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15 pages, 2327 KiB  
Article
The Novel Disease Vicia unijuga Caused by Colletotrichum tofieldiae in China: Implications for Host Growth, Photosynthesis, and Nutritional Quality
by Tong-Tong Wang, Hang Li and Yan-Zhong Li
J. Fungi 2025, 11(8), 567; https://doi.org/10.3390/jof11080567 - 29 Jul 2025
Viewed by 123
Abstract
Vicia unijuga, an important forage legume on China’s Qinghai–Tibetan Plateau, exhibited dark-brown sunken lesions on their stems at the Qingyang Experimental Station of Lanzhou University. The fungus isolated from the diseased tissues was identified as Colletotrichum tofieldiae via a multi-locus phylogeny (ITS- [...] Read more.
Vicia unijuga, an important forage legume on China’s Qinghai–Tibetan Plateau, exhibited dark-brown sunken lesions on their stems at the Qingyang Experimental Station of Lanzhou University. The fungus isolated from the diseased tissues was identified as Colletotrichum tofieldiae via a multi-locus phylogeny (ITS-ACT-Tub2-CHS-1-GADPH-HIS3). The pathogenicity was confirmed by Koch’s postulates. The inoculated plants showed significantly reduced (p < 0.05) growth parameters (height, root length, and biomass), photosynthetic indices (net rate, transpiration, and stomatal conductance), and nutritional quality (crude protein, crude fat, crude ash, and crude fiber) compared to the controls. C. tofieldiae additionally infected six legume species (V. sativa, Medicago sativa, Onobrychis viciifolia, Astragalus adsurgens, Trifolium pratense, and T. repens). Optimal in vitro growth occurred on oatmeal agar (mycelium) and cornmeal agar (spores), with D-sucrose and D-peptone as the best carbon and nitrogen sources. This first report of C. tofieldiae causing V. unijuga anthracnose advances the understanding of legume anthracnose pathogens. Full article
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19 pages, 2130 KiB  
Article
Isolation of ESBL-Producing Enterobacteriaceae in Food of Animal and Plant Origin: Genomic Analysis and Implications for Food Safety
by Rosa Fraccalvieri, Stefano Castellana, Angelica Bianco, Laura Maria Difato, Loredana Capozzi, Laura Del Sambro, Adelia Donatiello, Domenico Pugliese, Maria Tempesta, Antonio Parisi and Marta Caruso
Microorganisms 2025, 13(8), 1770; https://doi.org/10.3390/microorganisms13081770 - 29 Jul 2025
Viewed by 210
Abstract
Background: The spread of ESBL-producing Enterobacteriaceae (ESBL-PE) strains in food poses a potential risk to human health. The aim of the study was to determine the occurrence of ESBL-PE and to investigate their distribution on foods. Methods: A total of 1000 food [...] Read more.
Background: The spread of ESBL-producing Enterobacteriaceae (ESBL-PE) strains in food poses a potential risk to human health. The aim of the study was to determine the occurrence of ESBL-PE and to investigate their distribution on foods. Methods: A total of 1000 food samples, including both raw and ready-to-eat products, was analyzed for the presence of ESBL-producing Enterobacteriaceae using chromogenic selective agar. Antibiotic resistance in the isolated strains was assessed using conventional methods, while whole-genome sequencing was employed to predict antimicrobial resistance and virulence genes. Results: The overall occurrence of ESBL-PE strains was 2.8%, with the highest contamination in raw meat samples (10%). A total of 31 multidrug-resistant (MDR) strains was isolated, mainly Escherichia coli, followed by Klebsiella pneumoniae, Salmonella enterica, and Enterobacter hormaechei. All strains exhibited high levels of resistance to at least four different β-lactam antibiotics, as well as to other antimicrobial classes including sulfonamides, tetracyclines, aminoglycosides, and quinolones. Whole-genome sequencing identified 63 antimicrobial resistance genes, with blaCTX-M being the most prevalent ESBL gene. Twenty-eight (90%) isolates carried Inc plasmids, known vectors of multiple antimicrobial resistance genes, including those associated with ESBLs. Furthermore, several virulence genes were identified. Conclusions: The contamination of food with ESBL-PE represents a potential public health risk, underscoring the importance of the implementation of genomic surveillance to monitor and control the spread of antimicrobial resistance. Full article
(This article belongs to the Special Issue Food Microorganisms and Genomics, 2nd Edition)
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18 pages, 2560 KiB  
Article
In Vitro Insights into the Anti-Biofilm Potential of Salmonella Infantis Phages
by Jan Torres-Boncompte, María Sanz-Zapata, Josep Garcia-Llorens, José M. Soriano, Pablo Catalá-Gregori and Sandra Sevilla-Navarro
Antibiotics 2025, 14(8), 744; https://doi.org/10.3390/antibiotics14080744 - 24 Jul 2025
Viewed by 366
Abstract
Background/Objectives: As bacteriophage-based strategies to control bacterial pathogens continue to gain momentum, phage therapy is increasingly being explored across various fields. In the poultry industry, efforts to minimize the public health impact of Salmonella have spurred growing interest in phage applications, particularly [...] Read more.
