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29 pages, 6672 KiB  
Article
Discovery of a Novel Antimicrobial Peptide from Paenibacillus sp. Na14 with Potent Activity Against Gram-Negative Bacteria and Genomic Insights into Its Biosynthetic Pathway
by Nuttapon Songnaka, Adisorn Ratanaphan, Namfa Sermkaew, Somchai Sawatdee, Sucheewin Krobthong, Chanat Aonbangkhen, Yodying Yingchutrakul and Apichart Atipairin
Antibiotics 2025, 14(8), 805; https://doi.org/10.3390/antibiotics14080805 (registering DOI) - 6 Aug 2025
Abstract
Background/Objectives: Antimicrobial resistance (AMR) contributes to millions of deaths globally each year, creating an urgent need for new therapeutic agents. Antimicrobial peptides (AMPs) have emerged as promising candidates due to their potential to combat AMR pathogens. This study aimed to evaluate the antimicrobial [...] Read more.
Background/Objectives: Antimicrobial resistance (AMR) contributes to millions of deaths globally each year, creating an urgent need for new therapeutic agents. Antimicrobial peptides (AMPs) have emerged as promising candidates due to their potential to combat AMR pathogens. This study aimed to evaluate the antimicrobial activity of an AMP from a soil-derived bacterial isolate against Gram-negative bacteria. Method: Soil bacteria were isolated and screened for antimicrobial activity. The bioactive peptide was purified and determined its structure and antimicrobial efficacy. Genomic analysis was conducted to predict the biosynthetic gene clusters (BGCs) responsible for AMP production. Results: Genomic analysis identified the isolate as Paenibacillus sp. Na14, which exhibited low genomic similarity (61.0%) to other known Paenibacillus species, suggesting it may represent a novel species. The AMP from the Na14 strain exhibited heat stability up to 90 °C for 3 h and retained its activity across a broad pH range from 3 to 11. Structural analysis revealed that the Na14 peptide consisted of 14 amino acid residues, adopting an α-helical structure. This peptide exhibited bactericidal activity at concentrations of 2–4 µg/mL within 6–12 h, and its killing rate was concentration-dependent. The peptide was found to disrupt the bacterial membranes. The Na14 peptide shared 64.29% sequence similarity with brevibacillin 2V, an AMP from Brevibacillus sp., which also belongs to the Paenibacillaceae family. Genomic annotation identified BGCs associated with secondary metabolism, with a particular focus on non-ribosomal peptide synthetase (NRPS) gene clusters. Structural modeling of the predicted NRPS enzymes showed high similarity to known NRPS modules in Brevibacillus species. These genomic findings provide evidence supporting the similarity between the Na14 peptide and brevibacillin 2V. Conclusions: This study highlights the discovery of a novel AMP with potent activity against Gram-negative pathogens and provides new insight into conserved AMP biosynthetic enzymes within the Paenibacillaceae family. Full article
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18 pages, 4635 KiB  
Article
Nylon Affinity Networks Capture and Sequester Two Model Bacteria Spiked in Human Plasma
by Fatema Hashemi, Silvia Cachaco, Rocio Prisby, Weidong Zhou, Gregory Petruncio, Elsa Ronzier, Remi Veneziano, Barbara Birkaya, Alessandra Luchini and Luisa Gregori
Pathogens 2025, 14(8), 778; https://doi.org/10.3390/pathogens14080778 (registering DOI) - 6 Aug 2025
Abstract
Ensuring bacterial safety of blood transfusions remains a critical focus in medicine. We investigated a novel pathogen reduction technology utilizing nylon functionalized with synthetic dyes (nylon affinity networks) to capture and remove bacteria from plasma. In the initial screening process, we spiked phosphate [...] Read more.
