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Colistin resistance is a global concern warning for a one health approach to combat the challenge. Colistin resistant E. coli and their resistance determinants are widely distributed in the environment, and rats could be a potential source of these isolates and resistant determinants to a diverse environmental setting. This study was aimed to determine the presence of colistin resistant E. coli (CREC) in wild rats, their antimicrobial resistance (AMR) phenotypes, and genotypic analysis of mcr-1 CREC through whole genome sequencing (WGS). A total of 39 rats were examined and CREC was isolated from their fecal pellets onto MacConkey agar containing colistin sulfate (1 μg/ mL). AMR of the CREC was determined by disc diffusion and broth microdilution was employed to determine MIC to colistin sulfate. CREC were screened for mcr genes (mcr-1 to mcr-8) and phylogenetic grouping by PCR. Finally, WGS of one mcr-1 CREC was performed to explore its genetic characteristics especially resistomes and virulence determinants. 43.59% of the rats carried CREC with one (2.56%) of them carrying CREC with mcr-1 gene among the mcr genes examined. Examination of seventeen (17) isolates from the CREC positive rats (n = 17) revealed that majority of them belonging to the pathogenic phylogroup D (52.94%) and B2 (11.76%). 58.82% of the CREC were MDR on disc diffusion test. Shockingly, the mcr-1 CREC showed phenotypic resistance to 16 antimicrobials of 8 different classes and carried the ARGs in its genome. The mcr-1 gene was located on a 60 kb IncI2 plasmid. On the other hand, ARGs related to aminoglycosides, phenicols, sulfonamides, tetracyclines and trimethoprims were located on a 288 kb mega-plasmid separately. The mcr-1 CREC carried 58 virulence genes including genes related to adhesion, colonization, biofilm formation, hemolysis and immune-evasion. The isolate belonged to ST224 and closely related to E. coli from different sources including UPEC clinical isolates from human based on cgMLST analysis. The current research indicates that rats might be a possible source of CREC, and the presence of mcr-1 and other ARGs on plasmid increases the risk of ARGs spreading and endangering human health and other environmental components through this infamous pest. [ABSTRACT FROM AUTHOR]

In this study, the zinc ferrite decorated reduced graphene oxide (ZnFe 2 O 4 -rGO) was synthesised by using a simplified one-pot solvothermal technique. The synthesised nanomaterials were examined for structural, morphological and optical characteristics is greatly aided by the XRD, HRTEM, SAED, EDX spectroscopy, Raman spectroscopy and UV–Vis spectroscopy. The crystallite size calculated from XRD and HRTEM of the ZnFe 2 O 4 -rGO nanocomposite was around 10 nm. The presence of desired elements in the synthesised nanomaterials was confirmed using EDX spectroscopy. The Raman spectroscopy aided in identifying the active functional modes. The absorption maximum (λ max) was found to be approximately ∼310 nm and the optical bandgap of ZF was reduced from 2.1 eV to 1.97 eV in ZF-rG. The antibacterial activity of ZnFe 2 O 4 -rGO nanocomposite was discussed against Enterococcus faecalis, Staphylococcus aureus, Klebsiella pneumonia and Escherichia coli bacteria at different concentrations by agar gel diffusion method among which the highest bacterial growth inhibition was seen in Klebsiella pneumonia in all different concentrations in which 86% at 100 μg/ml was the most effective. [Display omitted] [ABSTRACT FROM AUTHOR]

Growing evidence suggests that biofilm formation in Pseudomonas aeruginosa (PA) results from quorum sensing, reducing bacterial sensitivity to antibiotics and fostering the development of antibiotic-resistant strains. Addressing the need for a compound that can inhibit quorum sensing signals to mitigate biofilm formation in PA, this study screened 40 essential oils for their potential as quorum sensing inhibitors against PA ATCC9027. Utilizing the agar well diffusion assay, the antibacterial activity of these essential oils was compared with levofloxacin, revealing that 25 out of 40 essential oils exhibited antibacterial activity against PA ATCC9027. The highest antibacterial activity was exhibited by essential oils from Cinnamomum burmannii (Cinnamon Bark Oil), Cananga odorata (Ylangylang Oil), and Eucalyptus globulus (Eucalyptus Oil), confirmed by respective MIC values of 0.09765% (v/v), 0.390625% (v/v), and 1.5625% (v/v), and MBC values of 0.09765% (v/v), > 1.5625% (v/v), and > 1.5625% (v/v). Growth curve inhibition assay followed by crystal violet biofilm formation assay revealed that only Ylang-ylang Oil exhibited biofilm formation inhibition at 1/4 MIC concentration. Docking analysis of compounds from these essential oils against quorum sensing enzymes (LasR, PqsR, and QscR) revealed that Ylang-ylang Oil compounds exhibited the highest binding affinity. In conclusion, among the 40 essential oils tested, 25 show potential as antibacterials against PA ATCC9027, with Ylang-ylang Oil standing out as a promising candidate for quorum sensing inhibitor development. The use and development of quorum sensing inhibitors from Ylang-ylang Oil compounds are expected to reduce the burden of antibiotic resistance, especially in Pseudomonas aeruginosa. [ABSTRACT FROM AUTHOR]

Agar, as a seaweed polysaccharide mainly extracted from Gracilariopsis lemaneiformis, has been commercially applied in multiple fields. To investigate factors indicating the agar accumulation in G. lemaneiformis, the agar content, soluble polysaccharides content, and expression level of 11 genes involved in the agar biosynthesis were analysed under 4 treatments, namely salinity, temperature, and nitrogen and phosphorus concentrations. The salinity exerted the greatest impact on the agar content. Both high (40‰) and low (10‰, 20‰) salinity promoted agar accumulation in G. lemaneiformis by 4.06%, 2.59%, and 3.00%, respectively. The content of agar as a colloidal polysaccharide was more stable than the soluble polysaccharide content under the treatments. No significant correlation was noted between the two polysaccharides, and between the change in the agar content and the relative growth rate of the algae. The expression of all 11 genes was affected by the 4 treatments. Furthermore, in the cultivar 981 with high agar content (21.30 ± 0.95%) compared to that (16.23 ± 1.59%) of the wild diploid, the transcriptional level of 9 genes related to agar biosynthesis was upregulated. Comprehensive analysis of the correlation between agar accumulation and transcriptional level of genes related to agar biosynthesis in different cultivation conditions and different species of G. lemaneiformis, the change in the relative expression level of glucose-6-phosphate isomerase II (gpiII), mannose-6-phosphate isomerase (mpi), mannose-1-phosphate guanylyltransferase (mpg), and galactosyltransferase II (gatII) genes was highly correlated with the relative agar accumulation. This study lays a basis for selecting high-yield agar strains, as well as for targeted breeding, by using gene editing tools in the future. [ABSTRACT FROM AUTHOR]

Individuals tend to move freely when there is enough room but would act collectively for their survival under external stress. In the case of living cells, for instance, when a drop of low-density flagellated bacterial solution is transferred onto the agar surface, the initially disordered movement of individual bacteria would be replaced with coordinated cell swarming after a lag phase of a few hours. Here, we study how such cooperation is established while overcoming the disorder at the onset of the lag phase with single nanoparticle tracking. Upon the spreading of the droplet, the bacteria in the solution cluster and align near the almost immobilized contact line confining the drop, forming a narrow ring of cells. As individual cells move in and out of the ring continuously, certain flow patterns emerge in the inter-bacterial fluid. We reveal high-speed long-distance unidirectional flows with definite chirality along the outside of the ring, along the inside of the ring and across the ring. We speculate that these flows enable the fast and efficient transport, facilitating the communication and unification of the bacterial community. [ABSTRACT FROM AUTHOR]

The postbiotic derived from Lactiplantibacillus plantarum bacteria was produced in three culture media: milk, MRS, and whey, and its antibacterial and antifungal properties were evaluated. To investigate the production efficiency of postbiotics, three methods, heating, sonication and centrifugation, were utilized to prepare postbiotics in MRS broth culture medium. The antibacterial potency of the postbiotic was evaluated using the agar well-diffusion method, and MIC and MBC tests were conducted for different treatments. The results of the study showed that the postbiotic prepared in food environments such as milk and cheese whey can have antibacterial and antifungal properties similar to the postbiotic prepared in the MRS culture medium. However, it is possible to enrich food matrices such as milk and cheese whey and make further adjustments in terms of pH settings. Additionally, the thermal process was able to create a nanoscale postbiotic, which is a significant achievement for the application of postbiotics in the food and pharmaceutical industries. The future outlook of postbiotics clearly indicates that the emergence of this generation of probiotics can have an attractive and functional position in the food and pharmaceutical industries. Therefore, future research focusing on this subject will contribute to the development of this generation of postbiotics. [ABSTRACT FROM AUTHOR]