Background/Objectives: As bacteriophage-based strategies to control bacterial pathogens continue to gain momentum, phage therapy is increasingly being explored across various fields. In the poultry industry, efforts to minimize the public health impact of Salmonella have spurred growing interest in phage applications, particularly as prophylactic and disinfecting agents. Although the disinfecting potential of bacteriophages has been recognized, in-depth studies examining their efficacy under varying environmental conditions remain limited. This study focused on evaluating the effectiveness of bacteriophages as disinfecting agents against biofilm-forming Salmonella Infantis under different environments. Methods: A comprehensive screening of biofilm-producing strains was conducted using Congo Red Agar and 96-well plate assays. Two strains with distinct biofilm-forming capacities were selected for further analysis under different environmental conditions: aerobic and microaerobic atmospheres at both 25 °C and 37 °C. The resulting biofilms were then treated with four phage preparations: three individual phages and one phage cocktail. Biofilm reduction was assessed by measuring optical density and CFU/well. Additionally, scanning electron microscopy was used to visualize both untreated and phage-treated biofilms. Results: The results demonstrated that all S. Infantis strains were capable of forming biofilms (21/21). All three phage candidates exhibited biofilm-disrupting activity and were able to lyse biofilm-embedded Salmonella cells. Notably, the lytic efficacy of the phages varied depending on environmental conditions, highlighting the importance of thorough phage characterization prior to application. Conclusions: These findings underscore that the effectiveness of bacteriophages as surface disinfectants can be significantly compromised if inappropriate phages are used, especially in the presence of biofilms. Full article
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25 pages, 8725 KiB  
Article
Novel 3D-Printed Replica Plate Device Ensures High-Throughput Antibacterial Screening of Halophilic Bacteria
by Kaloyan Berberov, Nikolina Atanasova, Nikolay Krumov, Boryana Yakimova, Irina Lazarkevich, Stephan Engibarov, Tsvetozara Damyanova, Ivanka Boyadzhieva and Lyudmila Kabaivanova
Mar. Drugs 2025, 23(8), 295; https://doi.org/10.3390/md23080295 - 23 Jul 2025
Viewed by 218
Abstract
Antibiotic resistance is one of the most significant public health issues today. As a consequence, there is an urgent need for novel classes of antibiotics. This necessitates the development of highly efficient screening methods for the rapid identification of antibiotic-producing bacteria. Here, we [...] Read more.
Antibiotic resistance is one of the most significant public health issues today. As a consequence, there is an urgent need for novel classes of antibiotics. This necessitates the development of highly efficient screening methods for the rapid identification of antibiotic-producing bacteria. Here, we describe a new method for high-throughput screening of antimicrobial compounds (AMC) producing halophilic bacteria. Our methodology used a newly designed 3D-printed Petri plate replicator used for drop deposition and colony replication. We employed this device in combination with a modified agar overlay assay to screen more than 7400 bacterial colonies. A total of 54 potential AMC producers were discovered at a success rate of 0.7%. Although 40% of them lost their antibacterial activity during the secondary screening, 22 strains retained inhibitory activity and were able to suppress the growth of one or more safe relatives of the ESKAPE group pathogens. The ethyl acetate extract from the most potent strain, Virgibacillus salarius POTR191, demonstrated moderate antibacterial activity against Enterococcus faecalis, Acinetobacter baumanii, and Staphylococcus epidermidis with minimal inhibitory concentrations of 128 μg/mL, 128 μg/mL, and 512 μg/mL, respectively. We propose that our replica plate assay could be used for target-based antimicrobial screening of various extremophilic bacteria. Full article
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23 pages, 8387 KiB  
Article
Solvent Fractionation of Polygonum cuspidatum Sieb. et Zucc. for Antioxidant, Biological Activity, and Chromatographic Characterization
by Yuchen Cheng, Yuri Kang and Woonjung Kim
Int. J. Mol. Sci. 2025, 26(14), 7011; https://doi.org/10.3390/ijms26147011 - 21 Jul 2025
Viewed by 266
Abstract
This study investigated the natural bioactive compounds in Polygonum cuspidatum Sieb. et Zucc. (P. cuspidatum) by fractionating a 70% ethanol extract using n-hexane, chloroform, ethyl acetate, n-butanol, and water. The total polyphenol and flavonoid contents of each fraction were [...] Read more.
