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23 pages, 5970 KiB  
Review
Practical Review on Aetio-Pathogenesis and Symptoms in Pigs Affected by Clinical and Subclinical Oedema Disease and the Use of Commercial Vaccines Under Field Conditions
by Juan Hernandez-Garcia, Isaac Ballarà Rodriguez, Ramon Jordà Casadevall, Sergi Bruguera, David Llopart and Emili Barba-Vidal
Animals 2025, 15(15), 2275; https://doi.org/10.3390/ani15152275 - 4 Aug 2025
Abstract
The impact of Oedema Disease produced by Shiga toxigenic Escherichia coli (STEC) in swine is increasing in some production countries due to increasing limitations on treatment with antimicrobials and zinc oxide, either because of the increased prevalence of multi-resistant strains or because of [...] Read more.
The impact of Oedema Disease produced by Shiga toxigenic Escherichia coli (STEC) in swine is increasing in some production countries due to increasing limitations on treatment with antimicrobials and zinc oxide, either because of the increased prevalence of multi-resistant strains or because of legal restrictions. The main pathological effect of Shiga toxin 2e is represented by damage to the endothelial cells of the blood vessel walls, leading to liquid extravasation and oedema formation in multiple tissues. These oedemas are generally easily identifiable in acute clinical cases. However, disease caused by Shiga toxin can occur without any externally visible oedema in the pigs, as observed in the subclinical presentation of Oedema Disease. It also causes productive losses, so it is important to identify and/or diagnose cases to set up control measures in order to optimize production and health. This article includes a comprehensive review of lesions and signs caused by Shiga toxin toxicosis in pigs, as well as other insights about the aetiology and epidemiology of STEC in pigs, and the effect of Shiga toxin recombinant toxoid vaccines in reducing these clinical and subclinical signs under field conditions. Full article
(This article belongs to the Section Pigs)
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14 pages, 1340 KiB  
Article
Exploring the Prevalence of Antimicrobial Resistance in the Environment Through Bonelli’s Eagles (Aquila fasciata) as Sentinels
by Barbara Martin-Maldonado, Ana Marco-Fuertes, Laura Montoro-Dasi, Laura Lorenzo-Rebenaque, Jose Sansano-Maestre, Jaume Jordá, Daniel Martín Solance, Fernando Esperón and Clara Marin
Antibiotics 2025, 14(8), 734; https://doi.org/10.3390/antibiotics14080734 - 22 Jul 2025
Viewed by 393
Abstract
Background/Objectives: Increasing levels of antimicrobial resistance (AMR) have recently been observed at the human–domestic animal–wildlife interface. Wild birds have been identified as carriers of antimicrobial-resistant bacteria and serve as excellent biomarkers for epidemiological studies. This study assessed the current AMR presence in Eastern [...] Read more.
Background/Objectives: Increasing levels of antimicrobial resistance (AMR) have recently been observed at the human–domestic animal–wildlife interface. Wild birds have been identified as carriers of antimicrobial-resistant bacteria and serve as excellent biomarkers for epidemiological studies. This study assessed the current AMR presence in Eastern Spain’s commensal Escherichia coli isolated from free-ranging Bonelli’s eagles (Aquila fasciata). Methods: Nestlings and their nests were intensively sampled between 2022 and 2024 to determine their AMR profile and characterize E. coli. AMR testing was conducted using the broth microdilution method, following the European Committee on Antimicrobial Susceptibility Testing guidelines. Additionally, the presence of eaeA (intimin gene) and stx-1 and stx-2 (shiga toxins) was analyzed by real-time PCR to classify E. coli strains into enteropathogenic (EPEC) and Shiga-toxigenic (STEC) pathotypes. Results: Of all E. coli isolates, 41.7% were resistant to at least one antimicrobial, and 30% were multidrug-resistant. Only two strains were classified as EPEC and none as STEC. The highest resistance rates were observed for amoxicillin and tetracycline (19.6% each). Alarmingly, resistance to colistin and meropenem, last-resort antibiotics in human medicine, was also detected. Conclusions: Although the mechanisms of resistance acquisition remain unclear, transmission is likely to occur through the food chain, with synanthropic prey acting as intermediary vectors. These results highlight the role of Bonelli’s eagles as essential sentinels of environmental AMR dissemination, even in remote ecosystems. Strengthening One Health-based surveillance is necessary to address AMR’s ecological and public health risks in wildlife. Full article
(This article belongs to the Special Issue Antimicrobial Resistance and Infections in Animals)
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12 pages, 1106 KiB  
Article
Antimicrobial Resistance and Virulence Determinants of Escherichia coli Isolates from Raw Milk of Dairy Cows with Subclinical Mastitis
by Ntelekwane George Khasapane, Olga de Smidt, Kgaugelo Edward Lekota, Jane Nkhebenyane, Oriel Thekisoe and Tsepo Ramatla
Animals 2025, 15(13), 1980; https://doi.org/10.3390/ani15131980 - 5 Jul 2025
Viewed by 292
Abstract
Subclinical mastitis (SCM) is a stealthy but devastating challenge in the dairy industry, leading to economic losses and hindering efforts to achieve milk self-sufficiency. This study investigated the prevalence of SCM, antimicrobial resistance, and virulence profiles of Escherichia coli. A total of [...] Read more.
