Downgrading in the yield of crop is due to the inadequate availability of water. The way out for this trouble is to construct synthetic resources dependent on natural polymers with great water absorption and preservation limits. The present study investigated the design of agar-agar (Agr) and gelatin (GE) copolymerized methyl acrylate (MA) and acrylic acid (AA) hydrogel (Agr/GE-co-MA/AA) as a soil conditioner for moisture maintenance in agriculture. Agr/GE-co-MA/AA hydrogel was prepared by utilizing microwave-assisted green synthesis following the most suitable reaction conditions to obtain a remarkable water swelling percentage. The fabricated Agr/GE-co-MA/AA hydrogel was investigated through field emission scanning electron microscopy (FESEM), fourier transform infrared spectroscopy (FTIR), and X-ray diffraction (XRD). The water holding capacity of the soil and sand was examined by mixing Agr/GE-co-MA/AA hydrogel with soil and sand. The result demonstrates that the water holding time extended from 10 to 30 days for soil and 6 to 10 days for sand by using Agr/GE-co-MA/AA hydrogel. This synthesized biodegradable, low-cost and non-toxic Agr/GE-co-MA/AA hydrogel shows novelty as soil water maintaining material for irrigation in agriculture.

There is a need for simple microbiology diagnostics to enable antimicrobial resistance surveillance in low- and middle-income countries.

The reference standard for fosfomycin antimicrobial susceptibility testing (AST) is agar dilution, but it is laborious and is not routinely used in diagnostic microbiology. In this study, we evaluated the performance of a ready-to-use commercially available agar dilution kit for fosfomycin AST (Liofilchem Diagnostics). We compared this kit with the reference standard agar dilution, performed according to the Clinical & Laboratory Standards Institute (CLSI) in 229 clinical isolates. The isolates were selected to represent both Gram-positive and Gram-negative microorganisms, with various MIC values. It consisted of 43 enterococci (E. faecalis n = 16, E. faecium n = 27), 13 methicillin-resistant S. aureus (MRSA), 118 Enterobacterales (Escherichia coli n = 94, Klebsiella pneumoniae n = 20, and Enterobacter cloacae complex n = 4), 55 Pseudomonas aeruginosa, and three ATCC isolates. Using CLSI breakpoints for enterococci for oral treatment of urinary tract infections, European Committee on Antimicrobial Susceptibility Testing (EUCAST) breakpoints for intravenous dosing for Enterobacterales and Staphylococci, and epidemiological cutoff value for P. aeruginosa, the essential agreement was 87.5%, and 99.6% after discrepancy resolution. There was no very major error, and 1.9% major error before, and 0.9% major error after resolution of discrepancies. The commercial test showed 100% reproducibility. In conclusion, in comparison to the reference standard, the ready-to-use commercially available agar dilution kit for fosfomycin AST showed excellent performance. IMPORTANCE Fosfomycin is increasingly needed to treat infection due to multidrug resistant microorganisms. Yet, the antimicrobial susceptibility test (AST) of fosfomycin is fraught with difficulties or often laborious to perform. An easy-to-use AST method for fosfomycin is thus needed. In this study, we showed that the ready-to-use commercially available agar dilution kit, in comparison to the reference standard, showed excellent performance.

Root system architecture and especially its plasticity in acclimation to variable environments play a crucial role in the ability of plants to explore and acquire efficiently soil resources and ensure plant productivity. Non-destructive measurement methods are indispensable to quantify dynamic growth traits. For closing the phenotyping gap, we have developed an automated phenotyping platform, GrowScreen-Agar, for non-destructive characterization of root and shoot traits of plants grown in transparent agar medium.

Agar have long been used as a growth media for plants. Here, we made agar media with embedded fluidic channels to study the effect of exposure to nutrient solution on root growth and pull-out force. Black Eye bean (Vigna Unguiculata) and Mung bean (Vigna Radiata) were used in this study due to their rapid root development. Agar media were fabricated using casting process with removable cores to form channels which were subsequently filled with nutrient solution. Upon germination, beans were transplanted onto the agar media and allowed to grow. Pull-out force was determined at 96, 120 and 144 h after germination by applying a force on the hypocotyl above the gel surface. The effect of nutrients was investigated by comparing corresponding data obtained from control plants which have not been exposed to nutrient solution. Pull-out force of Black Eye bean plantlets grown in agar with nutrient solution in channels was greater than those grown in gel without nutrients and was 110% greater after 144 h of germination. Pull-out force of Mung bean plantlets grown in agar with and without nutrient solution was similar. Tap root lengths of Black Eye bean and Mung Bean plantlets grown in agar with nutrient solution are shorter than those grown without nutrient.