Ensuring bacterial safety of blood transfusions remains a critical focus in medicine. We investigated a novel pathogen reduction technology utilizing nylon functionalized with synthetic dyes (nylon affinity networks) to capture and remove bacteria from plasma. In the initial screening process, we spiked phosphate buffer solution (PBS) and human plasma (1 mL each) with 10 or 100 colony forming units (cfu) of either Escherichia coli or Staphylococcus epidermidis, exposed the suspensions to affinity networks and assessed the extent of bacterial reduction using agar plate cultures as the assay output. Nineteen synthetic dyes were tested. Among these, Alcian Blue exhibited the best performance with both bacterial strains in both PBS and plasma. Next, bacterial suspensions of approximately 1 and 2 cfu/mL in 10 and 50 mL, respectively, were treated with Alcian Blue affinity networks in three sequential capture steps. This procedure resulted in complete bacterial depletion, as demonstrated by the lack of bacterial growth in the remaining fraction. The viability of the captured bacteria was confirmed by plating the post-treatment affinity networks on agar. Alcian Blue affinity networks captured and sequestered a few plasma proteins identified by liquid chromatography tandem mass spectrometry. These findings support the potential applicability of nylon affinity networks to enhance transfusion safety, although additional investigations are needed. Full article
(This article belongs to the Section Bacterial Pathogens)
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14 pages, 1033 KiB  
Systematic Review
Resistance of Gram-Negative Bacteria to Cefepime-Enmetazobactam: A Systematic Review
by Matthew E. Falagas, Laura T. Romanos, Dimitrios S. Kontogiannis, Katerina Tsiara and Stylianos A. Kakoullis
Pathogens 2025, 14(8), 777; https://doi.org/10.3390/pathogens14080777 - 6 Aug 2025
Abstract
Cefepime-enmetazobactam is a novel β-lactam/β-lactamase inhibitor combination showing good activity against multidrug-resistant (MDR) Gram-negative bacteria producing a variety of β-lactamases. In this systematic review, we aimed to evaluate the available data on resistance to this drug. We performed a thorough search of four [...] Read more.
Cefepime-enmetazobactam is a novel β-lactam/β-lactamase inhibitor combination showing good activity against multidrug-resistant (MDR) Gram-negative bacteria producing a variety of β-lactamases. In this systematic review, we aimed to evaluate the available data on resistance to this drug. We performed a thorough search of four databases (Embase, PubMed, Scopus, and Web of Science), as well as backward citation searching, to identify studies containing data on resistance to cefepime-enmetazobactam. The data were extracted and analyzed according to the breakpoints established by the European Committee on Antimicrobial Susceptibility Testing (EUCAST) and the Food and Drug Administration (FDA), or the specific breakpoints reported by the authors of the respective studies. Analysis based on the type of lactamases produced by the isolates was also performed. Ten studies reported in vitro susceptibility testing and mechanisms of antimicrobial resistance. The total number of isolates was 15,408. The activity of cefepime-enmetazobactam against β-lactamase-producing isolates was variable. The resistance of the studied extended-spectrum β-lactamase (ESBL)-producing and ampicillin C β-lactamase (AmpC)-producing isolates was low (0–2.8% and 0%, respectively). The resistance was higher among oxacillinase-48 β-lactamase (OXA-48)-producing and Klebsiella pneumoniae carbapenemase (KPC)-producing isolates (3.4–13.2% and 36.7–57.8%, respectively). High resistance was noted among metallo-β-lactamase (MBL)-producing isolates (reaching 87.5% in one study), especially those producing New Delhi metallo-β-lactamase (NDM) and Verona integron-encoded metallo-β-lactamase (VIM), which had the highest rates of resistance. The high activity of cefepime-enmetazobactam against Enterobacterales and selected lactose non-fermenting Gram-negative pathogens, including ESBL-producing and AmpC-producing isolates, makes it a potential carbapenem-sparing agent. The drug should be used after in vitro antimicrobial susceptibility testing in patients with infections caused by OXA-48, KPC, and MBL-producing isolates. Full article
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24 pages, 957 KiB  
Review
Biofilm and Antimicrobial Resistance: Mechanisms, Implications, and Emerging Solutions
by Bharmjeet Singh, Manju Dahiya, Vikram Kumar, Archana Ayyagari, Deepti N. Chaudhari and Jayesh J. Ahire
Microbiol. Res. 2025, 16(8), 183; https://doi.org/10.3390/microbiolres16080183 - 6 Aug 2025
Abstract
Biofilms are a spontaneously formed slimy matrix of extracellular polymeric substances (EPS) enveloping miniature bacterial colonies, which aid in pathogen colonization, shielding the bacteria from antibiotics, as well as imparting them resistance towards the same. Biofilms employ a robust communication mechanism called quorum [...] Read more.
