Bio-synthesis and characterization of silver nanoparticles from Trichoderma species against cassava root rot disease.

Cassava root rot disease caused by the fungal pathogens Fusarium solani and Lasiodiplodia theobromae produces severe damages on cassava production. This research was conducted to produce and assess silver nanoparticles (AgNPs) synthesized by Trichoderma harzianum for reducing root rot disease. The results revealed that using the supernatants of T. harzianum on a silver nitrate solution changed it to reddish color at 48 h, indicating the formation of AgNPs. Further characterization was identified using dynamic light scattering (DLS) and scanning electron microscope (SEM). DLS supported that the Z-average size is at 39.79 nm and the mean zeta potential is at - 36.5 mV. SEM revealed the formation of monodispersed spherical shape with a diameter between 60-75 nm. The antibacterial action of AgNPs as an antifungal agent was demonstrated by an observed decrease in the size of the fungal colonies using an increasing concentration of AgNPs until the complete inhibition growth of L. theobromae and F. solani at > 58 µg mL-1 and at ≥ 50 µg mL-1, respectively. At in vitro conditions, the applied AgNPs caused a decrease in the percentage of healthy aerial hyphae of L. theobromae (32.5%) and of F. solani (70.0%) compared to control (100%). The SR-FTIR spectra showed the highest peaks in the first region (3000-2800 cm-1) associated with lipids and fatty acids located at 2962, 2927, and 2854 cm-1 in the AgNPs treated samples. The second region (1700-1450 cm-1) consisting of proteins and peptides revealed the highest peaks at 1658, 1641, and 1548 cm-1 in the AgNPs treated samples. The third region (1300-900 cm-1), which involves nucleic acid, phospholipids, polysaccharides, and carbohydrates, revealed the highest peaks at 1155, 1079, and 1027 cm-1 in the readings from the untreated samples. Finally, the observed root rot severity on cassava roots treated with AgNPs (1.75 ± 0.50) was significantly lower than the control samples (5.00 ± 0.00).

Flagella Phenotypic Variations of ST34 Type Salmonella Typhimurium and Variants.

Salmonella enterica serovar Typhimurium and its variants are the most common serotypes of human salmonellosis cases. Serotyping Salmonella Typhimurium and its variants has always been challenging. Our previous work found that among 14 Salmonella Typhimurium and variant strains, some different antigenic formulas had 100% pulsed-field gel electrophoresis (PFGE) similarity. The 14 strains were sorted into 3 groups; in each group, the different antigenic formulas had the same PFGE patterns. This phenomenon suggested that different antigenic formula identification might originate from a common ancestor subtyped by PFGE. To assess whether the serotyping method on Salmonella Typhimurium and variant strains reflected the genetic relationship, we improved the discrimination for the phylogenetic relationship among the 14 Salmonella Typhimurium and variant strains using Fourier-transform infrared spectroscopy (FTIR) and whole-genome multilocus sequence typing (wgMLST). We compared the wgMLST assay of 14 Salmonella Typhimurium and variant strains from this study with 50 public ST34 strain data of Salmonella Typhimurium and variant strains. We also compared flagella (H antigen)-related genes based on the whole genome of 14 strains and the other 293 ST34 public database for further understanding of this question. The phylogenetic results (PFGE) showed no regularity between the antigenic formulas and genotypes. The results of the higher discrimination power assays (FTIR and whole-genome multilocus sequence typing) also showed no regularity between the antigenic formulas and genotypes (or phenotypes). The 58 flagella encoding genes of different antigenic formulas were sorted into 13 patterns. However, a similar phenomenon was found: the same flagella encoding gene patterns could express different antigenic formulas. In conclusion, there is no consistency between the antigenic formulas and phylogenetic relationships among ST34 Salmonella Typhimurium and variant strains, even in flagella antigenic formula and flagella encoding genes.

A comprehensive study of sous-vide cooked Korat chicken breast processed by various conditions: texture, compositional/structural changes, and consumer acceptance.