This study investigated the natural bioactive compounds in Polygonum cuspidatum Sieb. et Zucc. (P. cuspidatum) by fractionating a 70% ethanol extract using n-hexane, chloroform, ethyl acetate, n-butanol, and water. The total polyphenol and flavonoid contents of each fraction were determined, and their antioxidant activities were evaluated using DPPH, ABTS, and FRAP assays. Additionally, the anti-diabetic potential was assessed via α-glucosidase inhibitory activity, while anti-obesity activity was evaluated using lipase inhibitory activity. The fractions were also tested for tyrosinase and elastase inhibitory activities to assess their skin-whitening and anti-wrinkle potential, and their antibacterial activity against Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa was determined using the agar diffusion method. Finally, bioactive compounds were identified and quantified using HPLC and GC–MSD. The results showed that the ethyl acetate fraction possessed the highest total polyphenol content (0.53 ± 0.01 g GAE/g) and total flavonoid content (0.19 ± 0.02 g QE/g). It also exhibited strong antioxidant activity, with the lowest DPPH radical scavenging IC50 (0.01 ± 0.00 mg/mL), ABTS radical scavenging IC50 (0.06 ± 0.00 mg/mL), and the highest FRAP value (6.02 ± 0.30 mM Fe2+/mg). Moreover, it demonstrated potent enzyme inhibitory activities, including tyrosinase inhibitory activity (67.78 ± 2.50%), elastase inhibitory activity (83.84 ± 1.64%), α-glucosidase inhibitory activity (65.14 ± 10.29%), and lipase inhibitory activity (85.79 ± 1.04%). In the antibacterial activity, the ethyl acetate fraction produced a clear inhibitory zone of 19.50 mm against Staphylococcus aureus, indicating notable antibacterial activity. HPLC-PDA and GC–MSD analyses identified tannic acid and emodin as the major bioactive constituents. These findings suggest that the ethyl acetate fraction of P. cuspidatum extract, rich in polyphenol and flavonoid compounds, is a promising natural source of bioactive ingredients for applications in the food, pharmaceutical, and cosmetic industries. Further research is needed to explore its mechanisms and therapeutic applications. Full article
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23 pages, 4894 KiB  
Article
Evaluating Copper-Induced Oxidative Stress in Germinating Wheat Seeds Using Laser Photoacoustic Spectroscopy and EPR Techniques
by Mioara Petrus, Cristina Popa, Ana-Maria Bratu, Alexandra Camelia Joita and Vasile Bercu
Toxics 2025, 13(7), 604; https://doi.org/10.3390/toxics13070604 - 18 Jul 2025
Viewed by 367
Abstract
Copper is an essential micronutrient for plants, but excessive levels can induce toxicity and impair physiological functions. This study evaluates the toxic effects of copper sulfate (CuSO4) on the germination of common wheat (Triticum aestivum), with emphasis on the [...] Read more.
