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23 pages, 3128 KB  
Article
Antibacterial Activity of Bis(4-aminopyridinium) Compounds for Their Potential Use as Disinfectants
by Carolina Arriaza-Echanes, Claudio A. Terraza, Mateus Frazao, Sebastián Reyes-Cerpa, Loreto Sanhueza and Pablo A. Ortiz
Molecules 2025, 30(19), 3962; https://doi.org/10.3390/molecules30193962 - 2 Oct 2025
Abstract
The following study presents the initial evaluation (solubility, thermal stability, antibacterial activity, and cytotoxicity) of a series of previously described organic salts, derived from the bis(4-aminopyridinium) cation with different chain lengths, for their potential use as hospital disinfectants. Of the salts studied, those [...] Read more.
The following study presents the initial evaluation (solubility, thermal stability, antibacterial activity, and cytotoxicity) of a series of previously described organic salts, derived from the bis(4-aminopyridinium) cation with different chain lengths, for their potential use as hospital disinfectants. Of the salts studied, those with chain lengths between 2 and 10 carbon atoms (C2–C10) showed high solubility in water, methanol, and DMSO. All salts exhibited high thermal stability, showing a thermal decomposition temperature (T5%) above 330 °C. Antibiotic susceptibility testing of the studied E. coli, S. aureus, and S. typhimurium strains confirmed their resistance to different classes of commonly used clinical antibiotics, validating their selection. During the determination of antibacterial activity, the long-chain salts (C10 and C12) showed the greatest activity, with minimum inhibitory concentrations (MICs) from 31.2 μg/mL to 62.5 μg/mL in all the strains studied. Given the high activity of C10 and C12, their cytotoxicity was assessed in HeLa cells. They exhibited no cytotoxic effects after 12 h and only about 5% cytotoxicity after 24 h. Furthermore, the cell viability assay of the most active and water-soluble salt, C10, showed that this salt can interact with the bacterial cytoplasmic membrane, increasing its permeability in both Gram-positive and Gram-negative bacteria. However, these results cannot rule out the possibility that this salt may have more than one site of action within the bacterial cell. Full article
(This article belongs to the Special Issue Design, Synthesis and Applications of Bioactive Compounds)
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18 pages, 2457 KB  
Article
The Potential for Reusing Superabsorbent Polymer from Baby Diapers for Water Retention in Agriculture
by Kamilla B. Shishkhanova, Vyacheslav S. Molchanov, Ilya V. Prokopiv, Alexei R. Khokhlov and Olga E. Philippova
Gels 2025, 11(10), 795; https://doi.org/10.3390/gels11100795 - 2 Oct 2025
Abstract
Annually, about 2.4 million tons of superabsorbent polymers (SAPs) used in disposable diapers are thrown away, polluting our planet. This study aims to explore the potential for reusing SAPs removed from diapers to enhance soil water retention. To this end, the swelling and [...] Read more.
Annually, about 2.4 million tons of superabsorbent polymers (SAPs) used in disposable diapers are thrown away, polluting our planet. This study aims to explore the potential for reusing SAPs removed from diapers to enhance soil water retention. To this end, the swelling and water retention properties of SAP gels from three different types of diapers were compared to those of an agricultural gel, Aquasorb. Sand was used as a model for soil. When mixed with sand, diaper gels have a swelling degree of ca. 100 g per gram of dried polymer, and a swelling pressure of 12–26 kPa, which are similar to those of Aquasorb gel. Using a synthesized poly(acrylamide-co-sodium acrylate) gel as an example, the correlation between the swelling pressure and the compression modulus of the swollen gel was demonstrated. Soil-hydrological constants were estimated from water retention curves obtained by equilibrium centrifugation of gel/sand mixtures. It was observed that adding 0.3 vol% of diaper gels to sand leads to a 3–4-fold increase in water range available to plants, which is close to that provided by agricultural gel Aquasorb. The water-holding properties were shown to be maintained during several swelling/deswelling cycles in the sand medium. The addition of diaper gels to soil had a significant positive impact on mustard (Brassica juncea L.) seed germination and seedling growth, similar to the agricultural gel Aquasorb. This suggests high potential for the reuse of SAPs from diaper waste to improve soil water retention and water accessibility to plants. This would provide both economic and environmental benefits, conserving energy and raw materials to produce new agricultural gels and limiting the amount of waste. Full article
(This article belongs to the Special Issue Polymer Hydrogels and Networks)
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15 pages, 9569 KB  
Article
Cold–Temperate Betula platyphylla Sukaczev Forest Can Provide More Soil Nutrients to Increase Microbial Alpha Diversity and Microbial Necromass Carbon
by Yunbing Jiang, Mingliang Gao, Libin Yang, Zhichao Cheng, Siyuan Liu and Yongzhi Liu
Microorganisms 2025, 13(10), 2291; https://doi.org/10.3390/microorganisms13102291 - 1 Oct 2025
Abstract
Changes in vegetation type shape the soil microenvironment, thereby regulating the changes in the organic carbon pool by influencing microbial communities and the accumulation of microbial necromass carbon (MNC). This study investigated microbial biomass—via phospholipid fatty acids (PLFAs) analysis—and MNC accumulation across three [...] Read more.
