In Situ Antimicrobial Properties of Sabinene Hydrate, a Secondary Plant Metabolite.

The objective of this research was to investigate natural products for their potential against pathogenic microorganisms. Sabinene hydrate (SH), a monoterpenoid, is synthesised by numerous different plants as a secondary metabolite. At present, there is a lack of definite investigations regarding the antimicrobial activity of SH itself and its different isomers. The antimicrobial effects of commercially available SH (composed mainly of trans-isomer) were evaluated within a range of concentrations in three types of contact tests: solid and vapor diffusion and the macro-broth dilution method. Moreover, the effects of SH on the rate of linear growth and spore germination were also examined. Ethanolic SH solutions were tested against an array of microorganisms, including blue-stain fungi (Ceratocystis polonica, Ophiostoma bicolor, O. penicillatum), frequently originating from bark beetle galleries; three fungal strains (Musicillium theobromae, Plectosphaerella cucumerina, and Trichoderma sp.) isolated from a sapwood underneath bark beetle galleries (Ips typographus) on spruce (Picea abies) stems; Verticillium fungicola, isolated from diseased I. typographus larvae; two Gram-positive bacteria (Bacillus subtilis and Staphylococcus aureus), two Gram-negative bacteria (Escherichia coli and Pseudomonas aeruginosa); five yeasts (Candida albicans, C. krusei, C. parapsilosis, Saccharomyces cerevisiae, and Rhodotorula muscilaginosa), and two saprophytic fungi (Aspergillus niger and Penicillium notatum). In solid agar disc diffusion tests, Gram-positive bacteria exhibited greater susceptibility to SH than Gram-negative bacteria, followed by yeasts and fungi. The most resistant to SH in both the disc diffusion and broth macro-dilution methods were P. aeruginosa, A. niger, and Trichoderma sp. strains. Blue-stain fungi and fungi isolated from the Picea sapwood were the most resistant among the fungal strains tested. The minimum inhibition concentrations (MICs) generated by SH and determined using a disc volatilization method were dependent on the fungal species and played an important role in the development of microorganism inhibition. The two Gram-positive bacteria, B. subtilis and S. aureus (whose MICs were 0.0312 and 0.0625 mg/mL, respectively), were the organisms most susceptible to SH, followed by the Gram-negative bacterium, E. coli (MIC = 0.125 mg/mL) and two yeasts, C. albicans and C. kruei (MIC was 0.125 mg/mL and 0.25 mg/mL, respectively). C. parapsilosis (MIC = 0.75 mg/mL) was the yeast most resistant to SH. The investigation of antimicrobial properties of plant secondary metabolites is important for the development of a new generation of fungicides.

Fluorescent carbon dots for discriminating cell types: a review.

Carbon dots (CDs) are quasi-spherical carbon nanoparticles with excellent photoluminescence, good biocompatibility, favorable photostability, and easily modifiable surfaces. CDs, serving as fluorescent probes, have emerged as an ideal tool for cellular differentiation owing to their outstanding luminescence performance and tunable surface properties. In this review, we summarize the recent research progress with CDs in the differentiation of cancer/normal cells, Gram-positive/Gram-negative bacteria, and live/dead cells, as well as the cellular differences used for differentiation. Additionally, we summarize the preparation methods, raw materials, and properties of the CDs used for cell discrimination. The differentiation mechanisms and the advantages or limitations of the differentiation methods are also introduced. Finally, we propose several research challenges in this field and future research directions that require extensive investigation. It is hoped that this review will help researchers in the design of new CDs as ideal fluorescent probes for realizing diverse cell differentiation applications.

Phytochemical Screening and Antibacterial Activity of Commercially Available Essential Oils Combinations with Conventional Antibiotics against Gram-Positive and Gram-Negative Bacteria.