Subclinical mastitis (SCM) is a stealthy but devastating challenge in the dairy industry, leading to economic losses and hindering efforts to achieve milk self-sufficiency. This study investigated the prevalence of SCM, antimicrobial resistance, and virulence profiles of Escherichia coli. A total of 174 milk samples were analyzed using the California mastitis test (CMT), somatic cell counts (SCCs), bacteriological culture, MALDI-TOF MS, and polymerase chain reaction (PCR). The findings revealed that the SCM prevalence was 68/174 (39.08%) based on CMT and SCC. Among SCM-positive samples, 60/68 (88.23%) were identified as E. coli, confirmed by MALDI-TOF MS and PCR assay. The most frequently detected serogroups were 0113 (11.6%) and 0113 (3.3%). Additionally, the genes for Stx1 and Stx2 were also detected in nine (15%) and one (1.7%), respectively. Antimicrobial susceptibility tests showed widespread resistance, with E. coli isolates demonstrating resistance to penicillin in 43 (71.6%), followed by ciprofloxacin in 42 (70%) and gentamicin in 18 (30%). A larger proportion of the E. coli strains (100%) harbored the blaVIM gene, while 23 (38.3%), 20%, 20%, and 1.47% contained blaKPC, blaNMD, suli1, and msrA. Thirty (50%) isolates were considered multidrug-resistant (MDR). These findings underscore the urgent need for enhanced surveillance and antibiotic stewardship in dairy farming. The presence of MDR E. coli in SCM poses a dual threat of potential transmission to humans and treatment failures in mastitis management. This study highlights the importance of proactive control strategies to mitigate the spread of antimicrobial resistance in livestock and beyond. Full article
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18 pages, 989 KiB  
Review
Neurological Manifestations of Hemolytic Uremic Syndrome: A Comprehensive Review
by Una Tonkovic, Marko Bogicevic, Aarish Manzar, Nikola Andrejic, Aleksandar Sic, Marko Atanaskovic, Selena Gajić, Ana Bontić, Sara Helena Ksiazek, Ana Mijušković, Nikola M. Stojanović and Marko Baralić
Brain Sci. 2025, 15(7), 717; https://doi.org/10.3390/brainsci15070717 - 4 Jul 2025
Viewed by 702
Abstract
Hemolytic uremic syndrome (HUS), a thrombotic microangiopathy primarily affecting the kidneys, can also involve the central nervous system (CNS), often leading to significant morbidity and mortality. Neurologic manifestations are among the most severe extra-renal complications, particularly in children and during outbreaks of Shiga [...] Read more.
Hemolytic uremic syndrome (HUS), a thrombotic microangiopathy primarily affecting the kidneys, can also involve the central nervous system (CNS), often leading to significant morbidity and mortality. Neurologic manifestations are among the most severe extra-renal complications, particularly in children and during outbreaks of Shiga toxin-producing Escherichia coli (STEC)-associated HUS (typical (tHUS)). This review explores the clinical spectrum, pathophysiology, diagnostic workup, and age-specific outcomes of neurologic involvement in both typical (tHUS) and atypical (aHUS). Neurologic complications occur in up to 11% of pediatric and over 40% of adult STEC-HUS cases in outbreak settings. Presentations include seizures, encephalopathy, focal deficits, movement disorders, and posterior reversible encephalopathy syndrome (PRES). Magnetic resonance imaging (MRI) commonly reveals basal ganglia or parieto-occipital lesions, though subtle or delayed findings may occur. Laboratory workup typically confirms microangiopathic hemolytic anemia (MAHA), thrombocytopenia, and kidney damage, with additional markers of inflammation or metabolic dysregulation. Eculizumab is the first-line treatment for aHUS with CNS involvement, while its utility in STEC-HUS remains uncertain. Although many children recover fully, those with early CNS involvement are at greater risk of developing epilepsy, cognitive delays, or fine motor deficits. Adults may experience lingering neurocognitive symptoms despite apparent clinical recovery. Differences in presentation and imaging findings between age groups emphasize the need for tailored diagnostic and therapeutic strategies. Comprehensive neurorehabilitation and long-term follow-up are crucial for identifying residual deficits. Continued research into predictive biomarkers, neuroprotective interventions, and standardized treatment protocols is needed for improving outcomes in HUS patients with neurological complications. Full article
(This article belongs to the Special Issue New Advances in Neuroimmunology and Neuroinflammation)
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31 pages, 1686 KiB  
Review
Strategic Detection of Escherichia coli in the Poultry Industry: Food Safety Challenges, One Health Approaches, and Advances in Biosensor Technologies
by Jacquline Risalvato, Alaa H. Sewid, Shigetoshi Eda, Richard W. Gerhold and Jie Jayne Wu
Biosensors 2025, 15(7), 419; https://doi.org/10.3390/bios15070419 - 1 Jul 2025
Viewed by 985
Abstract
Escherichia coli (E. coli) remains a major concern in poultry production due to its ability to incite foodborne illness and public health crisis, zoonotic potential, and the increasing prevalence of antibiotic-resistant strains. The contamination of poultry products with pathogenic E. coli [...] Read more.