Biofilms are a spontaneously formed slimy matrix of extracellular polymeric substances (EPS) enveloping miniature bacterial colonies, which aid in pathogen colonization, shielding the bacteria from antibiotics, as well as imparting them resistance towards the same. Biofilms employ a robust communication mechanism called quorum sensing that serves to keep their population density constant. What is most significant about biofilms is that they contribute to the development of bacterial virulence by providing protection to pathogenic species, allowing them to colonize the host, and also inhibiting the activities of antimicrobials on them. They grow on animate surfaces (such as on teeth and intestinal mucosa, etc.) and inanimate objects (like catheters, contact lenses, pacemakers, endotracheal devices, intrauterine devices, and stents, etc.) alike. It has been reported that as much as 80% of human infections involve biofilms. Serious implications of biofilms include the necessity of greater concentrations of antibiotics to treat common human infections, even contributing to antimicrobial resistance (AMR), since bacteria embedded within biofilms are protected from the action of potential antibiotics. This review explores various contemporary strategies for controlling biofilms, focusing on their modes of action, mechanisms of drug resistance, and innovative approaches to find a solution in this regard. This review interestingly targets the extracellular polymeric matrix as a highly effective strategy to counteract the potential harm of biofilms since it plays a critical role in biofilm formation and significantly contributes to antimicrobial resistance. Full article
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17 pages, 3205 KiB  
Review
Microbiome–Immune Interaction and Harnessing for Next-Generation Vaccines Against Highly Pathogenic Avian Influenza in Poultry
by Yongming Sang, Samuel N. Nahashon and Richard J. Webby
Vaccines 2025, 13(8), 837; https://doi.org/10.3390/vaccines13080837 (registering DOI) - 6 Aug 2025
Abstract
Highly pathogenic avian influenza (HPAI) remains a persistent threat to global poultry production and public health. Current vaccine platforms show limited cross-clade efficacy and often fail to induce mucosal immunity. Recent advances in microbiome research reveal critical roles for gut commensals in modulating [...] Read more.
Highly pathogenic avian influenza (HPAI) remains a persistent threat to global poultry production and public health. Current vaccine platforms show limited cross-clade efficacy and often fail to induce mucosal immunity. Recent advances in microbiome research reveal critical roles for gut commensals in modulating vaccine-induced immunity, including enhancement of mucosal IgA production, CD8+ T-cell activation, and modulation of systemic immune responses. Engineered commensal bacteria such as Lactococcus lactis, Bacteroides ovatus, Bacillus subtilis, and Staphylococcus epidermidis have emerged as promising live vectors for antigen delivery. Postbiotic and synbiotic strategies further enhance protective efficacy through targeted modulation of the gut microbiota. Additionally, artificial intelligence (AI)-driven tools enable predictive modeling of host–microbiome interactions, antigen design optimization, and early detection of viral antigenic drift. These integrative technologies offer a new framework for mucosal, broadly protective, and field-deployable vaccines for HPAI control. However, species-specific microbiome variation, ecological safety concerns, and scalable manufacturing remain critical challenges. This review synthesizes emerging evidence on microbiome–immune crosstalk, commensal vector platforms, and AI-enhanced vaccine development, emphasizing the urgent need for One Health integration to mitigate zoonotic adaptation and pandemic emergence. Full article
(This article belongs to the Special Issue Veterinary Vaccines and Host Immune Responses)
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19 pages, 1579 KiB  
Article
Plasma-Treated Water Effect on Sporulating Bacillus cereus vs. Non-Sporulating Listeria monocytogenes Biofilm Cell Vitality
by Samantha Nestel, Robert Wagner, Mareike Meister, Thomas Weihe and Uta Schnabel
Appl. Microbiol. 2025, 5(3), 80; https://doi.org/10.3390/applmicrobiol5030080 - 5 Aug 2025
Abstract
Foodborne illness caused by bacterial pathogens is a global health concern and results in millions of infections annually. Therefore, food products typically undergo several processing stages, including sanitation steps, before being distributed in an attempt to remove pathogens. However, many sanitation methods have [...] Read more.
