Antifungal susceptibility testing following the CLSI M27 document, along with the measurement of MFC/MIC ratio, could be the optimal approach to detect amphotericin B resistance in Clavispora (Candida) lusitaniae. Susceptibility patterns of contemporary isolates of this species.

Antifungal susceptibility testing (AST) is crucial in clinical settings to guide appropriate therapy. Nevertheless, discrepancies between treatment response and some results still persist, particularly in detecting resistance to amphotericin B (AMB) in Clavispora (Candida) lusitaniae. This study aimed to assess the susceptibility patterns of 48 recent isolates of C. lusitaniae to 9 antifungal agents and explore the feasibility of using a CLSI reference-based method to identify AMB resistance. Microdilution techniques revealed a wide range of minimal inhibitory concentration (MIC) values for azole antifungals, while echinocandins and AMB exhibited a narrow range of MIC values, with all strains considered wild-type for the tested polyene and echinocandins. However, when agar diffusion (ellipsometry) was employed for AST, certain strains displayed colonies within the inhibition ellipse, indicating potential resistance. Interestingly, these strains did not respond to AMB treatment and were isolated during AMB treatment (breakthrough). Moreover, the evaluation of AMB minimum fungicidal concentrations (MFCs) indicated that only the strains with colonies inside the ellipse had MFC/MIC ratios ≥ 4, suggesting reduced fungicidal activity. In conclusion, this study confirms the effectiveness of ellipsometry with RPMI-1640 2% glucose agar for detecting AMB resistance in C. lusitaniae. Additionally, the proposed approach of culturing "clear" wells in the microdilution method can aid in uncovering resistant strains. The findings highlight the importance of appropriate AST methods to guide effective treatment strategies for deep-seated candidiasis caused by C. lusitaniae. Further collaborative studies are warranted to validate these findings and improve the detection of AMB clinical resistance.

Strain-specific features of Pleurotus ostreatus growth in vitro and some of its biological activities.

BACKGROUND: The production of Pleurotus ostreatus mycelium as a promising object for use in food and other industries is hampered by a lack of information about the strain-specificity of this fungus mycelium growth and its acquisition of various biological activities. Therefore, this research aimed to investigate mycelial growth of different P. ostreatus strains on varies solid and liquid media as well as to evaluate strains antagonistic, antibacterial, antiradical scavenging activities, and total phenolic content. RESULTS: Potato Dextrose Agar medium was suitable for all strains except P. ostreatus strain 2460. The best growth rate of P. ostreatus 2462 strain on solid culture media was 15.0 ± 0.8 mm/day, and mycelia best growth on liquid culture media-36.5 ± 0.2 g/l. P. ostreatus strains 551 and 1685 were more susceptible to positive effect of plant growth regulators Ivin, Methyur and Kamethur. Using of nutrient media based on combination of natural waste (amaranth flour cake and wheat germ, wheat bran, broken vermicelli and crumbs) has been increased the yield of P. ostreatus strains mycelium by 2.2-2.9 times compared to the control. All used P. ostreatus strains displayed strong antagonistic activity in co-cultivation with Aspergillus niger, Candida albicans, Issatchenkia orientalis, Fusarium poae, Microdochium nivale in dual-culture assay. P. ostreatus 2462 EtOAc mycelial extract good inhibited growth of Escherichia coli (17.0 ± 0.9 mm) while P. ostreatus 2460 suppressed Staphylococcus aureus growth (21.5 ± 0.5 mm) by agar well diffusion method. The highest radical scavenging effect displayed both mycelial extracts (EtOH and EtOAc) of P. ostreatus 1685 (61 and 56%) by DPPH assay as well as high phenolic content (7.17 and 6.73 mg GAE/g) by the Folin-Ciocalteu's method. The maximal total phenol content (7.52 mg GAE/g) demonstrated of P. ostreatus 2461 EtOH extract. CONCLUSIONS: It is found that the growth, antibacterial, antiradical scavenging activity as well as total phenolic content were dependent on studied P. ostreatus strains in contrast to antagonistic activity. The proposed culture mediums of natural waste could be an alternative to commercial mediums for the production mycelial biomass of P. ostreatus strains.