Changes in vegetation type shape the soil microenvironment, thereby regulating the changes in the organic carbon pool by influencing microbial communities and the accumulation of microbial necromass carbon (MNC). This study investigated microbial biomass—via phospholipid fatty acids (PLFAs) analysis—and MNC accumulation across three cold–temperate forest types: Larix gmelinii forest (L), Larix gmeliniiBetula platyphylla Sukaczev mixed forest (LB), and Betula platyphylla Sukaczev forest (B). The results showed that the L had the lowest contents of pH, water content (WC), soil organic carbon (SOC), total nitrogen (TN), available nitrogen (AN), and total phosphorus (TP), but the highest contents of dissolved organic carbon (DOC), available phosphorus (AP), and carbon to nitrogen ratio (C/N) (p < 0.05). LB had the lowest PLFAs content and the highest ratio of Gram-positive bacteria/Gram-negative bacteria (G+/G−), and total fungi/total bacteriai (F/B) of L was the highest. B had the highest alpha diversity index, and significantly positively correlated with pH, SOC, TN, AN, and TP. TP and C/N were the primary elements for significant differences in microbial community structure. The order of MNC content and its contribution to SOC was B > LB > L. MNC was significantly negatively correlated with PLFAs, DOC, and AP, and significantly positively correlated with pH, SOC, TN, AN, TP, Shannon–Wiener and Pielou indices. In conclusion, this study demonstrates that Betula platyphylla Sukaczev forest retains more carbon, nitrogen, and phosphorus, microbial alpha diversity, and acquires more MNC, which can provide a basis for subsequent forest management and carbon sequestration projects. Full article
(This article belongs to the Section Environmental Microbiology)
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14 pages, 279 KB  
Article
Molecular Epidemiology of Different Bacterial Pathogens and Their Antimicrobial Resistance Genes Among Patients Suffering from Surgical Site Infections in Lebanon
by Inass Kawtharani, Ghassan Ghssein, Ola Srour, Abdul Amir Chaaban and Pascale Salameh
Microbiol. Res. 2025, 16(10), 216; https://doi.org/10.3390/microbiolres16100216 - 1 Oct 2025
Abstract
Background: Antimicrobial resistance (AMR) is a major global health threat, particularly in surgical site infections (SSIs), where multidrug-resistant (MDR) pathogens complicate treatment. Objective: This study aimed to identify antimicrobial resistance genes and assess their prevalence in bacterial species causing SSIs in Lebanon. Materials [...] Read more.
Background: Antimicrobial resistance (AMR) is a major global health threat, particularly in surgical site infections (SSIs), where multidrug-resistant (MDR) pathogens complicate treatment. Objective: This study aimed to identify antimicrobial resistance genes and assess their prevalence in bacterial species causing SSIs in Lebanon. Materials and Methods: The present research is a multicenter and prospective study that included patients who developed SSIs after surgery in seven hospitals, within the period of January 2024–September 2024. Bacterial isolates from wound swabs or tissue samples were identified using standard microbiological methods. Antimicrobial susceptibility was tested by disk diffusion, and resistance genes were detected by PCR. Data were analyzed using Statistical Package for the Social Sciences (SPSS). Results: Among 6933 surgical patients, 63 developed SSIs (0.91%; 95% CI [0.70–1.15]). Gram-negative bacteria predominated (73%), mainly Escherichia coli and Pseudomonas aeruginosa, while Gram-positive isolates accounted for 27%, mostly Staphylococcus aureus. MDR was observed in 71% of Gram-positive and 61% of Gram-negative isolates. The most frequent genes were mecA in S. aureus (100%) and coagulase-negative staphylococci (83.3%); blaCTX-M in E. coli, Klebsiella pneumoniae, and Enterobacter cloacae (100%); and blaNDM in E. cloacae (100%) and Acinetobacter baumannii (60%). blaKPC was less common, and no isolates carried Imipenemase (IMP), Verona integron-encoded metallo-β-lactamase (VIM), and Oxacillinase-48-like β-lactamase (OXA-48). Conclusions: This study highlights the high prevalence of antibiotic resistance in agents causing SSIs in Lebanese hospitals. Resistance genes, particularly mecA, blaCTX-M, and blaNDM, were highly prevalent in SSI pathogens, underscoring the urgent need for surveillance and judicious antibiotic use in Lebanese hospitals. Full article
8 pages, 2184 KB  
Brief Report
X-Ray Crystal Structure of the N-Terminal Domain of Staphylococcus Aureus Cell-Cycle Protein GpsB
by Nathan I. Nicely, Thomas. M. Bartlett and Richard W. Baker
Crystals 2025, 15(10), 867; https://doi.org/10.3390/cryst15100867 - 30 Sep 2025
Abstract
GpsB is a conserved cell-cycle regulator in the Firmicute clade of Gram-positive bacteria that coordinates multiple aspects of envelope biogenesis. Recent studies demonstrate interactions between GpsB and the key division cytoskeleton FtsZ, suggesting that GpsB links cell division to various aspects of cell [...] Read more.