Although the pour plate method is widely employed in microbiological quality control, it has certain drawbacks, including having to melt the culture medium before seeding. In this study, the preparation of the culture medium was modified by using a lower concentration of agar (10 g/L), which was separated from the nutrients during sterilization. The new protocol was assessed in media frequently used in microbiological quality control of food, cosmetics, and pharmaceutical products, with tryptic soy agar (TSA), Sabouraud 4% dextrose agar (SDA), and violet red bile glucose agar (VRBG). In comparison with the conventionally produced media, the modifications significantly improved the growth of Saccharomyces cerevisiae in SDA, Staphylococcus aureus, Salmonella enterica subsp. enterica serovar Typhimurium, and Candida albicans in TSA and Escherichia coli ATCC 8739 and ATCC 25922 and S. Typhimurium in VRBG. The modified VRBG was also more selective for Pseudomonas aeruginosa. Regarding physicochemical properties, a significantly lower pH was observed in TSA and VRBG and lower strength values in TSA. Sterilizing agar separately from the other components of the medium and reducing the agar concentration to 10 g/L can improve microorganism growth and enhance the selectivity of differential media in the pour plate method. These modifications could facilitate the automation of this culture technique. IMPORTANCE In the era of rapid microbiological methods, there is a need to improve long-established culture techniques. Drawbacks of the pour plate method include having to melt each medium separately before seeding. For this technique, we demonstrate that separating the agar from the other components of commonly used media during sterilization and reducing the agar concentration to 10 g/L can enhance microbial growth. The new protocol could have advantages in routine laboratory practice because less agar is required and the same molten agar suspension can be used to prepare different media. Moreover, these modifications could facilitate the automation of the pour plate method.

Antibacterial surfaces have an enormous economic and social impact on the worldwide technological fight against diseases. However, bacteria develop resistance and coatings are often not uniform and not stable in time. The challenge is finding an antibacterial coating that is biocompatible, cost-effective, not toxic, and spreadable over large and irregular surfaces. Here we demonstrate an antibacterial cloak by laser printing of graphene oxide hydrogels mimicking the Cancer Pagurus carapace. We observe up to 90% reduction of bacteria cells. This cloak exploits natural surface patterns evolved to resist to microorganisms infection, and the antimicrobial efficacy of graphene oxide. Cell integrity analysis by scanning electron microscopy and nucleic acids release show bacteriostatic and bactericidal effect. Nucleic acids release demonstrates microorganism cutting, and microscopy reveals cells wrapped by the laser treated gel. A theoretical active matter model confirms our findings. The employment of biomimetic graphene oxide gels opens unique possibilities to decrease infections in biomedical applications and chirurgical equipment; our antibiotic-free approach, based on the geometric reduction of microbial adhesion and the mechanical action of Graphene Oxide sheets, is potentially not affected by bacterial resistance.

To develop agar derivatives with good antibacterial activity and decreased gelling and melting temperatures, two agar-gentamycin conjugates with 9.20% and 12.68% gentamicin immobilized were fabricated by oxidation, Schiff base and reduction reaction, and characterized by a Fourier Transform Infrared Spectrometer, 1H nuclear magnetic resonance and an elemental analyzer. It was found that the modifications changed the intermolecular interactions, leading to decreased gelling and melting temperatures for the oxidized agar and slightly increased gelling and melting temperatures for agar-gentamycin conjugates. Further studies of antimicrobial properties showed that the two agar-gentamycin conjugates possessed good antibacterial activity, which was positively correlated with the dosage and the immobilization rate of gentamicin. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of agar-gentamycin conjugates with higher immobilization rates of gentamicin against Escherichia coli were 39.1 μg/mL and 156.2 μg/mL, respectively, and the MICs and MBCs against Staphylococcus aureus were 19.5 μg/mL and 78.1 μg/mL, respectively. A biofilm test indicated that certain concentrations of agar-gentamycin conjugate could effectively inhibit the biofilm formation of Escherichia coli and Staphylococcus aureus. In summary, agar-gentamycin conjugates possess good antibacterial activities and may be applied as a new kind of antibacterial material.