Mutant Prevention Concentration, Frequency of Spontaneous Mutant Selection, and Mutant Selection Window-a New Approach to the In Vitro Determination of the Antimicrobial Potency of Compounds.

The analysis of antimicrobial activity is usually MIC- and minimal bactericidal concentration (MBC)-focused, though also crucial are resistance-related parameters, e.g., the frequency of spontaneous mutant selection (FSMS), the mutant prevention concentration (MPC), and the mutant selection window (MSW). In vitro-determined MPCs, however, are sometimes variable, poorly repeatable, and not always reproducible in vivo. We propose a new approach to the in vitro determination of MSWs, along with novel parameters: MPC-D, MSW-D (for dominant mutants, i.e., selected with a high frequency, without a fitness loss), and MPC-F, MSW-F (for inferior mutants, i.e., with an impaired fitness). We also propose a new method for preparing the high-density inoculum (>1011 CFU/mL). In this study, the MPC and MPC-D (limited by FSMS of <10-10) of ciprofloxacin, linezolid, and novel benzosiloxaborole (No37) were determined for Staphylococcus aureus ATCC 29213 using the standard agar method, while the MPC-D and MPC-F were determined by the novel broth method. Regardless of the method, MSWs1010 of linezolid and No37 were the same. However, MSWs1010 of ciprofloxacin in the broth method was narrower than in the agar method. In the broth method, the 24-h incubation of ~1010 CFU in a drug-containing broth differentiates the mutants that can dominate the cell population from those that can only be selected under exposure. We consider MPC-Ds in the agar method to be less variable and more repeatable than MPCs. Meanwhile, the broth method may decrease discrepancies between in vitro and in vivo MSWs. The proposed approaches may help establish MPC-D-related resistance-restricting therapies.

Interspecies interactions in mixed-species biofilms formed by Enterococcus faecalis and gram-negative bacteria isolated from polymicrobial diabetic foot ulcers.

Diabetic foot ulcers (DFU) are exacerbated by bacterial colonisation. Here, a high prevalence of Enterococcus faecalis was observed in DFU patients from an Argentinean hospital. E. faecalis was frequently co-isolated with Escherichia coli, Morganella morganii, and Pseudomonas aeruginosa. The effect of interspecies interactions on bacterial growth was investigated in mixed-species macrocolony biofilms developed in Lubbock-Glc-agar. Similar cell counts were found for E. faecalis and M. morganii growing in mixed and single-species biofilms. An E. faecalis strain showed 1 Log higher cell counts in mixed biofilms with E. coli. Remarkably, E. faecalis strains showed 2 to 4 Log higher cell counts in mixed biofilms with P. aeruginosa. This effect was not observed in planktonic growth or biofilms developed in tryptic soy agar. The present findings reveal bacterial interactions that benefit E. faecalis in mixed-species biofilms, mainly with P. aeruginosa, in a medium that partially mimics the nutrients found in DFU.

Nonuniform growth and surface friction determine bacterial biofilm morphology on soft substrates.

During development, organisms acquire three-dimensional (3D) shapes with important physiological consequences. While basic mechanisms underlying morphogenesis are known in eukaryotes, it is often difficult to manipulate them in vivo. To circumvent this issue, here we present a study of developing Vibrio cholerae biofilms grown on agar substrates in which the spatiotemporal morphological patterns were altered by varying the agar concentration. Expanding biofilms are initially flat but later undergo a mechanical instability and become wrinkled. To gain mechanistic insights into this dynamic pattern-formation process, we developed a model that considers diffusion of nutrients and their uptake by bacteria, bacterial growth/biofilm matrix production, mechanical deformation of both the biofilm and the substrate, and the friction between them. Our model shows quantitative agreement with experimental measurements of biofilm expansion dynamics, and it accurately predicts two distinct spatiotemporal patterns observed in the experiments-the wrinkles initially appear either in the peripheral region and propagate inward (soft substrate/low friction) or in the central region and propagate outward (stiff substrate/high friction). Our results, which establish that nonuniform growth and friction are fundamental determinants of stress anisotropy and hence biofilm morphology, are broadly applicable to bacterial biofilms with similar morphologies and also provide insight into how other bacterial biofilms form distinct wrinkle patterns. We discuss the implications of forming undulated biofilm morphologies, which may enhance the availability of nutrients and signaling molecules and serve as a "bet hedging" strategy.