GpsB is a conserved cell-cycle regulator in the Firmicute clade of Gram-positive bacteria that coordinates multiple aspects of envelope biogenesis. Recent studies demonstrate interactions between GpsB and the key division cytoskeleton FtsZ, suggesting that GpsB links cell division to various aspects of cell envelope biogenesis in Staphylococcus aureus and potentially other Firmicutes. We determined a 1.7 Å resolution crystal structure of the N-terminal domain of Staphylococcus aureus GpsB, revealing an asymmetric dimer with a bent conformation. This conformation is nearly identical to one of two conformations reported by Sacco et al., confirming the unique conformation of S. aureus GpsB compared to other Gram-positive bacteria. This structural agreement provides strong validation of the S. aureus GpsB fold and supports its proposed role in organizing the cell division machinery. Full article
(This article belongs to the Section Biomolecular Crystals)
15 pages, 2496 KB  
Article
Structures, Interactions, and Antimicrobial Activity of the Shortest Thanatin Peptide from Anasa tristis
by Swaleeha Jaan Abdullah, Jia Sheng Guan, Yuguang Mu and Surajit Bhattacharjya
Int. J. Mol. Sci. 2025, 26(19), 9571; https://doi.org/10.3390/ijms26199571 - 30 Sep 2025
Abstract
Antimicrobial peptides (AMPs), also referred to as host defense peptides, are promising molecules in the development of the next generation of antibiotics against drug-resistant bacterial pathogens. Thanatin comprises a family of naturally occurring cationic AMPs derived from several species of insects. The first [...] Read more.
Antimicrobial peptides (AMPs), also referred to as host defense peptides, are promising molecules in the development of the next generation of antibiotics against drug-resistant bacterial pathogens. Thanatin comprises a family of naturally occurring cationic AMPs derived from several species of insects. The first thanatin, 21 residues long, was identified from the spined soldier bug, and more thanatin peptides have been discovered in recent studies. The 16-residue thanatin from Anasa tristis, or Ana-thanatin, represents the shortest sequence in the family. However, the antimicrobial activity and mechanistic process underpinning bacterial cell killing have yet to be reported for Ana-thanatin peptide. In this work, we examined the antibacterial activity, structures, and target interactions of Ana-thanatin. Our results demonstrated that Ana-thanatin exerts potent antibiotic activity against strains of Gram-negative and Gram-positive bacteria. Biophysical studies demonstrated that Ana-thanatin interacts with LPS outer membrane and can permeabilize the OM barrier in the process. Atomic-resolution structures of the peptide in free solution and in complex with lipopolysaccharide (LPS) micelle were solved by NMR, determining canonical β-sheet structures. Notably, in complex with LPS, the β-sheet structure of the peptide was better defined in terms of the packing of amino acid residues. Further, MD simulations demonstrated rapid binding of the Ana-thanatin peptide with the LPS molecules within the lipid bilayers. These studies have revealed structural features which could be responsible for LPS-OM disruption of the Gram-negative bacteria. In addition, NMR heteronuclear single quantum coherence (HSQC) studies have demonstrated that Ana-thanatin can strongly interact with the LPS transport periplasmic protein LptAm, potentially inhibiting OM biogenesis. Taken together, we surmise that the Ana-thanatin peptide could serve as a template for the further development of novel antibiotics. Full article
(This article belongs to the Collection Feature Papers in Molecular Biophysics)
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19 pages, 1817 KB  
Article
Urinary Tract Infections in a Single-Center Bulgarian Hospital: Trends in Etiology, Antibiotic Resistance, and the Impact of the COVID-19 Pandemic (2017–2022)
by Milena Yancheva Rupcheva, Kostadin Kostadinov, Yordan Kalchev, Petya Gardzheva, Eli Hristozova, Zoya Rachkovska, Gergana Lengerova, Andreana Angelova, Marianna Murdjeva and Michael M. Petrov
Antibiotics 2025, 14(10), 982; https://doi.org/10.3390/antibiotics14100982 - 30 Sep 2025
Abstract
Background: Urinary tract infections (UTIs) are among the most common hospital- and community-acquired infections, creating a substantial healthcare burden due to recurrence, complications, and rising antimicrobial resistance. Accurate diagnosis and timely antimicrobial therapy are essential. This study aimed to identify trends in [...] Read more.