Introduction: Recent studies have shown that epithelial-stromal interactions could play a role in the development of colorectal cancer. Here, we investigated the role of fibroblasts in the transformation of normal colonocytes induced by 4-HNE. Methods: Normal Co colonocytes and nF fibroblasts from the same mouse colon were exposed, in monoculture (m) or coculture (c), to 4-HNE (5 μM) twice weekly for 3 weeks. Gene expression was then analysed and the ability of Co colonocytes to grow in anchorage-independent conditions was tested in soft agar. Fibroblasts previously treated or not with 4-HNE were also seeded in culture inserts positioned above the agar layers to allow paracrine exchanges with colonocytes. Results: First, 60% of the genes studied were modulated by coculture in Co colonocytes, with notably increased expression of BMP receptors. Furthermore, while 4-HNE increased the ability of monoculture-treated Co colonocytes to form colonies, this effect was not observed in coculture-treated Co colonocytes. Adding a selective BMPR1 inhibitor during the treatment phase abolished the protective effect of coculture. Conversely, addition of a BMP4 agonist to the medium of monoculture-treated Co colonocytes prevented phenotypic transformation by 4-HNE. Second, the presence of nF(m)-HNE fibroblasts during the soft agar assay increased the number and size of Co(m) colonocyte colonies, regardless of whether these cells had been previously treated with 4-HNE in monoculture. For soft agar assays performed with nF(c) and Co(c) cells initially treated in coculture, only the reassociation between Co(c)-HNE and nF(c)-HNE resulted in a small increase in the number of colonies. Conclusions: During the exposure phase, the epithelial-mesenchymal interaction protected colonocytes from 4-HNE-induced phenotypic transformation via activation of the BMP pathway. This intercellular dialogue also limited the ability of fibroblasts to subsequently promote colonocyte-anchorage-independent growth. In contrast, fibroblasts pre-exposed to 4-HNE in monoculture strongly increased the ability of Co(m) colonocytes to form colonies. [ABSTRACT FROM AUTHOR]

Non-degradable plastics of petrochemical origin are a contemporary problem of society. Due to the large amount of plastic waste, there are problems with their disposal or storage, where the most common types of plastic waste are disposable tableware, bags, packaging, bottles, and containers, and not all of them can be recycled. Due to growing ecological awareness, interest in the topics of biodegradable materials suitable for disposable items has begun to reduce the consumption of non-degradable plastics. An example of such materials are biodegradable biopolymers and their derivatives, which can be used to create the so-called bioplastics and biopolymer blends. In this article, gelatine blends modified with polysaccharides (e.g., agarose or carrageenan) were created and tested in order to obtain a stable biopolymer coating. Various techniques were used to characterize the resulting bioplastics, including Fourier-transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA)/differential scanning calorimetry (DSC), contact angle measurements, and surface energy characterization. The influence of thermal and microbiological degradation on the properties of the blends was also investigated. From the analysis, it can be observed that the addition of agarose increased the hardness of the mixture by 27% compared to the control sample without the addition of polysaccharides. In addition, there was an increase in the surface energy (24%), softening point (15%), and glass transition temperature (14%) compared to the control sample. The addition of starch to the gelatine matrix increased the softening point by 15% and the glass transition temperature by 6%. After aging, both compounds showed an increase in hardness of 26% and a decrease in tensile strength of 60%. This offers an opportunity as application materials in the form of biopolymer coatings, dietary supplements, skin care products, short-term and single-contact decorative elements, food, medical, floriculture, and decorative industries. [ABSTRACT FROM AUTHOR]

Although Hantzsch synthesis has been an established multicomponent reaction method for more than a decade, its derivative, whereby an aniline replaces ammonium acetate as the nitrogen source, has not been explored at great length. Recent studies have shown that the products of such a reaction, N-aryl-4-aryldihydropyridines (DHPs), have significant anticancer activity. In this study, we successfully managed to synthesize a wide range of DHPs (18 examples, 8 of which were novel) using a metal-free, mild, inexpensive, recoverable, and biopolymer-based heterogeneous catalyst, known as piperazine, which was supported in agar–agar gel. In addition, 8 further examples (3 novel) of such dihydropyridines were synthesized using isatin instead of aldehyde as a reactant, producing spiro-linked structures. Lastly, this catalyst managed to afford an unprecedented product that was derived using an innovative technique—a combination of multicomponent reactions. Essentially, the product of our previously reported aza-Friedel–Crafts multicomponent reaction could itself be used as a reactant instead of aniline in the synthesis of more complex dihydropyridines. [ABSTRACT FROM AUTHOR]

The fig‐milk dessert, a traditional and nutritionally rich treat infused with bioactive compounds, was subjected to a comprehensive analysis in this study. The novelty of this research lies in the investigation of the in vitro antioxidant, anticancer, and antimicrobial potential of the fig‐milk dessert. This was accomplished through the utilization of the 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) assay, Annexin/propidium iodide staining, microtiter plate‐based assay and agar well diffusion, respectively, for the first time. Additionally, the study assessed the total phenols and flavonoid content of the extract using the Folin–Ciocalteu assay and the aluminum chloride method, respectively. The findings revealed that the cooking method exerted a significant influence on the bioactive properties and nutritional composition of the dessert. Among the samples analyzed, CM1, consisting of figs steamed for 2 min and milk heated to 70°C, exhibited remarkable characteristics. This sample demonstrated the highest peptide concentration (1290 mg/L), superior antioxidant and anticancer activities, and favorable sensory attributes. Specifically, CM1 induced apoptosis in 84% of AGS cells and inhibited 68% of free radicals in the DPPH assay. It is noteworthy that the fig‐milk dessert did not exhibit any antibacterial properties. These discerning results carry substantial implications for the development of functional dairy products endowed with both nutritional and potential therapeutic properties. [ABSTRACT FROM AUTHOR]

Cyphostemma adenocaule is a therapeutic plant traditionally used to treat rabies, snake bite, diarrhea, and wound healing. To address the bioactive compounds exhibiting these activities, we performed a comprehensive study on the roots of the plant. Thus, the present study aims to inspect the in vitro antioxidant and antibacterial efficacies of compounds isolated from the combined dichloromethane : methanol (1 : 1) and methanol extracts of C. adenocaule along with the in silico study of their interaction with selected protein targets. The silica gel column chromatography technique was used for the isolation of compounds, and the antibacterial and antioxidant activities were evaluated using agar disc diffusion and DPPH radical scavenging assays, respectively. Furthermore, in silico molecular docking screening, pharmacokinetics, and toxicity protocols of the compound isolates were performed to offer the potential applications of the compounds in developing novel medications. A BIOVIA Discovery Studio in combination with AutoDock Vina 4.2 software, SwissADME, and ProTox-II prediction web tools were used to generate the molecular docking, pharmacokinetics, and toxicity profiles, respectively. Notably, the chromatographic separation of the combined extracts yielded six known compounds, namely, β-sitosterol (1), 3-hydroxyisoagatholactone (2), ε-viniferin (3), myricetin (4), tricuspidatol A (5), and parthenocissin A (6). The in vitro antibacterial activities revealed the highest inhibition zone by tricuspidatol A (5) (16.67 ± 0.47), showcasing its potent activity against S. aureus at 2 mg/mL, compared to ciprofloxacin (21.50 ± 0.41). ε-Viniferin (3) (IC50: 0.32 μg/mL) exhibited greater antioxidant activity than the others and displayed promising results compared to ascorbic acid (0.075 μg/mL). The molecular docking study revealed the highest binding affinity by ε-viniferin (3) (−9.9 kcal/mol) against topoisomerase II α. 3-Hydroxyisoagatholactone (2) and ε-viniferin (3) fulfilled Lipinski's rule with no violation, and the organ toxicity predictions revealed that all the compounds showed no cytotoxicity and hepatotoxicity effects. Thus, this study's combined in vitro and in silico outcomes suggest the potential use of the isolated compounds in drug discovery and support the traditional relevance of C. adenocaule. [ABSTRACT FROM AUTHOR]