The present study aims to evaluate the antibacterial activity of five commercially available essential oils (EOs), Lavender (LEO), Clove (CEO), Oregano (OEO), Eucalyptus (EEO), and Peppermint (PEO), against the most-known MDR Gram-positive and Gram-negative bacteria-Staphylococcus aureus (ATCC 25923), Escherichia coli (ATCC 25922), and Pseudomonas aeruginosa (ATCC 27853)-alone and in various combinations. Gas Chromatography-Mass Spectrometry (GC-MS) analysis established their complex compositions. Then, their antibacterial activity-expressed as the inhibition zone diameter (IZD) value (mm)-was investigated in vitro by the diffusimetric antibiogram method, using sterile cellulose discs with Ø 6 mm impregnated with 10 µL of sample and sterile borosilicate glass cylinders loaded with 100 µL; the minimum inhibitory concentration (MIC) value (µg/mL) for each EO was calculated from the IZD values (mm) measured after 24 h. The following EO combinations were evaluated: OEO+CEO, CEO+EEO, CEO+PEO, LEO+EEO, and EEO+PEO. Then, the influence of each dual combination on the activity of three conventional antibacterial drugs-Neomycin (NEO), Tetracycline (TET), and Bacitracin (BAC)-was investigated. The most active EOs against S. aureus and E. coli were LEO and OEO (IZD = 40 mm). They were followed by CEO and EEO (IZD = 20-27 mm); PEO exhibited the lowest antibacterial activity (IZD = 15-20 mm). EEO alone showed the highest inhibitory activity on P. aeruginosa (IZD = 25-35 mm). It was followed by CEO, LEO, and EEO (IZD = 7-11 mm), while PEO proved no antibacterial action against it (IZD = 0 mm). Only one synergic action was recorded (OEO+CEO against P. aeruginosa); EEO+PEO revealed partial synergism against S. aureus and CEO+PEO showed additive behavior against E. coli. Two triple associations with TET showed partial synergism against E. coli, and the other two (with NEO and TET) evidenced the same behavior against S. aureus; all contained EEO+PEO or CEO+PEO. Most combinations reported indifference. However, numerous cases involved antagonism between the constituents included in the double and triple combinations, and the EOs with the strongest antibacterial activities belonged to the highest antagonistic combinations. A consistent statistical analysis supported our results, showing that the EOs with moderate antibacterial activities could generate combinations with higher inhibitory effects based on synergistic or additive interactions.

Equinins as Novel Broad-Spectrum Antimicrobial Peptides Isolated from the Cnidarian Actinia equina (Linnaeus, 1758).

Sea anemones are valuable for therapeutic research as a diversified source of bioactive molecules, due to their diverse bioactive molecules linked to predation and defence mechanisms involving toxins and antimicrobial peptides. Acid extracts from Actinia equina tentacles and body were examined for antibacterial activity against Gram-positive, Gram-negative bacteria, and fungi. The peptide fractions showed interesting minimum inhibitory concentration (MIC) values (up to 0.125 µg/mL) against the tested pathogens. Further investigation and characterization of tentacle acid extracts with significant antimicrobial activity led to the purification of peptides through reverse phase chromatography on solid phase and HPLC. Broad-spectrum antimicrobial peptide activity was found in 40% acetonitrile fractions. The resulting peptides had a molecular mass of 2612.91 and 3934.827 Da and MIC ranging from 0.06 to 0.20 mg/mL. Sequencing revealed similarities to AMPs found in amphibians, fish, and Cnidaria, with anti-Gram+, Gram-, antifungal, candidacidal, anti-methicillin-resistant Staphylococcus aureus, carbapenemase-producing, vancomycin-resistant bacteria, and multi-drug resistant activity. Peptides 6.2 and 7.3, named Equinin A and B, respectively, were synthesized and evaluated in vitro towards the above-mentioned bacterial pathogens. Equinin B exerted interesting antibacterial activity (MIC and bactericidal concentrations of 1 mg/mL and 0.25 mg/mL, respectively) and gene organization supporting its potential in applied research.