Escherichia coli (E. coli) remains a major concern in poultry production due to its ability to incite foodborne illness and public health crisis, zoonotic potential, and the increasing prevalence of antibiotic-resistant strains. The contamination of poultry products with pathogenic E. coli, including avian pathogenic E. coli (APEC) and Shiga toxin-producing E. coli (STEC), presents risks at multiple stages of the poultry production cycle. The stages affected by E. coli range from, but are not limited to, the hatcheries to grow-out operations, slaughterhouses, and retail markets. While traditional detection methods such as culture-based assays and polymerase chain reaction (PCR) are well-established for E. coli detection in the food supply chain, their time, cost, and high infrastructure demands limit their suitability for rapid and field-based surveillance—hindering the ability for effective cessation and handling of outbreaks. Biosensors have emerged as powerful diagnostic tools that offer rapid, sensitive, and cost-effective alternatives for E. coli detection across various stages of poultry development and processing where detection is needed. This review examines current biosensor technologies designed to detect bacterial biomarkers, toxins, antibiotic resistance genes, and host immune response indicators for E. coli. Emphasis is placed on field-deployable and point-of-care (POC) platforms capable of integrating into poultry production environments. In addition to enhancing early pathogen detection, biosensors support antimicrobial resistance monitoring, facilitate integration into Hazard Analysis Critical Control Points (HACCP) systems, and align with the One Health framework by improving both animal and public health outcomes. Their strategic implementation in slaughterhouse quality control and marketplace testing can significantly reduce contamination risk and strengthen traceability in the poultry value chain. As biosensor technology continues to evolve, its application in E. coli surveillance is poised to play a transformative role in sustainable poultry production and global food safety. Full article
(This article belongs to the Special Issue Biosensors for Food Safety)
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33 pages, 1246 KiB  
Review
Shiga Toxin-Producing Escherichia coli (STEC) in Developing Countries: A 10-Year Review with Global Perspective
by Ali Nemati, Ali Dadvar, Mark Eppinger, Zohreh Karimpour, Soroush Saberi Kakhki, Alireza Sabeti Moghaddam Sabzevar, Mahdi Askari Badouei, Federica Gigliucci, Luis Fernando dos Santos, Keiji Nakamura, Hooman Javidi and Maryam Hafiz
Microorganisms 2025, 13(7), 1529; https://doi.org/10.3390/microorganisms13071529 - 30 Jun 2025
Cited by 1 | Viewed by 1034
Abstract
In the past two decades, Shiga toxin-producing Escherichia coli (STEC) has been responsible for multiple large-scale outbreaks worldwide, affecting thousands of individuals. While surveillance systems in developed countries such as the United States, the United Kingdom, Europe, Australia, Japan, and Canada are well-established, [...] Read more.