Copper is an essential micronutrient for plants, but excessive levels can induce toxicity and impair physiological functions. This study evaluates the toxic effects of copper sulfate (CuSO4) on the germination of common wheat (Triticum aestivum), with emphasis on the gas emission dynamics and oxidative stress biomarkers. Seeds were germinated in agar and exposed to CuSO4 at concentrations of 1 µM, 100 µM, 1 mM, and 10 mM; distilled water served as the control. Ethylene and ammonia emissions were quantified using CO2 laser photoacoustic spectroscopy, while electron paramagnetic resonance (EPR) spectroscopy was employed to detect free radicals and Cu2+ complexes. Exposure to Cu concentrations ≥ 1 mM significantly inhibited germination and biomass accumulation. Enhanced ethylene and ammonia emissions, particularly at 10 mM, indicated stress-related metabolic responses. The EPR spectra confirmed the presence of semiquinone radicals and Cu2+ complexes under higher Cu levels. These results demonstrate that photoacoustic and EPR techniques are effective tools for the early detection of metal-induced phytotoxicity and offer a non-invasive approach to environmental toxicity screening and plant stress assessment. Full article
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23 pages, 6606 KiB  
Article
Clove Oil-Based Nanoemulsion Containing Amphotericin B as a Therapeutic Approach to Combat Fungal Infections
by Marcel Lucas de Almeida, Ana Paula dos Santos Matos, Veronica da Silva Cardoso, Tatielle do Nascimento, Ralph Santos-Oliveira, Leandro Machado Rocha, Francisco Paiva Machado, Franklin Chimaobi Kenechukwu, Alane Beatriz Vermelho and Eduardo Ricci-Júnior
Pharmaceutics 2025, 17(7), 925; https://doi.org/10.3390/pharmaceutics17070925 - 17 Jul 2025
Viewed by 383
Abstract
Background/Objectives: Candidiasis, primarily caused by Candida albicans, and sporotrichosis, mainly caused by Sporothrix schenckii, are skin fungal infections that pose serious threats to global health. The Candida auris is a great concern in immunocompromised individuals, and while Sporothrix brasiliensis cause sporotrichosis, [...] Read more.
Background/Objectives: Candidiasis, primarily caused by Candida albicans, and sporotrichosis, mainly caused by Sporothrix schenckii, are skin fungal infections that pose serious threats to global health. The Candida auris is a great concern in immunocompromised individuals, and while Sporothrix brasiliensis cause sporotrichosis, an infection commonly found in cats, this disease can be transmitted to humans through scratches or bites. Existing treatments for these fungal infections often cause problems related to resistance and significant side effects. Consequently, development of alternative therapeutic approaches such as nanotechnology-based topical lipid-based formulations is interesting. Thus, the objectives of this study were to prepare clove oil (CO)-in-water nanoemulsions (NEs) containing amphotericin B (AmB) and characterize them with respect to stability, release profile, and in vitro cytotoxic activity against Candida and Sporothrix strains. As a future alternative for the treatment of fungal skin diseases. Methods: Chemical analysis of clove oil was obtained by GC-MS. The NEs were produced using an ultrasound (sonicator) method with varying proportions of CO, Pluronic® F-127, and AmB. The NEs were characterized by droplet size, morphology, stability and in vitro release profile. The antifungal and cytotoxic activity against C. albicans, C. auris, S. schenckii, and S. brasiliensis were ascertained employing agar diffusion and colorimetric MTT assay methods. A checkerboard assay was carried out using clove oil and amphotericin B against C. auris. Results: Eugenol was the major compound identified in CO at a concentration of 80.09%. AmB-loaded NEs exhibited particle sizes smaller than 50 nm and a polydispersity index below 0.25. The optimal Ne (NEMLB-05) remained stable after 150 days of storage at 4 °C. It exhibited rapid release within the first 24 h, followed by a slow and controlled release up to 96 h. NEMLB-05 more effectively inhibited C. auris compared to free AmB and also demonstrated greater activity against C. albicans, S. schenckii, and S. brasiliensis. Clove oil and amphotericin B presented synergism inhibiting the growth of C. auris. Conclusions: The selected CO-in-water NEs containing AmB demonstrated promising potential as a topical therapeutic alternative for treating fungal infections. Full article
(This article belongs to the Special Issue Nanotechnology in the Treatment of Neglected Parasitic Diseases)
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14 pages, 4862 KiB  
Article
Gastrointestinal Parasitic Infections in Macaca fascicularis in Northeast Thailand: A One Health Perspective on Zoonotic Risks
by Teputid Kuasit, Manachai Yingklang, Penchom Janwan, Wanchai Maleewong, Weerachai Saijuntha, Siriporn Kuanamon and Tongjit Thanchomnang
Animals 2025, 15(14), 2112; https://doi.org/10.3390/ani15142112 - 17 Jul 2025
Viewed by 783
Abstract
Gastrointestinal (GI) parasitic infections in non-human primates are of growing concern due to their implications for both veterinary and public health. Long-tailed macaques (Macaca fascicularis), commonly found in peri-urban and temple environments in Southeast Asia, may act as reservoirs for zoonotic [...] Read more.
