Gram-positive Micrococcus Luteus Research Articles

Comparison of automatic methods MALDI-TOF, VITEK2 and manual methods for the identification of intestinal microbial communities on the example of samples from alpacas (Vicugna pacos).

Universally, in microbiological diagnostics the detection of live bacteria is essential. Rapid identification of pathogens enables appropriate remedial measures to be taken. The identification of many bacteria simultaneously facilitates the determination of the characteristics of the accompanying microbiota and/or the microbiological complexity of a given environment. The effectiveness of the VITEK2 Compact automated microbial identification system and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS), analytical profile index (API) and Remel RapID tests were compared in identification of bacteria isolated from the alpaca gastrointestinal tract. Most isolates were Gram-positive, such as Bacillus cereus, Bacillus flexus, Bacillus licheniformis, Bacillus pumilus and Bacillus subtilis; Enterococcus faecium, Enterococcus gallinarum, Enterococcus hirae and Enterococcus casseliflavus; Staphylococcus aureus, Staphylococcus equorum, Staphylococcus lentus, Staphylococcus pseudintermedius and Staphylococcus sciuri; Paenibacillus amylolyticus; Cellulosimicrobium cellulans; Leuconostoc mesenteroides; Clostridium perfringens; Corynebacterium stationis, Corynebacterium xerosis, and Corynebacterium diphtheriae (the last only isolated manually by API Coryne and the VITEK2 system and Corynebacteria (CBC) card). Corynebacterium diphtheriae was misidentified by MALDI-TOF MS as Candida lipolytica (currently Yarrowia lipolytica). Gram-positive and Gram-variable Micrococcus luteus were also isolated. Gram-negative Enterobacter cloacae, Enterobacter gergoviae, Enterobacter hormaechei and Enterobacter ludwigii; E. coli; Klebsiella pneumoniae subsp. pneumoniae; Citrobacter braakii and Citrobacter freundii; Serratia liquefaciens, Serratia odorifera and Serratia marcescens; Morganella morganii subsp. morganii; Providencia alcalifaciens; Pseudomonas aeruginosa; Stenotrophomonas maltophilia; Moraxella osloensis; and Ochrobactrum intermedium were also found. The yeasts Candida albicans, Candida haemulonii and Candida ciferrii were also present. MALDI-TOF MS enabled the identification of pathogens and opportunistic pathogens from the alpaca gut which may represent a high risk to human and animal health.

Biosynthesis of silver nanoparticles with antioxidant and bactericidal activities using Capsicum annuum fruit extract

Metallic nanostructures, especially silver nanoparticles (AgNPs) have already found multiple applications in modern industry, science and medicine. Still, the production of nano-sized compounds often leads to the formation of toxic byproducts and possesses substantial environmental hazard. One of the promising solutions for the ecofriendly creation of nanomaterials predicts the use of a “green chemistry” approach using organisms, their parts or natural compounds to act as safe and effective producers of nanomaterials. Plant-mediated biosynthesis of silver nanoparticles includes the reduction of Ag+ to Ag0 by natural compounds, usually secondary metabolites which can be found in roots, leaves, cortex, fruits, flowers and seeds of various species. Aqueous extract of Capsicum annuum var. cv. (cultivar) Teja (S-17) fruits was used as a bioreducer for the reduction of AgNO3 solution to AgNPs. The formation of the AgNPs was confirmed by the presence of the Tyndall effect of light scattering combined with colour change of the solutions. The properties of the nanoparticles were assessed with UV-visible spectroscopy and scanning electron microscopy. In the present study, we report the experimental optimization of operating parameters needed for silver biotransformation by C. annuum. Biosynthesized nanoparticles were 13–22 nm in size and spherical in shape. Colloidal solutions of AgNPs were also confirmed to show antioxidant activity in vitro as analyzed by the reduction of DPPH radicals. Characterization and application of AgNPs as bactericidal agents on two Gram-positive (Micrococcus luteus, Staphylococcus aureus) and two Gram-negative (Escherichia coli, Pseudomonas aeruginosa) prokaryotic microorganisms demonstrated the prevalent influence on Gram-positive strains.