In the past two decades, Shiga toxin-producing Escherichia coli (STEC) has been responsible for multiple large-scale outbreaks worldwide, affecting thousands of individuals. While surveillance systems in developed countries such as the United States, the United Kingdom, Europe, Australia, Japan, and Canada are well-established, data on STEC prevalence in developing nations remain sparse, partly due to the absence of well-structured molecular diagnostic networks or surveillance systems. This review analyzed 250 studies published between 2014 and 2024 across 39 developing countries in Africa, Asia, Latin America, and the Caribbean, yielding 8986 STEC isolates. Detailed serogroup and serotype data were available for 55.9% of these, with O111, O157, and O26 being most common in humans. In animals, O157:H7 was most frequent, while food isolates mirrored global trends with O157 and O111 dominance. Notably, O145, a serogroup frequently reported in the U.S. and Europe, was absent from the ‘’Top Seven’’ serogroups. Shiga toxin subtypes stx1a and stx2a were most prevalent in human cases. In animal isolates, stx2e was the most prevalent subtype, while stx2c was most commonly found in food samples. We recommend establishing reference laboratories in these regions to improve data quality, strengthen monitoring efforts, and reduce the burden of STEC infections globally. Full article
(This article belongs to the Special Issue Advances in Human Infections and Public Health)
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25 pages, 980 KiB  
Review
Food Safety in Hydroponic Food Crop Production: A Review of Intervention Studies to Control Human Pathogens
by Melanie L. Lewis Ivey, Abigail Aba Mensah, Florian Diekmann and Sanja Ilic
Foods 2025, 14(13), 2308; https://doi.org/10.3390/foods14132308 - 29 Jun 2025
Viewed by 522
Abstract
The production of hydroponic fresh produce presents unique food safety and intervention challenges. A systematic approach was used to map and characterize the evidence on hydroponic food safety. Quantitative data describing the effectiveness of intervention studies were extracted, synthesized, and assessed for quality. [...] Read more.
The production of hydroponic fresh produce presents unique food safety and intervention challenges. A systematic approach was used to map and characterize the evidence on hydroponic food safety. Quantitative data describing the effectiveness of intervention studies were extracted, synthesized, and assessed for quality. A search of electronic databases yielded 131 relevant papers related to hydroponic food safety. Thirty-two studies focusing on food safety interventions reported 53 different interventions using chemical (n = 39), physical (n = 10), multiple-hurdle (n = 2), and biological (n = 2) approaches. Human pathogen indicators and surrogates were most often studied (n = 19), while pathogenic strains like Salmonella spp. (n = 9), Shiga toxin-producing Escherichia coli (STEC) (n = 5), Listeria monocytogenes (n = 2), and viruses (Hepatitis A virus (HAV), n = 1; norovirus (NoV), n = 1) were studied less frequently. Of fourteen articles (43.8%) investigating pre-harvest interventions, most (42.9%) did not specify the hydroponic system type. Gaps remain in the available evidence regarding the efficacy of interventions for controlling human pathogens in near-commercial hydroponic systems. The quality assessment revealed a significant lack of detailed reporting on methods and outcomes, making it difficult to translate the findings into practical recommendations for the industry; therefore, this review provides recommendations for the scientific community to improve future research design and reporting in this field. Full article
(This article belongs to the Section Food Quality and Safety)
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18 pages, 8365 KiB  
Article
Shedding of GPP130 by PC7 and Furin: Potential Implication in Lung Cancer Progression
by Priyanka Prabhala, Stephanie Duval, Alexandra Evagelidis, Maïlys Le Dévéhat, Vatsal Sachan and Nabil G. Seidah
Int. J. Mol. Sci. 2025, 26(13), 6164; https://doi.org/10.3390/ijms26136164 - 26 Jun 2025
Viewed by 427
Abstract
From a previously performed proteomics screen, GPP130, or Golgi phosphoprotein of 130 kDa, was identified as a potential substrate of the proprotein convertase 7 (PC7; PCSK7). GPP130 is a type-II transmembrane protein with a luminal domain containing endosomal and Golgi-retrieval determinants, enabling a [...] Read more.
From a previously performed proteomics screen, GPP130, or Golgi phosphoprotein of 130 kDa, was identified as a potential substrate of the proprotein convertase 7 (PC7; PCSK7). GPP130 is a type-II transmembrane protein with a luminal domain containing endosomal and Golgi-retrieval determinants, enabling a unique trafficking route. Most of the previous work on GPP130 relates to its binding and retrograde trafficking of the Shiga toxin. However, its cellular biology and its biochemical characterization remain understudied. Recently, GPP130 was reported to be implicated in cell cycle progression and cell proliferation in head and neck cancer cells. This led us to analyze the cBioPortal for Cancer Genomics, revealing that the GPP130/GOLIM4 gene is amplified in many cancers, including lung, ovarian, and cervical. This observation led us to use the A549 lung cancer cell line to investigate the growth-regulating roles of endogenous and overexpressed GPP130 and to analyze the impact of its cleavage/shedding by PC7 and/or Furin on cellular growth. Our cell-based assays suggest that GPP130 is a novel pro-protein convertase substrate that increases cell proliferation in A549, SKOV3, and HeLa cells, and that the latter activity is enhanced following its cleavage by PC7 and/or Furin into a membrane-bound N-terminal product and secreted C-terminal fragments. This novel work sheds light on the cell biology of the poorly characterized GPP130, its proliferative activity, and modulation upon its shedding by PC7 and Furin in lung cancer progression. Full article
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20 pages, 1308 KiB  
Article
Identification, Comparison, and Profiling of Selected Diarrhoeagenic Pathogens from Diverse Water Sources and Human and Animal Faeces Using Whole-Genome Sequencing
by Arinao Murei and Maggy Ndombo Benteke Momba
Microorganisms 2025, 13(6), 1373; https://doi.org/10.3390/microorganisms13061373 - 12 Jun 2025
Viewed by 494
Abstract
Consumption of contaminated drinking water is known to cause waterborne diseases such as diarrhoea, dysentery, typhoid, and hepatitis. This study applied whole-genome sequencing (WGS) to detect, identify, compare, and profile diarrhoeagenic pathogens (Vibrio cholerae, Shiga toxin-producing Escherichia coli, and Escherichia [...] Read more.