Potential of using medicinal plant extracts as photosensitizers for antimicrobial photodynamic therapy.

Antimicrobial photodynamic therapy (APDT) is a promising approach to overcome antimicrobial resistance. However, for widespread implementation of this approach, approved photosensitizers are needed. In this study, we used commercially available preparations (Calendulae officinalis floridis extract, Chamomillae recutitae floridis extract, Achillea millefolii herbae extract; Hypericum perforatum extract; Eucalyptus viminalis folia extract) as photosensitizers for inactivation of gram-negative (Pseudomonas aeruginosa) and gram-positive (Staphylococcus aureus) bacteria. Spectral-luminescent analysis has shown that the major chromophores are of chlorophyll (mainly chlorophyll a and b) and hypericin nature. The extracts are efficient generators of singlet oxygen with quantum yield (Î³Δ ) from 0.40 to 0.64 (reference compound, methylene blue with Î³Δ = 0.52). In APDT assays, bacteria before irradiation were incubated with extracts for 30 min. After irradiation and 24 h of incubation, colony-forming units (CFU) were counted. Upon exposure of P. aeruginosa to radiation of 405 nm, 590 nm, and 660 nm at equal energy dose of 30 J/cm2 (irradiance - 100 mW/cm2 , exposure time - 5 min), the most pronounced effect is observed with blue light (>3 log10 reduction); in case of S. aureus, the effect is approximately equivalent for light of indicated wavelengths and dose (>4 log10 reduction).

Anti Gram-Positive Bacteria Activity of Synthetic Quaternary Ammonium Lipid and Its Precursor Phosphonium Salt.

Organic ammonium and phosphonium salts exert excellent antimicrobial effects by interacting lethally with bacterial membranes. Particularly, quaternary ammonium lipids have demonstrated efficiency both as gene vectors and antibacterial agents. Here, aiming at finding new antibacterial devices belonging to both classes, we prepared a water-soluble quaternary ammonium lipid (6) and a phosphonium salt (1) by designing a synthetic path where 1 would be an intermediate to achieve 6. All synthesized compounds were characterized by Fourier-transform infrared spectroscopy and Nuclear Magnetic Resonance. Additionally, potentiometric titrations of NH3+ groups 1 and 6 were performed to further confirm their structure by determining their experimental molecular weight. The antibacterial activities of 1 and 6 were assessed first against a selection of multi-drug-resistant clinical isolates of both Gram-positive and Gram-negative species, observing remarkable antibacterial activity of both compounds against Gram-positive isolates of Enterococcus and Staphylococcus genus. Further investigations on a wider variety of strains of these species confirmed the remarkable antibacterial effects of 1 and 6 (MICs = 4-16 and 4-64 µg/mL, respectively), while 24 h-time-killing experiments carried out with 1 on different S. aureus isolates evidenced a bacteriostatic behavior. Moreover, both compounds 1 and 6, at the lower MIC concentration, did not show significant cytotoxic effects when exposed to HepG2 human hepatic cell lines, paving the way for their potential clinical application.

Development of Novel Chimeric Endolysin Conjugated with Chitosan-Zn-Metal-Organic Framework Nanocomposites with Antibacterial Activity.