Biosynthesis and Characterization of Gold Nanoparticles Produced Using Rhodococcus Actinobacteria at Elevated Chloroauric Acid Concentrations.

The growing industrial and medical use of gold nanoparticles (AuNPs) requires environmentally friendly methods for their production using microbial biosynthesis. The ability of actinobacteria of the genus Rhodococcus to synthesize AuNPs in the presence of chloroauric acid (HAuCl4) was studied. The effect of elevated (0.8-3.2 mM) concentrations of HAuCl4 on bacterial viability, morphology, and intracellular accumulation of AuNPs by different Rhodococcus species was shown. An increase in surface roughness, a shift of the zeta potential to the positive region, and the formation of cell aggregates of R. erythropolis IEGM 766 and R. ruber IEGM 1135 during nanoparticle synthesis were revealed as bacterial adaptations to toxic effects of HAuCl4. The possibility to biosynthesize AuNPs at a five times higher concentration of chloroauric acid compared to chemical synthesis, for example, using the citrate method, suggests greater efficiency of the biological process using Rhodococcus species. The main parameters of biosynthesized AuNPs (size, shape, surface roughness, and surface charge) were characterized using atomic force microscopy, dynamic and electrophoretic light scattering, and also scanning electron microscopy in combination with energy-dispersive spectrometry. Synthesized by R. erythropolis spherical AuNPs have smaller (30-120 nm) dimensions and are positively (12 mV) charged, unlike AuNPs isolated from R. ruber cells (40-200 nm and -22 mV, respectively). Such differences in AuNPs size and surface charge are due to different biomolecules, which originated from Rhodococcus cells and served as capping agents for nanoparticles. Biosynthesized AuNPs showed antimicrobial activity against Gram-positive (Micrococcus luteus) and Gram-negative (Escherichia coli) bacteria. Due to the positive charge and high dispersion, the synthesized by R. erythropolis AuNPs are promising for biomedicine, whereas the AuNPs formed by R. ruber IEGM 1135 are prone to aggregation and can be used for biotechnological enrichment of gold-bearing ores.

Novel Type of Tetranitrosyl Iron Salt: Synthesis, Structure and Antibacterial Activity of Complex [FeL'2(NO)2][FeL'L"(NO)2] with L'-thiobenzamide and L"-thiosulfate.

In this work a new donor of nitric oxide (NO) with antibacterial properties, namely nitrosyl iron complex of [Fe(C6H5C-SNH2)2(NO)2][Fe(C6H5C-SNH2)(S2O3)(NO)2] composition (complex I), has been synthesized and studied. Complex I was produced by the reduction of the aqueous solution of [Fe2(S2O3)2(NO)2]2− dianion by the thiosulfate, with the further treatment of the mixture by the acidified alcohol solution of thiobenzamide. Based on the structural study of I (X-ray analysis, quantum chemical calculations by NBO and QTAIM methods in the frame of DFT), the data were obtained on the presence of the NO…NO interactions, which stabilize the DNIC dimer in the solid phase. The conformation properties, electronic structure and free energies of complex I hydration were studied using B3LYP functional and the set of 6–31 + G(d,p) basis functions. The effect of an aquatic surrounding was taken into account in the frame of a polarized continuous model (PCM). The NO-donating activity of complex I was studied by the amperometry method using an “amiNO-700” sensor electrode of the “inNO Nitric Oxide Measuring System”. The antibacterial activity of I was studied on gram-negative (Escherichia coli) and gram-positive (Micrococcus luteus) bacteria. Cytotoxicity was studied using Vero cells. Complex I was found to exhibit antibacterial activity comparable to that of antibiotics, and moderate toxicity to Vero cells.

Tetrazolium-Based Visually Indicating Bacteria Sensor for Colorimetric Detection of Point of Contamination.