Consumption of contaminated drinking water is known to cause waterborne diseases such as diarrhoea, dysentery, typhoid, and hepatitis. This study applied whole-genome sequencing (WGS) to detect, identify, compare, and profile diarrhoeagenic pathogens (Vibrio cholerae, Shiga toxin-producing Escherichia coli, and Escherichia coli O157:H7) from 3168 water samples and 135 faecal samples (human and animal). Culture-based methods, MALDI-TOF mass spectrometry, and PCR were employed prior to WGS for identification of pathogens. Culture-based results revealed high presumptive prevalence of STEC (40.2%), V. cholerae (37.1%), and E. coli O157:H7 (22.7%). The MALDI-TOF confirmed 555 isolates with V. cholerae identified as Vibrio albensis. Shiga toxin-producing Escherichia coli (STEC) was more prevalent in wastewater (60%), treated water (54.1%), and groundwater (36.8%). PCR detected 46.4% of virulence genes from the water isolates and 66% of virulence genes from the STEC stool isolates. WGS also revealed STEC (92.9%) as the most prevalent species and found common virulence (e.g., hcp1/tssD1 and hlyE) and resistance (e.g., acrA and baeR) genes in all three types of samples. Five resistance and thirteen virulence genes overlapped among treated water and stool isolates. These findings highlight the diarrhoeagenic pathogens’ public health risk in water sources and underscore the need for better water quality monitoring and treatment standards. Full article
(This article belongs to the Special Issue Microbes in Wastewater Treatment)
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27 pages, 3028 KiB  
Article
Integrated Assessment of Antibacterial Activity, Polyphenol Composition, Molecular Docking, and ADME Properties of Romanian Oak and Fir Honeydew Honeys
by Calin Hulea, Diana Obistioiu, Anca Hulea, Mukhtar Adeiza Suleiman, Doris Floares (Oarga), Ersilia Alexa, Ilinca Merima Imbrea, Alina-Georgeta Neacșu, Marius Pentea, Cosmin Alin Popescu and Florin Imbrea
Antibiotics 2025, 14(6), 592; https://doi.org/10.3390/antibiotics14060592 - 8 Jun 2025
Viewed by 824
Abstract
Background: This study evaluated the polyphenolic composition, antibacterial activity, molecular docking interactions, and pharmacokinetic properties of Romanian oak and fir honeydew honeys. Methods: Spectrophotometric methods quantified total phenolic, flavonoid contents and antioxidant activity, and individual polyphenols were identified via HPLC-MS. Antibacterial efficacy against [...] Read more.
