Seaweed Extracts as an Effective Gateway in the Search for Novel Antibiofilm Agents against Staphylococcus aureus.

Treatment of biofilm-associated infections has become a major challenge in biomedical and clinical fields due to the failure of conventional treatments in controlling this highly complex and tolerant structure. Therefore, the search for novel antibiofilm agents with increased efficacy as those provided by natural products, presents an urgent need. The aim of this study was to explore extracts derived from three algae (green Ulva lactuca, brown Stypocaulon scoparium, red Pterocladiella capillacea) for their potential antibiofilm activity against Staphylococcus aureus, bacterium responsible for several acute and chronic infections. Seaweed extracts were prepared by successive maceration in various solvents (cyclohexane (CH), dichloromethane (DCM), ethyl acetate (EA), and methanol (MeOH)). The ability of the different extracts to inhibit S. aureus biofilm formation was assessed using colony-forming unit (CFU) counts method supported by epifluorescence microscopic analysis. Effects of active extracts on the biofilm growth cycle, as well as on S. aureus surface hydrophobicity were evaluated. Results revealed the ability of four extracts to significantly inhibit S. aureus biofilm formation. These findings were supported by microscopy analyses. The gradual increase in the number of adherent bacteria when the selected extracts were added at various times (t0, t2h, t4h, t6h, and t24h) revealed their potential effect on the initial adhesion and proliferation stages of S. aureus biofilm development. Interestingly, a significant reduction in the surface hydrophobicity of S. aureus treated with dichloromethane (DCM) extract derived from U. lactuca was demonstrated. These findings present new insights into the exploration of seaweeds as a valuable source of antibiofilm agents with preventive effect by inhibiting and/or delaying biofilm formation.

Seaweed Extracts: A Promising Source of Antibiofilm Agents with Distinct Mechanisms of Action against Pseudomonas aeruginosa.

The organization of bacteria in biofilms is one of the adaptive resistance mechanisms providing increased protection against conventional treatments. Thus, the search for new antibiofilm agents for medical purposes, especially of natural origin, is currently the object of much attention. The objective of the study presented here was to explore the potential of extracts derived from three seaweeds: the green Ulva lactuca, the brown Stypocaulon scoparium, and the red Pterocladiella capillacea, in terms of their antibiofilm activity against P. aeruginosa. After preparation of extracts by successive maceration in various solvents, their antibiofilm activity was evaluated on biofilm formation and on mature biofilms. Their inhibition and eradication abilities were determined using two complementary methods: crystal violet staining and quantification of adherent bacteria. The effect of active extracts on biofilm morphology was also investigated by epifluorescence microscopy. Results revealed a promising antibiofilm activity of two extracts (cyclohexane and ethyl acetate) derived from the green alga by exhibiting a distinct mechanism of action, which was supported by microscopic analyses. The ethyl acetate extract was further explored for its interaction with tobramycin and colistin. Interestingly, this extract showed a promising synergistic effect with tobramycin. First analyses of the chemical composition of extracts by GC-MS allowed for the identification of several molecules. Their implication in the interesting antibiofilm activity is discussed. These findings suggest the ability of the green alga U. lactuca to offer a promising source of bioactive candidates that could have both a preventive and a curative effect in the treatment of biofilms.

A multiresidue method for the analysis of pesticides, polycyclic aromatic hydrocarbons, and polychlorinated biphenyls in snails used as environmental biomonitors.

This paper reports an optimized multiresidue extraction strategy based on the Quick, Easy, Cheap, Effective, Rugged, and Safe (QuEChERS) extraction procedure and on solid-phase microextraction (SPME) for the simultaneous screening of 120 pesticides, 16 polycyclic aromatic hydrocarbons, and 22 polychlorinated biphenyls from the terrestrial snail Helix aspersa. The optimized extraction method was based on QuEChERS using acetonitrile, followed by dispersive-Solid-phase extraction clean-up using primary secondary amine and octadecyl (C18) sorbents. The obtained extracts were analyzed by liquid chromatography coupled with tandem mass spectrometry and gas chromatography coupled with tandem mass spectrometry. This latest technique was preceded by a pre-concentration step using SPME with appropriate fibers. Afterwards, the method was validated for its linearity, sensitivity, recovery, and precision. Results showed high sensitivity, accuracy, and precision, with limits of detection and quantification lower than 20 ng g - 1 for most considered pollutants. Both inter and intra-day analyses revealed low relative standard deviation (%), which was lower than 20% for most targeted compounds. Moreover, the obtained regression coefficient (R2) was higher than 0.98 and the recoveries were higher than 60% for the majority of the assessed pollutants.