Gastrointestinal (GI) parasitic infections in non-human primates are of growing concern due to their implications for both veterinary and public health. Long-tailed macaques (Macaca fascicularis), commonly found in peri-urban and temple environments in Southeast Asia, may act as reservoirs for zoonotic parasites, posing risks to humans and domestic animals. This study investigated the prevalence and species diversity of GI parasites in free-ranging macaques from four provinces in Northeast Thailand (Loei, Khon Kaen, Bueng Kan, and Sisaket). A cross-sectional study was conducted between April and May 2025. A total of 445 fecal samples were examined using two parasitological techniques: agar plate culture (APC) and the formalin–ethyl acetate concentration technique (FECT). The overall prevalence of parasitic infection was 86.5%, with Strongyloides sp. (65.2%) as the most prevalent helminth and Balantioides coli-like (29.5%) and Entamoeba histolytica-like (28.8%) as the predominant protozoa. Other parasites identified included helminths (Trichuris sp., Ascaris sp.) and protozoa (Blastocystis sp., Iodamoeba bütschlii, Entamoeba coli, and Chilomastix mesnili). Mixed infections were frequently observed, with both helminths and protozoa co-occurring in 37.3% of cases. The high infection rates and parasite diversity reflect substantial environmental contamination and sustained transmission cycles. These findings underscore the importance of integrated surveillance in wildlife populations and the need for One Health-based approaches to minimize zoonotic transmission risks at the human–animal–environment interface. Full article
(This article belongs to the Section Wildlife)
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9 pages, 1253 KiB  
Proceeding Paper
Effect of Far-UVC and Violet Irradiation on the Microbial Contamination of Spinach Leaves and Their Vitamin C and Chlorophyll Contents
by Alexander Gerdt, Anna-Maria Gierke, Petra Vatter and Martin Hessling
Biol. Life Sci. Forum 2025, 47(1), 1; https://doi.org/10.3390/blsf2025047001 - 16 Jul 2025
Viewed by 192
Abstract
Microbial contamination of food can lead to faster spoilage and infections. Therefore, disinfection processes are required that have a low detrimental effect on the nutritional content. Concerning radiation disinfection, two spectral ranges have recently become important. The Far-UVC spectral range, with a wavelength [...] Read more.
Microbial contamination of food can lead to faster spoilage and infections. Therefore, disinfection processes are required that have a low detrimental effect on the nutritional content. Concerning radiation disinfection, two spectral ranges have recently become important. The Far-UVC spectral range, with a wavelength below 230 nm and visible violet light. In this study, leaf spinach was used to investigate the extent to which these radiations inactivate Escherichia coli, but also to determine if the vitamin C or chlorophyll content was reduced. Frozen spinach leaves (Spinacia oleracea) were contaminated with E. coli × pGLO and irradiated with either a 222 nm krypton chloride lamp or 405 nm LEDs. The achieved bacterial reduction was determined by plating the irradiated samples on agar plates and subsequent colony counting. The vitamin C concentration was determined by means of redox titration, and the concentrations of chlorophyll a and chlorophyll b were determined using spectrometry. Both irradiations exhibited a strong antimicrobial impact on E. coli. The average log reduction doses were about 19 mJ/cm2 (222 nm) and 87 J/cm2 (405 nm), respectively. The vitamin C concentration decreased by 30% (222 nm) or 20% (405 nm), and the chlorophyll concentrations decreased by about 25%. Both irradiation approaches are able to substantially reduce microorganisms on spinach leaves by two orders of magnitude, but this is associated with a reduction in the nutrient content. Full article
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26 pages, 2712 KiB  
Article
[1,3]Thiazolo[3,2-b][1,2,4]triazolium Salts as Effective Antimicrobial Agents: Synthesis, Biological Activity Evaluation, and Molecular Docking Studies
by Mykhailo Slivka, Boris Sharga, Daryna Pylypiv, Hanna Aleksyk, Nataliya Korol, Maksym Fizer, Olena I. Fedurcya, Oleksandr G. Pshenychnyi and Ruslan Mariychuk
Int. J. Mol. Sci. 2025, 26(14), 6845; https://doi.org/10.3390/ijms26146845 - 16 Jul 2025
Viewed by 379
Abstract
This study focuses on the search for new effective synthetic antimicrobial compounds as a tool against the widespread presence of microorganisms resistant to existing drugs. Five derivatives of [1,3]thiazolo[3,2-b][1,2,4]triazoles were synthesized using an accessible protocol based on electrophilic heterocyclization and were characterized using [...] Read more.