Background: This study evaluated the polyphenolic composition, antibacterial activity, molecular docking interactions, and pharmacokinetic properties of Romanian oak and fir honeydew honeys. Methods: Spectrophotometric methods quantified total phenolic, flavonoid contents and antioxidant activity, and individual polyphenols were identified via HPLC-MS. Antibacterial efficacy against Gram-positive and Gram-negative bacteria was evaluated by determining the bacterial inhibition percentage and minimum inhibitory concentrations. The bioactive compounds identified via LC-MS analysis were used to further delineate the possible antibacterial activities in silico. Molecular docking was carried out to predict the binding interactions and complex formation of the identified compounds against protein crystal structures of the bacteria used in this study. Additionally, the pharmacokinetic profile of compounds with high inhibitory potential was assessed via ADMET (absorption, Distribution, Metabolism, Excretion, toxicity) predictors to ascertain their value. Results: Fir honeydew honey showed higher total phenolic (844.5 mg GAE/kg) and flavonoid contents (489.01 mg QUE/kg) compared to oak honeydew honey, correlating with more potent antioxidant activity (IC50 = 5.16 mg/mL). In vitro antimicrobial tests indicated a stronger inhibitory effect of fir honeydew honey, especially against Gram-positive strains like S. aureus, S. pyogenes, and L. monocytogenes, alongside certain Gram-negative strains such as E. coli and H. influenzae. Oak honeydew honey displayed selective antimicrobial action, particularly against P. aeruginosa and S. typhimurium. The docking outcomes showed rutin, rosmarinic acid, beta resorcylic acid, quercetin, ferulic acid, and p-coumaric acid have high inhibitory activities characterised by binding affinities and binding interactions against shiga toxin, riboflavin synthase, ATP-binding sugar transporter-like protein, undecaprenyl diphosphate synthase, putative lipoprotein, sortase A, and immunity protein, making them key contributors to the honey’s antimicrobial activity. Moreover, beta-resorcylic acid, quercetin, ferulic acid, and p-coumaric acid revealed interesting ADMET scores that qualify honey to serve as a good antimicrobial agent. Conclusions: These findings support their potential use as natural antibacterial agents and emphasise the value of integrating chemical, biological, and computational approaches for multidisciplinary characterisations. Full article
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22 pages, 3496 KiB  
Review
INM004: Polyclonal Neutralizing Antibodies Against Shiga Toxin as a Treatment for Hemolytic Uremic Syndrome
by Marta Rivas, Mariana Pichel, Vanesa Zylberman, Mariana Colonna, Marina Valerio, Carolina Massa, Romina Pardo, Andrés E. Ciocchini, Santiago Sanguineti, Ian Roubicek, Linus Spatz and Fernando Alberto Goldbaum
Toxins 2025, 17(6), 282; https://doi.org/10.3390/toxins17060282 - 5 Jun 2025
Viewed by 1413
Abstract
Hemolytic uremic syndrome (HUS) is a thrombotic microangiopathy characterized by microangiopathic hemolytic anemia, thrombocytopenia, and acute kidney injury (AKI). Shiga toxin (Stx)-producing Escherichia coli-associated HUS (STEC-HUS) is one of the leading causes of AKI in children. Approximately 1.5 to 3% of children [...] Read more.
Hemolytic uremic syndrome (HUS) is a thrombotic microangiopathy characterized by microangiopathic hemolytic anemia, thrombocytopenia, and acute kidney injury (AKI). Shiga toxin (Stx)-producing Escherichia coli-associated HUS (STEC-HUS) is one of the leading causes of AKI in children. Approximately 1.5 to 3% of children die during the acute phase, and about 30% experience long-term renal sequelae. Argentina has the highest incidence of STEC-HUS globally. Given the prominent role of Stx in its pathophysiology, STEC-HUS is considered more of a toxemia than a bacterial disease. Stx transport occurs before and after the STEC-HUS onset, allowing for the distinction between an early toxemia phase and an advanced toxemia phase. In this review, we present our efforts to develop INM004, an anti-Stx treatment aimed at ameliorating or preventing the clinical consequences of STEC-HUS. We describe the protein engineering that facilitated this development and the clinical path to demonstrate the safety and efficacy of INM004. This immunotherapy could represent a new step in the treatment of STEC-HUS, which could potentially prevent long-term damage. If phase 3 trials are successful, earlier and broader use of INM004 is envisioned. We also discuss the potential impact of INM004 therapy, targeted vaccination strategies, and new diagnostic tools for this disease. Full article
(This article belongs to the Special Issue Antibodies for Innovative Studies of Bacterial Toxins)
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25 pages, 3666 KiB  
Article
Validation of Core and Whole-Genome Multi-Locus Sequence Typing Schemes for Shiga-Toxin-Producing E. coli (STEC) Outbreak Detection in a National Surveillance Network, PulseNet 2.0, USA
by Molly M. Leeper, Morgan N. Schroeder, Taylor Griswold, Mohit Thakur, Krittika Krishnan, Lee S. Katz, Kelley B. Hise, Grant M. Williams, Steven G. Stroika, Sung B. Im, Rebecca L. Lindsey, Peyton A. Smith, Jasmine Huffman, Alyssa Kelley, Sara Cleland, Alan J. Collins, Shruti Gautam, Eishita Tyagi, Subin Park, João A. Carriço, Miguel P. Machado, Hannes Pouseele, Dolf Michielsen and Heather A. Carletonadd Show full author list remove Hide full author list
Microorganisms 2025, 13(6), 1310; https://doi.org/10.3390/microorganisms13061310 - 4 Jun 2025
Viewed by 1209
Abstract
Shiga-toxin-producing E. coli (STEC) is a leading causing of bacterial foodborne and zoonotic illnesses in the USA. Whole-genome sequencing (WGS) is a powerful tool used in public health and microbiology for the detection, surveillance, and outbreak investigation of STEC. In this study, we [...] Read more.