Poly(vinyl alcohol) (PVA) physical cryogels that contained the additives of o-, m-, and p-bis-phenols or phenol were prepared, and their physico-chemical characteristics and macroporous morphology and the solute release dynamics were evaluated. These phenolic additives caused changes in the viscosity of initial PVA solutions before their freeze–thaw processing and facilitated the growth in the rigidity of the resultant cryogels, while their heat endurance decreased. The magnitude of the effects depended on the interposition of phenolic hydroxyls in the molecules of the used additives and was stipulated by their H-bonding with PVA OH-groups. Subsequent rinsing of such "primary" cryogels with pure water led to the lowering of their rigidity. The average size of macropores inside these heterophase gels also depended on the additive type. It was found also that the release of phenolic substances from the additive-containing cryogels occurred via virtually a free diffusion mechanism; therefore, drug delivery systems such as PVA cryogels loaded with either pyrocatechol, resorcinol, hydroquinone, or phenol, upon the in vitro agar diffusion tests, exhibited antibacterial activity typical of these phenols. The promising biomedical potential of the studied nanocomposite gel materials is supposed. [ABSTRACT FROM AUTHOR]

Background: Poultry is one of the most prominent sources of Campylobacter jejuni, which is also a major means of transmission to people. Campylobacter jejuni contamination in chicken meat comes from chicken feces because it naturally exists in the intestines of chickens. Aim: The purpose of this study is to identify the antibiotic resistance patterns and genes of C. jejuni, which was found in chickens in Pasuruan, Indonesia. Methods: The samples used in this study were 200 contents of the small intestine of broiler chickens from 40 farms in Pasuruan Regency. The enriched sample was streaked on the selective media of modified charcoal cefoperazone deoxycholate agar containing the CCDA selective supplement. Antimicrobial susceptibility test utilizing the Kirby-Bauer diffusion test method in accordance with Clinical and Laboratory Standards Institute standards. The polymerase chain reaction (PCR) method was used to detect the (hipO), which encodes the C. jejuni strain, fluoroquinolone resistance (gyrA), beta-lactam resistance (blaOXA-61), and tetracycline resistance (tetO) genes. Results: The findings revealed a 14% (28/200) prevalence of C. jejuni in the small intestine of broiler chickens. These isolates showed high resistance to enrofloxacin (92.9%). All isolates (100%) were susceptible to amoxicillinclavulanate. The PCR results showed all C. jejuni isolates (100%) detected the gyrA gene, 96.4% detected the blaOXA-61 gene, and 50% detected the tetO gene. Conclusion: The findings of antimicrobial resistance at a high level from the small intestine of broiler chickens illustrate the potential threat to human health. To lessen the effects now and in the future, coordinated and suitable action is needed, as well as steps to guarantee the poultry industry's economic survival and public health insurance. [ABSTRACT FROM AUTHOR]

Amidst population growth and challenges with existing fertilizers, the development of smart and environmentally friendly agrochemicals is imperative. While 3D printing is widespread, its potential in slow-release agrochemicals remains unexplored. This proof-of-concept study employed solvent casting and 3D printing to develop agar–urea structures. These structures, comprising 2.5% (w/w) agar, incorporated either 7% (w/w) or 13% (w/w) urea as nitrogen nutrients. Rheological, mechanical, and morphological properties and sorption capabilities were explored. Rheological analysis revealed a substantial impact of urea, enhancing material resistance to deformation. In mechanical tests, inclusion of urea showed no significant impact on compressive strength. SEM analysis confirmed the successful entrapment of urea within the agar matrix. The inclusion of urea resulted in a diminished water sorption capacity, attributed to the urea–water interactions disrupting the hydrogen bonding ability of agar. Agar–urea inks were employed in 3D printing utilizing the direct-ink writing technique, and the nitrogen release behavior was investigated. Results revealed nearly complete urea release in the positive control within 48 h. In contrast, agar–urea formulations with 7% (w/w) and 13% (w/w) achieved nitrogen release rates of 88.8% and 94.4%, respectively, suggesting potential for 3D-printed agar formulations to modify the immediate release behavior seen in conventional urea fertilizers. [ABSTRACT FROM AUTHOR]

Bacteriocins, which have narrow-spectrum activity and limited adverse effects, are promising alternatives to antibiotics. In this study, we identified klebicin E (KlebE), a small bacteriocin derived from Klebsiella pneumoniae. KlebE exhibited strong efficacy against multidrug-resistant K. pneumoniae isolates and conferred a significant growth advantage to the producing strain during intraspecies competition. A giant unilamellar vesicle leakage assay demonstrated the unique membrane permeabilization effect of KlebE, suggesting that it is a pore-forming toxin. In addition to a C-terminal toxic domain, KlebE also has a disordered N-terminal domain and a globular central domain. Pulldown assays and soft agar overlay experiments revealed the essential role of the outer membrane porin OmpC and the Ton system in KlebE recognition and cytotoxicity. Strong binding between KlebE and both OmpC and TonB was observed. The TonB-box, a crucial component of the toxin-TonB interaction, was identified as the 7-amino acid sequence (E3ETLTVV9) located in the N-terminal region. Further studies showed that a region near the bottom of the central domain of KlebE plays a primary role in recognizing OmpC, with eight residues surrounding this region identified as essential for KlebE toxicity. Finally, based on the discrepancies in OmpC sequences between the KlebE-resistant and sensitive strains, it was found that the 91st residue of OmpC, an aspartic acid residue, is a key determinant of KlebE toxicity. The identification and characterization of this toxin will facilitate the development of bacteriocin-based therapies targeting multidrugresistant K. pneumoniae infections. [ABSTRACT FROM AUTHOR]

The prevention of COVID-19, caused by SARS-CoV-2, involves reducing transmission of infectious respiratory droplets and aerosols with different methods, including the use of face masks. There have been a variety of studies published about the effectiveness of face masks — they have been recognised as an inexpensive yet expedient prevention method. In this research we aimed to investigate the microbial contamination of used face masks and to compare the results based on mask usage time to determine the efficiency of facial mask use against COVID-19. Microbiological contamination of 51 used face masks was analysed by the imprinting method on suitable cultivation agars and colony forming units were determined. The majority of microorganisms found in our study were those of normal human skin and respiratory tract microbiota. A greater number of bacterial species was found on the outsides of the facial masks, which furthermore increased with prolonged usage time. We conclude that face mask use is effective in both detaining excreted microorganisms as well as protecting the wearer from microbes in the air, and therefore, should be implemented in the prevention strategies of respiratory diseases. Furthermore, our results show the importance of correct facial mask usage by their contamination with a spectrum of microorganisms. [ABSTRACT FROM AUTHOR]

In this study, antioxidant (TAS), oxidant (TOS) and antimicrobial activities of samples collected from Osmaniye province, which is located in the Eastern Mediterranean Region of Turkey and where Arum dioscoridis Sm. consumption is common, were determined. Water extracts of leaf parts of plant samples were obtained in a soxhlet device. TAS and TOS status were determined using Rel Assay kits. Antimicrobial activities of plant samples were determined against standard bacterial and fungal strains using agar dilution method. At the end of the studies, the highest TAS value among the plant samples was determined as 6.486±0.182. The highest TOS value was determined as 15.576±0.235. The highest OSI (oxidative stress index) value was determined as 0.285±0.007. In addition, A. dioscoridis extracts were determined to be effective against test microorganisms at 50-800 µg/mL concentrations. As a result, it was determined that the samples collected from different regions of the A. dioscoridis samples used in our study showed variability. In addition, it has been determined that A. dioscoridis may be an important antioxidant and antimicrobial source. [ABSTRACT FROM AUTHOR]

Mastitis is an inflammation of the mammary gland which has a particular importance in ruminants. The purpose of this study was to evaluate and compare two methods for cultivation of udder pathogens: classical laboratory cultivation on Columbia agar with 5% sheep blood versus MicroMast rapid plates. The results allowed assessing the incidence of mastitis and the prevalence of the pathogens. During the study have been investigated 227 cows in a dairy farm localized in the east of Slovakia. Subsequently, 141 quarter milk samples from the positive cows with California mastitis test score 1-4 have been undergone the laboratory culture on Columbia agar and MicroMast test in accordance with their respective steps. The values obtained from these tests showing sensitivity of positive samples using the MicroMast test at the level of 64.5%, and sensitivity of cultures on Columbia blood agar at the level of 61.7%. After biochemical identification of cultured isolates, the Columbia blood agar and MicroMast test identified both as the main pathogen present Staphylococcus aureus (S. aureus) with results of 13.4% and 14.9%, respectively. On the base of results, both tests are comparable and therefore a test that speeds up testing is more useful in practice. It will be a relevant importance in the following decades to succeed in the development of tests with directly detection of udder pathogens. [ABSTRACT FROM AUTHOR]