Effects of Temperature on Spore Germination and Mycelial Growth of Calosphaeria pulchella, Cytospora sorbicola, and Eutypa lata Isolates Associated with Sweet Cherry Canker Diseases.

Although fungal canker diseases constitute a limiting factor to orchard productivity and longevity, little is known about the effects of temperature on spore germination and mycelial growth of the fungal causal agents. Accordingly, the germination of spores and colony growth of Calosphaeria pulchella, Cytospora sorbicola, and Eutypa lata were evaluated after incubation on 2% water agar and 4% potato dextrose agar, respectively, at 5, 10, 15, 20, 25, 30, 35, and 40°C. Temperature optima for spore germination and mycelial growth were derived from nonlinear models fitted to germination rates and colony diameter data. The optimal temperatures for spore germination of Cal. pulchella were 28.5°C for ascospores and 29.2°C for conidia. The optimal temperatures for Cyt. sorbicola conidia and E. lata ascospore germination were 25.8 and 23.1°C, respectively. The germination of ascospores and conidia of Cal. pulchella at temperatures below 15°C required an incubation time of at least 72 h. Ascospores of E. lata and conidia of Cyt. sorbicola germinated at 10°C after 36 h. The optimal temperature for colony growth of Cal. pulchella was 24.6°C, whereas it was 21.7°C for both Cyt. sorbicola and E. lata. Our study indicates that temperature requirements for basic biological functions are higher for Cal. pulchella than for Cyt. sorbicola and E. lata. The overall higher temperatures of California relative to other cherry-producing regions in the United States or worldwide could explain the prevalence of Calosphaeria canker in the state. Conversely, Cyt. sorbicola and E. lata appear better adapted to cooler temperatures.

Biological properties of experimental dental alginate modified for self-disinfection using green nanotechnology.

OBJECTIVES: Disinfection of alginate impression materials is a mandatory step to prevent cross-infection in dental clinics. However, alginate disinfection methods are time-consuming and exert a negative impact on accuracy and mechanical properties. Thus, this study aimed to prepare disinfecting agents (CHX and AgNO3) and silver nanoparticles reduced by a natural plant extract to produce a self-disinfecting dental alginate. METHODS: Conventional alginate impression material was used in this study. Silver nitrate (0.2% AgNO3 group) and chlorohexidine (0.2% CHX group) solutions were prepared using distilled water, and these solutions were later employed for alginate preparation. Moreover, a 90% aqueous plant extract was prepared from Boswellia sacra (BS) oleoresin and used to reduce silver nitrate to form silver nanoparticles that were incorporated in the dental alginate preparation (BS+AgNPs group). The plant extract was characterized by gas chromatography/mass spectrometry (GC/MS) analysis while green-synthesized silver nanoparticles (AgNPs) were characterized by UV-visible (UV-vis) spectroscopy and scanning electron microscopy (SEM). An agar disc diffusion assay was used to test the antimicrobial activity against Candida albicans, Streptococcus mutans, Escherichia coli, methicillin-resistant and susceptible Staphylococcus aureus strains, and Micrococcus luteus. Agar plates were incubated at 37 ± 1 °C for 24 h to allow microbial growth. Diameters of the circular inhibition zones formed around each specimen were measured digitally by using ImageJ software. RESULTS: Chemical analysis of the plant extract revealed the presence of 41 volatile and semi-volatile active compounds. UV-Vis spectrophotometry, SEM, and EDX confirmed the formation of spherical silver nanoparticles using the BS extract. CHX, AgNO3, and the BS+AgNPs modified groups showed significantly larger inhibition zones than the control group against all tested strains. BS+AgNPs and CHX groups showed comparable efficacy against all tested strains except for Staphylococcus aureus, where the CHX-modified alginate had a significantly higher effect. CONCLUSIONS AND CLINICAL RELEVANCE: CHX, silver nitrate, and biosynthesized silver nanoparticles could be promising inexpensive potential candidates for the preparation of a self-disinfecting alginate impression material without affecting its performance. Green synthesis of metal nanoparticles using Boswellia sacra extract could be a very safe, efficient, and nontoxic way with the additional advantage of a synergistic action between metal ions and the phytotherapeutic agents of the plant extract.