Bacterial diseases have been considered the most crucial issue and are threatening human health all around the world. Also, resistance to antimicrobial drugs has become a big hurdle against efficient therapy. As a result, recombinant chimeric endolysin was produced in E. coli host to use as a potential antibacterial agent against bacteria resistance and replacement to conventional antibiotics in this study. Then, chitosan (C)-coated nanoscale metal-organic frameworks (CS-NMOFs) nanocomposite was synthesized as a novel nano delivery system to further improve the antibacterial activity of endolysin. After characterization of nanocomposite with analytical devices such as FT-IR, DLS, and TEM and determining the nanometric size of samples (30 nm to 90 nm), endolysin was covalently (endolysin-CS-NMOFs (C)) and non-covalently (endolysin-CS-NMOFs (NC)) conjugated to nanocomposite. Thereafter, the lytic ability, synergistic interaction, and biofilm reduction manner of endolysin-containing CS-NMOF nanocomposites were evaluated on E. coli, S. aureus, and P. aeruginosa strains. The results depicted an excellent lytic ability of nanocomposites after 24 h and 48 h of treatment, especially endolysin-CS-NMOFs (NC) on E. coli and P. aeruginosa strains. The synergistic interaction between nanocomposite and vancomycin did not attain for P. aeruginosa strain whereas the reverse was true for E. coli and S. aureus strains at 8 ng/mL concentration. Next, nanocomposites demonstrated potential biofilm reduction activities in various strains, especially in S. aureus and P. aeruginosa. Ultimately, our outputs demonstrate an efficient performance of the synthesized nanocomposite as an appropriate substitution for conventional antibiotics against bacteria.

Novel Chimeric Endolysin Conjugated Chitosan Nanocomplex as a Potential Inhibitor Against Gram-Positive and Gram-Negative Bacteria.

Resistance to antimicrobial agents has created potential problems in finding efficient treatments against bacteria. Thus, using new therapeutics, such as recombinant chimeric endolysin, would be more beneficial for eliminating resistant bacteria. The treatment ability of these therapeutics can be further improved if they are used with biocompatible nanoparticles like chitosan (CS). In this work, covalently conjugated chimeric endolysin to CS nanoparticles (C) and non-covalently entrapped endolysin in CS nanoparticles (NC) were effectively developed and, consequently, qualified and quantified using analytical devices, including FT-IR, dynamic light scattering, and TEM. Eighty to 150 nm and 100 nm to 200 nm in diameter were measured for CS-endolysin (NC) and CS-endolysin (C) using a TEM, respectively. The lytic activity, synergistic interaction, and biofilm reduction potency of nano-complexes were investigated on Escherichia coli (E. coli), Staphylococcus aureus (S. aureus), and Pseudomonas aeruginosa (P. aeruginosa) strains. The outputs revealed a good lytic activity of nano-complexes after 24 h and 48 h of treatment, especially in P. aeruginosa (approximately 40% cell viability after 48 h of treatment with 8 ng/mL), and potential biofilm reduction performance was attained in E. coli strains (about 70% reduction after treatment with 8 ng/mL). The synergistic interaction between nano-complexes and vancomycin was exhibited in E. coli, P. aeruginosa, and S. aureus strains at 8 ng/mL concentrations, while the synergistic effects of pure endolysin and vancomycin were not remarkable in E. coli strains. These nano-complexes would be more beneficial in suppressing the bacteria with a high level of antibiotic resistance.

Study of the Antimicrobial Potential of the Arthrospira platensis, Planktothrix agardhii, Leptolyngbya cf. ectocarpi, Roholtiella mixta nov., Tetraselmis viridis, and Nanofrustulum shiloi against Gram-Positive, Gram-Negative Bacteria, and Mycobacteria.

The incidence of diseases brought on by resistant strains of micro-organisms, including tuberculosis, is rising globally as a result of the rapid rise in pathogenic micro-organism resistance to antimicrobial treatments. Secondary metabolites with potential for antibacterial activity are produced by cyanobacteria and microalgae. In this study, gram-positive (S. aureus, E. faecalis) and gram-negative (K. pneumoniae, A. baumannii, P. aeruginosa) bacteria were isolated from pulmonary tuberculosis patients receiving long-term antituberculosis therapy. The antimicrobial potential of extracts from the cyanobacteria Leptolyngbya cf. ectocarpi, Planktothrix agardhii, Arthrospira platensis, Rohotiella mixta sp. nov., Nanofrustulum shiloi, and Tetraselmis (Platymonas) viridis Rouchijajnen was evaluated. On mouse splenocytes and peritoneal macrophages, extracts of cyanobacteria and microalgae had inhibitory effects. In vitro studies have shown that cyanobacteria and microalgae extracts suppress the growth of bacteria and mycobacteria. At the same time, it has been demonstrated that cyanobacterial and microalgal extracts can encourage bacterial growth in a test tube. Additionally, the enhanced fucoxanthin fraction significantly reduced the development of bacteria in vitro. In a mouse experiment to simulate tuberculosis, the mycobacterial load in internal organs was considerably decreased by fucoxanthin. According to the information gathered, cyanobacteria and microalgae are potential sources of antibacterial compounds that can be used in the manufacturing of pharmaceutical raw materials.