This work describes the design and synthesis of three series of hybrids of thienopyrimidines and sulfonamides. Dihydrofolate reductase enzyme was selected as a target for the in-silico screening of the synthesized thienopyrimidine–sulfonamide hybrid as an antibacterial, while squalene epoxidase was selected as an antifungal target protein. All screened compounds showed promising binding affinity ranges, with perfect fitting not exceeding 1.9 Å. The synthesized compounds were tested for their antimicrobial activity using agar well diffusion and minimum inhibitory concentration tests against six bacterial strains in addition to two Candida strains. Compounds 8iii and 12ii showed varying degrees of inhibition against Staphylococcus aureus and Escherichia coli bacterial strains, whereas the best antifungal activity against Candida was displayed by compound 8iii. Compound 12ii, the cyclohexathienopyrimidine coupled with sulfadiazine at position 3, has the best antibacterial activity, which is consistent with molecular docking results at the active site of the oxidoreductase protein. Interestingly, compound 12ii also has the highest docking binding energy at the antifungal squalene epoxidase active site. Investigating the physicochemical properties of the synthesized hybrids revealed their high tolerability with cell membranes, and moderate to poor oral bioavailability, and that all are drug-like candidates, among which 4i, the cyclohexathieno[2,3-d] pyrimidine core with sulphaguanidine incorporated at position 4, recorded the best score (1.58). [ABSTRACT FROM AUTHOR]

Objective(s): Cognizant of the harsh chemical method of nanoparticles synthesis, researchers are shifting towards the green phyto-mediated synthetic approaches. We herein report the green synthesis of Ag NPs and Co3O4 NPs using silver nitrate solution and cobalt nitrate solution as precursors added to aqueous extract of Croton Macrostachyus leaf extract to evaluate their antibacterial activity. Materials and Methods: The Characterization of the biosynthesized NPs were carried out by using spectroscopic techniques as X-ray diffraction (XRD), UV-Vis spectroscopy, Fourier Transform Infrared (FTIR), and scanning electron microscopy (SEM). Results: The average crystallite size of Ag NPs was found to be 12.62 nm from the XRD data, indicating a cubic crystal structure whereas that of Co3O4 NPs was found to be 12.75 nm, indicating a cubic spinel crystal structure. The energy band gap for Ag NPs and Co3O4 NPs were 3.38 eV, and 3.34 eV respectively. The SEM images showed non-homogeneity of particles distribution with irregular geometries attributable to their shape and size for Ag NPs whereas spherical as well as irregular geometries attributed to non-homogeneity of the particles for Co3O4 NPs. The FTIR identifies the functional groups of the bioactive molecules that were actively involved as stabilizing and capping agents to prevent agglomeration of Ag NPs and Co3O4 NPs. The agar-well diffusion method was employed to evaluate the antibacterial activities of the produced nanoparticles against gram-positive (S. aureus and E. faecalis) and gram-negative (E. coli, S. typhimrium) bacterial strains. Conclusion: The biosynthesized nanoparticles showed promising antibacterial activities with Ag NPs exhibiting the best inhibition activities towards all bacteria species. [ABSTRACT FROM AUTHOR]

The plant Russian knapweed is known as a blood sugar lowering drug, and the antimicrobial properties of its extracts and essential oils have been proven; Therefore, this study was conducted with the aim of extracting the aqueous extract of R. knapweed and investigating its antimicrobial activity. The aqueous extract of R. knapweed was extracted by the maceration method. The antibacterial activity of the extract against Escherichia coli, Salmonella typhi, Listeria monocytogenes, and Bacillus cereus was investigated using disk diffusion agar, well diffusion agar, minimum inhibitory concentration, and minimum bactericidal concentration methods. The results of this research showed that the antimicrobial activity of the extract was dependent on the concentration and the highest diameter of the inhibition zone was observed at the concentration of 100 mg/ml of the extract. Based on disk diffusion agar and well diffusion agar tests, Listeria monocytogenes bacteria had the highest and Escherichia coli, Salmonella typhi, and Bacillus cereus had the lowest inhibition zones, respectively. The minimum inhibitory concentration for Escherichia coli, Salmonella typhi, Listeria monocytogenes, and Bacillus cereus bacteria was obtained as 64, 64, 16, and 32 mg/ml, respectively, and the minimum bactericidal concentration values for these bacteria were 512, 512, 256, and 256 mg/ml, respectively. In generally, the Gram-positive bacteria were more sensitive to the extract compared to the Gram-negative types. The growth of pathogenic bacteria and the diseases transmitted by these bacteria can be controlled by using natural antimicrobial compounds such as R. knapweed aqueous extract. [ABSTRACT FROM AUTHOR]

Liquid marbles are widely known for their potential biomedical applications, especially due to their versatility and ease of preparation. In the present work, we prepared liquid marbles with various cores composed of water, agar-based hydrogels, magnetic fluids, or non-aqueous substances. As a coating material, we used biocompatible particles of plant origin, such as turmeric grains and Lycopodium pollen. Additionally, we provided marbles with magnetic properties by incorporating either magnetosomes or iron oxide nanoparticles as a powder or by injecting another magnetic fluid. Structures obtained in this way were stable and susceptible to manipulation by an external magnetic field. The properties of the magnetic components of our marbles were verified using electron paramagnetic resonance (EPR) spectroscopy and vibrating sample magnetometry (VSM). Our approach to encapsulation of active substances such as antibiotics within a protective hydrogel core opens up new perspectives for the delivery of hydrophobic payloads to the inherently hydrophilic biological environment. Additionally, hydrogel marbles enriched with magnetic materials showed promise as biocompatible heating agents under alternating magnetic fields. A significant innovation of our research was also the fabrication of composite structures in which the gel-like core was surrounded without mixing by a magnetic fluid covered on the outside by the particle shell. Our liquid marbles, especially those with a hydrogel core and magnetic content, due to the ease of preparation and favorable properties, have great potential for biomedical use. The fact that we were able to simultaneously produce, functionalize (by filling with predefined cargo), and manipulate (by means of an external magnetic field) several marbles also seems to be important from an application point of view. [ABSTRACT FROM AUTHOR]

This study was aimed to find out how common Listeria monocytogenes in a group of Iraqi women who had miscarriages. Also, investigates the dependability of molecular diagnosis, specifically evaluating the target hlyA gene as an alternate solution tool for identification. A total of 200 specimens comprising cervical swabs were collected from 176 women suffered from miscarriage and 24 women as control. The detection of L. monocytogenes in miscarriages women by using HiCrome™ Listeria Agar Base as both a selective and differential agar medium for quick and direct recognition of Listeria species. The biochemical tests were achieved for confirmation identification. In addition, virulenceassociated genes (hlyA gene) isolates were identified using a Real-time PCR approach. Results show out of the 200 (patients and control) specimens, 176 women with miscarriages were analyzed and it was found that 117 (66.5%) specimens from abortion cases were negative for L. monocytogenes, while 59 (33.5%) specimens were positive for Listeria species. All 24 women of control give negative result. Seventy-five (32.4%) isolates from the 59 positive women showed the presence of the hlyA gene in Real-time PCR. We found that molecular diagnostic through Real-Time PCR is a quick and accurate diagnostic method that can help minimize the time it takes to start a specific treatment and track its success. [ABSTRACT FROM AUTHOR]

Low-calorie orange marmalade was produced using sugar and reb A (0.08%) or reb A-sugar free recipes with pectin (1%) and agar agar (AA) (1%) stabilizers. The physical, chemical, and sensory properties of the marmalades were evaluated. AA stabilizers increased the viscosity of the marmalades compared to pectin, but they did not affect samples with reb A. Low-calorie samples had higher L* and b* values and lower a* values than sugar-containing samples. Total sugar content in reb A samples was 8.30 to 9.28 g/100 g, and it was lower in agar samples produced in parallel with pectin. The hydroxymethyl furfural (HMF) value was lower in sugarfree recipes (3.03 to 3.62) than in sugar-containing samples. Pectin-reb A samples had favorable taste, while AA-reb A samples favored consistency among reb A-containing samples. [ABSTRACT FROM AUTHOR]

The present study was conducted to investigate the effects of acid preservatives (AP) and food hydrocolloids (FH) on the shel f life of raw milk at room temperature (between 20 to 25°C). Raw milk samples were treated with different doses of acid preservatives (citric acid: E330 and ascorbic acid: E300) and food hydrocolloids (food agar-agar: E406 and starch: E1400) to determine the optimal dose that could prolong the shelf life of raw milk while maintaining its physicochemical, microbiological, and organoleptic quality. The results showed that the acid preservatives helped preserve the microbiological quality of the milk samples without significant effects on the final pH values. The food hydrocolloids prevented milk coagulation in the presence of acids only when applied at a dose ≤0.3 g in 100 mL of milk. Citric acid was more effective than ascorbic acid, while food agar-agar (E406) was the best food hydrocolloid. Acid preservatives and food hydrocolloids can be effective tools for prolonging the shelf life of raw milk at room temperature and improving food safety and nutrition in regions where raw milk preservation is a challenge. [ABSTRACT FROM AUTHOR]