The anti-adherence activity and bactericidal effect of GO against Streptococcus mutans from Iraqi dental patients.

The high rate of microbes and their biological activity in the patient's mouth is a concern in the domains of dental caries and periodontal disease. The study aimed to shed light on the relationship between graphene oxide's nanoparticles (nGOs) antimicrobial properties and the growth of dental pathogenic bacteria. The forty swab samples were frequently collected from the patient's cavity mouth between November 2019 and January 2020, from patients who visited dentist clinics in Baghdad by taking swabs from mouth cavities with various dental caries with two age groups (5-17) and (18-60) from male and female to streaking them on Brain-Heart Infusion (BHI) agar, then identified by re-streaking on Mitis Salivarius Bacitracin (MSB) agar. All isolates were confirmed as Streptococcus mutans after API 20 Strep method. As well as the Colony Forming Units (CFU) were then determined after diluting the bacterial cell suspensions to obtain cell samples containing 1.5 × 108 CFU/ ml. The collagen-binding adhesin (cnm) and glucosyltransferases (gtf) of S. mutans genes were identified using polymerase chain reaction (PCR) method before and after exposure to the nGOs, which were prepared in different pulse laser energy (500, 600, and 700 mJ) with presence and absence of the magnetic field, and the data have been analyzing. After counting the CFU, the nGOs shows high effectiveness inhibiting the growth of S. mutans. This research provides definitive answers about the relationship between nGOs, antibacterial caries, and periodontal disease.

[Species diversity of the genus Candida in the oral cavity of cancer patients in Barranquilla, Colombia]. / Diversidad de especies de Candida recuperadas de la cavidad bucal de pacientes oncológicos en Barranquilla, Colombia.

Candida spp. was characterized in the oral cavity of cancer patients in a health care center in Barranquilla, Colombia. This is a cross-sectional investigation including 60 oncological patients with oral candidiasis, selected by convenience sampling, from whom samples were subjected to culture in Sabouraud chloramphenicol agar, CHROMagar® Candida and Sabouraud dextrose agar were taken. The antifungal susceptibility profile was then identified and established. Descriptive statistics, Chi square test, and bivariate analysis were conducted using the Statgraphics Centurion XVII software with odds ratio (OR) for the probability of occurrence. A total of 107 Candida strains were identified belonging to 15 species, C. albicans with 23%, C. glabrata with 18%, C. tropicalis 13%, C. krusei 10%, C intermedia, and C. lipolytica with 1.5%. Species other than C. albicans were identified in 77% of the cases. A relationship between reproductive system cancer and C. guilliermondii was identified (p = 0.0001, <0.05) OR: 17.0. Between C. colliculosa and respiratory cancer (p = 0.0003, <0.05) OR 19.5. With regard to antifungal susceptibility, 99% of the identified Candida species were susceptible to the following antifungals: fluconazole, voriconazole, caspofungin, and micafungin. Only one strain of C. krusei was resistant. It is concluded that there was a diversity of Candida species, either single or mixed in cancer patients, which could determine that only one species is not responsible for fungal infection in the oral cavity.

Stochastic establishment of ß-lactam-resistant Escherichia coli mutants reveals conditions for collective resistance.

For antibiotic resistance to arise, new resistant mutants must establish in a bacterial population before they can spread via natural selection. Comprehending the stochastic factors that influence mutant establishment is crucial for a quantitative understanding of antibiotic resistance emergence. Here, we quantify the single-cell establishment probability of four Escherichia coli strains expressing ß-lactamase alleles with different activity against the antibiotic cefotaxime, as a function of antibiotic concentration in both unstructured (liquid) and structured (agar) environments. We show that concentrations well below the minimum inhibitory concentration (MIC) can substantially hamper establishment, particularly for highly resistant mutants. While the pattern of establishment suppression is comparable in both tested environments, we find greater variability in establishment probability on agar. Using a simple branching model, we investigate possible sources of this stochasticity, including environment-dependent lineage variability, but cannot reject other possible causes. Lastly, we use the single-cell establishment probability to predict each strain's MIC in the absence of social interactions. We observe substantially higher measured than predicted MIC values, particularly for highly resistant strains, which indicates cooperative effects among resistant cells at large cell numbers, such as in standard MIC assays.