Korat chicken (KC) is a slow-growing crossbreed renowned for its excellent growth and firm texture. This study investigated the effect of various sous-vide (SV) conditions (60 and 70°C, 1-3 h) on their texture, protein structure and degradation, as well as consumer acceptability, with the traditional boiling served as control. Texture showed significant improvement under all SV conditions compared to the control, as demonstrated by increased water holding capacity (WHC), cooking loss, and decreased shear force, hardness, and chewiness (P < 0.05). These changes corresponded to the higher sensory scores (P < 0.05). Among the SV samples, increased temperatures and longer cooking times led to higher degradation of myofibrils and connective tissue, as evidenced by a decrease in water-, salt-soluble proteins, and soluble collagen (P < 0.05). These findings aligned with the scanning electron microscopy (SEM) results, which showed a looser muscle structure in the meat under more intense cooking conditions. Based on synchrotron radiation-based Fourier transform infrared (SR-FTIR) results, a gradual increase in antiparallel forms within the amide I bands (1,700-1,600 cm-1) of the total spectra with higher temperature and longer cooking times was observed (P < 0.05), while the fluctuations were observed in the changes of α-helix, ß-sheet, and ß-turn structures. This suggested that the antiparallel structure represented a looser configuration developing during intense SV cooking. Combined with the principal component analysis (PCA) results, the findings indicated that the suitable SV condition for KC breast meat was 70°C for varying durations (1-3 h), as it showed the strongest correlation with sensory scores, particularly in terms of tenderness. In summary, these findings provided a better understanding of molecular changes and discovered SV conditions to enhance the texture quality of the KC meat.

Bacillus vallismortis TU-Orga21 blocks rice blast through both direct effect and stimulation of plant defense.

Beneficial microorganisms are an important strategy for sustainable plant production processes such as stimulate root exudation, stress tolerance, and yield improvement. This study investigated various microorganisms isolated from the rhizosphere of Oryza sativa L. in order to inhibit Magnaporthe oryzae cause of rice blast, by direct and indirect mode of action. The results indicated that Bacillus vallismortis strain TU-Orga21 significantly reduced M. oryzae mycelium growth and deformed the hyphal structures. The effects of biosurfactant TU-Orga21 was studied against M. oryzae spore development. The dose of ≥5% v/v biosurfactant significantly inhibited the germ tubes and appressoria formation. The biosurfactants were evaluated as surfactin and iturin A by Matrix-assisted laser desorption ionization dual time-of-flight tandem mass spectrometry. Under greenhouse conditions, priming the biosurfactant three times before M. oryzae infection significantly accumulated endogenous salicylic acid, phenolic compounds, and hydrogen peroxide (H2O2) during the infection process of M. oryzae. The SR-FT-IR spectral changes from the mesophyll revealed higher integral area groups of lipids, pectins, and proteins amide I and amide II in the elicitation sample. Furthermore, scanning electron microscope revealed appressorium and hyphal enlargement in un-elicitation leaves whereas appressorium formation and hyphal invasion were not found in biosurfactant-elicitation at 24 h post inoculation. The biosurfactant treatment significantly mitigated rice blast disease severity. Therefore, B. vallismortis can be a promising novel biocontrol agent which contains the preformed active metabolites for a rapid control of rice blast by a direct action against pathogen and by boosting plant immunity.

Investigating the Effect of Various Sous-Vide Cooking Conditions on Protein Structure and Texture Characteristics of Tilapia Fillet Using Synchrotron Radiation-Based FTIR.

The effects of various sous-vide (SV) cooking conditions (50-60℃, 30-60 min) on physicochemical properties related to the texture characteristics, protein structure/degradation, and sensory acceptability of tilapia fillet (Oreochromis niloticus) were investigated. With an increasing temperature and processing time of SV cooking, protein degradation (of both myofibrils and connective tissue) was more pronounced, as evaluated by the decrease in water- and salt-soluble proteins, total collagen, as well as the changes in the ratio of secondary protein structures (α-helix, ß-sheet, ß-turn, etc.), which were determined by synchrotron-FTIR (SR-FTIR). These degradations were associated with the improvement of meat tenderness, as estimated by shear force and texture profile analyzer (TPA) results. Among all SV conditions, using 60 ℃ for 45 min seems to be the optimal condition for tilapia meat, since it delivered the best results for texture characteristics and acceptability (p < 0.05). Moreover, principal component analysis (PCA) results clearly demonstrated that the highest texture-liking score of this condition was well associated with the intensity of ß-sheets, which seem to be the crucial component that affected the texture of SV-cooked tilapia more so than other parameters. The findings demonstrated the potential of SR-FTIR to decipher the biomolecular structure, particularly the secondary protein structure, of SV-cooked tilapia. This technique provided essential information for a better understanding of the changes in biomolecules related to the textural characteristics of this product.