The aim of the present study was to compare morphological and physiological responses to increasing salinity of the two subspecies of Mertensia maritima during in vitro propagation in two different systems, and subsequent ex vitro cultivation. Plants were brought into culture using seeds and further propagated on Murashige and Skoog medium supplemented with thidiazuron and naphthaleneacetic acid either on agar-solidified or liquid medium with a temporary immersion system (Plantform bioreactors). Salinity tolerance were tested both in tissue culture and with ex vitro greenhouse-cultivated plants. Low concentration of NaCl increased explant biomass in agar-solidified medium, with no negative consequences even at high concentration. However, the effect of salinity on liquid medium was genotype-specific. Explants in Plantform bioreactors showed several-fold higher biomass in comparison to that on agar-solidified medium, but the multiplication potential was not significantly affected. Ex vitro plants were negatively influenced by increasing NaCl concentration in substrate. In contrast to in vitro plants, accumulation potential of Na+ in ex vitro plants was low. One of adaptations to salinity at the whole plant level was maintenance of adequate tissue mineral nutrient homeostasis, as mineral nutrient concentrations were not negatively affected by increasing salinity. [ABSTRACT FROM AUTHOR]

The preparation of graphene materials from biomass resources is still a challenge, even more so if they are going to be employed as supports for electrocatalysts for water splitting. Herein, we describe the preparation and characterization of graphene oxides (GOs) from solid macroalgae waste obtained after processing an agar–agar residue. The structural and morphological characterization of the obtained GO confirm the presence of a lamellar material that is composed of few layers with an increased number of heteroatoms (including nitrogen) if compared with those observed in a GO obtained from graphite (reference). Three-dimensional electrodes were prepared from these GOs by depositing them onto a fibrous carbon paper, followed by electrodeposition of the catalyst, NiFe. The electrocatalytic performance of these hybrid systems for the oxygen evolution reaction (OER) showed a proactive effect of both graphene materials toward catalysis. Moreover, the electrode prepared from the algae-based graphene showed the highest electrocatalytic activity. This fact could be explained by the different structure of the algae-based graphene which, due to differences in the nucleation growth patterns and electroactive sites developed during the electrodeposition process, produced more reactive NiFe species (higher oxidation state). [ABSTRACT FROM AUTHOR]

Background: Agar is used as a gelling agent that possesses a variety of biological properties; it consists of the polysaccharides agarose and porphyrin. In addition, the monomeric sugars generated after agar hydrolysis can be functionalized for use in biorefineries and biofuel production. The main objective of this study was to develop a sustainable agar hydrolysis process for bioethanol production using nanotechnology. Peroxidase-mimicking Fe3O4-MNPs were applied for agar degradation to generate agar hydrolysate-soluble fractions amenable to Saccharomyces cerevisiae and Escherichia coli during fermentation. Results: Fe3O4-MNP-treated (Fe3O4-MNPs, 1 g/L) agar exhibited 0.903 g/L of reducing sugar, which was 21-fold higher than that of the control (without Fe3O4-MNP-treated). Approximately 0.0181% and 0.0042% of ethanol from 1% of agar was achieved using Saccharomyces cerevisiae and Escherichia coli, respectively, after process optimization. Furthermore, different analytical techniques (FTIR, SEM, TEM, EDS, XRD, and TGA) were applied to validate the efficiency of Fe3O4-MNPs in agar degradation. Conclusions: To the best of our knowledge, Fe3O4-MNP-treated agar degradation for bioethanol production through process optimization is a simpler, easier, and novel method for commercialization. [ABSTRACT FROM AUTHOR]

The development of a wound dressing with both antibacterial and healing-guiding functions is a major concern in the treatment of open and infected wounds. In this study, poly(hexamethylene biguanide) hydrochloride (PHMB) was loaded into a 3D silk fibroin (SF) scaffold based on electrostatic interactions between PHMB and SF, and PHMB/SF hybrid scaffolds were prepared via freeze-drying. The effects of the PHMB/SF ratio on the antibacterial activity and cytocompatibility of the hybrid scaffold were investigated. The results of an agar disc diffusion test and a bacteriostasis rate examination showed that when the mass ratio of PHMB/SF was greater than 1/100, the scaffold exhibited obvious antibacterial activity against E. coli and S. aureus. L-929 cells were encapsulated in the PHMB/SF scaffolds and cultured in vitro. SEM, laser scanning confocal microscopy, and CCK-8 assay results demonstrated that hybrid scaffolds with a PHMB/SF ratio of less than 2/100 significantly promoted cell adhesion, spreading, and proliferation. In conclusion, a hybrid scaffold with a PHMB/SF ratio of approximately 2/100 not only effectively inhibited bacterial reproduction but also showed good cytocompatibility and is expected to be usable as a functional antibacterial dressing for wound repair. [ABSTRACT FROM AUTHOR]

Natural products and analogues are a source of antibacterial drug discovery. Considering drug resistance levels emerging for antibiotics, identification of bacterial metalloenzymes and the synthesis of selective inhibitors are interesting for antibacterial agent development. Peptide nucleic acids are attractive antisense and antigene agents representing a novel strategy to target pathogens due to their unique mechanism of action. Antisense inhibition and development of antisense peptide nucleic acids is a new approach to antibacterial agents. Due to the increased resistance of biofilms to antibiotics, alternative therapeutic options are necessary. To develop antimicrobial strategies, optimised in vitro and in vivo models are needed. In vivo models to study biofilm-related respiratory infections, device-related infections: ventilator-associated pneumonia, tissue-related infections: chronic infection models based on alginate or agar beads, methods to battle biofilm-related infections are discussed. Drug delivery in case of antibacterials often is a serious issue therefore this review includes overview of drug delivery nanosystems. [ABSTRACT FROM AUTHOR]

Staphylococcus aureus is a bacterium that is both common and capable of causing harmful infections in humans. Staphylococcus aureus colonies around 30% of the human population. Staphylococcus aureus has a variety of virulence traits that contribute to its capacity to colonies and infiltrate host tissues, avoid the host's immune system, and collect nutrition. Methicillin-resistant Staphylococcus aureus (MRSA) outbreaks have arisen as a major concern in healthcare facilities around the world. Sample collection began from September 2022 to January 2023, during which 300 samples were isolated from patients, including wound swabs, medial urine samples, semen, upper vaginal swabs, and burn swabs. The samples were selected from the patients of Al-Furat Al-Awsat and Al-Najaf Teaching Hospitals in Al-Najaf Governorate in the Guidance Department, and the included age groups ranged from (1-60) years. After diagnosing the samples by Gram stain, catalase test, and Coagulase test and cultures on a mannitol agar, the diagnosis was confirmed by the Vitek device. DNA was extracted by extraction kit. In the next stage, virulence genes were detected by PCR technique, and the result was PVL (20%), while Nuc was (90%), Sea was (63.3%), and finally, icaA was (36.6%). Correlations of virulence genes were also made, namely between age, sex and gene coexistence in the isolates, with Sea and Nuc being the most prevalent, followed by Sea with icaA. [ABSTRACT FROM AUTHOR]

Increasing antimicrobial resistance to the action of existing antibiotics has prompted researchers to identify new natural molecules with antimicrobial potential. In this study, a green system was developed for biosynthesizing gold nanoparticles (BAuNPs) using sage (Salvia officinalis L.) leaf extract bioconjugated with non-toxic, eco-friendly, and biodegradable chitosan, forming chitosan/gold bioconjugates (Chi/BAuNPs). Characterization of the BAuNPs and Chi/BAuNPs conjugates takes place using transmission electron microscopy (TEM), X-ray spectra, Fourier transform infrared (FT-IR) spectroscopy, and zeta potential (Z-potential). The chemical composition of S. officinalis extract was evaluated via gas chromatography/mass spectrometry (GC/MS). This study evaluated the antioxidant and antimicrobial activities of human pathogenic multidrug-resistant (MDR) and multisensitive (MS) bacterial isolates using the agar diffusion method. Chi/BAuNPs showed inhibition of the MDR strains more effectively than BAuNPs alone as compared with a positive standard antibiotic. The cytotoxicity assay revealed that the human breast adenocarcinoma cancer cells (MCF7) were more sensitive toward the toxicity of 5-Fu + BAuNPs and 5-Fu + Chi/BAuNPs composites compared to non-malignant human fibroblast cells (HFs). The study shows that BAuNPs and Chi/BAuNPs, combined with 5-FU NPs, can effectively treat cancer at concentrations where the free chemical drug (5-Fu) is ineffective, with a noted reduction in the required dosage for noticeable antitumor action. [ABSTRACT FROM AUTHOR]

