10.3390/pathogens13111020 - 20 Nov 2024

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Abstract

Coxsackievirus A6 (CVA6) has become increasingly clinically relevant as a cause of Hand, Foot and Mouth Disease (HFMD) globally since 2008. However, most laboratories do not routinely determine the enteroviral type of positive samples. The non-pharmaceutical measures introduced to curb transmission during the [...] Read more.

Coxsackievirus A6 (CVA6) has become increasingly clinically relevant as a cause of Hand, Foot and Mouth Disease (HFMD) globally since 2008. However, most laboratories do not routinely determine the enteroviral type of positive samples. The non-pharmaceutical measures introduced to curb transmission during the COVID-19 pandemic may also have perturbed CVA6 epidemiology. We thus aimed to determine the prevalence, clinical presentation and genetic relationship of CVA6 across three complete epidemic seasons: one pre-SARS-CoV-2 emergence and two post-SARS-CoV-2 emergence in our regional healthcare setting. Surplus diagnostic nucleic acid from diagnosed enteroviral positives diagnosed between September and December of 2018 and between May 2021 and April of 2023 was subject to VP1 gene sequencing to determine the CVA6 cases and interrogate their phylogenetic relationship. The confirmed CVA6 cases were also retrospectively clinically audited. CVA6 infections were identified in 33 and 69 individuals pre- and post-pandemic, respectively, with cases peaking in November of 2018 and 2022, but in October of 2021. HFMD was the primary diagnosis in 85.5% of the post-pandemic cases, but only 69.7% of the pre-pandemic cases, where respiratory and neurological symptoms (45.5% and 12.1%, respectively) were significantly elevated. A complete VP1 sequence was retrieved for 94% of the CVA6 cases, revealing that studied infections were genetically diverse and suggestive of multiple local and international transmission chains. CVA6 presented a significant clinical burden in our regional U.K. hospital setting both pre- and post-pandemic and was subject to dynamic clinical and genetic epidemiology. Full article

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

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18 pages, 316 KiB

Open AccessReview

Auditory and Vestibular Involvement in Congenital Cytomegalovirus Infection

by Swetha G. Pinninti, William J. Britt and Suresh B. Boppana

Pathogens 2024, 13(11), 1019; https://doi.org/10.3390/pathogens13111019 - 20 Nov 2024

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Abstract

Congenital cytomegalovirus infection (cCMV) is a frequent cause of non-hereditary sensorineural hearing loss (SNHL) and developmental disabilities. The contribution of cCMV to childhood hearing loss has been estimated to be about 25% of all hearing loss in children at 4 years of age. [...] Read more.

Congenital cytomegalovirus infection (cCMV) is a frequent cause of non-hereditary sensorineural hearing loss (SNHL) and developmental disabilities. The contribution of cCMV to childhood hearing loss has been estimated to be about 25% of all hearing loss in children at 4 years of age. Although the vestibular insufficiency (VI) in cCMV has not been well-characterized and therefore, underestimated, recent studies suggest that VI is also frequent in children with cCMV and can lead to adverse neurodevelopmental outcomes. The pathogenesis of SNHL and VI in children with cCMV has been thought to be from direct viral cytopathic effects as well as local inflammatory responses playing a role. Hearing loss in cCMV can be of varying degrees of severity, unilateral or bilateral, present at birth or develop later (late-onset), and can progress or fluctuate in early childhood. Therefore, newborn hearing screening fails to identify a significant number of children with CMV-related SNHL. Although the natural history of cCMV-associated VI has not been well characterized, recent data suggests that it is likely that VI also varies considerably with respect to the laterality, timing of onset, degree of the deficit, and continued deterioration during early childhood. This article summarizes the current understanding of the natural history and pathogenesis of auditory and vestibular disorders in children with cCMV. Full article

(This article belongs to the Special Issue Understanding Human Cytomegalovirus Pathogenesis: Evidence from the Clinic and Laboratory Models)

10 pages, 1462 KiB

Open AccessArticle

The Leishmania Skin Test Predicts Clinic-Immunologic and Therapeutic Outcomes in Cutaneous Leishmaniasis

by Luiz H. Guimarães, Evelyn Zacarias, Sandra T. Nolasco, Almério N. Filho, Jamile Lago, Paulo R. L. Machado, Joyce Oliveira, Lucas P. Carvalho, Augusto Carvalho, Edgar M. Carvalho and Sérgio Arruda

Pathogens 2024, 13(11), 1018; https://doi.org/10.3390/pathogens13111018 - 19 Nov 2024

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Abstract

Cutaneous leishmaniasis (CL), caused by Leishmania braziliensis, is closely associated with a severe form of the disease, indicated by a positive Leishmania skin test (LST) that assesses and reflects the presence of immune T cells specific to Leishmania antigens. In this study, [...] Read more.

Cutaneous leishmaniasis (CL), caused by Leishmania braziliensis, is closely associated with a severe form of the disease, indicated by a positive Leishmania skin test (LST) that assesses and reflects the presence of immune T cells specific to Leishmania antigens. In this study, we compare the clinical, immunologic, and histopathologic features between Leishmania skin test-positive (LST+) and Leishmania skin test-negative (LST-) in CL. Compared to LST+ patients, LST- patients had larger lesions and had been sicker for longer, presented with more instances of therapeutic failure with meglumine antimonate, (MA) and the healing times were higher than LST+. While granulomas were less frequent and the parasite load was higher in LST-, there were more CD8+ T cells and an enhanced production of Granzyme B in the supernatants of biopsies from LST- subjects. This study shows that in LST-, an impairment in Th1 immune response is associated with a high parasite burden, and the pathology is mediated by CD8+ T cells and the enhanced production of Granzyme B. The abnormalities in the immunologic response in LST- patients lead to a more severe disease with a high rate of failure to therapy. Full article

(This article belongs to the Special Issue Host Immune Responses to Intracellular Pathogens)

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12 pages, 922 KiB

Open AccessArticle

Highly Pathogenic H5N1 Influenza A Virus (IAV) in Blue-Winged Teal in the Mississippi Flyway Is Following the Historic Seasonal Pattern of Low-Pathogenicity IAV in Ducks

by David E. Stallknecht, Deborah L. Carter, Lyndon Sullivan-Brügger, Paul Link, Emily Ferraro, Ciara McCarty, Bruce Davis, Lynda Knutsen, James Graham and Rebecca L. Poulson

Pathogens 2024, 13(11), 1017; https://doi.org/10.3390/pathogens13111017 - 19 Nov 2024

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Abstract

Highly pathogenic H5N1 (HP H5N1) influenza A virus (IAV) has been detected annually in North American ducks since its introduction during 2021, but it is unknown if this virus will follow the same seasonal and geographic patterns that have been observed with low-pathogenicity [...] Read more.