Flour on Gluten-Free Muffins from Different Edible Cassava Varieties in Thailand.

In Thailand new edible cassava varieties have been developed to be used in the food industry. The aim of this research was to analyze the difference between flour from three cassava varieties and to evaluate the suitability and quality of flour for gluten-free muffins. The physico-chemical properties of flour from three varieties were studied. The results showed the moisture content of flour was between 10.65 ± 0.01 and 10.85 ± 0.45%. Total protein content was highly significant with a difference of 1.97 ± 0.00%, 2.15 ± 0.01%, and 2.18 ± 0.01%, respectively. Moreover, ash and fat in each flour were highly significant. Amylose content was 19.93 ± 0.47%, and the viscosity was 6286.00 ± 1.52 mPa.s. The color of flour values of L* a* b* value was not statistically different in each variety of flour. Fourier transform infrared spectroscopy (FTIR) analysis was used for the biochemical change in flour. The PCA and cluster analysis results revealed that cassava flour from Pirun 6 was different from Pirun 2 and Pirun 4. After that, the test using selected cassava flour from Pirun 6 to test the physical properties and sensory attributes of gluten-free muffins compared with wheat flour found that gluten-free muffins were overall better than basic muffins.

Performance Evaluation of Focal Plane Array (FPA)-FTIR and Synchrotron Radiation (SR)-FTIR Microspectroscopy to Classify Rice Components.

The development of biochemical analysis techniques to study heterogeneous biological samples is increasing. These techniques include synchrotron radiation Fourier transform infrared (SR-FTIR) microspectroscopy. This method has been applied to analyze biological tissue with multivariate statistical analysis to classify the components revealed by the spectral data. This study aims to compare the efficiencies of SR-FTIR microspectroscopy and focal plane array (FPA)-FTIR microspectroscopy when classifying rice tissue components. Spectral data were acquired for mapping the same sample areas from both techniques. Principal component analysis and cluster imaging were used to investigate the biochemical variations of the tissue types. The classification was based on the functional groups of pectin, protein, and polysaccharide. Four layers from SR-FTIR microspectroscopy including pericarp, aleurone layer, sub-aleurone layer, and endosperm were classified using cluster imaging, while FPA-FTIR microspectroscopy could classify only three layers of pericarp, aleurone layer, and endosperm. Moreover, SR-FTIR microspectroscopy increased the image contrast of the biochemical distribution in rice tissue more efficiently than FPA-FTIR microspectroscopy. We have demonstrated the capability of the high-resolution synchrotron technique and its ability to clarify small structures in rice tissue. The use of this technique might increase in future studies of tissue characterization.

Resistance Induction by Salicylic Acid Formulation in Cassava Plant against Fusarium solani.

Fusarium root rot caused by the soil-borne fungus Fusarium solani is one of the most important fungal diseases of cassava in Thailand, resulting in high yield losses of more than 80%. This study aimed to investigate if the exogenous application of salicylic acid formulations (Zacha) can induce resistance in cassava against Fusarium root rot and observe the biochemical changes in induced cassava leaf tissues through synchrotron radiation based on Fourier-transform infrared (SR-FTIR) microspectroscopy. We demonstrated that the application of Zacha11 prototype formulations could induce resistance against Fusarium root rot in cassava. The in vitro experimental results showed that Zacha11 prototype formulations inhibited the growth of F. solani at approximately 34.83%. Furthermore, a significant reduction in the disease severity of Fusarium root rot disease at 60 days after challenge inoculation was observed in cassava plants treated with Zacha11 at a concentration of 500 ppm (9.0%). Population densities of F. solani were determined at 7 days after inoculation. Treatment of the Zacha11 at a concentration of 500 ppm resulted in reduced populations compared with the distilled water control and differences among treatment means at each assay date. Moreover, the SR-FTIR spectral changes of Zacha11-treated epidermal tissues of leaves had higher integral areas of lipids, lignins, and pectins (1,770-1,700/cm), amide I (1,700-1,600/cm), amide II (1,600-1,500/cm), hemicellulose, lignin (1,300-1,200/cm), and cellulose (1,155/cm). Therefore, alteration in defensive carbohydrates, lipids, and proteins contributed to generate barriers against Fusarium invasion in cassava roots, leading to lower the root rot disease severity.