Simple Summary: Artificial diets are one of the most critical components in insect mass-rearing facilities worldwide. Research on artificial diets to rear pestiferous tephritid flies dates back to the 1950s but is still a topic of great relevance, as fruit flies represent one of the ten most important pests of fruit in temperate and tropical regions of the world. In the past 15 years, much research has been performed to develop liquid- or gel-based diets more closely resembling the rearing environment a wild fruit fly larva experiences inside a fruit in nature. To identify new gelling agents that could perform better than already established ones such as agar and carrageenan, we assessed the performance of calcium alginate, a gelling agent widely used in the food industry, that had not been tested in the case of fruit fly mass rearing. We also examined larval density in gel diets as a critical factor that affects insect-rearing systems. Our results show that agar and carrageenan diets outperformed calcium alginate diets broadly and that the effects of larval density depended on the gel type used. We believe that the applications of calcium alginate in tephritid rearing should be analyzed in more depth in future studies. Research on larval rearing and nutrition of tephritid flies on artificial diets is key for the sterile insect technique. Here, we examined the effects of the type of gel (calcium alginate, agar, or carrageenan), at varying percentages in artificial diets for the polyphagous pest Anastrepha ludens, on the physicochemical and nutritional traits of the diets, and the effects of the type of gel, the gel content and the larval density (larvae/g of diet) used in production, quality parameters for mass-reared tephritids, diet removal (an indirect estimation of diet consumption), and nutritional traits of flies. Regardless of the gel content, calcium alginate diets were firmer and more resistant to penetration than the agar and carrageenan diets. The larval recovery, pupation, pupal weight, and flight ability of A. ludens were lower in calcium alginate diets than in agar and carrageenan diets. Diet removal was higher in calcium alginate diets; however, low levels of ammonium and high levels of uric acid in excretions from larvae on these diets suggest an alteration in protein metabolism. The firmness and penetration resistance characteristics of calcium alginate diets may have limited movement and feeding of larvae, but this could be overcome by the collective feeding of large groups of larvae. Our findings provide insights into the mechanism governing gel-diet rearing systems for A. ludens. [ABSTRACT FROM AUTHOR]

Nonomuraea gerenzanensis (N. gerenzanensis) is known for its ability to biosynthesize A40926, the precursor of the glycopeptide antibiotic (GPA) Dalbavancin. However, challenges and uncertainties related to the genetic manipulation of the rare actinomycetes remain. In order to improve the conjugation transfer of N. gerenzanensis, the crucial factors affecting conjugal transfer were evaluated, including agar medium, mycelial state, donor-recipient ratio, magnesium ion concentration, and antibiotic coverage time firstly. Additionally, γ-butyrolactone (GBL) for quorum sensing (QS) and antibiotics targeting bacterial walls were applied to evaluate their effects on conjugation transfer. As a result, the optimal conditions of 5%TSB of liquid medium, 24 h of the period time, V0.1 of agar medium, 30 mM of magnesium ion, the ratio 10:1 of donor-to-recipient, and 27 h of the overlaying time of antibiotic were determined. Furthermore, the results showed that autoinducer GBL and GPA teicoplanin had a synergetic effect on the conjugation transfer of N. gerenzanensis at a working concentration of 60 µM and 0.5 µg mL−1, respectively. The highest conjugation efficiency could reach about 1.3 depending on the optimal process conditions and the interference of QS and antibiotics. [ABSTRACT FROM AUTHOR]

Osteoarthritis (OA) is a degenerative disease. It is common in middle-aged and elderly people and is one of the main causes of disability. Currently, the etiology of OA is unclear, and no specific biomarkers for the diagnosis of OA have been identified. Therefore, finding a highly sensitive biomarker is essential for a proper diagnosis. TRNA-derived fragments (tRFs) and tRNA-derived stress-induced RNAs (tiRNAs) are newly discovered classes of noncoding RNAs. tRF has been proven in several studies to have significant associations with tumor diagnosis, making it a promising biomarker in cancer research. However, the diagnostic utility of tRF in OA patients and the correlation between OA progression and trf differential expression have yet to be elaborated. The purpose of this research was to identify tRFs with differential expression in OA to assess their potential as OA biomarkers. To determine the tRF-5022B expression level in this research, real-time fluorescence quantitative PCR has been employed. Agarose gel electrophoresis, Sanger sequencing, and other investigations have been employed for evaluating tRF-5022B's molecular properties. Receiver operating characteristic curve analysis has been utilized for assessing the diagnostic effectiveness of the tRF-5022B. The findings demonstrated that tRF-5022B expression was considerably lower in OA serum. The Kellgren–Lawrence grading scale was shown to correspond with serum expression levels. The ROC curve confirmed that tRF-5022B serum expression levels might differentiate OA cases from healthy individuals and RA patients. According to the aforementioned findings, tRF-5022B may be employed as a novel biomarker for OA diagnosis due to its excellent diagnostic value. [ABSTRACT FROM AUTHOR]

Atopic dermatitis (AD) is a common skin disease involving important immune mechanisms. There is an unmet need for a treatment for this condition. Herein, we focused on elucidating the role of Bi2-xMnxO3 nanospheres (BM) in alleviating skin inflammation in AD-like C57BL/6 mice. The BM was fabricated via sacrificial templates and its biosafety was systematically evaluated. The BM was applied topically to skin lesions of AD-like C57BL/6 mice. The phenotypic and histological changes in the skin were examined carefully. The responses of barrier proteins, inflammatory cytokines and cells to BM were evaluated in HaCaT cells and AD mouse models. The data demonstrated that BM treatment alleviated the AD phenotypes and decreased the level of inflammatory factors, while increasing the expression of the barrier proteins filaggrin/involucrin in the skin. BM effectively reduced the expression of phosphorylated STAT6, which in turn reduced the expression of GATA3, and further decreased the differentiation ratio of Th2 cells, thereby reducing the expression of IL-4. In conclusion, topical drug therapy with BM provides a safe and effective treatment modality for AD by reducing IL-4 and increasing barrier proteins. Highlights: This study was the first to explore the feasibility of Bi2-xMnxO3 nanospheres (BM) in agar gel as STAT phosphorylation regulators to improve the clinical manifestations in MC903-induced atopic dermatitis (AD)—like C57BL/6 mice, and to evaluate their immunomodulatory effects in vitro and in vivo. BM showed good skin penetration and system safety. BM reduced the expression of IL-4, IL-13 and p-STAT6 in TNF-α/IFN-γ-induced HaCaT inflammatory cells and in the MC903-induced mouse AD model, thereby effectively improving the expression of the barrier protein FLG/IVL and promoting the recovery of damaged skin. Moreover, the ratio of Th2 cells in the skin and spleen was considerably decreased after BM administration in MC903-induced AD mice, indicating that the as-prepared BM inhibits the Th2 cells response in the AD mouse model. BM has the potential to be an innovative therapeutic strategy for AD. [ABSTRACT FROM AUTHOR]

Flagella-driven motility is essential for Helicobacter pylori to colonize the human stomach, where it causes a variety of diseases, including chronic gastritis, peptic ulcer disease, and gastric cancer. H. pylori has evolved a high-torque-generating flagellar motor that possesses several accessories not found in the archetypical Escherichia coli motor. FlgV was one of the first flagellar accessory proteins identified in Campylobacter jejuni, but its structure and function remain poorly understood. Here, we confirm that deletion of flgV in H. pylori B128 and a highly motile variant of H. pylori G27 (G27M) results in reduced motility in soft agar medium. Comparative analyses of in-situ flagellar motor structures of wild-type, ΔflgV, and a strain expressing FlgV-YFP showed that FlgV forms a ring-like structure closely associated with the junction of two highly conserved flagellar components: the MS and C rings. The results of our studies suggest that the FlgV ring has adapted specifically in Campylobacterota to support the assembly and efficient function of the high-torque-generating motors. [ABSTRACT FROM AUTHOR]