Hidden bugs in a newly opened hospital: the distribution of skin microbiota among healthcare workers in a newly opened teaching hospital.

Background. Skin is a reservoir for millions of micro-organisms, all of which make up the skin microbiota. Hospitals have been identified as a favourable environment for transmitting micro-organisms and thus, it is important to know the distribution of skin microbiota among healthcare workers (HCWs), as such findings may provide baseline information for the distribution of skin microbiota in hospitals.Hypothesis. There is no significant association between the factors (age, gender, type of skin microenvironment, hand hygiene practices, usage of skin care products, current healthcare practices and previous workplace) and the distribution of the skin microbiota among HCWs.Aim. The study aims to identify type of skin microbiota and associated factors (age, gender, type of skin microenvironment, hand hygiene practices, use of skincare products, current healthcare practice, and previous workplace) that influence the growth of skin microbiota.Method. About 102 bacterial isolates were obtained from the skin of 63 healthcare workers in a newly opened teaching hospital, namely Hospital Pengajar Universiti Putra Malaysia (HPUPM). All isolated bacteria were subjected to phenotypic identification according to standard microbiological procedures.Results. The most common isolated skin microbiota were Gram-positive bacteria (84.3%), followed by Gram-negative bacteria (15.7%). A Chi-square test of independence was used to analyse the above factors and there was a significant association between the type of skin microenvironment and the distribution of skin microbiota (P=0.03) (type of skin microenvironment influences the distribution of skin microbiota).Conclusion. Coagulase-negative Staphylococcus spp. was the most common bacteria isolated from the skin of the healthcare workers. Even though coagulase-negative staphylococci (CoNS) are low pathogenic bacteria, but it may cause serious infection in high risk group of patients. Therefore, it is important to emphasize on the good hand hygiene practices and implement strict infection control measures to minimize the risk of HAI in newly opened hospitals.

Multiple Assays on Non-Target Organisms to Determine the Risk of Acute Environmental Toxicity in Tebuconazole-Based Fungicides Widely Used in the Black Sea Coastal Area.

The widespread use of Tebuconazole-based fungicides in phytosanitary treatments on a wide range of crops, on the one hand, and the lack of official reports on the amount of fungicide residues in nearby water basins, on the other hand, may lead to uncontrolled and hazardous contamination of water sources used by the resident population, and to serious effects on the environment and public health. Our study explores the acute toxicological risk of this fungicide on various organisms, from bacteria and yeast to fish, using a battery of tests (standardized Toxkit microbiotests and acute semi-static tests). By investigating the interaction between Tebuconazole and bacteria and yeast organisms, we observed that Gram-negative bacteria displayed a strong tolerance for Tebuconazole, while Gram-positive bacteria and yeasts proved to be very sensitive. The fish experiment was conducted on Chelon auratus juveniles exposed to five concentrations of the fungicide Tebustar EW (Tebuconazole, 250 g/L as active substance). After 96 h of exposure, the LC50 for C. auratus was 1.13 mg/L. In the case of the Toxkit microbiotests' application, the following results were recorded: Spirodela polyrhiza EC50 = 2.204 mg/L (after 72 h exposure), Thamnocephalus platyurus EC50 = 0.115 mg/L (after 24 h), and Daphnia magna EC50 = 2.37 mg/L (after 24-48 h). With the exception of bacteria and yeast, the same response pattern was observed for all non-target species tested; the response range expressed by concentrations causing growth inhibition or mortality was small, ranging between very close values that are quite low, thereby demonstrating the high toxicity of Tebuconazole-based fungicides to the environment.