Protective equipment for detecting bacterial contamination has been in high demand with increasing interest in public health and hygiene. Herein, a fiber-based visually indicating bacteria sensor (VIBS) embedded with iodonitrotetrazolium chloride is developed for the general purpose of detecting live bacteria, and its chromogenic effectiveness is investigated for Gram-negative Escherichia coli and Gram-positive Micrococcus luteus. The developed color intensity is measured by the light absorption coefficient to the scattering coefficient (K/S) based on the Kubelka-Munk equation, and the colorimetric sensitivities of different membranes are examined by calculating the limit of detection (LOD) and the limit of quantification (LOQ). The results demonstrate that the interactions between VIBS and bacteria depend on the wetting properties of membranes. A hydrophobic membrane shows excessive interactions at high concentrations of Gram-negative E. coli bacteria, whose cell membrane is lipophilic. The membrane blended with hydrophobic and hydrophilic polymers displays linear colorimetric responses for both Gram-negative and Gram-positive bacteria strains, demonstrating a reliable sensing capability in the range of the tested bacteria concentration. This study is significant in that explorative experimentations are performed to conceive a proof of concept of a fiber-based bacteria sensor, which is readily applicable in various fields where bacteria pose a threat.

Dandelion seeds as a new and valuable source of bioactive extracts obtained using the supercritical fluid extraction technique

The present study was intended to explore dandelion seeds as a new source of biologically active constituents using an environmentally friendly extraction technique. The process-function relationships were determined by following the influence of temperature (40 and 60 °C) and pressure (from 100 to 450 bar) on the amount of separated extract, its density, composition, and bioactivity. It was shown that the supercritical CO2-assisted technique enables the separation of 0.6–25.0% extract having a density in a range of 827–941 kg/m3. Extracts contained a high amount of unsaturated fatty acids (89.9–90.6%) including linoleic acid (71%) and α-linolenic acid (up to 0.9%), as well as phenolic (5.5–12.1 mg GAE/g) and flavonoid compounds (208.6–564.5 μg QE/g). The antioxidant activity test showed that 20 mg/mL extract solutions inhibit 26.0–64.4% DPPH radical. In addition, extracts showed significant antimicrobial activity, especially against Gram-positive Bacillus subtilis and Micrococcus luteus. There was no cytotoxic effect on normal green monkey kidney cells and human skin fibroblasts nor on hypopharyngeal cancer, cervical adenocarcinoma, and colon cancer cells. The study indicated the possible safe application of bioactive dandelion seeds extract in the pharmaceutical, food, and cosmetic industry.

Antibiotic profiling of wild-type bacilli led to the discovery of new lanthipeptide subtilin-producing Bacillus spizizenii strains whose 16S rDNA sequences differ from the B. spizizenii typing strain

Two dozen field-collected Bacillus and a dozen Bacillus spizizenii wild-type strains from strain collections were selected on the basis of their antagonistic properties against the Gram-positive strain Micrococcus luteus. Based on their genetic and antibiotic profiles, they were characterized (subtilin encoding spaS gene sequences, mass spectrometric, and quantitative-reversed phase liquid chromatographic analyses, as well as the presence of the lanthionine cyclase protein SpaC by western blotting), seven novel producers of the lanthipeptide subtilin. Phylogenetic analyses of the subtilin-producing wild-type strains based on their 16S rRNA sequences showed that all seven strains could be classified as B. spizizenii: The field-collected strains HS and N5, as well as strains DSM 618, 1087, 6395, 6405, and 8439 from the German Collection of Microorganisms and Cell Cultures. To the best of our knowledge, all B. spizizenii strains described so far are characterized by the fact that they can produce a lanthipeptide of the subtilin family. Both the lanthipeptide structures and the organization and sequences of the 16S rRNA-encoding genes suggest a subdivision of B. spizizenii into subspecies: The subtilin-producing B. spizizenii strains are distinctly different from the entianin-producing B. spizizenii typing strain TU-B-10 T (DSM 15029 T).

A Novel Antimicrobial Metabolite Produced by Paenibacillus apiarius Isolated from Brackish Water of Lake Balkhash in Kazakhstan.