Background: Urinary tract infections (UTIs) are among the most common hospital- and community-acquired infections, creating a substantial healthcare burden due to recurrence, complications, and rising antimicrobial resistance. Accurate diagnosis and timely antimicrobial therapy are essential. This study aimed to identify trends in the etiology, treatment, and resistance patterns of UTIs through a retrospective analysis of urine isolates processed at the Laboratory of Microbiology at University Hospital St. George in Plovdiv, the largest tertiary care and reference microbiology center in Bulgaria, between 2017 and 2022. Materials and Methods: A retrospective single-center study was performed at the hospital’s Microbiology Laboratory. During the study period, 74,417 urine samples from 25,087 hospitalized patients were screened with the HB&L UROQUATTRO system. Positive specimens were cultured on blood agar, Eosin-Methylene Blue, and chromogenic media. Identification was performed using biochemical assays, MALDI-TOF MS, and the Vitek 2 Compact system. Antimicrobial susceptibility testing included disk diffusion, MIC determination, broth microdilution (for colistin), and Vitek 2 Compact, interpreted according to EUCAST standards. Descriptive analysis and temporal resistance trends were evaluated with regression models, and interrupted time-series analysis was applied to assess COVID-19-related effects. Results: Out of 10,177 isolates, Gram-negative bacteria predominated (73%), with Escherichia coli, Klebsiella pneumoniae, and Proteus mirabilis as the leading pathogens. Among Gram-positives, Enterococcus faecalis was the most frequent. In the post-COVID-19 period, ESBL production increased in E. coli (34–38%), K. pneumoniae (66–77%), and P. mirabilis (13.5–24%). Carbapenem resistance rose in K. pneumoniae (to 40.6%) and P. aeruginosa (to 24%), while none was detected in E. coli. Colistin resistance increased in K. pneumoniae but remained absent in E. coli and P. aeruginosa. High-level aminoglycoside resistance in E. faecalis was stable (~70%), and vancomycin resistance in E. faecium rose from 4.6% to 8.9%. Conclusions: Both community- and hospital-acquired UTIs in Southeastern Bulgaria are increasingly linked to multidrug-resistant pathogens, particularly ESBL-producing and carbapenem-resistant Enterobacterales. Findings from the region’s largest referral center highlight the urgent need for continuous surveillance, rational antibiotic use, and novel therapeutic approaches. Full article
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21 pages, 4240 KB  
Article
Development and Characterization of Effective Hemostatic Composites Based on Polyvinyl Alcohol/Kaolinite/Chitosan
by Aruzhan Alimbek, Bayansulu Otegenova, Zhanar Bekissanova, Balzhan Savdenbekova, Nailya Ibragimova, Renata Nemkayeva, Myroslav Sprynskyy and Alyiya Ospanova
Polymers 2025, 17(19), 2637; https://doi.org/10.3390/polym17192637 - 30 Sep 2025
Abstract
In this study, hemolytically safe and antibacterial polyvinyl alcohol/kaolinite/chitosan (PVA/KAO/CS) hydrogels were obtained using the freeze–thaw method. The structure of the chemical bonds present in the developed hydrogels was investigated by Fourier transform infrared spectroscopy (FTIR). Scanning electron microscopy (SEM) and optical microscopy [...] Read more.