Phospholipid-Mimetic Aggregation-Induced Emission Luminogens for Specific Elimination of Gram-Positive and Gram-Negative Bacteria.

Precise elimination of both Gram-positive and Gram-negative bacteria greatly contributes to the fight against bacterial infection but remains challenging. Herein, we present a series of phospholipid-mimetic aggregation-induced emission luminogens (AIEgens) that selectively kill bacteria by capitalizing on both the different structure of two bacterial membrane and the regulated length of substituted alkyl chains of AIEgens. Because of the positive charges that they contain, these AIEgens are able to kill bacteria by anchoring onto the bacterial membrane. For AIEgens with short alkyl chains, they could combine with the membrane of Gram-positive bacteria other than Gram-negative bacteria, because of their complicated outer layers, thus exhibiting selective ablation to Gram-positive bacteria. On the other hand, AIEgens with long alkyl chains have strong hydrophobicity with bacterial membranes, as well as large sizes. This inhibits the combination with Gram-positive bacterial membrane but destroys the membranes of Gram-negative bacteria, resulting in selective ablation to Gram-negative bacteria. Moreover, the combined processes to two bacteria are clearly observed by fluorescent imaging, and in vitro and in vivo experiments show the extraordinary antibacterial selectivity toward a Gram-positive and Gram-negative bacterium. This work could facilitate the development of species-specific antibacterial agents.

Pyrrolidine derivatives as antibacterial agents, current status and future prospects: a patent review.

Bacterial infections are increasingly epitomizing major global health concerns, with rising death rates. Since the most complete assessment of the worldwide impact of antimicrobial resistance to date, with over 1.2 million people dead in 2019 as a direct result of antibiotic-resistant bacterial infections. The majority of antimicrobial drugs have been associated with a multitude of adverse effects including financial costs as well. Pyrrolidine derivatives have sparked the interest of researchers to create novel synthetic molecules with minimal side effect and drawbacks. To close the research gap, the current review discusses the synthetic compounds with active pyrrolidine scaffolds, critical findings and most crucially the structure-activity relationship that affects the activity of the ring over the last one and half decade.

Stibonic acids and related stibonate-phosphonate clusters: Synthesis, characterization and bioactivity evaluation.

The reaction of 4-(azobenzene)phenylstibonic acid with t-butylphosphonic acid led to the isolation of the tetranuclear oxo-hydroxo antimony cluster of formulae [(4-azobenzene-C6H4Sb)4(OH)4(tBuPO3)6] (C1). The reaction of (p-t-butyl phenyl stibonic acid with phenyl phosphonic acid resulted in the isolation of complex with formulae [(p-t-BuC6H4Sb)4(O)2(PhPO3)4(PhPO3H)4] (C2). Based upon the initial results from docking studies, parent stibonic acids, t-butyl-phenylstibonic acid, p-isopropylphenylstibonic acid, 4-azobenzenephenylstibonic acid, and the derived tetranuclear organoantimonate-phosphonate clusters were screened against different cancer cell lines, various Gram-positive and Gram-Negative bacteria and mycobacteria for possible bioactivity profile.

On Gram-Positive- and Gram-Negative-Bacteria-Associated Canine and Feline Skin Infections: A 4-Year Retrospective Study of the University Veterinary Microbiology Diagnostic Laboratory of Naples, Italy.