Foodborne illness caused by bacterial pathogens is a global health concern and results in millions of infections annually. Therefore, food products typically undergo several processing stages, including sanitation steps, before being distributed in an attempt to remove pathogens. However, many sanitation methods have compounding effects on the color, texture, flavor, and nutritional quality of the product or do not effectively reduce the pathogens that food can be exposed to. Some bacterial pathogens particularly possess traits and tactics that make them even more difficult to mitigate such as biofilm formation. Non-thermal plasma sanitation techniques, including plasma-treated water (PTW), have proven to be promising methods that significantly reduce pathogenic bacteria that food is exposed to. Published work reveals that PTW can effectively mitigate both gram-positive and gram-negative bacterial biofilms. This study presents a novel analysis of the differences in antimicrobial effects of PTW treatment between biofilm-forming gram-positive bacteria, commonly associated with foodborne illness, that are sporulating (Bacillus cereus) and non-sporulating (Listeria monocytogenes). After treatment with PTW, the results suggest the following hypotheses: (1) that the non-sporulating species experiences less membrane damage but a greater reduction in metabolic activity, leading to a possible viable but non-culturable (VBNC) state, and (2) that the sporulating species undergoes spore formation, which may subsequently convert into vegetative cells over time. PTW treatment on gram-positive bacterial biofilms that persist in food processing environments proves to be effective in reducing the proliferating abilities of the bacteria. However, the variance in PTW’s effects on metabolic activity and cell vitality between sporulating and non-sporulating species suggest that other survival tactics might be induced. This analysis further informs the application of PTW in food processing as an effective sanitation method. Full article
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18 pages, 3033 KiB  
Article
Mathematical Modelling of Upper Room UVGI in UFAD Systems for Enhanced Energy Efficiency and Airborne Disease Control: Applications for COVID-19 and Tuberculosis
by Mohamad Kanaan, Eddie Gazo-Hanna and Semaan Amine
Math. Comput. Appl. 2025, 30(4), 85; https://doi.org/10.3390/mca30040085 (registering DOI) - 5 Aug 2025
Abstract
This study is the first to investigate the performance of ultraviolet germicidal irradiation (UVGI) in underfloor air distribution (UFAD) systems. A simplified mathematical model is developed to predict airborne pathogen transport and inactivation by upper room UVGI in UFAD spaces. The proposed model [...] Read more.
This study is the first to investigate the performance of ultraviolet germicidal irradiation (UVGI) in underfloor air distribution (UFAD) systems. A simplified mathematical model is developed to predict airborne pathogen transport and inactivation by upper room UVGI in UFAD spaces. The proposed model is substantiated for the SARS-CoV-2 virus as a simulated pathogen through a comprehensive computational fluid dynamics methodology validated against published experimental data of upper room UVGI and UFAD flows. Simulations show an 11% decrease in viral concentration within the upper irradiated zone when a 15 W louvered germicidal lamp is utilized. Finally, a case study on Mycobacterium tuberculosis (M. tuberculosis) bacteria is carried out using the validated simplified model to optimize the use of return air and UVGI implementation, ensuring acceptable indoor air quality and enhanced energy efficiency. Results reveal that the UFAD-UVGI system may consume up to 13.6% less energy while keeping the occupants at acceptable levels of M. tuberculosis concentration and UV irradiance when operated with 26% return air and a UVGI output of 72 W. Full article
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17 pages, 1027 KiB  
Review
Chimeric Antigen Receptor Immunotherapy for Infectious Diseases: Current Advances and Future Perspectives
by Maria Kourti, Paschalis Evangelidis, Emmanuel Roilides and Elias Iosifidis
Pathogens 2025, 14(8), 774; https://doi.org/10.3390/pathogens14080774 - 5 Aug 2025
Abstract
Chimeric antigen receptor (CAR)-T immunotherapy has revolutionized the management of patients with relapsed/refractory B-cell hematological malignancies. There is emerging evidence that CAR-engineered cells—not only T cells, but also natural killers and macrophages—might have a crucial role in the treatment of autoimmune disorders and [...] Read more.