Highly pathogenic H5N1 (HP H5N1) influenza A virus (IAV) has been detected annually in North American ducks since its introduction during 2021, but it is unknown if this virus will follow the same seasonal and geographic patterns that have been observed with low-pathogenicity (LP) IAV in this reservoir. We monitored blue-winged teal in the Mississippi flyway prior to the detection of HP H5N1 and during two post-introduction migration cycles from spring 2022 to spring 2024, testing birds for infection and antibodies to IAV nucleoprotein (NP), hemagglutinin subtype H5, and neuraminidase subtype N1. Antigens representing clade 2.3.4.4b HP H5 and LP North American H5 were used for hemagglutination inhibition (HI) and virus neutralization (VN) tests for H5 antibodies. Virologic results were consistent with historic seasonal and geographic patterns reported for LP IAV with peak infections occurring in pre-migration staging areas in Minnesota during fall 2022. However, the high prevalence of the H5 subtype was exceptional compared to historic prevalence estimates at this same site and for the Mississippi flyway. HP H5N1 was detected on wintering areas in Louisiana and Texas during the fall of that same year and this was followed by an increase in estimated antibody prevalence to NP, H5, and N1 with no HP H5N1 detections during the wintering or spring migration periods of 2022/2023. HP H5N1 was not detected in Minnesota during fall 2023 but was detected from a single bird in Louisiana. However, a similar increase in antibody prevalence was observed during the winter and spring period of 2023 and 2024. Over the two migration cycles, there was a temporal shift in observed prevalence and relative titers against the H5 antigens with a higher proportion of ducks testing positive to the 2.3.4.4b H5 antigen and higher relative titer to that antigen compared to the representative LP North American H5 antigen. The seasonal and geographic patterns observed appear to be driven by population immunity during the migration cycle. Results support an initial high infection rate of HP H5N1 in blue-winged teal in the Mississippi flyway followed by a high prevalence of antibodies to NP, H5, and N1. Although prevalence was much reduced in the second migration cycle following introduction, it is not known if this pattern will persist in the longer term or affect historic patterns of subtype diversity in this reservoir. Full article

(This article belongs to the Special Issue Pathogenesis, Epidemiology, and Control of Animal Influenza Viruses)

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9 pages, 3789 KiB

Open AccessArticle

Optimizing Red Light-Based Photodynamic Therapy for Effective Bactericidal Action Against Fusobacterium nucleatum Subspecies

by Jianglan Li, Takayuki Nambu, Chao Wang, Hiroki Takigawa, Hugo Maruyama, Chiho Mashimo and Toshinori Okinaga

Pathogens 2024, 13(11), 1016; https://doi.org/10.3390/pathogens13111016 - 19 Nov 2024

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Abstract

Fusobacterium nucleatum (F. nucleatum), a key pathogen implicated in periodontal disease, contributes to oral biofilm maturation and is linked to development of systemic diseases like colorectal cancer and liver cirrhosis. Photodynamic therapy (PDT) combined with 5-aminolevulinic acid (5-ALA) treatment (ALA-PDT) selectively [...] Read more.

Fusobacterium nucleatum (F. nucleatum), a key pathogen implicated in periodontal disease, contributes to oral biofilm maturation and is linked to development of systemic diseases like colorectal cancer and liver cirrhosis. Photodynamic therapy (PDT) combined with 5-aminolevulinic acid (5-ALA) treatment (ALA-PDT) selectively targets F. nucleatum by inducing porphyrin accumulation. The bactericidal effect of red light-based PDT on F. nucleatum has not been evaluated previously. This study investigates the effect of ALA-PDT using red light-emitting diode (LED) light on F. nucleatum subspecies and their porphyrin accumulation. F. nucleatum subspecies were cultured with varying concentrations of 5-ALA under anaerobic conditions. Porphyrin accumulation was measured via fluorescence spectroscopy, and colony-forming units were measured to determine bacterial viability post-treatment. Additionally, other subspecies responded well to 0.01% 5-ALA, and uroporphyrin I accumulation correlated with bacterial death, revealing optimal bactericidal conditions. These results suggest that optimizing light intensity and 5-ALA concentration can significantly enhance the therapeutic potential of ALA-PDT in oral healthcare. Full article

(This article belongs to the Special Issue Oral Microbiome and Human Systemic Health)

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19 pages, 4881 KiB

Open AccessArticle

A Novel Tax-Responsive Reporter T-Cell Line to Analyze Infection of HTLV-1

by Stefanie Heym, Pauline Krebs, Kristin Ott, Norbert Donhauser, Laura M. Kemeter, Florian Simon, Sebastian Millen and Andrea K. Thoma-Kress

Pathogens 2024, 13(11), 1015; https://doi.org/10.3390/pathogens13111015 - 19 Nov 2024

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Abstract

Human T-cell leukemia virus type 1 (HTLV-1) infects CD4⁺ T-cells through close cell–cell contacts. The viral Tax-1 (Tax) protein regulates transcription by transactivating the HTLV-1 U3R promoter in the 5′ long terminal repeat of the integrated provirus. Here, we generated a clonal [...] Read more.

Human T-cell leukemia virus type 1 (HTLV-1) infects CD4⁺ T-cells through close cell–cell contacts. The viral Tax-1 (Tax) protein regulates transcription by transactivating the HTLV-1 U3R promoter in the 5′ long terminal repeat of the integrated provirus. Here, we generated a clonal Tax-responsive T-cell line to track HTLV-1 infection at the single-cell level using flow cytometry, bypassing intracellular viral protein staining. Jurkat T-cells stably transduced with the SMPU vector carrying green fluorescent protein (GFP) under control of 18 × 21 bp Tax-responsive element repeats of the U3R were evaluated. Among 40 clones analyzed for Tax responsiveness, the top two were characterized. Upon overexpression of Tax, over 40% of the cells showed GFP positivity, and approximately 90% of the Tax-positive cells were GFP-positive, indicating efficient reporter activity. However, with CREB-deficient Tax mutant M47, both total GFP-positive cell counts and those within the Tax-positive group significantly decreased. Co-culture with chronically HTLV-1-infected MT-2 or C91-PL cells led to an average of 0.9% or 2.4% GFP-positive cells, respectively, confirming the suitability to monitor HTLV-1 transmission and that HTLV-1 infection is very low. Thus, the novel Tax-responsive reporter T-cell line is a suitable tool to monitor infection of HTLV-1 on the single-cell level. Full article

(This article belongs to the Special Issue New Directions in HTLV-1 Research)

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9 pages, 2040 KiB

Open AccessArticle

Evaluating the Effectiveness of External Molecular Proficiency Testing in the Global Polio Laboratory Network, 2021–2022

by Nancy Gerloff and Cara C. Burns

Pathogens 2024, 13(11), 1014; https://doi.org/10.3390/pathogens13111014 - 19 Nov 2024

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Abstract

In the Global Poliovirus Laboratory Network (GPLN), participation and successful completion in annual proficiency test (PT) panels has been a part of the WHO accreditation process for decades. The PT panel is a molecular external quality assessment (mEQA) that evaluates laboratory preparedness, technical [...] Read more.