Shiga-toxin-producing E. coli (STEC) is a leading causing of bacterial foodborne and zoonotic illnesses in the USA. Whole-genome sequencing (WGS) is a powerful tool used in public health and microbiology for the detection, surveillance, and outbreak investigation of STEC. In this study, we applied three WGS-based subtyping methods, high quality single-nucleotide polymorphism (hqSNP) analysis, whole genome multi-locus sequence typing using chromosome-associated loci [wgMLST (chrom)], and core genome multi-locus sequence typing (cgMLST), to isolate sequences from 11 STEC outbreaks. For each outbreak, we evaluated the concordance between subtyping methods using pairwise genomic differences (number of SNPs or alleles), linear regression models, and tanglegrams. Pairwise genomic differences were highly concordant between methods for all but one outbreak, which was associated with international travel. The slopes of the regressions for hqSNP vs. allele differences were 0.432 (cgMLST) and 0.966 wgMLST (chrom); the slope was 1.914 for cgMLST vs. wgMLST (chrom) differences. Tanglegrams comprised of outbreak and sporadic sequences showed moderate clustering concordance between methods, where Baker’s Gamma Indices (BGIs) ranged between 0.35 and 0.99 and Cophenetic Correlation Coefficients (CCCs) were ≥0.88 across all outbreaks. The K-means analysis using the Silhouette method showed the clear separation of outbreak groups with average silhouette widths ≥0.87 across all methods. This study validates the use of cgMLST for the national surveillance of STEC illness clusters using the PulseNet 2.0 system and demonstrates that hqSNP or wgMLST can be used for further resolution. Full article
(This article belongs to the Special Issue The Molecular Epidemiology of Infectious Diseases)
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21 pages, 1815 KiB  
Systematic Review
The Global Prevalence of Antibiotic Resistance and Shiga Toxin-Producing Escherichia coli in Chickens: A Systematic Review and Meta-Analysis (2011–2024)
by Tsepo Ramatla, Nkhebenyane Jane, Mohapi Dineo, Tawana Mpho, Motlhaoloa Tshegofatso and Ntelekwane George Khasapane
Antibiotics 2025, 14(6), 568; https://doi.org/10.3390/antibiotics14060568 - 31 May 2025
Viewed by 833
Abstract
Background: Shiga toxin-producing E. coli (STEC) are important foodborne pathogens that cause serious public health consequences worldwide. This study conducted a systematic review and meta-analysis of the global prevalence of antibiotic resistance and STEC in chickens. Methods: The assessment of previous study records [...] Read more.
Background: Shiga toxin-producing E. coli (STEC) are important foodborne pathogens that cause serious public health consequences worldwide. This study conducted a systematic review and meta-analysis of the global prevalence of antibiotic resistance and STEC in chickens. Methods: The assessment of previous study records was carried out following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Heterogeneity between studies was assessed using Cochrane’s Q test and I2 test statistics based on the random effects model, and comprehensive meta-analysis (CMA) software v4.0 was used to analyse the pooled prevalence estimate (PPE) of antibiotic resistance and STEC in chickens. Results: A total of 61 studies comprising 823 STEC from 18 countries were included in this study. The overall pooled prevalence of STEC was 8.9% (95% CI: 0.620–0.126). m-PCR assay showed the highest PPE of 21.0% (95%: 0.088–0.420). stx1 had the higher PPE of 12.9% (95%: 0.081–0.199), while stx2 had a PPE of 11.8% (95%: 0.077–0.176). Furthermore, the serotype O157 had the higher PPE of 80.5% (95%: 0.520–0.940). The isolates were resistant to the following antibiotics: amoxicillin and clavulanic acid, chloramphenicol, tetracycline, ciprofloxacin, gentamycin, ampicillin, neomycin, and amoxicillin. Conclusions: These findings may assist in the prevention and control of STEC in chickens globally. To minimise the spread of STEC and antibiotic resistance, future foodborne pathogen prevention and control programmes should prioritise increasing laboratory capacity for the early identification of antibiotic resistance. Full article
(This article belongs to the Special Issue Antibiotics Resistance in Animals and the Environment, 2nd Edition)
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13 pages, 272 KiB  
Article
The Effect of Mitomycin C on Induction of Shiga Toxin Production in Clinical STEC Isolates
by Surangi H. Thilakarathna, Brendon Parsons and Linda Chui
Toxins 2025, 17(6), 267; https://doi.org/10.3390/toxins17060267 - 27 May 2025
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Abstract
Early determination of the Shiga toxin type of Shiga toxin-producing Escherichia coli (STEC) is crucial for guiding STEC-infected patients for proper and timely treatment and patient care. Most diagnostic microbiology laboratories rely on PCR assays to detect the presence of stx1 and/or stx2 [...] Read more.