Chimeric antigen receptor (CAR)-T immunotherapy has revolutionized the management of patients with relapsed/refractory B-cell hematological malignancies. There is emerging evidence that CAR-engineered cells—not only T cells, but also natural killers and macrophages—might have a crucial role in the treatment of autoimmune disorders and solid tumors. Moreover, given the burden of chronic infectious diseases, the mortality and morbidity of infections in immunocompromised individuals, and the development of multidrug-resistant pathogens, including bacteria, fungi, and mycobacteria, a need for novel and personalized therapeutics in this field is emerging. To this end, the development of CAR cells for the management of chronic infections has been reported. In this literature review, we summarize the ongoing clinical and pre-clinical data about CAR cell products in the field of infectious diseases. Currently, clinical studies on CAR immunotherapy for infections mainly concern human immunodeficiency virus infection treatment, and data regarding other infections largely originate from preclinical in vitro and in vivo models. In the era of personalized medicine, effective and safe therapies for the management of chronic infections and infectious complications in immunocompromised patients are crucial. Full article
(This article belongs to the Special Issue Bacterial Resistance and Novel Therapeutic Approaches)
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24 pages, 3027 KiB  
Article
Resisting the Final Line: Phenotypic Detection of Resistance to Last-Resort Antimicrobials in Gram-Negative Bacteria Isolated from Wild Birds in Northern Italy
by Maria Cristina Rapi, Joel Filipe, Laura Filippone Pavesi, Stefano Raimondi, Maria Filippa Addis, Maria Pia Franciosini and Guido Grilli
Animals 2025, 15(15), 2289; https://doi.org/10.3390/ani15152289 - 5 Aug 2025
Abstract
Antimicrobial resistance (AMR) is a growing global health threat, with wild birds increasingly recognized as potential reservoirs of resistant pathogens and as sentinels of environmental AMR. This study investigated the occurrence and AMR profiles of Gram-negative bacteria isolated from wild birds that died [...] Read more.
Antimicrobial resistance (AMR) is a growing global health threat, with wild birds increasingly recognized as potential reservoirs of resistant pathogens and as sentinels of environmental AMR. This study investigated the occurrence and AMR profiles of Gram-negative bacteria isolated from wild birds that died at the Wildlife Rescue Center in Vanzago, Lombardy, in 2024. Cloacal swabs were collected from 112 birds representing various ecological categories. A total of 157 Gram-negative bacteria were isolated and identified, including clinically relevant genera and species, such as Escherichia coli, Klebsiella pneumoniae, Enterobacter spp., Salmonella spp., Pseudomonas aeruginosa, and Acinetobacter baumannii. Antimicrobial susceptibility testing revealed resistance to first-line and critically important antimicrobials, including those exclusively authorized for human use. Notably, a phenotype compatible with Extended-Spectrum Beta-Lactamase (ESBL) production was detected in four out of ten (40%) K. pneumoniae isolates. In addition, 20 out of the 157 (12.7%) isolated bacteria phenotypically exhibited a resistance profile indicative of AmpC beta-lactamase (AmpC) production, including Enterobacter spp. and P. aeruginosa. Resistance patterns were particularly interesting in birds with carnivorous, scavenging, or migratory-associated behaviors. These findings highlight the role of wild birds in the ecology and dissemination of antimicrobial-resistant bacteria (ARB) and highlight the need for wildlife-based AMR monitoring programs as part of a One Health approach. Full article
(This article belongs to the Section Birds)
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43 pages, 1183 KiB  
Review
Harnessing Legume Productivity in Tropical Farming Systems by Addressing Challenges Posed by Legume Diseases
by Catherine Hazel Aguilar, David Pires, Cris Cortaga, Reynaldo Peja, Maria Angela Cruz, Joanne Langres, Mark Christian Felipe Redillas, Leny Galvez and Mark Angelo Balendres
Nitrogen 2025, 6(3), 65; https://doi.org/10.3390/nitrogen6030065 - 5 Aug 2025
Abstract
Legumes are among the most important crops globally, serving as a major food source for protein and oil. In tropical regions, the cultivation of legumes has expanded significantly due to the increasing demand for food, plant-based products, and sustainable agriculture practices. However, tropical [...] Read more.