In the Global Poliovirus Laboratory Network (GPLN), participation and successful completion in annual proficiency test (PT) panels has been a part of the WHO accreditation process for decades. The PT panel is a molecular external quality assessment (mEQA) that evaluates laboratory preparedness, technical proficiency, the accuracy of data interpretation, and result reporting. Using the Intratypic Differentiation (ITD) real-time RT-PCR kits from CDC, laboratories run screening assays and report results in accordance with the ITD algorithm to identify and type polioviruses. The mEQA panels consisted of 10 blinded, non-infectious lyophilized RNA transcripts, including programmatically relevant viruses and targets contained in the real-time PCR assays. Sample identities included wildtype, vaccine-derived (VDPV), Sabin-like polioviruses, enterovirus, and negatives, as well as categories of invalid and indeterminate. The performance of individual laboratories was assessed based on the laboratory’s ability to correctly detect and characterize the serotype/genotype identities of each sample. The scoring scheme assessed the laboratory readiness following GPLN guidelines. Laboratories receiving mEQA scores of 90 or higher passed the assessment, scores of less than 90 failed and required remedial actions and re-evaluation. In 2021 and 2022, 123 and 129 GPLN laboratories were invited to request the annual PT panel, and 118 and 127 laboratories submitted results, respectively. The overall results were good, with 86% and 91.5% of laboratories passing the PT panel on their first attempt in 2021 and 2022, respectively. Most labs scored the highest score of 100, and less than one quarter scored between 90 and 95. Less than 10% of submitting laboratories failed the PT, resulting in in-depth troubleshooting to identify root causes and remediations. Most of these laboratories were issued a second PT panel for repeat testing, and almost all laboratories passed the repeat PT panel. The results of the 2021 and 2022 annual mEQA PTs showed that, despite the COVID-19 pandemic, the performance remained high in the GPLN, with most labs achieving the highest score. For these labs, the real-time PCR assay updates that were implemented during 2021–2022 were carried out with full adherence to procedures and algorithms. Even initially failing labs achieved passing scores after remediation. Full article

(This article belongs to the Special Issue Human Poliovirus)

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11 pages, 1050 KiB

Open AccessCase Report

Tick-Borne Encephalitis Virus Infection in a Two-Year-Old Child Returning from Switzerland (July–August 2023): Is It Time for TBE Immunization of Serbian Travelers?

by Verica Simin, Ognjen Ležakov, Ivana Bogdan, Dragana Mijatović, Dragana Gazibara, Lidija Popović-Dragonjić, Gordana Vijatov Đurić, Borko Milanović, Jelena Kesić, Andrea Đuretić, Ágota Ábrahám, Zsófia Lanszki, Gábor Kemenesi, Brigitta Zana, Krisztián Bányai, Zbigniew Zając, Dejan Jakimovski, Alejandro Cabezas-Cruz and Pavle Banović

Pathogens 2024, 13(11), 1013; https://doi.org/10.3390/pathogens13111013 - 18 Nov 2024

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Abstract

Tick-borne encephalitis (TBE) is a vaccine-preventable viral infection that poses significant public health challenges, particularly in regions where tick-borne diseases are endemic. This case report describes a 2-year-old boy with confirmed abortive TBEV infection following a tick bite during travel to Switzerland. The [...] Read more.

Tick-borne encephalitis (TBE) is a vaccine-preventable viral infection that poses significant public health challenges, particularly in regions where tick-borne diseases are endemic. This case report describes a 2-year-old boy with confirmed abortive TBEV infection following a tick bite during travel to Switzerland. The patient developed fever and mild symptoms but did not exhibit central nervous system involvement. The case underscores the importance of raising awareness among healthcare providers and travelers from non-endemic areas, such as Serbia, about TBE risk and the potential benefits of preventive vaccination. Strategic immunization campaigns could mitigate the public health impact of travel-related TBE. Full article

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

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16 pages, 847 KiB

Open AccessReview

The Mechanisms of Developing Fungicide Resistance in Fusarium graminearum Causing Fusarium Head Blight and Fungicide Resistance Management

by Malini Anudya Jayawardana and Wannakuwattewaduge Gerard Dilantha Fernando

Pathogens 2024, 13(11), 1012; https://doi.org/10.3390/pathogens13111012 - 18 Nov 2024

Viewed by 348

Abstract

Fusarium head blight (FHB), primarily caused by Fusarium graminearum, is one of the economically significant diseases in small grains. FHB causes severe damage to wheat production and grain quality. Several management strategies have been developed to control FHB, and chemical control through [...] Read more.

Fusarium head blight (FHB), primarily caused by Fusarium graminearum, is one of the economically significant diseases in small grains. FHB causes severe damage to wheat production and grain quality. Several management strategies have been developed to control FHB, and chemical control through fungicides plays a significant role. Although fungicides have effectively controlled F. graminearum in the field, the continuous exposure causes a selection pressure in the pathogen population towards fungicide resistance. Several studies have identified fungicide-resistant F. graminearum isolates and fungicide-resistance mechanisms. Although new fungicides with a new mode of action can be introduced into the market, developing a new fungicide is time-consuming, and extra efforts are needed for testing, approvals, and registrations. Therefore, it is essential to strategize the methods to delay the fungicide resistance. This review focuses on the impact of several fungicide applications currently used on FHB, focusing on Fusarium graminearum, the status of the fungicide sensitivity for fungicide classes, the resistance mechanisms against fungicides, and the mitigation strategies to delay the development of fungicide resistance in the pathogen population. Studying the fungicide resistance mechanisms and the mitigation strategies will be helpful in the future to use the available fungicides against F. graminearum without losing its effectiveness. Full article

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

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22 pages, 2243 KiB

Open AccessArticle

Molecular Analysis of Aggregatibacter actinomycetemcomitans ApiA, a Multi-Functional Protein

by Sera Jacob, Luciana Gusmao, Dipti Godboley, Senthil Kumar Velusamy, Nisha George, Helen Schreiner, Carla Cugini and Daniel H. Fine

Pathogens 2024, 13(11), 1011; https://doi.org/10.3390/pathogens13111011 - 18 Nov 2024

Viewed by 481

Abstract

Aggregatibacter actinomycetemcomitans ApiA is a trimeric autotransporter outer membrane protein (Omp) that participates in multiple functions, enabling A. actinomycetemcomitans to adapt to a variety of environments. The goal of this study is to identify regions in the apiA gene responsible for three of [...] Read more.

Aggregatibacter actinomycetemcomitans ApiA is a trimeric autotransporter outer membrane protein (Omp) that participates in multiple functions, enabling A. actinomycetemcomitans to adapt to a variety of environments. The goal of this study is to identify regions in the apiA gene responsible for three of these functions: auto-aggregation, buccal epithelial cell binding, and complement resistance. Initially, apiA was expressed in Escherichia coli. Finally, wild-type A. actinomycetemcomitans and an apiA-deleted version were tested for their expression in the presence and absence of serum and genes related to stress adaptation, such as oxygen regulation, catalase activity, and Omp proteins. Sequential deletions in specific regions in the apiA gene as expressed in E. coli were examined for membrane proteins, which were confirmed by microscopy. The functional activity of epithelial cell binding, auto-aggregation, and complement resistance were then assessed, and regions in the apiA gene responsible for these functions were identified. A region spanning amino acids 186–217, when deleted, abrogated complement resistance and Factor H (FH) binding, while a region spanning amino acids 28–33 was related to epithelial cell binding. A 13-amino-acid peptide responsible for FH binding was shown to promote serum resistance. An apiA deletion in a clinical isolate (IDH781) was created and tested in the presence and/or absence of active and inactive serum and genes deemed responsible for prominent functional activity related to A. actinomycetemcomitans survival using qRT-PCR. These experiments suggested that apiA expression in IDH781 is involved in global regulatory mechanisms that are serum-dependent and show complement resistance. This is the first study to identify specific apiA regions in A. actinomycetemcomitans responsible for FH binding, complement resistance, and other stress-related functions. Moreover, the role of apiA in overall gene regulation was observed. Full article