In this study, hemolytically safe and antibacterial polyvinyl alcohol/kaolinite/chitosan (PVA/KAO/CS) hydrogels were obtained using the freeze–thaw method. The structure of the chemical bonds present in the developed hydrogels was investigated by Fourier transform infrared spectroscopy (FTIR). Scanning electron microscopy (SEM) and optical microscopy study results showed the morphological and structural characteristics of the hydrogels’ surface. The thermal stability and phase transitions of the obtained hydrogel samples were determined by thermogravimetric analysis (TGA). Porosity, swelling, gel fractions, and mechanical properties were also examined. Biomedical properties of the samples were evaluated using in vitro and in vivo tests such as hemolytic activity, inhibition of protein denaturation, antimicrobial activity, and hemostatic activity. The obtained hydrogels demonstrated safe hemolytic activity, pronounced hemostatic activity, the ability to prevent thermal denaturation of albumin, as well as antimicrobial activity against Gram-positive bacteria Staphylococcus aureus ATCC BAA-39 and Streptococcus pyogenes ATCC 19615 and Gram-negative bacteria Pseudomonas aeruginosa ATCC 9027 and Escherichia coli ATCC 8739. All the obtained characteristics confirmed the promising biomedical applications of the obtained hydrogels. Full article
(This article belongs to the Section Polymer Composites and Nanocomposites)
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15 pages, 3880 KB  
Article
Effect of Riboflavin and Blue Light-Emitting Diode Irradiation on Microbial Inactivation and the Physicochemical Properties of Betel Leaves
by Rattanaporn Rinpan, Vethaga Panudta, Rawisara Phongkhedkham, Siriyakorn Janpitu, Suphat Phongthai, Wannaporn Klangpetch and Tabkrich Khumsap
Processes 2025, 13(10), 3130; https://doi.org/10.3390/pr13103130 - 29 Sep 2025
Abstract
This study evaluated the effectiveness of photodynamic treatment (PDT) using riboflavin (Rbf) and blue light-emitting diode (BL) irradiation for microbial inactivation and quality preservation in fresh betel leaves (Piper betle L.). Non-pathogenic surrogates Escherichia coli K-12 and Listeria innocua were used to [...] Read more.
This study evaluated the effectiveness of photodynamic treatment (PDT) using riboflavin (Rbf) and blue light-emitting diode (BL) irradiation for microbial inactivation and quality preservation in fresh betel leaves (Piper betle L.). Non-pathogenic surrogates Escherichia coli K-12 and Listeria innocua were used to model Gram-negative and Gram-positive bacteria. The combined Rbf-BL treatment significantly reduced microbial populations by up to 5.3 log CFU/g for E. coli and 6.2 log CFU/g for L. innocua on leaf surfaces (p < 0.05) and 1.3–1.5 log CFU/mL in broth cultures. Treated samples showed significantly higher total soluble solids (12.0 ± 0.0 °Brix), total phenolic content (0.17 ± 0.02 mmol GAE/g, p < 0.05), and antioxidant activity (62.0 ± 3.1% DPPH inhibition, p < 0.05), with minimal color alteration after treatment (ΔE = 4.68). The total fluence measured at the leaf surface was approximately 11.72 J/cm2. As a mild thermal treatment utilizing a GRAS photosensitizer, riboflavin-assisted PDT presents a promising strategy for enhancing microbial safety and promoting phytochemical quality in betel leaves. Full article
(This article belongs to the Special Issue Food Processing and Ingredient Analysis)
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10 pages, 480 KB  
Article
Low Seroprevalence of Bovine Brucellosis in Communal Areas of Limpopo Province, South Africa
by Karabelo Madiba, Nomakorinte Gcebe, Carin Boshoff, Mohamed Sirdar, Ngoako Ramaselela and Tiny Hlokwe
Vet. Sci. 2025, 12(10), 942; https://doi.org/10.3390/vetsci12100942 - 29 Sep 2025
Abstract
Brucellosis is caused by Gram-negative coccobacilli of the Brucella genus, with cattle mainly infected with Brucella abortus. The disease burden is a threat to socioeconomic development (agriculture/tourism) as well as to animal health, biodiversity and to human health due to the zoonotic [...] Read more.
Brucellosis is caused by Gram-negative coccobacilli of the Brucella genus, with cattle mainly infected with Brucella abortus. The disease burden is a threat to socioeconomic development (agriculture/tourism) as well as to animal health, biodiversity and to human health due to the zoonotic nature of this pathogen. In South Africa (S.A), the prevalence of the disease in cattle and livestock in general is mostly unknown in communal farms. A cross-sectional study with a multistage sampling strategy was applied in communal areas from three district municipalities, i.e., Mopani, Capricorn and Sekhukhune of Limpopo province, South Africa. Sera (n = 1133) were collected and screened for antibodies against the Brucella species using the Rose Bengal Test (RBT) and confirmation of positive reactors with a Complement Fixation Test (CFT). The brucellosis seroprevalence was found to be 0.79% (95% CI: 0.38–1.45) by a CFT. Univariate analysis indicated that only the frequency of birth was significantly associated with CFT positivity (OR = 20; 95% Cl: 1.61–247.99; p = 0.039). The multivariable logistic regression model revealed that the frequency of birth, age, breed, gender, municipality and district were not statistically significant predictors at 0.05 level. However, some variables like cattle aged more than five years, had higher odds of CFT positivity compared to those younger than five years (OR = 5.66; 95% CI: 0.36–87.97), although the association was not statistically significant (p = 0.215). All positive reactors detected originated from the Mopani district municipality. Overall, the findings reveal a much lower seroprevalence of brucellosis in the communal farms of Limpopo province than previously assumed. We are of the opinion that the low seroprevalence is attributed to effective control strategies implemented by the Limpopo provincial veterinary services and hence provide important information to assist the regulatory bodies in the control and eradication of the disease. Full article
(This article belongs to the Section Veterinary Microbiology, Parasitology and Immunology)
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21 pages, 2804 KB  
Article
The PTS EIIB Component Drives Strain-Specific Virulence in Listeria monocytogenes: Divergent Regulation of Biofilm Formation and Host Infection in High- and Low-Virulence Strains
by Lu Liu, Caixia Liu, Ruixuan Qian, Yatao Qi, Zhongke Yin, Ruifeng Luo, Dongdong Du, Zengqi Liu, Lichao Kang and Jing Wang
Microorganisms 2025, 13(10), 2274; https://doi.org/10.3390/microorganisms13102274 - 28 Sep 2025
Abstract
Listeria monocytogenes (L. monocytogenes) is a Gram-positive intracellular pathogen capable of causing severe infections. The Listeria pathogenicity island 4 (LIPI-4) encodes a phosphotransferase system (PTS) with its EIIB component playing a critical role in carbohydrate phosphorylation and virulence. However, the precise [...] Read more.