Legumes are among the most important crops globally, serving as a major food source for protein and oil. In tropical regions, the cultivation of legumes has expanded significantly due to the increasing demand for food, plant-based products, and sustainable agriculture practices. However, tropical environments pose unique challenges, including high temperatures, erratic rainfall, soil infertility, and a high incidence of pests and diseases. Indeed, legumes are vulnerable to infections caused by bacteria, fungi, oomycetes, viruses, and nematodes. This review highlights the importance of legumes in tropical farming and discusses major diseases affecting productivity and their impact on the economy, environment, and lives of smallholder legume farmers. We emphasize the use of legume genetic resources and breeding, and biotechnology innovations to foster resistance and address the challenges posed by pathogens in legumes. However, an integrated approach that includes other cultivation techniques (e.g., crop rotation, rational fertilization, deep plowing) remains important for the prevention and control of diseases in legume crops. Finally, we highlight the contributions of plant genetic resources to smallholder resilience and food security. Full article
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24 pages, 3176 KiB  
Article
Influence of Seasonality and Pollution on the Presence of Antibiotic Resistance Genes and Potentially Pathogenic Bacteria in a Tropical Urban River
by Kenia Barrantes-Jiménez, Bradd Mendoza-Guido, Eric Morales-Mora, Luis Rivera-Montero, José Montiel-Mora, Luz Chacón-Jiménez, Keilor Rojas-Jiménez and María Arias-Andrés
Antibiotics 2025, 14(8), 798; https://doi.org/10.3390/antibiotics14080798 - 5 Aug 2025
Abstract
Background/Objectives: This study examines how seasonality, pollution, and sample type (water and sediment) influence the presence and distribution of antibiotic resistance genes (ARGs), with a focus on antibiotic resistance genes (ARGs) located on plasmids (the complete set of plasmid-derived sequences, including ARGs) in [...] Read more.
Background/Objectives: This study examines how seasonality, pollution, and sample type (water and sediment) influence the presence and distribution of antibiotic resistance genes (ARGs), with a focus on antibiotic resistance genes (ARGs) located on plasmids (the complete set of plasmid-derived sequences, including ARGs) in a tropical urban river. Methods: Samples were collected from three sites along a pollution gradient in the Virilla River, Costa Rica, during three seasonal campaigns (wet 2021, dry 2022, and wet 2022). ARGs in water and sediment were quantified by qPCR, and metagenomic sequencing was applied to analyze chromosomal and plasmid-associated resistance profiles in sediments. Tobit and linear regression models, along with multivariate ordination, were used to assess spatial and seasonal trends. Results: During the wet season of 2021, the abundance of antibiotic resistance genes (ARGs) such as sul-1, intI-1, and tetA in water samples decreased significantly, likely due to dilution, while intI-1 and tetQ increased in sediments, suggesting particle-bound accumulation. In the wet season 2022, intI-1 remained low in water, qnrS increased, and sediments showed significant increases in tetQ, tetA, and qnrS, along with decreases in sul-1 and sul-2. Metagenomic analysis revealed spatial differences in plasmid-associated ARGs, with the highest abundance at the most polluted site (Site 3). Bacterial taxa also showed spatial differences, with greater plasmidome diversity and a higher representation of potential pathogens in the most contaminated site. Conclusions: Seasonality and pollution gradients jointly shape ARG dynamics in this tropical river. Plasmid-mediated resistance responds rapidly to environmental change and is enriched at polluted sites, while sediments serve as long-term reservoirs. These findings support the use of plasmid-based monitoring for antimicrobial resistance surveillance in aquatic systems. Full article
(This article belongs to the Special Issue Origins and Evolution of Antibiotic Resistance in the Environment)
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11 pages, 972 KiB  
Article
Rapid and Accurate Detection of the Most Common Bee Pathogens; Nosema ceranae, Aspergillus flavus, Paenibacillus larvae and Black Queen Cell Virus
by Simona Marianna Sanzani, Raied Abou Kubaa, Badr-Eddine Jabri, Sabri Ala Eddine Zaidat, Rocco Addante, Naouel Admane and Khaled Djelouah
Insects 2025, 16(8), 810; https://doi.org/10.3390/insects16080810 (registering DOI) - 5 Aug 2025
Abstract
Honey bees are essential pollinators for the ecosystem and food crops. However, their health and survival face threats from both biotic and abiotic stresses. Fungi, microsporidia, and bacteria might significantly contribute to colony losses. Therefore, rapid and sensitive diagnostic tools are crucial for [...] Read more.