(This article belongs to the Special Issue Aggregatibacter actinomycetemcomitans)

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15 pages, 2874 KiB

Open AccessArticle

Kinetics of RPR Decline in Pregnant Persons Treated for Syphilis in Pregnancy and Their Infants

by Danielle Schwartz, Alena Tse-Chang, Joan Robinson, Jennifer Gratrix, Petra Smyczek and Michael T. Hawkes

Pathogens 2024, 13(11), 1010; https://doi.org/10.3390/pathogens13111010 - 17 Nov 2024

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Abstract

Congenital syphilis is a re-emerging infectious threat in areas of North America. The purpose of this study was to quantitatively describe the rate of decline of nontreponemal (rapid plasma reagin, RPR) titers in pregnant persons with syphilis and their infants. In a retrospective [...] Read more.

Congenital syphilis is a re-emerging infectious threat in areas of North America. The purpose of this study was to quantitatively describe the rate of decline of nontreponemal (rapid plasma reagin, RPR) titers in pregnant persons with syphilis and their infants. In a retrospective review, we included 120 pregnant persons with 563 reactive RPR measurements (median 5, range 2 to 11 per person) and 35 infants with 81 RPR measurements (median 2, range 2 to 6 per infant). First-order decay, second-order decay, and a mathematical model representing functional FcRn-mediated antibody recycling were fitted to individual patient RPR trajectories. The RPR titers decreased with a median half-life of 39 days (IQR 28–59) and 27 days (IQR 17–41) in birthing parents and infants, respectively. The half-life varied with the initial RPR titer, suggesting that the kinetics of RPR decline was not first-order. A mathematical model accounting for saturable antibody recycling explained the longevity of RPR reactivity, predicted the observed non-linear kinetics, and fit the empiric data well. In summary, RPR titers decline with a half-life of roughly one month; however, the elimination does not follow first-order kinetics. Saturable antibody recycling may explain the prolonged and non-linear elimination of nontreponemal antibodies. Full article

(This article belongs to the Special Issue Updates on Pediatric Infectious Diseases)

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22 pages, 7188 KiB

Open AccessReview

In Silico Genomic Analysis of Avian Influenza Viruses Isolated From Marine Seal Colonies

by Klaudia Chrzastek and Darrell R. Kapczynski

Pathogens 2024, 13(11), 1009; https://doi.org/10.3390/pathogens13111009 - 16 Nov 2024

Viewed by 773

Abstract

Genetically diverse avian influenza viruses (AIVs) are maintained in wild aquatic birds with increasingly frequent spillover into mammals, yet these represent a small proportion of the overall detections. The isolation of AIVs in marine mammals, including seals, has been reported sporadically over the [...] Read more.

Genetically diverse avian influenza viruses (AIVs) are maintained in wild aquatic birds with increasingly frequent spillover into mammals, yet these represent a small proportion of the overall detections. The isolation of AIVs in marine mammals, including seals, has been reported sporadically over the last 45 years. Prior to 2016, all reports of AIVs detected in seals were of low-pathogenicity AIVs. In spite of this, the majority of reported AIV outbreaks caused fatal respiratory diseases, with harbor seals particularly susceptible to infection. The H5 clade 2.3.4.4b highly pathogenic AIV (HPAIV) was detected in seals for the first time in 2016. Recently, many cases of mass seal die-offs have occurred because of 2.3.4.4b HPAIV and are attributed to spillover from wild bird species. The potential for seal-to-seal transmission has been considered after the mass mortality of southern elephant seals off the coast of Argentina. Close contact between seals and wild birds, the rapid evolution of H5N1 AIVs, and the possibility of efficient mammal-to-mammal transmission are increasing concerns due to the potential for the establishment of a marine mammal reservoir and public health risks associated with the pandemic potential of the virus. This manuscript details the detection of AIVs in the seal population, comparing interesting features of various subtypes with an emphasis on avian-to-mammal-to-mammal transmission. Phylogenetic characterizations of the representative seal isolates were performed to demonstrate the relationships within the different virus isolates. Furthermore, we demonstrate that the reassortment events between different LPAIVs occurred before and after the viruses reached the seal population. The reassortment of viral segments plays an important role in the evolution of influenza viruses. Taken together, these data report on the 45 year history between seals and AIVs. Full article

(This article belongs to the Special Issue Pathogenesis, Epidemiology, and Control of Animal Influenza Viruses)

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14 pages, 246 KiB

Open AccessArticle

Quantifying the Molecular Properties of the Elk Chronic Wasting Disease Agent with Mass Spectrometry

by Christopher J. Silva, Melissa L. Erickson-Beltran, Eric D. Cassmann and Justin J. Greenlee

Pathogens 2024, 13(11), 1008; https://doi.org/10.3390/pathogens13111008 - 16 Nov 2024

Viewed by 467

Abstract

Chronic wasting disease (CWD) is a prion disease afflicting wild and farmed elk. CWD prions (PrP^Sc) are infectious protein conformations that replicate by inducing a natively expressed prion protein (PrP^C) to refold into the prion conformation. Mass spectrometry was [...] Read more.

Chronic wasting disease (CWD) is a prion disease afflicting wild and farmed elk. CWD prions (PrP^Sc) are infectious protein conformations that replicate by inducing a natively expressed prion protein (PrP^C) to refold into the prion conformation. Mass spectrometry was used to study the prions resulting from a previously described experimental inoculation of MM132, ML132, and LL132 elk with a common CWD inoculum. Chymotryptic digestion times and instrument parameters were optimized to yield a set of six peptides, TNMK, MLGSAMSRPL, LLGSAMSRPL, ENMYR, MMER, and VVEQMCITQYQR. These peptides were used to quantify the amount, the M132 and L132 polymorphic composition, and the extent of methionine oxidation of elk PrP^Sc. The amount (ng/g brain tissue) of PrP^Sc present in each sample was determined to be: MM132 (5.4 × 10² ± 7 × 10¹), ML132 (3.3 × 10² ± 6 × 10¹ and 3.6 × 10² ± 3 × 10¹) and LL132 (0.7 × 10² ± 1 × 10¹, 0.2 × 10² ± 0.2 × 10¹, and 0.2 × 10² ± 0.5 × 10¹). The proportion of L132 polymorphism in ML132 (heterozygous) PrP^Sc from CWD-infected elk was determined to be 43% ± 2% or 36% ± 3%. Methionine oxidation was detected and quantified for the M132 and L132 polymorphisms in the samples. In this way, mass spectrometry can be used to characterize prion strains at a molecular level. Full article

(This article belongs to the Special Issue Advances in Chronic Wasting Disease)

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18 pages, 323 KiB

Open AccessReview

Group A Streptococcus Infections in Children: Epidemiological Insights Before and After the COVID-19 Pandemic

by Eleni Karapati, Andreas G. Tsantes, Zoi Iliodromiti, Theodora Boutsikou, Styliani Paliatsiou, Aglaia Domouchtsidou, Petros Ioannou, Vasileios Petrakis, Nicoletta Iacovidou and Rozeta Sokou

Pathogens 2024, 13(11), 1007; https://doi.org/10.3390/pathogens13111007 - 15 Nov 2024

Viewed by 447

Abstract

Group A streptococcus (GAS) is the cause of both mild and invasive infections in humans with a high morbidity and mortality rate. The transmission of disease usually occurs via droplets, so the implementation of infection mitigation strategies (IMS) during the COVID-19 pandemic altered [...] Read more.