In vitro gastrointestinal digestion of Lentinus squarrosulus powder and impact on human fecal microbiota.

Humans have long-used mushrooms as food and medicine, but digestion and colonic fermentation of most mushrooms, including Lentinus squarrosulus is markedly unknown. Here, nutritional profile, digestion and colonic fermentation of L. squarrosulus powder (LP) were determined. The powder contained mainly carbohydrate and protein. SEM and F-TIR analysis of the resistant hydrolysate (RH) revealed that the structure and ratio of carbohydrate and protein components were altered, and released known immunomodulation agents; beta-glucans and mannose. Both LP and RH promoted selected probiotic bacteria, especially Bifidobacterium strains. Using fecal microbiota of five volunteers (V1, V2, V3, V4 and V5), RH stimulated the microbiota of all used volunteers, via decreasing the ratio of Firmicutes/Bacteroidetes ranging from 1.3 to 8.2 times. Also, RH increased the relative abundance of vital immunomodulators; Bacteroides, Bifidobacterium, Clostridium cluster XIVa and IV, and Sutterella. Additionally, RH fermentation enriched the content of branch-chain fatty acids (BCFA) and short-chain fatty acids (SCFA), indicating protein and carbohydrate usage. Notably, propionic and butyric acids were abundant in V1, V2 and V3, while in V4 and V5, acetic and butyric acids were most enriched. Suggesting L. squarrosulus as functional mushroom to improve health and prevent diseases by enhancing gut health.

Development of Natural Luminescent Powder for the Detection of Latent Fingerprint.

<b>Background and Objective:</b> Nowadays, Dyes is widely used to improve fingerprints identification test. Natural dyes are another interesting way that can be used instead of chemical dyes because of its non-toxicity and lower cost. In this research, the development of rust powder from <i>Plumeria</i> tree was applied for fingerprints identification due to its fluorescence property under UV. Rust and Small Particle Reagent (SPR), containing ZnCO₃were applied to detect hidden fingerprints on non-porous surfaces in both dried and wet condition. <b>Materials and Methods:</b> Yellowish Rust from <i>Plumeria</i> tree was extracted with ethanol, grinded, dried and then mixed with ZnCO₃. Powder slurry was sprayed over fingerprint mark on different surfaces and monitored in both dried and wet condition. Visualization of fingerprint under UV was observed. Scanning microscope (SEM), UV-visible spectroscopy (UV-VIS), Fourier-Transform Infrared Spectroscopy (FTIR) and Energy-Dispersive X-ray (EDX) were also used to characterize physical and chemical properties of rust powder. <b>Results:</b> Fingerprints identification by dust technique using <i>Plumeria</i> rust powder as ingredient, provide best quality enhancement of fingerprints under UV light due to its fluorescent property, whereas a conventional technique of Small Particle Reagent technique (SPR) doesn't show fluorescent under UV. Data from SEM and FTIR show slight adhesion between zinc carbonate particles and rust powder. <b>Conclusion:</b> Fluorescence properties of rust powder is still interesting. Further improvement in powder recipe will be further investigated.

Identification of a Chitooligosaccharide Mechanism against Bacterial Leaf Blight on Rice by In Vitro and In Silico Studies.

This study focuses on a commercial plant elicitor based on chitooligosaccharides (BIG®), which aids in rice plant growth and disease resistance to bacterial leaf blight (BLB). When the pathogen (Xoo) vigorously attacks rice that has suffered yield losses, it can cause damage in up to 20% of the plant. Furthermore, Xoo is a seed-borne pathogen that can survive in rice seeds for an extended period. In this study, when rice seeds were soaked and sprayed with BIG®, there was a significant increase in shoot and root length, as well as plant biomass. Furthermore, BIG®-treated rice plants showed a significant reduction in BLB severity of more than 33%. Synchrotron radiation-based Fourier transform infrared (SR-FTIR) analysis was used to characterize BIG®'s mechanism in the chemical structure of rice leaves. The SR-FTIR results at 1650, 1735, and 1114 cm-1 indicated changes in biochemical components such as pectins, lignins, proteins, and celluloses. These findings demonstrated that commercial BIG® not only increased rice growth but also induced resistance to BLB. The drug's target enzyme, Xoo 1075 from Xanthomonas oryzae (PDB ID: 5CY8), was analyzed for its interactions with polymer ingredients, specifically chitooligosaccharides, to gain molecular insights down to the atomic level. The results are intriguing, with a strong binding of the chitooligosaccharide polymer with the drug target, revealing 10 hydrogen bonds between the protein and polymer. Overall, the computational analysis supported the experimentally demonstrated strong binding of chitooligosaccharides to the drug target.