Four aerobic bacteria with bacteriolytic capabilities were isolated from the brackish water site Strait Uzynaral of Lake Balkhash in Kazakhstan. The morphology and physiology of the bacterial isolates have subsequently been analyzed. Using matrix assisted laser desorption ionization-time of flight mass spectrum and partial 16S rRNA gene sequence analyses, three of the isolates have been identified as Pseudomonas veronii and one as Paenibacillus apiarius. We determined the capability of both species to lyse pre-grown cells of the Gram-negative strains Pseudomonas putida SBUG 24 and Escherichia coli SBUG 13 as well as the Gram-positive strains Micrococcus luteus SBUG 16 and Arthrobacter citreus SBUG 321 on solid media. The bacteriolysis process was analyzed by creating growth curves and electron micrographs of co-cultures with the bacteriolytic isolates and the lysis sensitive strain Arthrobacter citreus SBUG 321 in nutrient-poor liquid media. One metabolite of Paenibacillus apiarius was isolated and structurally characterized by various chemical structure determination methods. It is a novel antibiotic substance.

Brain Natriuretic Peptide (BNP) Affects Growth and Stress Tolerance of Representatives of the Human Microbiome, Micrococcus luteus C01 and Alcaligenes faecalis DOS7.

Simple SummaryThe body of an average person weighing 70 kg contains approximately 39 trillion bacterial cells, which densely inhabit the gastrointestinal tract, skin, mucous membranes, etc. Bacteria respond to the signaling molecules in the human body, regulate the expression of the necessary genes, and thus adapt to the physiology of the host. Signaling molecules include hormones, neurotransmitters, immune system molecules, as well as natriuretic peptides, which are involved in the regulation of the circulatory system, water and electrolyte metabolism, and adipose tissue metabolism. Brain natriuretic peptide (BNP) is secreted by the ventricles during congestion and signals heart failure. This study showed that the presence of BNP in the growth medium of human symbiont bacteria affects their growth characteristics, survival, and stress resistance, including antibiotic resistance. It was concluded that bacterial populations that develop in a healthy person at a BNP level of up to 250 pg/mL will be more stress resistant than in a person suffering from heart failure. Our findings are promising to be used both in clinical medical practice and in the production of bacterial preparations for cosmetology, agriculture, and waste management.Brain natriuretic peptide (BNP) is secreted by the ventricles of the heart during overload to signal heart failure. Slight bilateral skin itching induced by BNP has been associated with response activity of the skin microbiota. In this work, we studied the effect of 25–250,000 pg BNP/mL on the growth, long-term survival, and stress (H2O2, antibiotics, salinity, heat and pH shock) resistance of human symbiont bacteria: Gram-positive Micrococcus luteus C01 and Gram-negative Alcaligenes faecalis DOS7. The effect of BNP turned out to be dose-dependent. Up to 250 pg BNP/mL made bacteria more stress resistant. At 2500 pg BNP/mL (heart failure) the thermosensitivity of the bacteria increased. Almost all considered BNP concentrations increased the resistance of bacteria to the action of tetracycline and ciprofloxacin. Both bacteria survived 1.3–1.7 times better during long-term (up to 4 months) storage. Our findings are important both for clinical medical practice and for practical application in other areas. For example, BNP can be used to obtain stress-resistant bacteria, which is important in the collection of microorganisms, as well as for the production of bacterial preparations and probiotics for cosmetology, agriculture, and waste management.

Biogenic Synthesis and Characterization of Chitosan-CuO Nanocomposite and Evaluation of Antibacterial Activity against Gram-Positive and -Negative Bacteria.