This study focuses on the search for new effective synthetic antimicrobial compounds as a tool against the widespread presence of microorganisms resistant to existing drugs. Five derivatives of [1,3]thiazolo[3,2-b][1,2,4]triazoles were synthesized using an accessible protocol based on electrophilic heterocyclization and were characterized using infrared (FTIR) and nuclear magnetic resonance (NMR) spectroscopies, and their in vitro antimicrobial and antifungal activities were evaluated using the agar plate diffusion method and the microdilution plate procedure. Both antibacterial (Gram-positive and Gram-negative) and antifungal activities were found for the examined samples. The minimum inhibitory concentration (MIC) varied from 0.97 to 250 µg/mL, and the minimum bactericidal concentration (MBC) from 1.95 to 500 µg/mL. Compound 2a showed good antifungal action against Candida albicans and Saccharomyces cerevisiae with minimum fungicidal concentration (MFC) 125 and MIC 31.25 µg/mL. The molecular docking revealed that the 2-heptyl-3-phenyl-6,6-trimethyl-5,6-dihydro-3H-[1,3]thiazolo[3,2-b][1,2,4]triazol-7-ium cation stands out as a highly promising candidate for further investigation due to a wide range of interactions, including conventional hydrogen bonds, π–σ, π–π T-shaped, and hydrophobic alkyl interactions. The synthesis and preliminary evaluation of [1,3]thiazolo[3,2-b][1,2,4]triazoles yielded promising antimicrobial and antifungal candidates. The diverse interaction profile of the 2-heptyl derivative salt allows this compound’s selection for further biological studies. Full article
(This article belongs to the Section Materials Science)
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19 pages, 12207 KiB  
Case Report
Dermatophytoses Caused by Trichophyton indotineae: The First Case Reports in Malaysia and the Global Epidemiology (2018–2025)
by Yi Xian Er, Kin Fon Leong, Henry Boon Bee Foong, Anis Amirah Abdul Halim, Jing Shun Kok, Nan Jiun Yap, Yuong Chin Tan, Sun Tee Tay and Yvonne Ai-Lian Lim
J. Fungi 2025, 11(7), 523; https://doi.org/10.3390/jof11070523 - 15 Jul 2025
Viewed by 533
Abstract
Trichophyton indotineae is emerging globally from its origin in India, presenting with a terbinafine resistance and causing significant clinical burden. We report herein the first four confirmed cases of T. indotineae dermatophytoses in Malaysia, which were diagnosed based on the microscopic examination of [...] Read more.
Trichophyton indotineae is emerging globally from its origin in India, presenting with a terbinafine resistance and causing significant clinical burden. We report herein the first four confirmed cases of T. indotineae dermatophytoses in Malaysia, which were diagnosed based on the microscopic examination of skin scrapings using potassium hydroxide (KOH) wet mount, followed by confirmation via culture and Internal Transcribed Spacer (ITS1) sequencing. In contrast to conventional Trichophyton infections, T. indotineae dermatophytoses demonstrate extensive cutaneous involvement and marked inflammation with erythematous lesions. All cases exhibited a chronic course lasting more than three months, with evidence of person-to-person transmission. Although one patient reported a travel to Singapore, three had no recent travel history, suggesting possible local transmission. The isolates produced characteristic white, cottony colonies with radial mycelial growth on Mycosel agar after incubation at 30 °C for four days. Three patients responded well to oral itraconazole (200 mg daily), with reduced inflammation and erythematous lesions observed two weeks after treatment initiation. The occurrence of T. indotineae particularly among patients without a travel history, suggests a potential endemic establishment. This fungal pathogen warrants consideration in cases of extensive or recalcitrant dermatophytoses. Further investigations into the diagnostic methods, antifungal susceptibility profiles, and epidemiological risk factors of Malaysian strains are warranted to enhance clinical management and inform public health interventions. Full article
(This article belongs to the Section Fungal Pathogenesis and Disease Control)
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20 pages, 3297 KiB  
Article
Streptococcus equi subsp. zooepidemicus Supernatant Containing Streptolysin S Alters the Equine Nasal and Vaginal Mucosa, Modulating Equine Herpesvirus 1, 3 and 4 Infections
by Eslam Mohamed, Jolien Van Cleemput, Burak Şahin, Wim Van den Broeck, Filip Boyen and Hans Nauwynck
Viruses 2025, 17(7), 980; https://doi.org/10.3390/v17070980 - 14 Jul 2025
Viewed by 398
Abstract
The equine respiratory and reproductive tract microbiomes are complex and subject to constant fluctuations. Among the microbial inhabitants, Streptococcus equi subsp. zooepidemicus (SEZ) is recognized as the dominant bacterium. It is an opportunistic pathogen that may occasionally lead to various types of infections. [...] Read more.