Listeria monocytogenes (L. monocytogenes) is a Gram-positive intracellular pathogen capable of causing severe infections. The Listeria pathogenicity island 4 (LIPI-4) encodes a phosphotransferase system (PTS) with its EIIB component playing a critical role in carbohydrate phosphorylation and virulence. However, the precise function of EIIB in virulence regulation across diverse pathogenic strains remains unclear. Here, we generated an EIIB deletion mutant (LM873ΔEIIB) and its complemented strain (CLM873ΔEIIB) from the low-virulence strain LM873, and performed comparative analyses with the high-virulence strain LM928 and its corresponding mutants. Deletion of EIIB differentially modulated biofilm formation: suppressing it in LM928 while enhancing it in LM873, accompanied by corresponding transcriptional changes in biofilm-associated and virulence genes. Both mutants exhibited impaired hemolytic activity, whereas motility attenuation was specific to LM928ΔEIIB. At the cellular level, LM873ΔEIIB enhanced adhesion to and invasion of Caco-2 but impaired intracellular proliferation in JEG-3; In contrast, LM928ΔEIIB promoted Caco-2 invasion while attenuating JEG-3 adhesion, invasion, and intracellular replication, as well as reducing invasion and proliferation in RAW264.7 macrophage. Animal experiments demonstrated that EIIB deletion attenuated LM928 colonization in the liver and spleen, but had no significant impact on LM873. Collectively, our findings establish EIIB as a strain-dependent virulence regulator in L. monocytogenes, particularly modulating biofilm formation and host–pathogen interactions. Full article
(This article belongs to the Section Molecular Microbiology and Immunology)
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25 pages, 3491 KB  
Article
Selective Targeting and Enhanced Photodynamic Inactivation of Methicillin-Resistant Staphylococcus aureus (MRSA) by a Decacationic Vancomycin–Mesochlorin Conjugate
by He Yin, Xiaojing Liu, Min Wang, Ying Wang, Tianhong Dai and Long Y. Chiang
Antibiotics 2025, 14(10), 978; https://doi.org/10.3390/antibiotics14100978 - 28 Sep 2025
Abstract
Background/Objectives: Covalent conjugation of an antibiotic vancomycin (VCM) moiety and a photosensitizing mesochlorin (mChlPd) unit into one molecular entity may present the potential to produce the combinatorial effect of both antibacterial photodynamic therapeutic (aPDT) and antibiotic activities. Our recent [...] Read more.