Chitosan-copper oxide (CHT-CuO) nanocomposite was synthesized using olive leaf extract (OLE) as reducing agent and CuSO4⋅5H2O as precursor. CHT-CuO nanocomposite was prepared using an in situ method in which OLE was added to a solution of chitosan and CuSO4⋅5H2O mixture in the ratio of 1:5 (v/v) and heated at a temperature of 90 °C. The obtained CHT-CuO nanocomposite was characterized using field emission scanning electron microscopy (FE-SEM), X-ray diffraction (XRD), ultraviolet-visible (UV-Vis) spectrophotometry, energy-dispersive X-ray spectroscopy (EDAX), Fourier transform infrared spectroscopy (FTIR), and high-resolution transmission electron microscopy (TEM). TEM results indicated that CHT-CuO nanocomposite are spherical in shape with size ranging from 3.5 to 6.0 nm. Antibacterial activity of the synthesized nanocomposites was evaluated against Gram-positive (Bacillus cereus, Staphyloccous haemolytica and Micrococcus Luteus) and Gram-negative (Escherichia coli, Pseudomonas citronellolis, Pseudomonas aeruginosa, kliebisella sp., Bradyrhizobium japonicum and Ralstonia pickettii) species by cup platting or disc diffusion method. Overall, against all tested bacterial strains, the diameters of the inhibition zone of the three nanocomposites fell between 6 and 24 mm, and the order of the antimicrobial activity was as follows: CuO-1.0 > CuO-0.5 > CuO-2.0. The reference antibiotic amoxicillin and ciprofloxacin showed greater activity based on the diameter of zones of inhibition (between 15–32 mm) except for S. heamolytica and P. citronellolis bacteria strains. The nanocomposites MIC/MBC were between 0.1 and 0.01% against all tested bacteria, except S. heamolityca (>0.1%). Based on MIC/MBC values, CuO-0.5 and CuO-1.0 were more active than CuO-2.0, in line with the observations from the disc diffusion experiment. The findings indicate that these nanocomposites are efficacious against bacteria; however, Gram-positive bacteria were less susceptible. The synthesized CHT-CuO nanocomposite shows promising antimicrobial activities and could be utilized as an antibacterial agent in packaging and medical applications.

Assessment of Ocimum basilicum Essential Oil Anti-Insect Activity and Antimicrobial Protection in Fruit and Vegetable Quality.

Basil (Ocimum basilicum) is a commonly used herb; it also contains essential oils and other valuable compounds. The basil oil obtained has a pleasant aroma, but also a broad spectrum of biological activity. This work reports on the chemical composition, antioxidant, antimicrobial and anti-insect activity in vitro and in situ of Ocimum basilicum essential oil (OBEO) obtained by steam distillation of fresh flowering plants. Gas chromatography–mass spectrometry, DPPH, agar and disc diffusion and vapor phase methods were used to analyze the OBEO properties. The analysis of the chemical composition of OBEO showed that its main components were methyl chavicol (88.6%), 1,8-cineole (4.2%) and α-trans-bergamotene (1.7%). A strong antioxidant effect was demonstrated at the level of 77.3%. The analysis of antimicrobial properties showed that OBEO exerts variable strength of inhibiting activity against various groups of microorganisms. The growth inhibition zones ranged from 9.67 to 15.33 mm in Gram-positive (G+) and Gram-negative (G−) bacteria and from 5.33 to 7.33 mm in yeast. The lowest measured minimal inhibition concentration (MIC) was 3.21 µL/mL against Gram-negative Azotobacter chrococcum and Gram-positive Micrococcus luteus. The antimicrobial activity of in situ vapor phase of OBEO was also confirmed on apples, pears, potatoes and kohlrabi. The highest insecticidal activity against Pyrrhocoris apterus, observed at the concentration of 100%, caused the death of 80% of individuals. Due to its broad spectrum of activity, OBEO seems an ideal candidate for preserving fruit and vegetables.

Synthesis, structure and antibacterial activity of dinitrosyl iron complexes (DNICs) dimers functionalized with 5-(nitrophenyl) -4-H-1,2,4-triazole-3-thiolyls

This work aims at the synthesis and investigation of a new series of antibacterial agents, viz dimers of dinitrosyl iron complexes (DNICs) of [Fe2(SR)2(NO)4] composition, where R is 5-(2-nitrophenyl)-4-H-1,2,4-triazole-3-thiolyl (I), 5-(3-nitrophenyl)-4-H-1,2,4-triazole-3-thiolyl (II), 5-(4-nitrophenyl)-4-H-1,2,4-triazole-3-thiolyl (III), that are nitric oxide (NO) donors, easily obtained by replacement of thiosulfate ligands in [Fe(S2O3)2(NO)2]2- anion. Based on investigation of the structures of I and its isostructural analogs II and III (X-ray analysis, quantum-chemical calculations in the frame of DFT using NBO and QTAIM methods), data have been obtained on molecular interactions stabilizing DNIC dimer of µ–N–C–S structural type functionalized by 5-(2-nitrophenyl)-4-H-1,2,4-triazole-3-thiolyl. NBO calculations provided the assessment of bond energy for the Fe-S and Fe-N bonds incorporated in the µ–N–C–S bridge. From QTAIM analysis, detailed information on the nature and covalence of the forming bonds has been obtained. Assessment of antibacterial activity of I-III with respect to gram-negative (Escherichia coli) and gram-positive (Micrococcus luteus) bacteria has been performed.