Group A streptococcus (GAS) is the cause of both mild and invasive infections in humans with a high morbidity and mortality rate. The transmission of disease usually occurs via droplets, so the implementation of infection mitigation strategies (IMS) during the COVID-19 pandemic altered the incidence of GAS infection. This review aims to provide an overview of the influence of the COVID-19 pandemic on the incidence of GAS infection in children (invasive or non-invasive). A surge in the incidence of invasive GAS infection was noted in December 2022 after the reversal of IMS. A global uprise in GAS infection (invasive and non-invasive) was noted, especially concerning the pediatric population. Children younger than 5 years old were mostly affected, with complicated pneumonia being the leading clinical manifestation, causing many deaths worldwide. Emm1, specifically M1UK, was recognized as the dominant lineage in Europe and correlated with invasive disease. Healthcare professionals need to be alert about the severity of GAS-related infections, leading to early identification and treatment. Full article

(This article belongs to the Special Issue Antimicrobial Resistance in the Post-COVID Era: A Silent Pandemic)

18 pages, 6479 KiB

Open AccessArticle

Antimicrobial Resistance Profile of Zoonotic Clinically Relevant WHO Priority Pathogens

by Elaine Meade, Mark Anthony Slattery and Mary Garvey

Pathogens 2024, 13(11), 1006; https://doi.org/10.3390/pathogens13111006 - 15 Nov 2024

Viewed by 384

Abstract

The World Health Organization announced critically important bacterial and fungal pathogens displaying alarming levels of antimicrobial resistance, which currently represent difficult-to-treat cases of morbidity. Within this grouping, the ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, [...] Read more.

The World Health Organization announced critically important bacterial and fungal pathogens displaying alarming levels of antimicrobial resistance, which currently represent difficult-to-treat cases of morbidity. Within this grouping, the ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) are causative of significant morbidity and mortality. Studies described herein demonstrate the presence of critically important fungal and ESKAPE bacterial species in companion animals which are zoonotic in nature. The relationship between the environment, animals, and human infectious disease has long been recognized as part of One Health. This research investigates the resistance patterns of isolated zoonotic pathogens using recognized in vitro methodologies, namely disk diffusion, minimum inhibitory concentration testing, and genetic screening. Antibiotic susceptibility testing and gene analysis demonstrated an association between multi-drug resistance and extended beta spectrum lactamase production in critical-priority bacteria. Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii, and Pseudomonas aeruginosa exhibit great levels of multi-drug resistance. Fungal isolates demonstrated high levels of resistance, with Amphotericin B proving the most effective antifungal agent investigated. The level of antimicrobial resistance present in clinically relevant bacterial and fungal pathogens isolated from animal cases of morbidity in this study is alarming. In conclusion, this study shows that animals can act as a reservoir facilitating the transmission of antibiotic-resistant pathogens and genes zoonotically. Full article

(This article belongs to the Section Bacterial Pathogens)

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25 pages, 958 KiB

Open AccessReview

The Intriguing Connection Between the Gut and Lung Microbiomes

by Magdalena Druszczynska, Beata Sadowska, Jakub Kulesza, Nikodem Gąsienica-Gliwa, Ewelina Kulesza and Marek Fol

Pathogens 2024, 13(11), 1005; https://doi.org/10.3390/pathogens13111005 - 15 Nov 2024

Viewed by 321

Abstract

Recent advances in microbiome research have uncovered a dynamic and complex connection between the gut and lungs, known as the gut–lung axis. This bidirectional communication network plays a critical role in modulating immune responses and maintaining respiratory health. Mediated by immune interactions, metabolic [...] Read more.

Recent advances in microbiome research have uncovered a dynamic and complex connection between the gut and lungs, known as the gut–lung axis. This bidirectional communication network plays a critical role in modulating immune responses and maintaining respiratory health. Mediated by immune interactions, metabolic byproducts, and microbial communities in both organs, this axis demonstrates how gut-derived signals, such as metabolites and immune modulators, can reach the lung tissue via systemic circulation, influencing respiratory function and disease susceptibility. To explore the implications of this connection, we conducted a systematic review of studies published between 2001 and 2024 (with as much as nearly 60% covering the period 2020–2024), using keywords such as “gut–lung axis”, “microbiome”, “respiratory disease”, and “immune signaling”. Studies were selected based on their relevance to gut–lung communication mechanisms, the impact of dysbiosis, and the role of the gut microbiota in respiratory diseases. This review provides a comprehensive overview of the gut–lung microbiome axis, emphasizing its importance in regulating inflammatory and immune responses linked to respiratory health. Understanding this intricate pathway opens new avenues for microbiota-targeted therapeutic strategies, which could offer promising interventions for respiratory diseases like asthma, chronic obstructive pulmonary disease, and even infections. The insights gained through this research underscore the potential of the gut–lung axis as a novel target for preventative and therapeutic approaches in respiratory medicine, with implications for enhancing both gut and lung health. Full article

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11 pages, 543 KiB

Open AccessArticle

Molecular Exploration of Mycoplasma fermentans and Mycoplasma genitalium in Mexican Women with Cervicitis

by Abraham David Bustos-López, Marcos R. Escobedo-Guerra, Marcela López-Hurtado, Jesús Roberto Villagrana-Zesati, Martha Valdés-Ramírez, Silvia Giono-Cerezo and Fernando M. Guerra-Infante

Pathogens 2024, 13(11), 1004; https://doi.org/10.3390/pathogens13111004 - 15 Nov 2024

Viewed by 383

Abstract

Genital Mycoplasmas are implicated in adverse pregnancy outcomes and the development of infertility. However, the role of Mycoplasma fermentans in these outcomes has not been adequately studied; therefore, its participation in these sufferings requires further investigation. This study aimed to evaluate the prevalence [...] Read more.