Polyacrylic Acid grafted Agar-agar (AAc-graf-Agar), and polyacrylamide grafted Agar-Agar (AAm-graf-Agar) have been synthesised by free radical polymerisation route initiated by ammonium peroxodisulphate (APS), the grafted polymers were characterised by FTIR, TGA and SEM methods. The swelling properties were studied in deionised water and saline solution at room temperature. The prepared hydrogels were examined by removing cationic methylene blue (MB) dye from the aqueous solution, in which the adsorption kinetics and isotherms models were also investigated. It was found that the pseudo-second-order and Langmuir equations are the most suitable for the different sorption processes. The maximum dye adsorption capacity was 1035.96 mg∙g-1 for AAc-graf-Agar in pH medium 12 and 1015.7 mg∙g-1 for AAm-graf-Agar in neutral pH medium. This indicates that the AAc-graf-Agar hydrogel could be an excellent adsorbent for removing MB from aqueous solutions.

Hydrogels are three-dimensional soft polymeric materials that can entrap huge amounts of water. They are widely attractive in the biomedicine area because of their outstanding applications such as biosensors, drug delivery vectors, or matrices for cell scaffolds. Generally, the low mechanical strength and fragile structure of the hydrogels limit their feasibility, but this is not the case. In this work, acrylic acid-agar hydrogels with excellent mechanical properties were synthesized using gamma radiation as a crosslinking promoter. The obtained hydrogels exhibited a water absorption capacity up to 6000% in weight without breaking and keeping their shape; additionally, they showed a noticeable adhesion to the skin. The synthesized materials were characterized by infrared spectroscopy (FTIR-ATR), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), scanning electron microscopy (SEM), and mechanical testing. Additionally, their water uptake capacity and critical pH were studied. Net(Agar/AAc) hydrogel exhibited a noticeable capacity to load silver nanoparticles (AgNPs), which endowed it with antimicrobial activity that was demonstrated when challenged against Escherichia coli and methicillin-resistant Staphylococcus aureus (MRSA) on in vitro conditions.

Linezolid is an alternative treatment option for infections with multidrug-resistant Gram-positive bacteria including vancomycin-resistant enterococci. Some countries report an increasing number of isolates with resistance to linezolid. The recent publication of the Commission for Hospital Hygiene in Germany on enterococci/VRE recommends screening for linezolid-resistant enterococci (LRE). However, a suitable selective medium or a genetic test is not available. Our aim was to establish a selective screening agar for LRE detection and validate its application with a comprehensive collection of clinical LRE and linezolid-susceptible enterococci.

In this study, agar/κ-carrageenan/montmorillonite (MMT) hydrogels were prepared to examine their usability as wound dressing materials and to see the effect of MMT amount on some properties of agar/κ-carrageenan hydrogel materials. Hydrogels were characterized by SEM-EDX, TEM and DSC analyses. By increasing the MMT content within hydrogel matrix from 0% to 5%, the decomposition temperature of the hydrogel material was increased from 256.6 °C to 262.1 °C. Swelling amount of hydrogels in d-glucose solution (2682%) was found to be much higher compared with other physiological solutions such as physiological saline solution (937%), synthetic urine solution (746%) and simulated wound fluid (563%). The release studies of analgesic lidocaine hydrochloride (LDC) and antibiotic chloramphenicol (CLP) drugs from hydrogel systems demonstrated that the release amount of LDC and CLP from hydrogels could be controlled by MMT amount within hydrogel matrix. The concentrations of drugs within hydrogel sample stored at 4 °C for 6 months did not exhibit a significant change. Hydrogel materials containing CLP exhibited good antibacterial activity against E. coli and S. aureus. Cytotoxicity test results indicated that hydrogels were biocompatible with MG-63 cells. The ultimate compressive stress of agar/κ-carrageenan hydrogel with LDC and CLP and agar/κ-carrageenan/MMT hydrogel including 5% MMT with LDC and CLP was measured as 38.30 kPa and 47.70 kPa, respectively. The experimental results revealed that prepared agar/κ-carrageenan and agar/κ-carrageenan/MMT hydrogels have great potential for wound care applications.

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.

Manual counting of bacterial colony forming units (CFUs) on agar plates is laborious and error-prone. We therefore implemented a colony counting system with a novel segmentation algorithm to discriminate bacterial colonies from blood and other agar plates.A colony counter hardware was designed and a novel segmentation algorithm was written in MATLAB. In brief, pre-processing with Top-Hat-filtering to obtain a uniform background was followed by the segmentation step, during which the colony images were extracted from the blood agar and individual colonies were separated. A Bayes classifier was then applied to count the final number of bacterial colonies as some of the colonies could still be concatenated to form larger groups. To assess accuracy and performance of the colony counter, we tested automated colony counting of different agar plates with known CFU numbers of S. pneumoniae, P. aeruginosa and M. catarrhalis and showed excellent performance.