ROOT PENETRATION INDEX 3, a major quantitative trait locus associated with root system penetrability in Arabidopsis.

Soil mechanical impedance precludes root penetration, confining root system development to shallow soil horizons where mobile nutrients are scarce. Using a two-phase-agar system, we characterized Arabidopsis responses to low and high mechanical impedance at three root penetration stages. We found that seedlings whose roots fail to penetrate agar barriers show a significant reduction in leaf area, root length, and elongation zone and an increment in root diameter, while those capable of penetrating show only minor morphological effects. Analyses using different auxin-responsive reporter lines, exogenous auxins, and inhibitor treatments suggest that auxin responsiveness and PIN-mediated auxin distribution play an important role in regulating root responses to mechanical impedance. The assessment of 21 Arabidopsis accessions revealed that primary root penetrability varies widely among accessions. To search for quantitative trait loci (QTLs) associated to root system penetrability, we evaluated a recombinant inbred population derived from Landsberg erecta (Ler-0, with a high primary root penetrability) and Shahdara (Sha, with a low primary root penetrability) accessions. QTL analysis revealed a major-effect QTL localized in chromosome 3, ROOT PENETRATION INDEX 3 (q-RPI3), which accounted for 29.98% (logarithm of odds=8.82) of the total phenotypic variation. Employing an introgression line (IL-321) with a homozygous q-RPI3 region from Sha in the Ler-0 genetic background, we demonstrated that q-RPI3 plays a crucial role in root penetrability. This multiscale study reveals new insights into root plasticity during the penetration process in hard agar layers, natural variation, and genetic architecture behind primary root penetrability in Arabidopsis.

The identification of phage vB_1086 of multidrug-resistant Klebsiella pneumoniae and its synergistic effects with ceftriaxone.

BACKGROUND: The continued rise of Klebsiella pneumoniae resistance to antibiotics is precipitating a medical crisis. Bacteriophages have been hailed as one possible therapeutic option to enhance the efficacy of antibiotics. This study describes the genomic characterization and biological property of a new bacteriophage vB_1086 and its potential for phage therapy application against Klebsiella pneumoniae. METHODS: In our study, the double-layer agar plate method isolated a lytic bacteriophage named vB_1086. Besides, we analyzed its biological characteristics and genetic background. Then the antibacterial ability of the bacteriophage vB_1086 combined with antibiotics were analyzed by the combined checkerboard method. The impact on the formation of biofilms was analyzed by crystal violet staining method. RESULTS: vB_1086 is a lytic bacteriophage with stable biological characteristics and clear genetic background, showing good antibacterial activity in combination with ceftriaxone, and the combination of phage and meropenem can effectively inhibit the formation of biofilm. Besides, the combination of bacteriophage and antimicrobials can effectively alleviate the generation of bacterial resistance and reduce the dosage of antimicrobials. CONCLUSION: vB_1086 is a novel phage. To some extent, these results provide valuable information that phage vB_1086 can be combined with antibiotics to reduce the dosage of antimicrobials and alleviate the generation of bacterial resistance.

Mapping sex differences in the effects of protein and carbohydrates on lifespan and reproduction in Drosophila melanogaster: is measuring nutrient intake essential?