The equine respiratory and reproductive tract microbiomes are complex and subject to constant fluctuations. Among the microbial inhabitants, Streptococcus equi subsp. zooepidemicus (SEZ) is recognized as the dominant bacterium. It is an opportunistic pathogen that may occasionally lead to various types of infections. A key virulence factor of SEZ is the streptolysin S (SLS) toxin, which is responsible for the characteristic β-hemolysis on blood agar and tissue damage. Viruses and bacteria may interact and aggravate lesions and disease. This study aimed to evaluate the impact of an SLS-containing supernatant from SEZ on the nasal and vaginal mucosa and the subsequent replication of equine herpesviruses. The SLS-containing supernatant was prepared, and three 10-fold dilutions (optical density “OD” 10−2, 10−3, 10−4) were applied to equine nasal and vaginal explants. Untreated and EGTA-treated explants served as controls. Epithelial integrity was assessed by measuring the thickness and intercellular spaces. Nasal explants were inoculated with EHV-1 and EHV-4, while vaginal explants received EHV-1 and EHV-3. Viral replication was estimated via immunofluorescence staining and confocal microscopy. SLS-containing supernatants 10−2 and 10−3 compromised epithelial integrity. Viral replication increased in explants treated with SLS 10−3, demonstrating SLS’s damaging effects on the epithelium, facilitating equine herpesvirus replication. Full article
(This article belongs to the Section Animal Viruses)
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13 pages, 3978 KiB  
Article
Agar–Agar Gels Carrying Curative and Preventive Agents Against Helminths: An In Vitro Compatibility Evaluation
by Izaro Zubiría, Inês Abreu, David Boso, Gustavo Pérez, Cristiana Cazapal, Rita Sánchez-Andrade, María Sol Arias, Adolfo Paz-Silva, José Ángel Hernández and Mercedes Camiña
Gels 2025, 11(7), 542; https://doi.org/10.3390/gels11070542 - 12 Jul 2025
Viewed by 241
Abstract
The global market size of animal parasiticides was valued at USD 12.9 billion in 2024. Animal deworming only results in temporary cures with little to no preventive effects; therefore, a strategy that combines animal deworming with prevention is essential in improving the control [...] Read more.
The global market size of animal parasiticides was valued at USD 12.9 billion in 2024. Animal deworming only results in temporary cures with little to no preventive effects; therefore, a strategy that combines animal deworming with prevention is essential in improving the control of helminths. The effectiveness of co-administrating curative and preventive agents and their compatibility were considered based on the parasitophagous fungus Mucor circinelloides, which was developed in edible agar–agar (red seaweed)-carrying dewormers. Accordingly, Petri dishes were prepared with either a biopolymer alone (control, G-C) or with the anthelmintic piperazine (550, 1102, 2210, and 5500 mg/plate) or levamisole (37.5, 75, 150, and 300 mg/plate) and were used to culture the fungus Mucor circinelloides. Strong fungal growth and high numbers of spores were observed in the presence of the anthelmintics. No differences were measured between the control plates and those containing parasiticide drugs. Similar mycelial growth patterns and sporogenesis rates were recorded for different amounts of each anthelmintic. In conclusion, this novel formulation based on biopolymers containing anthelmintics and enriched with the parasitophagous fungus represents a highly promising tool to consider for jointly deworming animals and minimizing the risks of helminth infection. Further studies are in progress to confirm these in vitro results. Full article
(This article belongs to the Special Issue Advances in Functional Hydrogels and Their Applications)
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12 pages, 1380 KiB  
Article
Halicin: A New Approach to Antibacterial Therapy, a Promising Avenue for the Post-Antibiotic Era
by Imane El Belghiti, Omayma Hammani, Fatima Moustaoui, Mohamed Aghrouch, Zohra Lemkhente, Fatima Boubrik and Ahmed Belmouden
Antibiotics 2025, 14(7), 698; https://doi.org/10.3390/antibiotics14070698 - 11 Jul 2025
Viewed by 629
Abstract
Background: The global spread of antibiotic-resistant bacteria presents a major public health challenge and necessitates the development of innovative antimicrobial agents. Artificial intelligence (AI)-driven drug discovery has recently enabled the repurposing of existing compounds with novel therapeutic potential. Halicin, originally developed as an [...] Read more.