Chemical Composition, Antioxidant, In Vitro and In Situ Antimicrobial, Antibiofilm, and Anti-Insect Activity of Cedar atlantica Essential Oil.

The present study was designed to evaluate commercial cedar essential oil (CEO), obtained by hydrodistillation from cedar wood, in relationship to its chemical composition and antioxidant, in vitro and in situ antimicrobial, antibiofilm, and anti-insect activity. For these purposes, gas chromatography–mass spectrometry, DPPH radical-scavenging assay, agar and disc diffusion, and vapor phase methods were used. The results from the volatile profile determination showed that δ-cadinene (36.3%), (Z)-β-farnesene (13.8%), viridiflorol (7.3%), and himachala-2,4-diene (5.4%) were the major components of the EO chemical constitution. Based on the obtained results, a strong antioxidant effect (81.1%) of the CEO was found. CEO is characterized by diversified antimicrobial activity, and the zones of inhibition ranged from 7.33 to 21.36 mm in gram-positive and gram-negative bacteria, and from 5.44 to 13.67 mm in yeasts and fungi. The lowest values of minimal inhibition concentration (MIC) were noted against gram-positive Micrococcus luteus (7.46 µL/mL) and against yeast Candida krusei (9.46 µL/mL). It seems that the vapor phase of CEO can inhibit the growth of the microscopic filamentous fungi of the genus Penicillium according to in situ antifungal analysis on bread, carrots, and celery. This finding confirms the impact of CEO on the change in the protein structure of older biofilms of Pseudomonas fluorescens and Salmonella enterica subsp. enterica. Insecticidal activity of a vapor phase has also been demonstrated against Pyrrhocoris apterus. CEO showed various advantages on antimicrobial activity, and it is an ideal substitute for food safety.

Isolation and Characterization of Antimicrobial Peptides Isolated from Fagonia bruguieri.

The majority of pathogenic microorganisms have developed resistance to commercial antibiotics. It causes the risk of illness relapse with current antimicrobial therapy regimens; additional and/or different antibacterial drugs are needed to treat diseases caused by these pathogenic microorganisms. The applied analysis in the present study was purification and characterization of plant peptides isolated from the leaves of Fagonia bruguieri as well as their antibacterial activities against Gram-positive bacteria, Staphylococcus aureus, Bacillus subtilis, Micrococcus luteus, and methicillin-resistant Staphylococcus aureus, in addition to Gram-negative bacteria, Escherichia coli and Pseudomonas aeruginosa. The minimum inhibitory concentration for the isolated peptide ranges from 25 to 62.5mg/mL. The methanolic solvent was used for the extraction followed by reversed-phase high-performance liquid chromatography for purification of peptides. Eventually, the peptide characterization and identification were also determined by MALDI-TOF/TOF and SEM analysis. This study paves a way to the effective antimicrobials from the plant resources.

Feeding-Induced Changes of Bacteriolytic Activity and the Pattern of Bacteriolytic Compounds in the Stomach and Small Intestine of the Haematophagous Bug Triatoma infestans (Klug, 1834) (Reduviidae, Triatominae)

Intestinal homeostasis mechanisms of the haematophagous triatomines regulate the development of mutualistic symbionts and other gut bacteria. Investigating antimicrobial compounds of these insects, we have determined spectrophotometrically that the bacteriolytic activity is between pH 3 and pH 9 using homogenates of fifth instar Triatoma infestans stomachs and small intestines from unfed bugs and up to 50 days after feeding. The activity against Gram-positive Micrococcus luteus was strongest at pH 4 and pH 7 and was higher in the stomach than in the small intestine. Symbiotic Rhodococcus triatomae were not lysed. Lysis of Gram-negative Escherichia coli showed a maximum at pH 7 in the stomach and at pH 5 in the small intestine. Bacteriolytic activity against both M. luteus and E. coli was reduced 24 h after feeding, then increased, and at 50 days after feeding was strongly reduced. In zymographs, the activity against M. luteus was mainly correlated to proteins of about 16 kDa. At different periods of time after feeding, seven bands of lysis appeared between 15 and 40 kDa and more bands using extracts of the small intestine than those of the stomach. This is the first proof for the synthesis of antibacterial proteins of 22–40 kDa in triatomines.