Understanding how diet affects reproduction and survival is a central aim in evolutionary biology. Although this relationship is likely to differ between the sexes, we lack data relating diet to male reproductive traits. One exception to this general pattern is Drosophila melanogaster, where male dietary intake was quantified using the CApillary FEeder (CAFE) method. However, CAFE feeding reduces D. melanogaster survival and reproduction, so may distort diet-fitness outcomes. Here, we use the Geometric Framework of Nutrition to create nutrient landscapes that map sex-specific relationships between protein, carbohydrate, lifespan and reproduction in D. melanogaster. Rather than creating landscapes with consumption data, we map traits onto the nutrient composition of forty agar-based diets, generating broad coverage of nutrient space. We find that male and female lifespan was maximised on low protein, high carbohydrate blends (~ 1P:15.9C). This nutrient ratio also maximised male reproductive rates, but females required more protein to maximise daily fecundity (1P:1.22C). These results are consistent with CAFE assay outcomes. However, the approach employed here improved female fitness relative to CAFE assays, while effects of agar versus CAFE feeding on male fitness traits depended on the nutrient composition of experimental diets. We suggest that informative nutrient landscapes can be made without measuring individual nutrient intake and that in many cases, this may be preferable to using the CAFE approach. The most appropriate method will depend on the question and species being studied, but the approach adopted here has the advantage of creating nutritional landscapes when dietary intake is hard to quantify.

Establishment of a reference panel of Helicobacter pylori strains for antimicrobial susceptibility testing.

BACKGROUND: Eradication treatment for Helicobacter pylori gastritis is covered by national health insurance since 2013 in Japan. However, eradication failure due to the increase of antimicrobial resistance has become a serious problem. The present study aims to establish a reference panel of Japanese H. pylori strains for antimicrobial susceptibility testing. METHOD: A total of 28 strains were collected from 4 medical facilities in Japan. Antimicrobial susceptibility tests (ASTs) to clarithromycin (CLR), amoxicillin (AMX), and metronidazole (MNZ), were used to select standard reference strains. Complete genome sequences were also determined. RESULTS: Three H. pylori strains (JSHR3, JSHR6 and JSHR31) were selected as standard reference strains by the Japanese Society for Helicobacter Research (JSHR). The minimum inhibitory concentrations (MICs) of the antibiotics against these 3 strains by agar dilution method with Brucella-based horse-serum-containing agar medium were as follows: JSHR3 (CLR 16 µg/ml, AMX 0.032 µg/ml and MNZ 4 µg/ml), JSHR6 (CLR 0.016 µg/ml, AMX 0.032 µg/ml and MNZ 4 µg/ml), and JSHR31 (CLR 16 µg/ml, AMX 1 µg/ml and MNZ 64 µg/ml). CONCLUSIONS: A reference panel of H. pylori JSHR strains was established. The panel consisted of JSHR6, which was antibiotic-susceptible, JSHR3, which was CLR-resistant, and JSHR31, which was multi-resistant. This reference panel will be essential for standardized ASTs before the optimal drugs are selected for eradication treatment.

Oligosaccharides from agar extends lifespan through activation of unfolded protein response via SIR-2.1 in Caenorhabditis elegans.

PURPOSE: Agaro-oligosaccharides (AGO), hydrolysis products of agarose, is known to have antioxidant and anti-inflammatory properties. Speculating that AGO is effective for preventing aging, we investigated the longevity-supporting effects of AGO and their mechanisms using Caenorhabditis elegans. METHODS: Caenorhabditis elegans were fed AGO from young adulthood. The lifespan, locomotory activity, lipofuscin accumulation, and heat stress resistance of the worms were examined. To elucidate mechanisms of AGO-mediated longevity, we conducted comprehensive expression analysis using microarrays. Moreover, we used quantitative real-time PCR (qRT-PCR) to verify the genes showing differential expression levels. Furthermore, we measured the lifespan of loss-of-function mutants to determine the genes related to AGO-mediated longevity. RESULTS: AGO extended the lifespan of C. elegans, reduced lipofuscin accumulation, and maintained vigorous locomotion. The microarray analysis revealed that the endoplasmic reticulum-unfolded protein response (ER-UPR) and insulin/insulin-like growth factor-1-mediated signaling (IIS) pathway were activated in AGO-fed worms. The qRT-PCR analysis showed that AGO treatment suppressed sir-2.1 expression, which is a negative regulator of ER-UPR. In loss-of-function mutant of sir-2.1, AGO-induced longevity and heat stress resistance were decreased or cancelled completely. Furthermore, the pro-longevity effect of AGO was decreased in loss-of-function mutants of abnormal Dauer formation (daf) -2 and daf-16, which are IIS pathway-related genes. CONCLUSION: AGO delays the C. elegans aging process and extends their lifespan through the activations of ER-UPR and the IIS pathway.