Mesoporous silica nanoparticles (MSNs) are amongst the most used nanoparticles in biomedicine. However, the potentially toxic effects of MSNs have not yet been fully evaluated, being a controversial matter in research. In this study, bare MSNs, PEGylated MSNs (MSNs-PEG), and galacto-oligosaccharide-functionalized MSNs (MSNs-GAL) are synthesized and characterized to assess their genotoxicity and transforming ability on human lung epithelial BEAS-2B cells in short- (48 h) and long-term (8 weeks) exposure scenarios. Initial short-term treatments show a dose-dependent increase in genotoxicity for MSNs-PEG-treated cells but not oxidative DNA damage for MSNs, MSNs-PEG, or for MSNs-GAL. In addition, after 8 weeks of continuous exposure, neither induced genotoxic nor oxidative DNA is observed. Nevertheless, long-term treatment with MSNs-PEG and MSNs-GAL, but not bare MSNs, induces cell transformation features, as evidenced by the cell's enhanced ability to grow independently of anchorage, to migrate, and to invade. Further, the secretome from cells treated with MSNs and MSNs-GAL, but not MSNs-PEG, shows certain tumor-promoting abilities, increasing the number and size of HeLa cell colonies formed in the indirect soft-agar assay. These results show that MSNs, specifically the functionalized ones, provoke some measurable adverse effects linked to tumorigenesis. These effects are in the order of other nanomaterials, such as carbon nanotubes or cerium dioxide nanoparticles, but they are lower than those provoked by some approved drugs, such as doxorubicin or dexamethasone. [ABSTRACT FROM AUTHOR]

Recovering a sufficient amount of microbial DNA from extremely low-biomass specimens, such as human skin, to investigate the community structure of the microbiome remains challenging. We developed a sampling solution containing agar to increase the abundance of recovered microbial DNA. Quantitative PCR targeting the 16S rRNA gene revealed a significant increase in the amount of microbial DNA recovered from the developed sampling solution compared with conventional solutions from extremely low-biomass skin sites such as the volar forearm and antecubital fossa. In addition, we confirmed that the developed sampling solution reduces the contamination rate of probable non-skin microbes compared to the conventional solutions, indicating that the enhanced recovery of microbial DNA was accompanied by a reduced relative abundance of contaminating microbes in the 16S rRNA gene amplicon sequencing data. In addition, agar was added to each step of the DNA extraction process, which improved the DNA extraction efficiency as a co-precipitant. Enzymatic lysis with agar yielded more microbial DNA than conventional kits, indicating that this method is effective for analyzing microbiomes of low-biomass specimens. [ABSTRACT FROM AUTHOR]

Hypervirulent Klebsiella pneumoniae strains (hvKp) can cause invasive community-acquired infections in healthy patients of all ages. In this study, the prevalence of putative hvKp in a German tertiary center was investigated and hvKp were characterized by phenotypic and molecular assays. All K. pneumoniae isolates in routine microbiological diagnostics from a single center were screened by string-testing over a period of 6 months. String-test positive (≥ 0.5 mm) isolates were re-evaluated on different media and under various conditions (aerobe, anaerobe). For string-test positive isolates, genes (magA, iutA, rmpA and rmpA2) associated with hypermucoviscosity and hypervirulence were amplified by multiplex PCR. PCR-positive isolates were subjected to whole-genome sequencing and sedimentation and biofilm formation assays. From 1310 screened K. pneumoniae isolates in clinical routine 100 isolates (7.6%) were string test positive. From these, 9% (n = 9) were defined as putative hvKp (string-test+/PCR+). Highest rate of string-test-positive isolates was observed on MacConkey agar under aerobic conditions. Amongst these nine putative hvKp isolates, the international lineage ST23 carrying hvKp-plasmid pKpVP-1 was the most common, but also a rare ST86 with pKpVP-2 was identified. All nine isolates showed hypermucoviscosity and weak biofilm formation. In conclusion, 9% of string-positive, respectively 0.69% of all K. pneumoniae isolates from routine were defined as putative hypervirulent. MacConkey agar was the best medium for hvKp screening. [ABSTRACT FROM AUTHOR]

This study aimed to evaluate the antifungal effect of Iranian oak Jaft extract on Penicillium italicum and Penicillium digitatum, as an indicator of mold spoilage after citrus fruits harvest. Different extraction methods were used to obtain tannins and flavonoids from the Oak Jaft. The study investigated the Minimum Inhibitory Concentration (MIC), Minimum Fungicidal Concentration (MFC), and non-growth diameter of the molds using three different methods: tube dilution, surface culture, and diffusion in agar well. The results showed that different concentrations of aqueous extract had varying effects on the non-growth diameter of the molds. The antifungal effect was significantly increased with a concentration of 40 and 80 mg/ml of aqueous extract. In comparison, the control group, which used Fludioxonil 25% poison, showed a significant difference in mold growth. Therefore, the extract of Iranian oak Jaft can be used as an alternative to control fungal spoilage of citrus fruits. [ABSTRACT FROM AUTHOR]

Although many protocols have been previously developed for genomic DNA (gDNA) extraction from S. cerevisiae, to take advantage of recent advances in laboratory automation and DNA-barcode sequencing, there is a need for automated methods that can provide high-quality gDNA at high efficiency. Here, we describe and demonstrate a fully automated protocol that includes five basic steps: cell wall and RNA digestion, cell lysis, DNA binding to magnetic beads, washing with ethanol, and elution. Our protocol avoids the use of hazardous reagents (e.g., phenol, chloroform), glass beads for mechanical cell disruption, or incubation of samples at 100°C (i.e., boiling). We show that our protocol can extract gDNA with high efficiency both from cells grown in liquid culture and from colonies grown on agar plates. We also show results from gel electrophoresis that demonstrate that the resulting gDNA is of high quality. [ABSTRACT FROM AUTHOR]

Simple Summary: Overuse of antibiotics has resulted in antibiotic-resistant bacteria and adverse changes in aquaculture ecology. Thus, natural bioproducts have been sourced for use in aquaculture as substitutes for antibiotics. This study investigated bacterial exopolysaccharide (EPS) as an antioxidant and a stimulant for fish immune systems. Agro-industrial biowaste carbon sources that are more economical than sugars are preferred to mitigate the production cost of EPS for commercial applications. In this study, the EPS produced from riceberry broken rice and soybean meal by Bacillus tequilensis PS21 bacteria from milk kefir grain within 72 h displayed antioxidant activity and antimicrobial activity toward pathogenic bacteria Streptococcus agalactiae. In Nile tilapia (Oreochromis niloticus), EPS supplementation at a high dose of 2.0 g EPS/kg feed significantly increased fish survival post-challenge with S. agalactiae EW1 and also boosted the fish's immune system. The study suggested a method for utilizing agro-industrial biowaste as a value-added EPS source for a bio-circular green economy model to preserve a healthy environment while also enabling sustainable aquaculture. Overuse of antibiotics in aquaculture has generated bacterial resistance and altered the ecology. Aquacultural disease control requires an environmentally sustainable approach. Bacterial exopolysaccharides (EPSs) as bioimmunostimulants have not been extensively explored in aquaculture. This study investigated EPS produced from 5% w/v riceberry broken rice as a carbon source and 1% w/v soybean meal as a nitrogen source by Bacillus tequilensis PS21 from milk kefir grain for its immunomodulatory, antioxidant activities and resistance to pathogenic Streptococcus agalactiae in Nile tilapia (Oreochromis niloticus). The FTIR spectrum of EPS confirmed the characteristic bonds of polysaccharides, while the HPLC chromatogram of EPS displayed only the glucose monomer subunit, indicating its homopolysaccharide feature. This EPS (20 mg/mL) exhibited DPPH scavenging activity of 65.50 ± 0.31%, an FRAP value of 2.07 ± 0.04 mg FeSO4/g DW, and antimicrobial activity (14.17 ± 0.76 mm inhibition zone diameter) against S. agalactiae EW1 using the agar disc diffusion method. Five groups of Nile tilapia were fed diets (T1 (Control) = 0.0, T2 = 0.1, T3 = 0.2, T4 = 1.0, and T5 = 2.0 g EPS/kg diet) for 90 days. Results showed that EPS did not affect growth performances or body composition, but EPS (T4 + T5) significantly stimulated neutrophil levels and serum lysozyme activity. EPS (T5) significantly induced myeloperoxidase activity, catalase activity, and liver superoxide dismutase activity. EPS (T5) also significantly increased the survival of fish at 80.00 ± 5.77% at 14 days post-challenge with S. agalactiae EW1 compared to the control (T1) at 53.33 ± 10.00%. This study presents an efficient method for utilizing agro-industrial biowaste as a prospective source of value-added EPS via a microbial factory to produce a bio-circular green economy model that preserves a healthy environment while also promoting sustainable aquaculture. [ABSTRACT FROM AUTHOR]