The use of vegetation, bees, and snails as important tools for the biomonitoring of atmospheric pollution-a review.

The continuous discharge of diverse chemical products in the environment is nowadays of great concern to the whole world as some of them persist in the environment leading to serious diseases. Several sampling techniques have been used for the characterization of this chemical pollution, although biomonitoring using natural samplers has recently become the technique of choice in this field due to its efficiency, specificity, and low cost. In fact, several living organisms known as biomonitors could accumulate the well-known persistent environmental pollutants allowing their monitoring in the environment. In this work, a review on environmental biomonitoring is presented. The main sampling techniques used for monitoring environmental pollutants are first reported, followed by an overview on well-known natural species used as passive samplers and known as biomonitors. These species include conifer needles, lichen, mosses, bees and their byproducts, and snails, and were widely used in recent research as reliable monitors for environmental pollution.

Determination of 16 PAHs and 22 PCBs in honey samples originated from different region of Lebanon and used as environmental biomonitors sentinel.

In order to assess the air quality in some Lebanese regions, a biomonitoring study based on honey as biomonitor candidate was conducted. For this, 18 samples of honey collected from four regions in Lebanon, were analyzed for their contamination by 16 polycyclic aromatic hydrocarbons (PAHs) and 22 polychlorinated biphenyls (PCBs). Samples were first extracted using a multi-residue extraction method based on the quick, easy, cheap, effective, rugged, and safe extraction method (QuEChERS) followed by a concentration step using Solid-phase microextraction (SPME) procedures. The extraction was then followed by chromatographic analysis by gas chromatography-ion-trap tandem mass spectrometry (GC-MS/MS). After PAHs samples assessment, different ratios of founded PAHs were calculated in order to estimate the sources of the pollution by these compounds. The obtained results showed that the four analyzed regions were contaminated with PAHs originated from both pyro and petro genic sources while none of them was found to be contaminated by any of the 22 assessed PCBs. The results of this study show that honey can be used as potential biomonitor candidate allowing the assessment of the pollution statement of a given environment.

A multiresidue method for the analysis of 90 pesticides, 16 PAHs, and 22 PCBs in honey using QuEChERS-SPME.

An optimized analytical method was developed for the simultaneous analysis of 90 pesticides, 16 polycyclic aromatic hydrocarbons, and 22 polychlorinated biphenyls. The method was based on quick, easy, cheap, effective, rugged, and safe (QuEChERS) extraction using acetonitrile followed by a dispersive solid-phase extraction cleanup using primary-secondary amine and octadecyl (C18). The extract obtained was concentrated by evaporation and then reconstituted with acetonitrile to prepare it for chromatographic analysis by liquid chromatography-triple-quadrupole tandem mass spectrometry and gas chromatography-ion-trap tandem mass spectrometry, which was preceded by a preconcentration step using solid-phase microextraction with appropriate fibers. The combination of the two extraction steps ensured efficient extract cleanup. The use of the two analytical instruments allowed the analysis of a large number of pollutants with a high reliability rate. The method developed was validated for linearity, which was studied with use of matrix-matched calibration curves in the concentration range between 10 and 3000 ng g-1. The correlation coefficient (R 2) obtained was higher than 0.98 for most of the target compounds, with a relative standard deviation lower than 20% for repeatability and reproducibility. The limits of detection and quantification were lower than 20 and 60 ng g-1 respectively for the compounds analyzed, and the recoveries were between 60% and 103% for most compounds. Finally, the method was tested for its efficiency on real samples by the analysis of three honey samples in which seven pesticides and nine polycyclic aromatic hydrocarbons were determined. Graphical Abstract ᅟ.