Genital Mycoplasmas are implicated in adverse pregnancy outcomes and the development of infertility. However, the role of Mycoplasma fermentans in these outcomes has not been adequately studied; therefore, its participation in these sufferings requires further investigation. This study aimed to evaluate the prevalence of M. fermentans in pregnant and non-pregnant women. End-point PCR was used to analyze two hundred and twenty-eight endocervical samples for M. hominis, M. genitalium, M. fermentans, M. pirum, Ureaplasma urealyticum, and U. parvum diagnoses. The prevalence of Mycoplasma spp. was as follows: U. parvum was found in 83 samples (36.4%), U. urealyticum in 39 instances (17.1%), M. hominis in 36 (15.7%), M. fermentans in 32 (14%), M. genitalium in 15 (6.6%), and M. pirum in 0 samples. No association was found between the Mycoplasma spp. and some infertility conditions or adverse pregnancy. However, M. fermentans and M. hominis were found to be associated with bacterial vaginosis (RR = 3.4 CI 95% 1.85–6.3, p < 0.005). In conclusion, M. fermentans and M. hominis were isolated more often in women with bacterial vaginosis, which suggests that these bacteria could contribute to the development of this pathology. Full article

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

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18 pages, 5753 KiB

Open AccessArticle

Mycoplasma bovis Invades Non-Phagocytic Cells by Clathrin-Dependent Endocytic Pathways and Escapes from Phagocytic Vesicles

by Bin Li, Yabin Lu, Yaru Feng, Xiaolong Jiao, Qiuyu Zhang, Mengting Zhou, Yuyu Zhang, Jian Xu, Yuefeng Chu and Duoliang Ran

Pathogens 2024, 13(11), 1003; https://doi.org/10.3390/pathogens13111003 - 15 Nov 2024

Viewed by 356

Abstract

Mycoplasma bovis (M. bovis) is capable of causing pneumonia, arthritis, mastitis, and various other ailments in cattle of all age groups, posing a significant threat to the healthy progression of the worldwide cattle industry. The invasion of non-phagocytic host cells serves [...] Read more.

Mycoplasma bovis (M. bovis) is capable of causing pneumonia, arthritis, mastitis, and various other ailments in cattle of all age groups, posing a significant threat to the healthy progression of the worldwide cattle industry. The invasion of non-phagocytic host cells serves as a pivotal mechanism enabling M. bovis to evade the immune system and penetrate mucosal barriers, thereby promoting its spread. To investigate the differences in M. bovis invasion into four types of non-phagocytic cells (Madin–Darby bovine kidney (MDBK) cells, embryonic bovine lung (EBL) cells, bovine embryo tracheal (EBTr) cells and bovine turbinate (BT) cells) and further elucidate its invasion mechanism, this study first optimized the experimental methods for M. bovis invasion into cells. Utilizing laser scanning confocal microscopy, transmission electron microscopy, and high-content live-cell imaging systems, the invasion process of M. bovis into four types of non-phagocytic cells was observed. The invasion rates of three different strains of M. bovis (PG45, 07801, 08M) were quantified through the plate counting method. In order to clarify the specific pathway of M. bovis invasion into cells, chlorpromazine (CPZ), amiloride (AMI), and methyl-β-cyclodextrin (M-β-CD) were used to inhibit CLR-mediated clathrin-dependent endocytosis (CDE) pathway, macropinocytosis, and lipid raft pathway, respectively. Subsequently, the invasion rates of PG45 into these four types of cells were measured. Using siRNA technology, the expression of clathrin (CLR) in EBL cells was knocked down to further verify the role of CLR in the invasion process of M. bovis. The results showed that the optimal conditions for M. bovis to invade non-phagocytic cells were a multiplicity of infection (MOI) of 1000 and an optimal invasion time of 4 h. All three strains of M. bovis have the ability to invade the four types of non-phagocytic cells, yet their invasion abilities vary significantly. Observations from transmission electron microscopy further confirmed that at 120 min post-infection, PG45 had successfully invaded EBL cells and was present within endocytic vesicles. It is noteworthy that almost all PG45 successfully escaped from the endocytic vesicles after 240 min of infection had passed. Through chemical inhibition experiments and CLR protein knockdown experiments, it was found that when the CDE and lipid raft pathways were blocked or CLR protein expression was reduced, the invasion rates of PG45, 07801, and 08M in MDBK, EBL, EBTr, and BT cells were significantly decreased (p < 0.05). The above results indicate that M. bovis can invade all types of non-phagocytic cells through endocytic pathways involving CDE (clathrin-dependent endocytosis) or lipid raft-mediated endocytosis, and possesses the ability to escape from phagosomes. Full article

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12 pages, 1424 KiB

Open AccessBrief Report

Intestinal Carriage of Two Distinct stx_2f-Carrying Escherichia coli Strains by a Child with Uncomplicated Diarrhea

by Florence Crombé, Angela H. A. M. van Hoek, Heleen Nailis, Frédéric Auvray, Toon Janssen and Denis Piérard

Pathogens 2024, 13(11), 1002; https://doi.org/10.3390/pathogens13111002 - 15 Nov 2024

Viewed by 371

Abstract

Two distinct stx_2f-carrying Escherichia coli (E. coli) strains, isolated from a child with uncomplicated diarrhea fifteen weeks apart, were characterized by combining short- and long-read sequencing to compare their genetic relatedness. One strain was characterized as Shiga toxin-producing E. [...] Read more.

Two distinct stx_2f-carrying Escherichia coli (E. coli) strains, isolated from a child with uncomplicated diarrhea fifteen weeks apart, were characterized by combining short- and long-read sequencing to compare their genetic relatedness. One strain was characterized as Shiga toxin-producing E. coli (STEC)/typical enteropathogenic E. coli (tEPEC) O63:H6 with a repertoire of virulence genes including stx_2f, eae (α2-subtype), cdt, and bfpA. The other STEC with serotype O157:H16, reported for the first time as stx_2f-carrying Escherichia coli in this study, possessed, in addition, eae (ε-subtype) and cdt, amongst other virulence-related genes. BLAST comparison showed that the stx_2f-harboring prophage sequences of both strains were highly homologous (99.6% identity and 96.1% coverage). These results were corroborated by core Stx2f phage Multilocus Sequence Typing (cpMLST) as the stx_2f-harboring prophages of both isolates clustered together when compared to those of 167 other human stx_2f-carrying Escherichia coli. Overall, the stx_2f-harboring prophages of the two distinct E. coli strains isolated from the present case were highly similar, suggesting that the stx_2f-harboring phage might have been transferred from the STEC/tEPEC O63:H6 strain to the atypical EPEC (aEPEC) O157:H16 strain in the gut of the child. Full article

(This article belongs to the Section Bacterial Pathogens)

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30 pages, 3020 KiB

Open AccessArticle

The Evolution of Wisteria Vein Mosaic Virus: A Case Study Approach to Track the Emergence of New Potyvirus Threats

by Massimiliano Morelli, Giusy D’Attoma, Pasquale Saldarelli and Angelantonio Minafra

Pathogens 2024, 13(11), 1001; https://doi.org/10.3390/pathogens13111001 - 15 Nov 2024

Viewed by 351

Abstract

Wisteria vein mosaic virus (WVMV, Potyvirus wisteriae), a virus belonging to the genus Potyvirus, is responsible for Wisteria vein mosaic disease (WMD), a severe disease that affects Wisteria, a genus of garden plants acclaimed worldwide. Although probably originating in the [...] Read more.