Evaluation of distinct modes of oxidative secondary injury generated in heat-treated cells of Escherichia coli with solid/liquid and complex/semi-synthetic media sets.

AIMS: To characterize and evaluate oxidative secondary injury generated in heat-treated Escherichia coli cells during recovery cultivation either on agar or in a broth of a semi-synthetic enriched M9 (EM9) medium and a complex Luria broth (LB) medium with different types of antioxidants. METHODS AND RESULTS: E. coli cells grown in the EM9 and LB broth were heated at 50°C in a buffer (pH 7.0). Heated cells were recovered on the same kind of agar medium as that used for growth, with or without different antioxidants. Although these antioxidants mostly protected the cells from oxidative secondary injury on the recovery media, sodium thiosulphate and sodium pyruvate were most protective on EM9 and LB agars, respectively. Determination of viability using the most probable number and growth delay analysis methods showed significant reductions in the protective effects of antioxidants in the EM9 and LB media. CONCLUSION: Oxidative secondary injury generated in heated E. coli cells was found to be qualitatively and quantitatively diverse under cellular and environmental conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results suggest that different modes of oxidation should be considered in viability determination and injured cell enumeration of heat-treated cells.

Modified low-temperature synthesis of graphene oxide nanosheets: Enhanced adsorption, antibacterial and antioxidant properties.

Herein, we report a simple, low-temperature, ecofriendly synthesis of graphene oxide nanosheets (GONs). Graphite powder was treated with KMnO4 and a concentrated H2SO4/H3PO4 mixture to synthesize GONs. The effects of various reaction conditions such as reaction time, temperature, amounts of cleaving agents (H2SO4/H3PO4), and oxidant (KMnO4) were investigated. The synthesized GONs were examined by various techniques in order to investigate their characteristics. The best results of the synthesized GONs were observed at 35 °C within 10 h of reaction time having 8:2 ratios of H2SO4/H3PO4 acid mixture. The main absorption peak in the UV-vis spectra of GONs was at 258 nm, which is due to the π-π* transition of the atomic CC bonds. The existence of stretching vibrations of C꞊O, O-H, C-H, and C-O in the Fourier transform infrared (FTIR) spectra verified the formation of GONs. Presence of a sharp peak at 2θ = 10° with an interlayer spacing distance of 0.88 nm in the observed XRD pattern revealed that the synthesized GONs were totally oxidized and that the interlayer spacing increased. The morphological investigations confirmed the formation of ultrathin, transparent, curly, and homogenous GONs. The synthesized GONs were applied as an adsorbent for the rapid uptake of four different pesticides viz.; Profenofos, Ethion, Cypermethrin, Thiamethoxam (TMX) from the pesticides spiked water samples. About 86% adsorption of Profenofos + Cypermethrin, and 50% adsorption of ethion and thiamethoxam took place within 20 min in presence of 10 mg GONs. In addition to this, the prepared GONs were tested for the antibacterial activity against four bacterial strains by agar well diffusion method. The synthesized GONs provide a significant inhibition for gram -positive (Bacillus subtilis, and Staphylococcus aureus) and gram-negative (Escherichia coli and Pseudomonas aeruginosa) bacterial strains. Moreover, the radical scavenging activities (RSA) of GONs were also checked and compared with Gallic acid as a standard. The obtained RSA of GONs was 60% in comparison to the 80% as of the standard Gallic acid at 1000 µg/mL concentration.

Anti-obesity effects of agar (Gelidium amansii)-derived oligosaccharides in high-fat diet-treated C57BL/6N mice due to differential regulations of lipogenesis and lipolysis.