Effect of Salicylic AcidFormulations on Induced Plant Defense against Cassava Anthracnose Disease.

This study was to investigate defense mechanisms on cassava induced by salicylic acid formulation (SA) against anthracnose disease. Our results indicated that the SA could reduce anthracnose severity in cassava plants up to 33.3% under the greenhouse condition. The ß-1,3-glucanase and chitinase enzyme activities were significantly increased at 24 hours after inoculation (HAI) and decrease at 48 HAI after Colletotrichum gloeosporioides challenge inoculation, respectively, for cassava treated with SA formulation. Synchrotron radiation-based Fourier-transform infrared microspectroscopy spectra revealed changes of the C=H stretching vibration (3,000-2,800 cm-1), pectin (1,740-1,700 cm-1), amide I protein (1,700-1,600 cm-1), amide II protein (1,600-1,500 cm-1), lignin (1,515 cm-1) as well as mainly C-O-C of polysaccharides (1,300-1,100 cm-1) in the leaf epidermal and mesophyll tissues treated with SA formulations, compared to those treated with fungicide carbendazim and distilled water after the challenged inoculation with C. gloeosporioides. The results indicate that biochemical changes in cassava leaf treated with SA played an important role in the enhancement of structural and chemical defense mechanisms leading to reduced anthracnose severity.

Identification of Salicylic Acid Mechanism against Leaf Blight Disease in Oryza sativa by SR-FTIR Microspectroscopic and Docking Studies.

The present study was to investigate the application and mechanism of salicylic acid (SA) as SA-Ricemate for the control of leaf blight disease using a Synchrotron Radiation-based Fourier-Transform Infra-Red (SR-FTIR) microspectroscopy and docking studies. After treating rice plants cv. KDML 105 with SA-Ricemate, the leaves were inoculated with Xanthomonas oryzae pv. oryzae, the causal agent of leaf blight, and disease severity were assessed. The leaves were also used to detect changes in endogenous SA content. The results indicated that SA-Ricemate, as an activated compound, reduced disease severity by 60% at three weeks post-inoculation and increased endogenous content by 50%. The SR-FTIR analysis of changes in the mesophyll of leaves (treated and untreated) showed that the groups of lipids, pectins, and proteins amide I and amide II occurred at higher values, and polysaccharides were shown at lower values in treated compared to untreated. Besides, docking studies were used to model a three-dimensional structure for Pathogenesis-related (PR1b) protein and further identify its interaction with SA. The results showed that ASP28, ARG31, LEU32, GLN97, and ALA93 are important residues that have strong hydrogen bonds with SA. The docking results showed that SA has a good interaction, confirming its role in expression.

Risk of Antimicrobial Resistant Non-Typhoidal Salmonella during Asymptomatic Infection Passage between Pet Dogs and Their Human Caregivers in Khon Kaen, Thailand.