Background: The global spread of antibiotic-resistant bacteria presents a major public health challenge and necessitates the development of innovative antimicrobial agents. Artificial intelligence (AI)-driven drug discovery has recently enabled the repurposing of existing compounds with novel therapeutic potential. Halicin, originally developed as an anti-diabetic molecule, has been identified through AI screening as a promising antibiotic candidate due to its broad-spectrum activity, including efficacy against multidrug-resistant pathogens. Methods: In this study, the antibacterial activity of halicin was evaluated against a range of clinically relevant multidrug-resistant bacterial strains. Bacterial isolates were first characterized using the agar disk diffusion method with a panel of 22 conventional antibiotics to confirm resistance profiles. The minimum inhibitory concentration (MIC) of halicin was then determined for selected isolates, including Escherichia coli ATCC® 25922™ and Staphylococcus aureus ATCC® 29213™, using broth microdilution according to Clinical and Laboratory Standards Institute (CLSI) guidelines. Results: Halicin demonstrated notable antibacterial activity, with MIC values of 16 μg/mL and 32 μg/mL against E. coli ATCC® 25922™ and S. aureus ATCC® 29213™, respectively. A dose-dependent inhibition of bacterial growth was observed for the majority of tested isolates, except for Pseudomonas aeruginosa, which exhibited intrinsic resistance. This lack of susceptibility is likely related to reduced outer membrane permeability, limiting the intracellular accumulation of halicin. Conclusions: Our findings support the potential of halicin as a novel antimicrobial agent for the treatment of infections caused by antibiotic-resistant bacteria. However, further investigations, including pharmacokinetic, pharmacodynamic, and toxicity studies, are essential to assess its clinical safety and therapeutic applicability. Full article
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Communication
Repeated Detection of Bartonella DNA in Feline Placenta: Potential Implications for Placental and Fetal Development
by Charlotte O. Moore, Ricardo Maggi, Kelli Ferris and Edward B. Breitschwerdt
Animals 2025, 15(14), 2041; https://doi.org/10.3390/ani15142041 - 11 Jul 2025
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Abstract
The domestic cat is the primary reservoir host of three flea-borne Bartonella species, one of which (Bartonella henselae) causes reduced fertility and reproductive failure in experimentally infected cats. Vertical transmission of Bartonella has been documented only in B-cell deficient mice, but [...] Read more.
The domestic cat is the primary reservoir host of three flea-borne Bartonella species, one of which (Bartonella henselae) causes reduced fertility and reproductive failure in experimentally infected cats. Vertical transmission of Bartonella has been documented only in B-cell deficient mice, but not immunocompetent animals. As many free-roaming cats are chronically infected with Bartonella and may be immunocompromised by environmental stress or coinfection, we attempted to isolate Bartonella from the fetal and placental tissues of pregnant queens spayed during trap–neuter–release. Four samples from each tissue (ovary, uterus, fetus, and placenta) were split for direct DNA extraction, liquid culture, and culture on a blood agar plate. Samples from infected queens were inoculated into liquid media and sampled weekly for three weeks for DNA extraction and plating. Bartonella DNA was sequenced directly from 28% (5/18) of the free-roaming queens. For these five queens, liquid enrichment culture was attempted in duplicate for fetal and placental samples. Bartonella clarridgeiae DNA was amplified using qPCR liquid enrichment cultures from the placentas of two cats. These findings suggest that viable Bartonella organisms are present in feline reproductive tissue. Additional studies are needed to assess the transplacental transmission of Bartonella spp. and Bartonella’s influence on fetal development. Full article
(This article belongs to the Section Companion Animals)
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