Wisteria vein mosaic virus (WVMV, Potyvirus wisteriae), a virus belonging to the genus Potyvirus, is responsible for Wisteria vein mosaic disease (WMD), a severe disease that affects Wisteria, a genus of garden plants acclaimed worldwide. Although probably originating in the Far East, WVMV infection was first reported in the US, and subsequently in numerous countries. Following the first molecular detection of an Italian isolate, WVMV Bari, its full-length genome was achieved using NGS barcoding technology. A PhyML phylogenetic analysis, supported by clustering algorithm validation, identified a clear separation between two phylogroups. One major clade comprised WVMV strains isolated from Wisteria spp. A second clade grouped three highly divergent strains, at the borderline species threshold, all found in non-wisteria hosts. Relying on a Relative Time Dated Tips (RTDT) molecular clock, the first emergence of WVMV clades has been traced back to around the 17th century. A network inference analysis confirmed the sharp separation between the two host-related phylogroups, also highlighting the presence of potential intermediate variants. Inter-population genetic parameters revealed a very high genetic differentiation in both populations, which was made reliable by statistically significant permutation tests. The migrant number (Nm) and fixation index (F_ST) evidenced a restricted gene flow and strong population structures. According to the dN/dS ratio and negative neutrality tests, it was derived that purifying selection at the expense of non-silent variants is underway within WVMV populations. Targeting WVMV evolutionary traits, the present effort raised interesting questions about the underestimated potential of this culpably neglected species to spread in economically relevant crops. The main intention of our study is, therefore, to propose an evolution-based analysis approach that serves as a case study to investigate how other potyviruses or newly emerging viruses may spread. Full article

(This article belongs to the Special Issue The Plant RNA Viruses)

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19 pages, 1002 KiB

Open AccessReview

Advances on Bioactive Metabolites with Potential for the Biocontrol of Plant Pathogenic Bacteria

by Pierluigi Reveglia, Gaetano Corso and Antonio Evidente

Pathogens 2024, 13(11), 1000; https://doi.org/10.3390/pathogens13111000 - 15 Nov 2024

Viewed by 473

Abstract

The increase in the world population, which will be almost 10 billion by 2050, will require considerable efforts to significantly increase food production. Despite the considerable progress made in agriculture, this need is becoming an emergency due to desertification, environmental pollution and climate [...] Read more.

The increase in the world population, which will be almost 10 billion by 2050, will require considerable efforts to significantly increase food production. Despite the considerable progress made in agriculture, this need is becoming an emergency due to desertification, environmental pollution and climate changes. Biotic stresses, such as pathogenic bacteria and fungi, primarily contribute to significant losses in agricultural productivity and compromise food safety. These harmful agents are predominantly managed using large quantities of synthetic pesticides. However, this widespread use has led to substantial environmental pollution, increased pest resistance and toxic residues in agricultural produce, which subsequently enter the food supply, posing severe health risks to humans and animals. These challenges have significantly driven the advancement of integrated pest management strategies to reduce or eliminate synthetic pesticides. A practical and viable alternative lies in biopesticides—methods developed from natural products that are safe for human and animal health. This approach aligns with the strong demand from consumers and public authorities for safer pest control solutions. This review was focused on the isolation, chemical and biological characterization of natural products for the biocontrol of phytopathogenic bacteria and, in some cases, fungi with potential eco-friendly applications. Full article

(This article belongs to the Special Issue Control of Plant Pathogenic Bacteria with the Lowest Environmental Impact Volume II)

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13 pages, 817 KiB

Open AccessArticle

Salmonella Infantis Adhesion to Various Surfaces and In Vitro Antimicrobial Efficacy of Commercial Disinfectants

by Katja Kranjc, Jana Avberšek, Neva Šemrov, Olga Zorman-Rojs and Darja Barlič-Maganja

Pathogens 2024, 13(11), 999; https://doi.org/10.3390/pathogens13110999 - 14 Nov 2024

Viewed by 375

Abstract

Salmonella Infantis poses a significant challenge in poultry production due to its persistence and resistance to disinfectants. This study investigated the survival of the S. Infantis strain on different surfaces and evaluated the efficacy of disinfectants in both preventing and treating biofilms. The [...] Read more.

Salmonella Infantis poses a significant challenge in poultry production due to its persistence and resistance to disinfectants. This study investigated the survival of the S. Infantis strain on different surfaces and evaluated the efficacy of disinfectants in both preventing and treating biofilms. The survival of the tested S. Infantis strain was assessed on plastic and stainless steel surfaces after 24 and 48 h. The minimum inhibitory concentrations (MICs) of five disinfectants were determined, and their antiadhesion effectiveness was evaluated using crystal violet. The efficacy of biofilm treatment was evaluated by cell culturability. The results showed that the adhesion of S. Infantis was significantly higher on the plastic surface. The disinfectants were effective at reducing biofilm formation only within the first 24 h. Fresh solutions of disinfectants based on quaternary ammonium compounds exhibited the highest antimicrobial efficacy, while chlorocresol was the most effective for both the prevention and treatment of biofilms. The study results suggest that the presence of plastic surfaces may contribute to the dissemination of Salmonella. Additionally, the effectiveness of disinfectants varied based on storage conditions and contact time, while biofilms demonstrated reduced susceptibility compared to planktonic cells. However, given the laboratory scale of this study, further validation on a commercial scale is necessary to confirm these findings. Full article

(This article belongs to the Special Issue Development and Evaluation of Detection Method for Pathogenic Bacteria from Food)

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14 pages, 290 KiB

Open AccessArticle

Foodborne Pathogens Across Different Food Matrices in Sicily (Southern Italy)

by Valeria Blanda, Ilenia Giacchino, Valeria Vaglica, Vanessa Milioto, Sergio Migliore, Santina Di Bella, Francesca Gucciardi, Carmelo Bongiorno, Giuseppina Chiarenza, Cinzia Cardamone, Isabella Mancuso, Maria Luisa Scatassa, Vincenza Cannella, Annalisa Guercio, Giuseppa Purpari and Francesca Grippi

Pathogens 2024, 13(11), 998; https://doi.org/10.3390/pathogens13110998 - 14 Nov 2024

Viewed by 378

Abstract

Foodborne diseases result from the consumption of foods contaminated with pathogens or their toxins and represent a serious public health problem worldwide. This study aimed to assess the presence of Rotavirus (RoV), Adenovirus (AdV), Norovirus (NoV), Hepatitis A and Hepatitis E viruses (HAV [...] Read more.