Honey bees are essential pollinators for the ecosystem and food crops. However, their health and survival face threats from both biotic and abiotic stresses. Fungi, microsporidia, and bacteria might significantly contribute to colony losses. Therefore, rapid and sensitive diagnostic tools are crucial for effective disease management. In this study, molecular assays were developed to quickly and efficiently detect the main honey bee pathogens: Nosema ceranae, Aspergillus flavus, Paenibacillus larvae, and Black queen cell virus. In this context, new primer pairs were designed for use in quantitative Real-time PCR (qPCR) reactions. Various protocols for extracting total nucleic acids from bee tissues were tested, indicating a CTAB-based protocol as the most efficient and cost-effective. Furthermore, excluding the head of the bee from the extraction, better results were obtained in terms of quantity and purity of extracted nucleic acids. These assays showed high specificity and sensitivity, detecting up to 250 fg of N. ceranae, 25 fg of P. larvae, and 2.5 pg of A. flavus DNA, and 5 pg of BQCV cDNA, without interference from bee DNA. These qPCR assays allowed pathogen detection within 3 h and at early stages of infection, supporting timely and efficient management interventions. Full article
(This article belongs to the Section Insect Behavior and Pathology)
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13 pages, 1769 KiB  
Article
Antimicrobial Photodynamic Activity of the Zn(II) Phthalocyanine RLP068/Cl Versus Antimicrobial-Resistant Priority Pathogens
by Ilaria Baccani, Sara Cuffari, Francesco Giuliani, Gian Maria Rossolini and Simona Pollini
Int. J. Mol. Sci. 2025, 26(15), 7545; https://doi.org/10.3390/ijms26157545 - 5 Aug 2025
Abstract
The emergence and spread of antimicrobial resistance among pathogens are significantly reducing available therapeutic options, highlighting the urgent need for novel and complementary treatment strategies. Antimicrobial photodynamic therapy (aPDT) is a promising alternative approach that can overcome antimicrobial resistance through a multitarget mechanism [...] Read more.
The emergence and spread of antimicrobial resistance among pathogens are significantly reducing available therapeutic options, highlighting the urgent need for novel and complementary treatment strategies. Antimicrobial photodynamic therapy (aPDT) is a promising alternative approach that can overcome antimicrobial resistance through a multitarget mechanism of action, exerting direct bactericidal and fungicidal effects with minimal risk of resistance development. Although aPDT has shown efficacy against a variety of pathogens, data on its activity against large collections of clinical multidrug-resistant strains are still limited. In this study, we assessed the antimicrobial activity of the photosensitizer RLP068/Cl combined with a red light-emitting LED source at 630 nm (Molteni Farmaceutici, Italy) against a large panel of Gram-negative and Gram-positive bacterial strains harboring relevant resistance traits and Candida species. Our results demonstrated the significant microbicidal activity of RLP068/Cl against all of the tested strains regardless of their resistance phenotype, with particularly prominent activity against Gram-positive bacteria (range of bactericidal concentrations 0.05–0.1 µM), which required significantly lower exposure to photosensitizer compared to Candida and Gram-negative species (range 5–20 µM). Overall, these findings support the potential use of RLP068/Cl-mediated aPDT as an effective therapeutic strategy for the management of localized infections caused by MDR organisms, particularly when conventional therapeutic options are limited. Full article
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21 pages, 3536 KiB  
Article
Gold(III) Complexes with Aromatic Cyano-Substituted Bisdithiolate Ligands as Potential Anticancer and Antimicrobial Agents
by Dulce Belo, Sandra Rabaça, Sara G. Fava, Sílvia A. Sousa, Diogo Coelho, Jorge H. Leitão, Teresa Pinheiro, Célia Fernandes and Fernanda Marques
Molecules 2025, 30(15), 3270; https://doi.org/10.3390/molecules30153270 - 4 Aug 2025
Abstract
Cancer and infectious diseases are major causes of global morbidity and mortality stressing the need to find novel drugs with promising dual anticancer and antimicrobial efficacy. Gold complexes have been studied for the past years due to their anticancer properties, with a few [...] Read more.