A 4-year retrospective study (2016-2019) of selected routine bacteriological examinations of the veterinary microbiology laboratory of the University Veterinary Teaching Hospital of Naples (Italy) was carried out. A total of 189 bacteriological samples were collected from 171 dogs and 18 cats suffering from skin infections. In dogs, the most common cutaneous infection was otitis externa, while pyoderma was found to be prevalent in cats. The number of recorded Gram-positive strains over the study period did not vary considerably from year to year and was always significantly higher (p-value = 0.0007) in comparison with Gram-negative bacterial isolations. In dogs, Staphylococcus pseudintermedius was the most common identified Gram-positive bacterium (65%), while Pseudomonas aeruginosa (36%) was the one among the isolated Gram-negative bacteria. In cats, coagulase-negative staphylococci were the most predominant isolated bacteria (47%). The phenotypic profiles of antibiotic resistance showed that most of the strains were resistant to amoxicillin-clavulanate, penicillin, clindamycin, and trimethoprim-sulfamethoxazole. Several multi-drug-resistant strains (35%) were detected in canine isolates. An updating of antibiotic resistance profiles of the main Gram-positive and Gram-negative bacteria principally associated with skin infections of pet animals is necessary to improve stewardship programs of veterinary hospitals and clinics.

Broad-Spectrum Bactericidal Activity of a Synthetic Random Copolymer Based on 2-Methoxy-6-(4-Vinylbenzyloxy)-Benzylammonium Hydrochloride.

Low-molecular-weight organic ammonium salts exert excellent antimicrobial effects by interacting lethally with bacterial membranes. Unfortunately, short-term functionality and high toxicity limit their clinical application. On the contrary, the equivalent macromolecular ammonium salts, derived from the polymerization of monomeric ammonium salts, have demonstrated improved antibacterial potency, a lower tendency to develop resistance, higher stability, long-term activity, and reduced toxicity. A water-soluble non-quaternary copolymeric ammonium salt (P7) was herein synthetized by copolymerizing 2-methoxy-6-(4-vinylbenzyloxy)-benzylammonium hydrochloride monomer with N, N-di-methyl-acrylamide. The antibacterial activity of P7 was assessed against several multidrug-resistant (MDR) clinical isolates of both Gram-positive and Gram-negative species. Except for colistin-resistant Pseudomonas aeruginosa, most isolates were susceptible to P7, also including some Gram-negative bacteria with a modified charge in the external membrane. P7 showed remarkable antibacterial activity against isolates of Enterococcus, Staphylococcus, Acinetobacter, and Pseudomonas, and on different strains of Escherichia coli and Stenotrophomonas maltophylia, regardless of their antibiotic resistance. The lowest minimal inhibitory concentrations (MICs) observed were 0.6-1.2 µM and the minimal bactericidal concentrations (MBC) were frequently overlapping with the MICs. In 24-h time-kill and turbidimetric studies, P7 displayed a rapid non-lytic bactericidal activity. P7 could therefore represent a novel and potent tool capable of counteracting infections sustained by several bacteria that are resistant to the presently available antibiotics.

Diversity of microbiota in Slovak summer ewes' cheese "Bryndza".

"Bryndza" cheese is an important Slovak traditional regional product. New knowledge on the role of microorganisms involved the "Bryndza" ripening process may provide valuable data on its quality and safety. In our study, the "Bryndza" made from pasteurized ewes milk was studied towards total count of bacteria, coliforms bacteria, enterococci, lactic acid bacteria, and microscopic filamentous fungi. All those groups of microbiota were detected using classical microbiological methods and identified using mass spectrometry. A total of 3,758 isolates were identified with score higher than 2.00. Altogether, 13 families, 24 genus, and 44 species of microbiota were identified in Slovak cheese "Bryndza." The most often isolated species were yeasts Yarrowia lipolitica and Dipodascus geotrichum and the lactic acid bacteria Lactobacillus paracasei subsp. paracasei.

Electrochemical cell lysis of gram-positive and gram-negative bacteria: DNA extraction from environmental water samples.