Paper is one of the most promising materials for food packaging and wrapping due to its low environmental impact, but surface treatments are often needed to improve its performance, e.g., the resistance to fats and oils. In this context, this research is focused on the formulation of a new paper bio-coating. Paper was coated with liquids containing poly(hexano-6-lactone) (PCL), glycerol and variable percentages of starch (5–10% w/w PCL dry weight), agar-agar (0–1.5% w/w PCL dry weight), and polyethylene glycol (PEG) (5% or 15% w/w PCL dry weight) to improve coating uniformity and diffusion. A design of experiments approach was implemented to find statistically reliable results in terms of the best coating formulation. Coated paper was characterized through mechanical and physical properties. Results showed that agar content (1.5% w/w PCL dry weight) has a beneficial effect on increasing the resistance to oil. Furthermore, the best coating composition has been calculated, and it is 10% w/w PCL dry weight of starch, 1.5% w/w PCL dry weight of agar, and 15% w/w PCL dry weight of PEG. [ABSTRACT FROM AUTHOR]

The performance of Bioactive Mesoporous Glasses (MBGs) as tissue regeneration scaffolds can be enhanced by the addition of therapeutic inorganic ions (TIIs) or organic molecules. In order to maximise the benefits conferred by different TIIs and organic molecules on bioactive glasses, we investigated the behavior of a 1:1 mixture (wt.%) of MBGs doped with cerium and loaded separately with gentamicin (Gen) and polyphenols (POLY). The resulting admixed material preserves bioactivity of the unloaded cerium-doped MBGs, assessed by evaluating the formation of an apatitic layer. Furthermore, it shows both antioxidant properties, evaluated by the reduction of Reactive Oxygen Species (ROS) measured by enzymatic assays, conferred by cerium POLY loadings and antibacterial properties conferred by Gen, and estimated by a modification of the Kirby-Bauer agar diffusion method. Ce-MBGs can thus be applied as a useful dual drug delivery system (DDS) with hard tissue regeneration capabilities. [ABSTRACT FROM AUTHOR]

Aims and Objectives Mupirocin is a widely used topical antibiotic for the treatment of skin and soft tissue infections. This has resulted in resistance leading to treatment failure. Hence, the present study aimed to determine the prevalence of mupirocin resistance among staphylococcal isolates obtained from the skin and soft tissue infections. Also, comparison of disc diffusion and agar dilution method in detecting mupirocin resistance was done. Materials and Methods This cross-sectional study was conducted in the Department of Microbiology of a tertiary health care center in Karnataka from January to December, 2018. Clinical samples such as wound swabs, tissues, and pus were included in the study. All staphylococcal isolates were screened for mupirocin resistance using 5 µg and 200 µg discs for low-level (MuL) and high-level mupirocin resistance (MuH), respectively. Minimum inhibitory concentration (MIC) was determined using the agar dilution method. Results Out of 100 staphylococcal isolates, 68 were Staphylococcus aureus and 32 were CoNS. MuH was detected in 11 isolates. MuH was more common in CoNS (10/11) compared with S. aureus (1/11). MuL was not found in the study. Discussion In our study, 10 out of 11 mupirocin-resistant isolates were methicillin resistant, which is statistically significant (p < 0.05). The correlation between results of disc diffusion and MIC were appropriate in this study. Conclusion Judicial prescription of mupirocin after knowing the susceptibility report should become the standard practice. Screening for mupirocin resistance can be done by disc diffusion in resource-limited settings. [ABSTRACT FROM AUTHOR]

Nowadays, the increasing demand for healthy foods has led to application of natural ingredients such as fruits in the formulation of food products. In this study, date waste (grade two) has been used in the formulation of a kind of nutritious jelly candy (up to 30%) with substitution of agar gum instead of gelatin (0, 15, 25 and 35%). Then physicochemical properties (moisture, dry matter, total ash, water activity, and color indices), texture characteristics (hardness, adhesiveness, springiness, stringiness and fracturability) and sensorial properties of the samples were analyzed and one-way ANOVA test at significant level of 95% was used for statistical analysis. The result showed that addition of date puree into jelly candy formula decreased moisture content, L* and b*, significantly (p<0.05) while total ash, a*, adhesiveness and stringiness showed an increasing manner (p<0.05). But effect of adding date puree on water activity and other texture characteristics including hardness, springiness and fracturability was not significant (p>0.05). Substitution of gelatin with agar (to 35%) resulted to decrease in moisture content and increase of total ash, color parameters and texture characteristics (p<0.05) while its effect on the other texture characteristics and water activity was not significant (p>0.05). The result of sensory evaluation indicated that the jelly candy containing 100% gelatin (agar free) recognized as the best specimen because of obtaining the suitable physicochemical properties and desirable texture. [ABSTRACT FROM AUTHOR]

The effect of 44 agrochemicals (fungicides, insecticides and adjuvants) on the mycelial growth and germination conidia of Trichoderma koningiopsis Th003 was determined by seeding of 5 mm discs of fungal mycelium in Sabouraud agar supplemented with each agrochemical and seeding of conidia suspended in solutions of agrochemicals in water agar. For the adjuvants, their effect on the phyllospheric establishment of T. koningiopsis Th003 was determined by following their population in cape gooseberry leaflets inoculated with the fungus suspended in them. Eight fungicides did not inhibit the fungus mycelial growth or the conidia germination. Fenhexamid - Tebuconazole, Flutriafol and Kasugamicina inhibited it between 34 and 48% without affecting germination, Thiram - Pyrimethanil, Prochloraz, Tiabendazol, Spiroxamina and Triadimenol - Tebuconazole inhibited the growth and Thiram - Pyrimethanil and Dodine did not allow conidia germination. Insecticides and adjuvants presented an inhibition of up to 70% but did not affect the conidia germination. No negative effects of the adjuvants on the phyllosphere establishment of T. koningiopsis Th003 were observed, recovering from the treatments approximately 1x103 CFU g-1. [ABSTRACT FROM AUTHOR]

The genus Armillaria has high edible and medical values, with zones of antagonism often occurring when different species are paired in culture on agar media, while the antagonism-induced metabolic alteration remains unclear. Here, the metabolome of mycelial exudates of two Chinese Armillaria biological species, C and G, co-cultured or cultured separately was analysed to discover the candidate biomarkers and the key metabolic pathways involved in Armillaria antagonists. A total of 2,377 metabolites were identified, mainly organic acids and derivatives, lipids and lipid-like molecules, and organoheterocyclic compounds. There were 248 and 142 differentially expressed metabolites between group C-G and C, C-G, and G, respectively, and fourteen common differentially expressed metabolites including malate, uracil, Leu-Gln-Arg, etc. Metabolic pathways like TCA cycle and pyrimidine metabolism were significantly affected by C-G co-culture. Additionally, 156 new metabolites (largely organic acids and derivatives) including 32 potential antifungal compounds, primarily enriched into biosynthesis of secondary metabolites pathways were identified in C-G co-culture mode. We concluded that malate and uracil could be used as the candidate biomarkers, and TCA cycle and pyrimidine metabolism were the key metabolic pathways involved in Armillaria antagonists. The metabolic changes revealed in this study provide insights into the mechanisms underlying fungal antagonists. [ABSTRACT FROM AUTHOR]

In recent years, the antimicrobial resistance in Escherichia coli has gradually developed into a global problem. These resistant bacteria could be transmitted to humans through animal feces in the environment or direct contact with pets, leading to a problem in bacterial treatment for humans and animals. Now, the antibiotic resistance of oral and intestinal microbiota from dog origins remains unclear in China. Therefore, this study first analyzed the current colistin resistance of oral and intestinal microbiota from dog origins in mainland China. A total of 536 samples were collected from dogs in mainland China and, respectively, cultured on the SS and MacConkey agar plate containing colistin (4 μg/mL) to obtain bacteria, and the antibiotic-resistance phenotype of Escherichia coli was investigated for nine antibiotics. Results showed that a total of 2259 colistin-resistant bacteria were isolated from samples and identified, and among them, the isolated rate of Escherichia coli (34.01%, 769/2259) was relatively higher than that of other bacteria. Subsequently, it was found that the resistance of these Escherichia coli was very severe by exploring its resistance to different antibiotics, particularly to three common antibiotics in a clinic which were ceftriaxone, ampicillin and trimethoprim/sulfamethoxazole, with the resistance rates of 60.60% (466/769), 57.22% (440/769), and 53.06% (408/769), respectively. Moreover, the simultaneous resistance of Escherichia coli to one or more antibiotics was determined, and 69.96% (538/769) strains have defined the resistance to both two or more antibiotics, and even 13 of Escherichia coli strains that were resistant to all nine antibiotics, indicating that the Escherichia coli from dog origins has severe antibiotic resistance in the clinic. In conclusion, this study guided the use of antibiotics and could draw attention to antibiotic resistance in veterinary clinical treatment for animals in the future. [ABSTRACT FROM AUTHOR]