Aminothiazole-Linked Metal Chelates: Synthesis, Density Functional Theory, and Antimicrobial Studies with Antioxidant Correlations.

During the current study, the new aminothiazole Schiff base ligands (S1) and (S2) were designed by reacting 1,3-thiazol-2-amine and 6-ethoxy-1,3-benzothiazole-2-amine separately with 3-methoxy-2-hydroxybenzaldehyde in good yields (68–73%). The ligands were characterized through various analytical, physical, and spectroscopic (FT-IR, UV–Vis, 1H and 13C NMR, and MS) methods. The ligands were exploited in lieu of chelation with bivalent metal (cobalt, nickel, copper, and zinc) chlorides in a 1:2 (M:L) ratio. The spectral (UV–Vis, FT-IR, and MS), as well as magnetic, results suggested their octahedral geometry. The theoretically optimized geometrical structures were examined using the M06/6-311G+(d,p) function of density function theory. Their bioactive nature was designated by global reactivity parameters containing a high hardness (η) value of 1.34 eV and a lower softness (σ) value of 0.37 eV. Different microbial species were verified for their potency (in vitro), revealing a strong action. The Gram-positive Micrococcus luteus and Gram-negative Escherichia coli gave the highest activities of 20 and 21 mm for compounds (8) and (7), respectively. The antifungal activity against the Aspergillus niger and Aspergillus terreus species gave the highest activities of 20 and 18 mm for compounds (7) and (6), respectively. The antioxidant activity, evaluated as DPPH and ferric reducing power, gave the highest inhibition (%) as 72.0 ± 0.11% (IC50 = 144 ± 0.11 μL) and 66.3% (IC50 = 132 ± 0.11 μL) for compounds (3) and (8), respectively. All metal complexes were found to be more biocompatible than free ligands due to their chelation phenomenon. The energies of LUMOs had a link with their activities.

Improving the Activity of Antimicrobial Peptides Against Aquatic Pathogen Bacteria by Amino Acid Substitutions and Changing the Ratio of Hydrophobic Residues.

With the increasing number of drug-resistant bacteria, there is an urgent need for new antimicrobial agents, and antimicrobial peptides (AMPs), which exist in the human non-specific immune system, are one of the most promising candidates. It is an effective optimization strategy to modify antimicrobial peptides (AMPs) according to the distribution of amino acids and hydrophobic characteristics. The addition of bacterial pheromones to the N short peptide can increase the ability to recognize bacteria. In this study, we designed and synthesized AMP1–6 by amino acid substitution of mBjAMP1. Additionally, P-6, S-6, and L-6 were designed and synthesized by adding bacterial pheromones based on 1–6. Functional tests showed that the four AMPs had the ability to kill Gram-negative Vibrio anguillarum, Pseudomonas mendocina, and Vibrio parahaemolyticus, and Gram-positive Micrococcus luteus and Listeria monocytogenes. Additionally, all four AMPs induced permeabilization and depolarization of bacterial cell membranes and increased intracellular reactive oxygen species (ROS) levels. Importantly, they had little or no mammalian cytotoxicity. At the same time, 1–6 and L-6 protected the stability of intestinal flora in Sebastes schlegelii and increased the relative abundance of Lactobacillaceae. In summary, our results indicate that the designed AMPs have broad application prospects as a new type of polypeptide antimicrobial agent.

Characterization and functional analysis of a Relish gene from the Asian corn borer, Ostrinia furnacalis (Guenée).