Cell lysis is an essential step for the nucleic acid-based surveillance of bacteriological water quality. Recently, electrochemical cell lysis (ECL), which is based on the local generation of hydroxide at a cathode surface, has been reported to be a rapid and reagent-free method for cell lysis. Herein, we describe the development of a milliliter-output ECL device and its performance characterization with respect to the DNA extraction efficiency for gram-negative bacteria (Escherichia coli and Salmonella Typhi) and gram-positive bacteria (Enterococcus durans and Bacillus subtilis). Both gram-negative and gram-positive bacteria were successfully lysed within a short but optimal duration of 1 min at a low voltage of ∼5 V. The ECL method described herein, is demonstrated to be applicable to various environmental water sample types, including pond water, treated wastewater, and untreated wastewater with DNA extraction efficiencies similar to a commercial DNA extraction kit. The ECL system outperformed homogeneous chemical lysis in terms of reaction times and DNA extraction efficiencies, due in part to the high pH generated at the cathode surface, which was predicted by simulations of the hydroxide transport in the cathodic chamber. Our work indicates that the ECL method for DNA extraction is rapid, simplified and low-cost with no need for complex instrumentation. It has demonstrable potential as a prelude to PCR analyses of waterborne bacteria in the field, especially for the gram-negative ones.

Natural drug physcion encapsulated zeolitic imidazolate framework, and their application as antimicrobial agent.

In this study, we have encapsulated natural physcion (PHY) drug loading into metal-organic frameworks MOFs, zeolitic imidazolate frameworks (ZIFs) through straight-forward nano-precipitation technique. The synthesized PHY@ZIF-8 indicated high drug loading encapsulation efficiency i.e. 88%, whereas, drug loading capacity was found to be 11.49%. The characterization of PHY loaded-ZIF 8 (PHY@ZIF-8) was carried out by powder x-ray diffraction (PXRD), transmission electron microscopy (TEM), scanning electron microscopy (SEM), thermal gravimetric analysis (TGA), and FT-IR methods. The release of PHY loaded in ZIF-8 was 88.72% at pH 5.0 which is approximately three time higher than its release in physiological system with pH 7.4 (27.61%). The remarkable stability of PHY@ZIF-8 NPs even after 25 days stem it as an effective and stable candidate. Furthermore, the antibacterial activity of pure PHY, ZIF-8 and PHY@ZIF-8 were investigated against gram negative strains and gram positive strain. The PHY@ZIF-8 showed maximum growth inhibition zones against all microorganism as compare to pure PHY. We hope that this model drug could have the potential ability for treatment of various infectious diseases.

New and Potent Quinuclidine-Based Antimicrobial Agents.

Developing new antibiotics is currently very important since antibiotic resistance is one of the biggest problems of global health today. In the search for a new class of potential antimicrobial agents, ten new compounds were designed and synthesized based on the quinuclidinium heterocyclic core and the oxime functional group. The antimicrobial activity was assessed against a panel of representative gram-positive and gram-negative bacteria. All compounds demonstrated potent activity against the tested microorganisms, with the minimum inhibitory concentration (MIC) values ranging from 0.25 to 256.00 µg/mL. Among the tested compounds, two quaternary compounds, para-N-chlorobenzyl and meta-N-bromobenzyl quinuclidinium oximes, displayed the most potent and broad-spectrum activity against both gram-positive and gram-negative bacterial strains (MIC values from 0.25 to 4.00 µg/mL), with the lowest value for the important multidrug resistant gram-negative pathogen Pseudomonas aeruginosa. In the case of Klebsiella pneumoniae, activity of those compounds are 256-fold and 16-fold better than gentamicin, respectively. MTT assays showed that compounds are nontoxic for human cell lines. Multi-way analysis was used to separately reduce dimensionality of quantum chemical data and biological activity data to obtain a regression model and the required parameters for the enhancement of biological activity.