Cancer and infectious diseases are major causes of global morbidity and mortality stressing the need to find novel drugs with promising dual anticancer and antimicrobial efficacy. Gold complexes have been studied for the past years due to their anticancer properties, with a few of them displaying antimicrobial properties, which support their pharmacological interest. Within this scope, we investigated six gold bisdithiolate complexes [Au (bdt)2] (1), [Au (dcbdt)2] (2), [Au (3-cbdt)2] (3), [Au (4-cbdt)2] (4), [Au (pdt)2] (5) and [Au (dcdmp)2] (6), and) against the ovarian cancer cell lines A2780 and A2780cisR, the Gram-positive bacteria Staphylococcus aureus Newman, the Gram-negative bacteria Escherichia coli ATCC25922 and Burkholderia contaminans IST408, and the pathogenic yeasts Candida glabrata CBS138 and Candida albicans SC5134. Complexes 2 and 6, with ligands containing aromatic pyrazine or phenyl rings, substituted with two cyanonitrile groups, showed after 24 h of incubation high anticancer activities against A2780 ovarian cancer cells (IC50~5 µM), being also able to overcome cisplatin resistance in A2780cisR cells. Both complexes induced the formation of ROS, activated caspase-3/7, and induced necrosis (LDH release) in a dose-dependent way, in a greater extent in the case of 6. Among the bacterial and fungal strains tested, only complex 6 presented antimicrobial activity against S. aureus Newman, indicating that this complex is a potential novel anticancer and antibacterial agent. These results delve into the structure-activity relationship of the complexes, considering molecular alterations such as replacing a phenyl group for a pyrazine group, and the inclusion of one or two cyanonitrile appendage groups, and their effects on biological activity. Overall, both complexes were found to be promising leads for the development of future anticancer drugs against low sensitive or cisplatin resistant tumors. Full article
(This article belongs to the Special Issue 10th Anniversary of the Bioorganic Chemistry Section of Molecules)
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17 pages, 6882 KiB  
Article
Development and Evaluation of a Solar Milk Pasteurizer for the Savanna Ecological Zones of West Africa
by Iddrisu Ibrahim, Paul Tengey, Kelci Mikayla Lawrence, Joseph Atia Ayariga, Fortune Akabanda, Grace Yawa Aduve, Junhuan Xu, Robertson K. Boakai, Olufemi S. Ajayi and James Owusu-Kwarteng
Solar 2025, 5(3), 38; https://doi.org/10.3390/solar5030038 - 4 Aug 2025
Abstract
In many developing African countries, milk safety is often managed through traditional methods such as fermentation or boiling over firewood. While these approaches reduce some microbial risks, they present critical limitations. Firewood dependency contributes to deforestation, depletion of agricultural residues, and loss of [...] Read more.
In many developing African countries, milk safety is often managed through traditional methods such as fermentation or boiling over firewood. While these approaches reduce some microbial risks, they present critical limitations. Firewood dependency contributes to deforestation, depletion of agricultural residues, and loss of soil fertility, which, in turn, compromise environmental health and food security. Solar pasteurization provides a reliable and sustainable method for thermally inactivating pathogenic microorganisms in milk and other perishable foods at sub-boiling temperatures, preserving its nutritional quality. This study aimed to evaluate the thermal and microbial performance of a low-cost solar milk pasteurization system, hypothesized to effectively reduce microbial contaminants and retain milk quality under natural sunlight. The system was constructed using locally available materials and tailored to the climatic conditions of the Savanna ecological zone in West Africa. A flat-plate glass solar collector was integrated with a 0.15 cm thick stainless steel cylindrical milk vat, featuring a 2.2 cm hot water jacket and 0.5 cm thick aluminum foil insulation. The system was tested in Navrongo, Ghana, under ambient temperatures ranging from 30 °C to 43 °C. The pasteurizer successfully processed up to 8 L of milk per batch, achieving a maximum milk temperature of 74 °C by 14:00 GMT. Microbial analysis revealed a significant reduction in bacterial load, from 6.6 × 106 CFU/mL to 1.0 × 102 CFU/mL, with complete elimination of coliforms. These results confirmed the device’s effectiveness in achieving safe pasteurization levels. The findings demonstrate that this locally built solar pasteurization system is a viable and cost-effective solution for improving milk safety in arid, electricity-limited regions. Its potential scalability also opens avenues for rural entrepreneurship in solar-powered food and water treatment technologies. Full article
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