To investigate the effects of agar oligosaccharides (AO) on lipid metabolism, changes in obesity phenotypes and related molecular factors were evaluated in C57BL/6N mice fed a high-fat diet (HFD). When HFD-induced obese mice were fed AO, they lost weight. Also, fat accumulation in abdominal and liver tissues was lower in the AO groups than in the Vehicle group. Lipid droplet sizes in tissue sections were reduced by AO, and these observations were mirrored by serum lipid contents. To evaluate the effects of AO on lipid metabolism, lipogenesis and lipolysis-related factors were analyzed. The mRNA expressions of genes involved in lipogenesis, such as adipocyte-protein 2 (aP2) and fatty acid synthase (FAS), were reduced by AO administration, and the expressions of lipolysis-associated proteins, including perilipin, hormone-sensitive lipase (HSL), and fat triglyceride lipase (ATGL), were increased. Taken together, our results suggest that AO should be considered a valuable natural agent that inhibits obesity.

Characterization of a new Bacillus velezensis as a powerful biocontrol agent against tomato gray mold.

Biocontrol microbes are environment-friendly and safe for humans and animals. To seek biocontrol microbes effective in suppressing tomato gray mold is important for tomato production. Therefore, serial experiments were conducted to characterize the antagonism of Bacillus velezensis HY19, a novel self-isolated biocontrol bacterium, against Botrytis cinerea in vitro and the control on tomato gray mold in greenhouse. This bacterium produced extracellular phosphatase, protease, cellulose and siderophores, and considerably inhibited the growth of B. cinerea. A liquid chromatography-mass spectrometry (LC-MS) detected salicylic acid and numerous antifungal substances present in B. velezensis HY19 fermentation liquid (BVFL). When B. cinerea was grown on potato glucose agar, BVFL crude extract remarkably suppressed the fungal growth and reduced protein content and the activities of catalase (CAT), superoxide dismutase (SOD), and peroxidase (POD). Transcriptome studies showed that BVFL crude extract significantly induced different expression of numerous genes in B. cinerea, most of which were down-regulated. Theses differently expressed genes were involved in the biological process, cell compartment, molecular functions, and metabolisms of glycine, serine, threonine, and sulfur in pathogen hyphae. Thus, this biocontrol bacterium antagonized B. cinerea in multiple ways due to the production of numerous antifungal substances that acted on multiple targets in the cells. BVFL significantly increased antioxidant enzyme activities in tomato leaves and decreased the incidence of tomato gray mold, with the control efficacies of 73.12-76.51%. Taken together, B. velezensis HY19 showed a promising use potential as a powerful bioagent against tomato gray mold.

Effects of sodium hexametaphosphate microparticles or nanoparticles on the growth of saliva-derived microcosm biofilms.

OBJECTIVES: This study evaluated the effects of sodium hexametaphosphate microparticles (HMPmicro) or nanoparticles (HMPnano) on the growth of saliva-derived microcosm biofilms MATERIALS AND METHODS: Saliva-derived biofilms were formed on glass coverslips for 24 h. Thereafter, Streptococcus mutans (C180-2) was incorporated or not into the biofilms. From that time point onwards, solutions containing 0.2% HMPmicro or HMPnano, combined or not with 220 ppm F, were constantly present in the culture medium. In addition, 220 ppm F alone (220F) and McBain medium without any compound were also tested as positive and negative controls (CTL), respectively. After 96 h, the biofilms were plated on anaerobic blood agar or sucrose agar bacitracin for total and S. mutans CFU-counting, respectively. Biofilms' lactic acid production was analysed spectrophotometrically. Data were submitted to ANOVA or Kruskal-Wallis' tests, followed by Student-Newman-Keuls' test (p<0.05; n=12). RESULTS: HMPmicro or HMPnano led to significantly lower lactic acid production, and significant reductions in total CFU-counting in microcosm biofilms, supplemented or not with S. mutans, in comparison to both controls, with significant differences between 220F and CTL. No significant differences were observed among the groups treated with HMPmicro or HMPnano (with or without F). The same trend was seen for S. mutans CFU-counting, in biofilms supplemented with S. mutans. CONCLUSIONS: HMP significantly reduced total and S. mutans CFU counts, as well as lactic acid production by saliva-derived microcosm biofilms. CLINICAL RELEVANCE: These findings in saliva-derived microcosm biofilms suggest that HMP stands as a promising alternative for the control of cariogenic biofilms.