To explore the risk of antimicrobial resistant (AMR) non-typhoidal Salmonella during asymptomatic infection passage between pet dogs and human caregivers in Khon Kaen, Thailand, one hundred forty paired fecal samples (n = 280) were obtained from companion dogs and their human caregivers, interviewed from 140 households during 2019-2020. The purified Salmonella isolates were serotype-identified and tested for antimicrobial resistance against ampicillin, ciprofloxacin, chloramphenicol, nalidixic acid, streptomycin, sulfamethoxazole-trimethoprim, and tetracycline. The homologous Salmonella isolate pairs (suggesting Salmonella infections may have been due to passage between each one of the pair, or derived from the same source) were subsequently characterized by serotype screening, pulsed field gel electrophoresis (PFGE), and Synchrotron Fourier transform infrared spectroscopy (SR-FTIR). The Salmonella prevalence observed in dogs, 12.86% (18/140), was not significantly different from that observed in humans, 17.86% (25/140) using McNemar's test. The AMR patterns (the patterns among the isolates of pet dogs and caregivers) and the serotypes (thirteen serotypes with 18 isolates from pet dogs plus thirteen serotypes with 25 isolates from humans) between pet dogs and humans were not significantly different using Pearson's chi-squared test. The homologous Salmonella isolates from the Salmonella-present households was 5.13% (2/39). This study demonstrated that the hypothesis that there is a high risk of Salmonella infection passage between dogs and humans with close contact in Khon Kaen is doubtful. Only 5.13% of homologous Salmonella isolates from Salmonella-present households were found in Khon Kaen, Thailand, although the prevalence of Salmonella-positive samples, serotypes, and antimicrobial resistance patterns were quite similar among the two populations.

Pupae protein extracts exert anticancer effects by downregulating the expression of IL-6, IL-1ß and TNF-α through biomolecular changes in human breast cancer cells.

The Pupae of Bombyx mori and Samia ricini are a source of high-quality proteins and essential nutrient elements for human. Recent studies revealed that protein extracted from pupae possessed therapeutic benefits for the treatment of many diseases. However, the anticancer activity of protein extracts from the pupae of B. mori and S. ricini has been rarely reported. Our objective was to study the effect of protein extracts from the pupae of B. mori and S. ricini on cytotoxicity and expression of pro-inflammatory cytokines; IL-6, IL-1ß and TNF-α, in breast cancer cells (MCF-7). Additionally, anticancer action of protein extracted from the pupae was further investigated through biomolecular changes in MCF-7 cells using Fourier transform infrared (FTIR) spectroscopy. Pupae protein extracts of B. mori exhibited cytotoxic effects with an IC50 value of 15.23 + 0.4 µg/mL with higher selectivity than doxorubicin on MCF-7 cells. Fourier transform infrared (FTIR) spectroscopy revealed that lipid contents in MCF-7 cells treated with pupae protein extracts of B. mori were higher than untreated cells. Treatment with protein extracts from pupae of B. mori or S. ricini caused significantly reduced protein and nucleic acid contents of MCF-7 cells. The expression of IL-6, IL-1ß and TNF-α in MCF-7 treated cells was investigated using RT-qPCR and ELISA. Our results revealed that protein extracts from the pupae of B. mori or S. ricini significantly decreased IL-6, IL-1ß and TNF-α in MCF-7 cells both at mRNA and protein levels. Expression of IL-6 and IL-1ß in MCF-7 treated cells, especially IL-6, was strongly reduced compared to untreated cells, while TNF-α expression was slightly decreased. These findings suggest that pupae protein extracted from B. mori or S. ricini may play a role in breast cancer through a down-regulatory action on the expression of IL-6, IL-1ß and TNF-α, and may also exert anticancer effects by causing biochemical changes of lipids, proteins and nucleic acids. These findings indicate that pupae protein extracted from B. mori or S. ricini may provide a potential novel therapeutic target for breast cancer.

The synergy and mode of action of quercetin plus amoxicillin against amoxicillin-resistant Staphylococcus epidermidis.

BACKGROUND: Staphylococcus epidermidis is one of the most multiple resistances to antibiotics in the recent years. Therefore, practically-prescribed antibiotics in the treatment of these strains are not effective. Plant-derived antibacterial is one of the most interesting sources of new therapeutics. The present study was to investigate antibacterial, synergy and modes of action of quercetin and amoxicillin against amoxicillin-resistant Staphylococcus epidermidis (ARSE). METHODS: The MICs, checkerboard assay, viability curves, cytoplasmic membrane (CM) permeability, enzyme assay, transmission electron microscopy, confocal microscopy and FT-IR microspectroscopy measurement was performed. RESULTS: The MICs of amoxicillin, penicillin, quercetin and kaempferol against all ARSE strains were 16, 200, 256-384 and >1024 µg/mL respectively. Synergistic effects were exhibited on amoxicillin plus quercetin and penicillin plus kaempferol against these strains at FIC index 0.50 and <0.38 respectively. The synergistic activity of quercetin plus amoxicillin was confirmed by the viable count. This combination increased CM permeability, caused marked morphological, peptidoglycan and cytoplasmic membrane damage, increased protein amide I and II, but decreased fatty acid in bacterial cells. The quercetin had an inhibitory activity against ß-lactamase. CONCLUSIONS: So, these findings are the first report that quercetin has the synergistic effect with amoxicillin against ARSE via four modes of actions, inhibit peptidoglycan synthesis and ß-lactamases activity, increase CM permeability and protein amide I and II but decrease fatty acid in bacterial cells. Of course, this flavonol has the dominant potential to develop a brand-new collateral phytochemical agent plus amoxicillin to treat ARSE. Future work should focus on the bioavailability, efficacy and toxicity in animal and human studies, as well as, the synergistic effect on blood and tissue should be evaluated and achieved.