Background/Objectives: Covalent conjugation of an antibiotic vancomycin (VCM) moiety and a photosensitizing mesochlorin (mChlPd) unit into one molecular entity may present the potential to produce the combinatorial effect of both antibacterial photodynamic therapeutic (aPDT) and antibiotic activities. Our recent study indicated that a short linkage of <4 (C−C/or C−N) bond distances between these two moieties resulted in significant steric hindrance due to the bulky VCM, which greatly reduces the accessibility of the agent to the cell surface of methicillin-resistant Staphylococcus aureus (MRSA). The observed aPDT efficacy was found to be minimal. Here, we report that the revision of this linkage, via an EG10 unit using identical synthetic procedures, was able to resolve the issue. Methods: Accordingly, the corresponding combinatorial aPDT−antibiotic compound, consisting of two covalently bonded quaternary ammonium pentacationic arms on the mesochlorin chromophore core, designated as VCMe-mChlPd-N10+ (LC40e+), was prepared for applications in antibacterial photodynamic inactivation (aPDI) activity. It was selected to investigate its enhanced binding and targeting ability to the surface of Gram-positive MRSA cells. Subsequent antibacterial photodynamic therapeutic (aPDT) activity to inactivate MRSA was investigated to substantiate the corresponding cell-surface binding effect on the efficacy of aPDT. Results: We found that the covalent combination of 10 positive charges and an MRSA-targeting vancomycin (VCM) moiety in a conjugated structure, functioning as an antibiotic–decacationic photosensitizing agent (Abx-dcPS), was capable of largely improving the MRSA cell-targeting efficiency. Importantly, variation in the chain length of the oligo(ethylene glycol) linker of VCMe-mChlPd-N10+, which was sufficiently long enough to properly separate the photoactive mesochlorin ring moiety from the VCM moiety within the molecular structure, resulted in significantly enhanced aPDT activity. The new conjugate provided nearly complete eradication (>6.5-log10 colony-forming units (CFU) reduction) of MRSA cells in vitro. The aPDT efficacy followed the order Abx-dcPS (combinatorial decacationic) > dcPS (decacationic) >> nPS (nonionic). This order was also verified by the relative physical binding trend of these PSs using either nPS-, dcPS-, or Abx-dcPS-pretreated and pre-fixed MRSA cells in investigations of fluorescent confocal microscopy, UV–vis fluorescence spectroscopy, and transmission electron microscopy (TEM). Conclusions: Furthermore, the molecular conjugate of Abx-dcPS may provide covalent co-delivery of two drug components concurrently, which might also serve as an effective antibiotic agent after aPDT and potentially prevent the reoccurrence of MRSA-induced infection. Full article
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12 pages, 1038 KB  
Article
Rapid Identification of Carbapenemase Genes Directly from Blood Culture Samples
by Ghada A. Ziad, Deena Jalal, Mohamed Hashem, Ahmed A. Sayed, Sally Mahfouz, Ahmed Bayoumi, Maryam Lotfi, Omneya Hassanain, May Tolba, Youssef Madney, Lobna Shalaby and Mervat Elanany
Diagnostics 2025, 15(19), 2480; https://doi.org/10.3390/diagnostics15192480 - 28 Sep 2025
Abstract
Background/Objectives: The rapid identification of carbapenemase genes directly from positive blood culture (BC) samples shortens the time needed to initiate optimal antimicrobial therapy for Carbapenemase-Producing Enterobacterales (CPE) infections. Several commercial automated PCR systems are available for detecting CPE resistance genes but are expensive. [...] Read more.
Background/Objectives: The rapid identification of carbapenemase genes directly from positive blood culture (BC) samples shortens the time needed to initiate optimal antimicrobial therapy for Carbapenemase-Producing Enterobacterales (CPE) infections. Several commercial automated PCR systems are available for detecting CPE resistance genes but are expensive. The Xpert® Carba-R assay (Cepheid GeneXpert System) has high sensitivity and specificity for the detection of carbapenamase genes from bacterial colonies or rectal swabs, with an affordable price. This assay was not used for positive BC testing of CPE resistance genes. Whole-Genome Sequencing (WGS) for resistance genes can be used as the gold standard at a research level. In this study, we evaluated the performance of the Xpert® Carba-R assay for the early detection of carbapenamase genes directly from positive BCs, using WGS as the gold standard. Methods: A prospective observational study was conducted at Children’s Cancer Hospital-Egypt (CCHE-57357). All positive BCs underwent direct gram staining and conventional cultures. A total of 590 positive BCs containing Gram-negative rods (GNRs) were identified. The Xpert® Carba-R assay was used to detect carbapenemase genes directly from the positive BC bottle compared with WGS results. Results: Among the 590 GNR specimens, 178 were found to carry carbapenemase genes using the Xpert® Carba-R assay, with results obtained in approximately one hour. The main genotypes detected were blaNDM, blaOXA-48-like, and dual blaNDM/blaOXA-48-like at 27%, 29%, and 33%, respectively. The agreement between Xpert® Carba-R assay and WGS results was almost perfect for the genotype resistance pattern of isolates and individual gene detection. Conclusions: The use of the Xpert® Carba-R assay directly from BC bottles was an easy-to-use, time-saving, affordable tool with high accuracy in identifying carbapenemase genes and, thus, shortens the time needed to initiate optimal antimicrobial therapy for CPE infections. Full article
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25 pages, 4994 KB  
Article
In Vitro Antibacterial Efficacy of Recombinant Phage-Derived Endolysin LysTAC1 Against Carbapenem-Resistant Acinetobacter baumannii
by Inam Ullah, Song Cui, Qiulong Yan, Hayan Ullah, Shanshan Sha and Yufang Ma
Antibiotics 2025, 14(10), 975; https://doi.org/10.3390/antibiotics14100975 - 26 Sep 2025
Abstract
Background: The rapid emergence of antibiotic resistance in Acinetobacter baumannii has led the World Health Organization (WHO) to designate it as a “high priority” pathogen. The emergence of multidrug-resistant (MDR) and pandrug-resistant (PDR) strains poses considerable treatment challenges. As antimicrobial resistance (AMR) escalates [...] Read more.