Early determination of the Shiga toxin type of Shiga toxin-producing Escherichia coli (STEC) is crucial for guiding STEC-infected patients for proper and timely treatment and patient care. Most diagnostic microbiology laboratories rely on PCR assays to detect the presence of stx1 and/or stx2 and enzymatic immunoassays (EIA) to detect the presence of the Shiga toxins 1 and/or 2 in STEC-positive stool samples. Occasionally, the stool samples test positive for STEC by PCR assays but test negative for the presence of Shiga toxins. Insufficient toxin production under laboratory conditions is the main culprit of this discordance. To test whether EIA-based STEC detection could be improved, various clinical STEC strains were treated with mitomycin C, which is a commonly used inducer of Shiga toxin production. A dose-dependent increase in Shiga toxin production, in response to mitomycin C doses of up to 500 ng/mL, was observed without any bactericidal effects. Depending on the serotype, 5–50 times more Shiga toxin 2 was produced than Shiga toxin 1. Shiga toxin production was not induced by the mitomycin C treatment in certain STEC serotypes carrying the toxin subtypes stx1a, stx2a, 2b, 2f, or 2h. This diversity in toxin production indicates that other factors may determine toxin expression in certain STEC strains, which warrant further exploration. Full article
(This article belongs to the Special Issue Multi Methods for Detecting Natural Toxins)
15 pages, 1501 KiB  
Article
Curcumin Reverses Antibiotic Resistance and Downregulates Shiga Toxin Expression in Enterohemorrhagic E. coli
by Martin Zermeño-Ruiz, Mirian Cobos-Vargas, Mauro Donaldo Saucedo-Plascencia, Rafael Cortés-Zárate, Leonardo Hernandez-Hernandez, Teresa Arcelia Garcia-Cobian, Teresa Estrada-Garcia and Araceli Castillo-Romero
Diseases 2025, 13(5), 154; https://doi.org/10.3390/diseases13050154 - 17 May 2025
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Abstract
Background: Enterohemorrhagic Escherichia coli (EHEC) is a considerable public health concern associated with several foodborne outbreaks of bloody diarrhea (BD) and the potentially lethal hemolytic uremic syndrome (HUS), the pathophysiology of which is attributable to the Shiga toxin (Stx) produced by this bacterium. [...] Read more.
Background: Enterohemorrhagic Escherichia coli (EHEC) is a considerable public health concern associated with several foodborne outbreaks of bloody diarrhea (BD) and the potentially lethal hemolytic uremic syndrome (HUS), the pathophysiology of which is attributable to the Shiga toxin (Stx) produced by this bacterium. In most patients, supportive treatment will be sufficient; however, in some cases, antibiotic treatment may be necessary. Most antibiotics are not recommended for EHEC infection treatment, particularly those that kill the bacteria, since this triggers the release of Stx in the body, inducing or worsening HUS. Azithromycin, which prevents the release of Stx and is a weaker inducer of the SOS system, has been successfully used to reduce EHEC shedding. It is necessary to identify compounds that eliminate EHEC without inducing Stx release. The use of natural compounds such as curcumin (CUR), a polyphenol derived from turmeric, has been highlighted as an alternative bactericidal treatment approach. Objective: The objective of this study was to establish the effect of CUR and its interactions with selected antibiotics on resistant EHEC O157/H7/EDL933. Methods: Bacterial cultures were exposed to CUR at three different concentrations (110, 220, and 330 µg/mL) and 1.2% DMSO, and the antimicrobial activity of CUR was assessed by measuring the optical density at 600 nm (OD600). The synergy of CUR and the antibiotics was determined with the FIC method. RT-PCR was performed to determine the expression levels of the blaCTX-M-15, catA1, acrAB-tolC stx2A, and stx2B genes. Results: Our data indicate that CUR did not affect the growth of EHEC, but when combined with the antibiotics, it acted as a bacterial resistance breaker. Synergistic combinations of CUR and cefotaxime or chloramphenicol significantly reduced colony counts. Conclusions: Our findings support the potential of CUR as a sensitizer or in combination therapy against EHEC. Full article
(This article belongs to the Section Infectious Disease)
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