Synergy and Mode of Action of Ceftazidime plus Quercetin or Luteolin on Streptococcus pyogenes.

Streptococcus pyogenes causes streptococcal toxic shock syndrome. The recommended therapy has been often failure through the interfering of beta-lactamase-producing bacteria (BLPB). The present study was to investigate antibacterial activity, synergy, and modes of action of luteolin and quercetin using alone and plus ceftazidime against S. pyogenes. The MICs of ceftazidime, luteolin, and quercetin against all S. pyogenes were 0.50, 128, and 128 µg mL(-1), respectively. A synergistic effect was exhibited on luteolin and quercetin plus ceftazidime against these strains at fractional inhibitory concentration indices 0.37 and 0.27, respectively, and was confirmed by the viable count. These combinations increased cytoplasmic membrane (CM) permeability, caused irregular cell shape, peptidoglycan, and CM damage, and decreased nucleic acid but increased proteins in bacterial cells. Enzyme assay demonstrated that these flavonoids had an inhibitory activity against ß-lactamase. In summary, this study provides evidence that the inhibitory mode of action of luteolin and quercetin may be mediated via three mechanisms: (1) inhibiting of peptidoglycan synthesis, (2) increasing CM permeability, and (3) decreasing nucleic acid but increasing the protein contents of bacterial cells. So, luteolin and quercetin propose the high potential to develop adjunct to ceftazidime for the treatment of coexistence of the BLPB and S. pyogenes infections.

Reversing ß-lactam antibiotic resistance of Staphylococcus aureus with galangin from Alpinia officinarum Hance and synergism with ceftazidime.

The purpose of this investigation was to extract and identify the bioactive phytochemicals from smaller galanga (Alpinia officinarum Hance). The antibacterial, synergy effects and primary mechanism of action of galangin and ceftazidime against S. aureus DMST 20651 are also investigated by minimum inhibitory concentration (MIC), checkerboard, killing curve determinations, enzyme assay and electronmicroscopy method. The rhizomes chloroform extract of this plant showed that these compounds were galangin, kaempferide and kaempferide-3-O-ß-D-glucoside, which had not been previously reported in this species. Synergistic FIC indices were observed in the combination of test flavonoids (galangin, quercetin and baicalein) and all selected ß-lactams (methicillin, ampicillin, amoxicillin, cloxacillin, penicillin G and ceftazidime) (FIC index, <0.02-0.11). The combination of ceftazidime at 5 µg/ml and 5 µg/ml of test flavonoids (galangin, quercetin and baicalein) exhibited synergistic effect by reduced the cfu/ml of this strain to 1×10(3) over 6 and throughout 24 h. Galangin showed marked inhibitory activity against penicillinase and ß-lactamase. Electronmicroscopy clearly showed that the combination of galangin and ceftazidime caused damage to the ultrastructures of the cells of this strain. It was concluded that galangin, quercetin and baicalein exhibited the potential to reverse bacterial resistance to ß-lactam antibiotics against penicillin-resistant S. aureus (PRSA). This may involve three mechanisms of action that galangin inhibit protein synthesis and effect on PBP 2a, interact with penicillinase and cause cytoplasmic membrane damage. These findings lead us to develop a new generation of phytopharmaceuticals that may use galangin, quercetin and baicalein in combination with ceftazidime to treat PRSA that currently almost untreatable microorganism. The anti-PRSA activity and mode of action of galangin is reported for the first time. These in vitro results have to be still confirmed in an animal test or in humans.