Background: The rapid emergence of antibiotic resistance in Acinetobacter baumannii has led the World Health Organization (WHO) to designate it as a “high priority” pathogen. The emergence of multidrug-resistant (MDR) and pandrug-resistant (PDR) strains poses considerable treatment challenges. As antimicrobial resistance (AMR) escalates toward a post-antibiotic era, innovative therapeutic solutions are urgently needed. Objectives: To clone, over-express, and characterize a novel endolysin, LysTAC1, from Acinetobacter phage TAC1 for its antibacterial efficacy against multidrug-resistant bacteria. Methods: A 24 kDa endolysin featuring a glycoside hydrolase Family 19 chitinase domain was tested against carbapenem-resistant Acinetobacter baumannii clinical isolates and various Escherichia coli strains following outer membrane permeabilization with Ethylenediaminetetraacetic acid (EDTA). Stability assays and molecular docking studies were performed. Results: LysTAC1 demonstrated potent lytic activity against Gram-negative bacteria but showed no activity against Gram-positive bacteria (Staphylococcus aureus ATCC 29213 and Enterococcus gallinarum HCD 28-1). LysTAC1 maintained activity across pH 6–9 and temperatures 4–65 °C, with differential sensitivity to metal ions where K+ showed no inhibitory effect at any concentration (0.1–100 mM), and Fe2+ was non-inhibitory at lower concentrations (0.1–1 mM), while Mg2+ and Ca2+ demonstrated concentration-dependent inhibition across the tested range (0.1–100 mM). Molecular docking revealed LysTAC1 interactions with chitinase substrates 4-nitrophenyl N-acetyl-β-D-glucosaminide and 4-nitrophenyl N, N-Diacetyl-β-D-chitobioside, with binding energies of −5.82 and −6.85 kcal/mol, respectively. Conclusions: LysTAC1 shows significant potential as a targeted therapeutic agent against A. baumannii with robust stability under physiological conditions. Full article
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20 pages, 1217 KB  
Article
Pylephlebitis: A Rare but Redoubtable Complication of Intra-Abdominal Infections—A Series of 15 Cases
by Serban Nicolae Benea, Teodora Deaconu, Dragos Florea, Ruxandra Moroti, Gabriela Oprica, Alina Nae, Raluca Elena Patrascu, Eliza Militaru, Habip Gedik and Ilinca Savulescu-Fiedler
Life 2025, 15(10), 1525; https://doi.org/10.3390/life15101525 - 26 Sep 2025
Abstract
Pylephlebitis is the suppurative thrombosis of the portal vein system. Mainly reported as a severe complication of diverticulitis or appendicitis, it is an uncommon intra-abdominal infection: approximately 200 cases have been reported in the English literature, mostly from surgical wards. Our study aims [...] Read more.
Pylephlebitis is the suppurative thrombosis of the portal vein system. Mainly reported as a severe complication of diverticulitis or appendicitis, it is an uncommon intra-abdominal infection: approximately 200 cases have been reported in the English literature, mostly from surgical wards. Our study aims to assess the role of an infectious disease setting in managing pylephlebitis. We reviewed medical records from 2001 to 2024 at a tertiary infectious diseases hospital and identified 15 cases. The median age was 58 years [IQR = 28], with a male-to-female ratio of 4:1. Along with portal vein thrombosis (PVT), liver abscess(es) was/were the main radiological finding (n = 12), representing 80% of cases. The liver abscesses appear as secondary events in the case of pylephlebitis. In seven of 15 cases, we found the primary event associated with pylephlebitis. Blood cultures were positive in eight cases, with Gram-negative aerobic bacteria being commonly isolated (n = 5), followed by anaerobes (n = 3); in half of the cases, more than one pathogen was involved. All patients received broad-spectrum antibiotics containing beta-lactams, including eight who received carbapenems. Anticoagulation therapy was used in 14 cases. Two deaths were recorded, and four patients required surgical intervention, highlighting the importance of prompt diagnosis and swift antibiotic and anticoagulant treatment. Full article
(This article belongs to the Special Issue Diagnosis and Management of Microbial Infections)
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