Analysis of Dithiocarbamate Fungicides in Vegetable Matrices Using HPLC-UV Followed by Atomic Absorption Spectrometry.

A simple method combining ion-pair methylation, high-performance liquid chromatography (HPLC) analysis with detection at 272 nm and atomic absorption spectrometry was developed in order to determine 10 dithiocarbamate fungicides (Dazomet, Metam-sodium, Ferbam, Ziram, Zineb, Maneb, Mancozeb, Metiram, Nabam and Propineb) and distinguish ethylenbisdithiocarbamates (EBDTCs) Zineb, Maneb and Mancozeb in diverse matrices. This method associates reverse phase analysis by HPLC analysis with detection at 272 nm, with atomic absorption spectrometry in order to distinguish, with the same extraction protocol, Maneb, Mancozeb and Zineb. The limits of detection (0.4, 0.8, 0.5, 1.25 and 1.97) and quantification (1.18, 2.5, 1.52, 4.2 and 6.52) calculated in injected nanogram, respectively, for Dazomet, Metam-Na, dimethyldithiocarbamates (DMDTCs), EBDTCs and propylenebisdithiocarbamates (PBDTCs) justify the sensitivity of the method used. The coefficients of determination R2 were 0.9985, 0.9978, 0.9949, 0.988 and 0.9794, respectively, for Dazomet, Metam-Na, DMDTCs, EBDTCs and PBDTCs, and the recovery from fortified apple and leek samples was above 90%. Results obtained with the atomic absorption method in comparison with spectrophotometric analysis focus on the importance of the atomic absorption as a complementary specific method for the distinction between different EBDTCs fungicides.

The use of conifer needles as biomonitor candidates for the study of temporal air pollution variation in the Strasbourg region.

The continuous emission of polluting chemicals into the atmosphere requires the implementation of monitoring of ambient air quality. The use of vegetation for environmental monitoring can be considered as a simple monitoring technique by providing a cheap and accessible matrix. In this study, needles of two conifers (Pinus nigra and Cedrus atlantica), were used for the consecutive biomonitoring of multipollutants such as pesticides, polychlorinated biphenyls, and polyaromatic hydrocarbons (PAHs) in an urban area in Strasbourg (France). The extraction was performed by accelerated solvent extraction, solid-phase extraction, and solid-phase microextraction and was followed by gas chromatography coupled to tandem mass spectrometry and liquid chromatography coupled to tandem mass spectrometry analyses. The results obtained for conifer samples collected in 5 successive weeks (April 09-May 07, 2015) show a similar variation of the different types of pollutants. A pollution peak was observed during the second week of analysis, and the concentration of all pollutants then decreased to complete disappearance at the end of the sampling period. PAHs were the most concentrated with a total concentration of about 35.87 ng g-1, and naphthalene was, among these pollutants, the most concentrated with a total concentration of about 15.1 ng g-1. The analysis of meteorological data during this period suggests that the results correlated with climatic conditions that widely vary during this period of the year. The results show that the concentration peak was obtained when no precipitation was detected.

Contribution to the food products' analysis: A research and evaluation on the hemolytic effect of some pesticides used in Lebanon.

Pesticides are a real concern for the society as their use has become critical, leading sometimes to their accumulation as residues in fruits and vegetables. After examining the pesticides sold in Northern Lebanon, this study is focused on the analysis and identification of pesticides residues in fruits and vegetables that are harvested in this region and treated with the locally sold pesticides. Results show: first, (i) a use of Zineb by the name of another pesticide Micronized Sulfur to avoid prosecution; (ii) a significant presence of Metalaxyl in lemons and oranges; (iii) a significant presence of Trifluralin in strawberries; and (iv) a significant presence of Zineb in lemons and tomatoes. Second, with the use of hemolytic tests on human blood results show: (i) a critical concentration and a significant hemolytic effect of some pesticides used in Lebanon; and (ii) an absence of hemolytic effect in the collected fractions of the different analyzed fruit extracts containing pesticides. Finally, this work is the first step for pesticides' analysis in vegetables and fruits in Lebanon, initiating a wider analytical study in order to control and examine the use of pesticides which, according to our results, could have an adverse effect on human health over a long term.