Foodborne diseases result from the consumption of foods contaminated with pathogens or their toxins and represent a serious public health problem worldwide. This study aimed to assess the presence of Rotavirus (RoV), Adenovirus (AdV), Norovirus (NoV), Hepatitis A and Hepatitis E viruses (HAV and HEV, respectively), Toxoplasma gondii, Coxiella burnetii and Leptospira spp. across various food matrices in Sicily. The analysis concerned 504 samples, including mussels, farmed meat, game meat, vegetables and bulk milk. Following appropriate pre-treatment, acid nucleic extraction was carried out and amplification of pathogen nucleic acids was carried out by molecular methods. The mussels tested positive for NoVs (3/51, 5.9%) and farm meat resulted positive for T. gondii (1/34, 2.9%). The game offal samples tested positive for HEV, which was detected in 17 out of 222 samples (7.7%), and T. gondii (18/318, 5.7%) and Leptospira spp. (2/318, 0.6%). The milk samples tested positive for C. burnetii (15/85, 17.6%), T. gondii (2/85, 2.4%) and Leptospira spp. (1/85, 1.2%). This study highlights the variability in the risk of contamination of different food matrices, confirming the importance of vigilance in the consumption of potentially contaminated food products. Full article

(This article belongs to the Section Viral Pathogens)

18 pages, 3173 KiB

Open AccessArticle

Interrelation Between Pathoadaptability Factors and Crispr-Element Patterns in the Genomes of Escherichia coli Isolates Collected from Healthy Puerperant Women in Ural Region, Russia

by Yulia Mikhaylova, Marina Tyumentseva, Konstantin Karbyshev, Aleksandr Tyumentsev, Anna Slavokhotova, Svetlana Smirnova, Andrey Akinin, Andrey Shelenkov and Vasiliy Akimkin

Pathogens 2024, 13(11), 997; https://doi.org/10.3390/pathogens13110997 - 14 Nov 2024

Viewed by 328

Abstract

Escherichia coli is a commensal and opportunistic bacterium widely distributed around the world in different niches including intestinal of humans and animals, and its extraordinary genome plasticity led to the emergence of pathogenic strains causing a wide range of diseases. E. coli is [...] Read more.

Escherichia coli is a commensal and opportunistic bacterium widely distributed around the world in different niches including intestinal of humans and animals, and its extraordinary genome plasticity led to the emergence of pathogenic strains causing a wide range of diseases. E. coli is one of the monitored species in maternity hospitals, being the main etiological agent of urogenital infections, endometriosis, puerperal sepsis, and neonatal diseases. This study presents a comprehensive analysis of E. coli isolates obtained from the maternal birth canal of healthy puerperant women 3–4 days after labor. According to whole genome sequencing data, 31 sequence types and six phylogenetic groups characterized the collection containing 53 isolates. The majority of the isolates belonged to the B2 phylogroup. The data also includes phenotypic and genotypic antibiotic resistance profiles, virulence factors, and plasmid replicons. Phenotypic and genotypic antibiotic resistance testing did not demonstrate extensive drug resistance traits except for two multidrug-resistant E. coli isolates. The pathogenic factors revealed in silico were assessed with respect to CRISPR-element patterns. Multiparametric and correlation analyses were conducted to study the interrelation of different pathoadaptability factors, including antimicrobial resistance and virulence genomic determinants carried by the isolates under investigation. The data presented will serve as a valuable addition to further scientific investigations in the field of bacterial pathoadaptability, especially in studying the role of CRISPR/Cas systems in the E. coli genome plasticity and evolution. Full article

(This article belongs to the Special Issue Detection and Epidemiology of Drug-Resistant Bacteria)

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17 pages, 6838 KiB

Open AccessArticle

Bibliometric Analysis of Research Trends and Prospective Directions of Lung Microbiome

by Chunjing Du, Yi Zhang, Hanwen Zhang, Hua Zhang, Jingyuan Liu and Ning Shen

Pathogens 2024, 13(11), 996; https://doi.org/10.3390/pathogens13110996 - 14 Nov 2024

Viewed by 491

Abstract

The lung microbiome has emerged as a pivotal area of research in human health. Despite the increasing number of publications, there is a lack of research that comprehensively and objectively presents the current status of lung microbiome-related studies. Thus, this study aims to [...] Read more.

The lung microbiome has emerged as a pivotal area of research in human health. Despite the increasing number of publications, there is a lack of research that comprehensively and objectively presents the current status of lung microbiome-related studies. Thus, this study aims to address this gap by examining over two decades of publications through bibliometric analysis. The original bibliographic data of this study were obtained from the Web of Science Core Collection, focusing on publications from 2003 to 2023. The analysis included the data extraction and examination of authors, affiliations, countries, institutions, abstracts, keywords, references, publication dates, journals, citations, H-indexes, and journal impact factors. A total of 845 publications were identified, showing an increasing trend in both publications and citations over the years, particularly in the last decade. The analysis highlighted the most productive authors, institutions, and countries/regions, and identified potential partners for interested researchers. Co-citation analysis revealed that lung microbiome- and infectious/pulmonary disease-related studies are at the forefront of the field. The hotspots and frontiers of the lung microbiome field have progressed from basic composition to exploring specific mechanisms and the clinical value of diseases. In conclusion, this study provides a comprehensive overview of the current research status and trends in the field of the lung microbiome over the past two decades and highlights the areas that need more attention and research efforts. It offers valuable insights for researchers and institutions and identifies key hotspots and frontiers, which can serve as references for related researchers and future research. Full article

(This article belongs to the Special Issue Reviews of Infectious Diseases)

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8 pages, 217 KiB

Open AccessBrief Report

Infections with Soil-Transmitted Helminths in BaAka Pygmies Inhabiting the Rain Forests in the Central African Republic

by Wanesa Wilczyńska and Krzysztof Korzeniewski

Pathogens 2024, 13(11), 995; https://doi.org/10.3390/pathogens13110995 - 14 Nov 2024

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Abstract

Poor sanitation, improper food handling, limited access to safe drinking water sources, and limited access to healthcare services contribute to a high prevalence of infections caused by soil-transmitted helminths (STHs) among the BaAka Pygmies, an indigenous community living in Central Africa. The aim [...] Read more.

Poor sanitation, improper food handling, limited access to safe drinking water sources, and limited access to healthcare services contribute to a high prevalence of infections caused by soil-transmitted helminths (STHs) among the BaAka Pygmies, an indigenous community living in Central Africa. The aim of this study was to determine the rates of STH infections in the BaAka people inhabiting the rain forests of the south-western parts of the Central African Republic (CAR) as well as to examine the validity of biannual deworming with a single dose of albendazole 400 mg in high-risk communities exposed to extreme environmental conditions. The study was conducted in August 2021 and involved a sample of 49 BaAka Pygmies inhabiting the rain forest of the Sangha-Mbaéré Prefecture, CAR. The study consisted of collecting single stool samples from each participant and examining the samples for intestinal parasites by light microscopy methods. The collected samples were fixed in SAF fixative and next transported from Africa to Europe, where they were analyzed by light microscopy using three different diagnostic methods (direct smear in Lugol’s solution, the Fülleborn’s flotation, the Kato–Katz thick smear) at the Department of Epidemiology and Tropical Medicine in Gdynia, Poland. Microscopic examination found that 61.2% of the study group were infected with at least one helminthic species. The parasitological screening found invasions with four different species of nematodes, of which hookworm invasions were the most prevalent. The study results demonstrated that although the WHO-recommended mass deworming, which is provided to the BaAka Pygmies in healthcare centers set up on the premises of catholic missions, can effectively reduce the number of infections with soil-transmitted helminths, the prevalence of STH infections remains high in the region. The study findings suggest that in order to contain the spread of STHs in the local community, it will be necessary to implement additional preventive measures, apart from only conducting mass deworming programs. Full article

(This article belongs to the Special Issue Parasitic Diseases in the Contemporary World)

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Pathogens, Volume 13, Issue 11 (November 2024) – 108 articles

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