Pathogen-induced host immune responses reduce the efficacy of pathogens used to control pests. However, compared to the well-deciphered immunity system of Drosophila melanogaster, the immunity system of agricultural pests is largely unconfirmed through functional analysis. Beginning to unveil mechanisms of transcription regulation of immune genes in the Asian corn borer, Ostrinia furnacalis, we cloned the complementary DNA (cDNA) of a transcription factor Relish by rapid amplification of cDNA ends. The 3164 bp cDNA, designated Of-Relish, encodes a 956-residue protein. Bioinformatic analysis showed that Of-Relish had a Rel homology domain, a predicted cleavage site between Q409 and L410 , six ankyrin repeats, and a death domain. The response of Of-Relish expression to the Gram-negative bacteria Pseudomonas aeruginosa was sooner and stronger than to the Gram-positive Micrococcus luteus. The antimicrobial peptide genes Attacin and Gloverin had similar expression patterns in response to the infections. Knockdown of Of-Relish led to a decrease in Attacin and Gloverin messenger RNA levels, suggesting that Attacin and Gloverin were regulated by Of-Relish. Together, the results suggested that Of-Relish is a key component of the IMD pathway in O. furnacalis, involved in defense against P. aeruginosa through activation of Attacin and Gloverin.

A Short-Type Peptidoglycan Recognition Protein 1 (PGRP1) Is Involved in the Immune Response in Asian Corn Borer, Ostrinia furnacalis (Guenée).

The insect immune response is initiated by the recognition of invading microorganisms. Peptidoglycan recognition proteins (PGRPs) function primarily as pattern recognition receptors by specifically binding to peptidoglycans expressed on microbial surfaces. We cloned a full-length cDNA for a PGRP from the Asian corn borer Ostrinia furnacalis (Guenée) and designated it as PGRP1. PGRP1 mRNA was mainly detected in the fat bodies and hemocytes. Its transcript levels increased significantly upon bacterial and fungal challenges. Purified recombinant PGRP1 exhibited binding activity to the gram-positive Micrococcus luteus, gram-negative Escherichia coli, entomopathogenic fungi Beauveria bassiana, and yeast Pichia pastoris. The binding further induced their agglutination. Additionally, PGRP1 preferred to bind to Lys-type peptidoglycans rather than DAP-type peptidoglycans. The addition of recombinant PGRP1 to O. furnacalis plasma resulted in a significant increase in phenoloxidase activity. The injection of recombinant PGRP1 into larvae led to a significantly increased expression of several antimicrobial peptide genes. Taken together, our results suggest that O. furnacalis PGRP1 potentially recognizes the invading microbes and is involved in the immune response in O. furnacalis.

SUSTENANCE, THE SAFETY OF PLANTS AND FEED OF Galega orientalis (Lam.)

Conducted a comprehensive comparative study of the nutritional and biological value Galega orientalis (Lam.): the solubility of the protein in vitro - Galega’s seeds, leaves, hay, which was harvested in the phase of budding - beginning of flowering. It is believed to be the essential influence of feed type on the solubility of the protein. According to the conducted studies it has revealed that the solubility of the feed protein is in the range from 37.0% to 50.0%. The rate of solubility of Galega orientalis (Lam.) seed protein is 50.0%, litestock - 40.0%, and hay - 37.0% from its total. The research results indicate that the protein of Galega orientalis (Lam.) refers to the feed with the average level of punct solubility. It has researched qualitative assessment of the biologically active substances content in organs of Galega orientalis’ (Lam.) plants. It has identified the highest content of flavonoids and saponins in rosette leaves and ascorbic acid in the organs of plants and petals. There is the high content of tannins found in the petals of Galega orientalis (Lam.). There is a high content of coumarins and water-soluble polysaccharides in the roots of the studied plants. Phytochemical studies of the plant Galega orientalis (Lam.) indicates the absence in it of cardiac glycosides and antrahlikozydiv. It was first conducted testing on antimicrobial activity of a plant Galega orientalis (Lam.) on the growth pure cultural of bacteria Gram-positive (Micrococcus luteus), Gram-negative (Escherichia coli XL1, DH5) and yeasts (Saccharomyces cerevisiae W303). The material for the study was dried vegetative mass Galega orientalis (Lam.) in the phase of budding and early flowering. As a result of the experiment was revealed that 20% concentration of aqueous extract of this plant had inhibitory effects on the growth of pure cultures of bacteria and yeast. It has grounded reasonably possible relationship between the antimicrobial activity of the extract of this plant and the lack of bloating in cows. It has outlined prospects of further researches of this problem