The physical interactions of growing bacterial cells with each other and with their surroundings significantly affect the structure and dynamics of biofilms. Here a 3D agent-based model is formulated to describe the establishment of simple bacterial colonies expanding by the physical force of their growth. With a single set of parameters, the model captures key dynamical features of colony growth by non-motile, non EPS-producing E. coli cells on hard agar. The model, supported by experiment on colony growth in different types and concentrations of nutrients, suggests that radial colony expansion is not limited by nutrients as commonly believed, but by mechanical forces. Nutrient penetration instead governs vertical colony growth, through thin layers of vertically oriented cells lifting up their ancestors from the bottom. Overall, the model provides a versatile platform to investigate the influences of metabolic and environmental factors on the growth and morphology of bacterial colonies.

Ralstonia solanacearum invades plants through their roots and causes devastating bacterial diseases in multiple crops. Here, we present a versatile inoculation assay in Arabidopsis thaliana seedlings grown in sterile agar plates. We describe steps for plant preparation, bacterial inoculation, tissue sampling, and bacterial quantification. This protocol can be used for accurate assessment of bacterial colonization and observation of plant response to infection. For complete details on the use and execution of this protocol, please refer to Dindas et al. (2022).1.

The fibroblast mitogen platelet-derived growth factor -BB (PDGF-BB) induces a transient expression of the orphan nuclear receptor NR4A1 (also named Nur77, TR3 or NGFIB). The aim of the present study was to investigate the pathways through which NR4A1 is induced by PDGF-BB and its functional role. We demonstrate that in PDGF-BB stimulated NIH3T3 cells, the MEK1/2 inhibitor CI-1040 strongly represses NR4A1 expression, whereas Erk5 downregulation delays the expression, but does not block it. Moreover, we report that treatment with the NF-κB inhibitor BAY11-7082 suppresses NR4A1 mRNA and protein expression. The majority of NR4A1 in NIH3T3 was found to be localized in the cytoplasm and only a fraction was translocated to the nucleus after continued PDGF-BB treatment. Silencing NR4A1 slightly increased the proliferation rate of NIH3T3 cells; however, it did not affect the chemotactic or survival abilities conferred by PDGF-BB. Moreover, overexpression of NR4A1 promoted anchorage-independent growth of NIH3T3 cells and the glioblastoma cell lines U-105MG and U-251MG. Thus, whereas NR4A1, induced by PDGF-BB, suppresses cell growth on a solid surface, it increases anchorage-independent growth.

Adsorption of methylene blue (MB) on agar was investigated as a function of temperature (308-328 K), different concentrations of NaCl and HCl and various weight percentages of binary mixtures of ethanol with water. It was observed that the maximum experimental adsorption capacity, qm, exp, in water is up to 50 mg g-1 and decreases with increase in weight percentage of ethanol and NaCl and HCl concentration compared to that of water. Analysis of data using ARIAN model showed that MB adsorbs as monomer and dimer on the surface of agar. Binding constants of MB to agar were calculated using the Temkin isotherm. The process is exothermic in water and other solutions. The mean adsorption energy (E) value indicated binding of MB to agar is chemical adsorption. Kinetics of this interaction obeys from the pseudo-second-order model and diffusion of the MB molecules into the agar is the main rate-controlling step.

Plant roots sense salt gradients in soil to avoid saline environments through halotropism. Here, we present a protocol to study halotropism with an optimized split-agar system that simulates the salt gradient in soil. We describe steps for preparation of the split-agar system, measurement of Na+, and observation of root bending. We then detail segmentation of root cells and visualization of microtubules and cellulose synthases. This system is simple to operate and has broader applications, such as hydrotropism and chemotropism. For complete details on the use and execution of this protocol, please refer to Yu et al. (2022).1.

Animal models for host-microbial interactions have proven valuable, yielding physiologically relevant data that may be otherwise difficult to obtain. Unfortunately, such models are lacking or nonexistent for many microbes. Here, we introduce organ agar, a straightforward method to enable the screening of large mutant libraries while avoiding physiological bottlenecks. We demonstrate that growth defects on organ agar were translatable to bacterial colonization deficiencies in a murine model. Specifically, we present a urinary tract infection agar model to interrogate an ordered library of Proteus mirabilis transposon mutants, with accurate prediction of bacterial genes critical for host colonization. Thus, we demonstrate the ability of ex vivo organ agar to reproduce in vivo deficiencies. Organ agar was also useful for identifying previously unknown links between biosynthetic genes and swarming motility. This work provides a readily adoptable technique that is economical and uses substantially fewer animals. We anticipate this method will be useful for a wide variety of microorganisms, both pathogenic and commensal, in a diverse range of model host species.