Antibacterial activity of exopolysaccharide produced by bee gut-resident Enterococcus sp. BE11 against marine fish pathogens.

BACKGROUND: In recent years, the demand for innovative antimicrobial agents has grown, considering the growing problem of antibiotic resistance in aquaculture. Adult Apis mellifera honeybees' gut represents an outstanding habitat to isolate novel lactic acid bacteria (LAB) able to produce prominent antimicrobial agents. METHODS: In the current study, twelve LAB were isolated and purified from the gut of adult Apis mellifera. The isolates were screened for exopolysaccharide (EPS) production. The most promising isolate BE11 was identified biochemically and molecularly using 16 S rRNA gene sequence analysis as Enterococcus sp. BE11 was used for the mass production of EPS. The partially purified BE11-EPS features were disclosed by its physicochemical characterization. Moreover, the antimicrobial activity of BE11 cell free supernatant (CFS) and its EPS was investigated against some fish pathogens namely, Pseudomonas fluorescens, Streptococcus agalactiae, Aeromonas hydrophila, Vibrio sp. and Staphylococcus epidermidis using well-cut diffusion method. RESULTS: The physicochemical characterization of BE11-EPS revealed that the total carbohydrate content was estimated to be ~ 87%. FTIR and NMR analysis ascertained the presence of galactose and glucose residues in the EPS backbone. Moreover, the GC-MS analysis verified the heterogeneous nature of the produced BE11-EPS made up of different monosaccharide moieties: galactose, rhamnose, glucose, arabinose sugar derivatives, and glucuronic acid. BE11 CFS and its EPS showed promising antimicrobial activity against tested pathogens as the inhibition zone diameters (cm) ranged from 1.3 to 1.7 and 1.2-1.8, respectively. CONCLUSION: The bee gut-resident Enterococcus sp. BE11, CFS, and EPS were found to be promising antimicrobial agents against fish pathogens and biofilm producers affecting aquaculture. To the best of our knowledge, this is the first study to purify and make a chemical profile of an EPS produced by a member of the bee gut microbiota as a potential inhibitor for fish pathogens.

Lipid-membranes interaction, structural assessment, and sustainable production of polyhydroxyalkanoate by Priestia filamentosa AZU-A6 from sugarcane molasses.

This study presented for the first time the PHA-lipid interactions by circular dichroism (CD) spectroscopy, besides a sustainable PHA production strategy using a cost-effective microbial isolate. About 48 bacterial isolates were selected from multifarious Egyptian sites and screened for PHAs production. The Fe(AZU-A6) was the most potent isolate, and identified genetically as Priestia filamentosa AZU-A6, while the intracellular PHA granules were visualized by TEM. Sugarcane molasses (SCM) was used an inexpensive carbon source and the production conditions were optimized through a Factor-By-Factor strategy and a Plackett-Burman statistical model. The highest production (6.84 g L-1) was achieved at 8.0 % SCM, pH 8.0, 35 °C, 250 rpm, and 0.5 g L-1 ammonium chloride after 72 h. The complementary physicochemical techniques (e.g., FTIR, NMR, GC-MS, DSC, and TGA) have ascertained the structural identity as poly-3-hydroxybutyrate (P3HB) with a characteristic melting temperature of 174.5 °C. The circular dichroism analysis investigated the existence of interactions between the PHB and the different lipids, particularly 1,2-dimyristoyl-sn-glycero-3-phosphocholine. The ATR technique for the lipid-PHB films suggested that both the hydrophobic and electrostatic forces control the lipid-PHB interactions that might induce changes in the structuration of PHB.

Complementary spectroscopy studies and potential activities of levan-type fructan produced by Bacillus paralicheniformis ND2.

This study aimed at the production of marine bacterial exopolysaccharides (EPS) as biodegradable and nontoxic biopolymers, competing the synthetic derivatives, with detailed structural and conformational analyses using spectroscopy techniques. Twelve marine bacterial bacilli were isolated from the seawater of Mediterranean Sea, Egypt, then screened for EPS production. The most potent isolate was identified genetically as Bacillus paralicheniformis ND2 by16S rRNA gene sequence of ~99 % similarity. Plackett-Burman (PB) design identified the optimization conditions of EPS production, which yielded the maximum EPS (14.57 g L-1) with 1.26-fold increase when compared to the basal conditions. Two purified EPSs namely NRF1 and NRF2 with average molecular weights (Mw¯) of 15.98 and 9.70 kDa, respectively, were obtained and subjected for subsequent analyses. FTIR and UV-Vis reflected their purity and high carbohydrate contents while EDX emphasized their neutral type. NMR identified the EPSs as levan-type fructan composed of ß-(2-6)-glycosidic linkage as a main backbone, and HPLC explained that the EPSs composed of fructose. Circular dichroism (CD) suggested that NRF1 and NRF2 had identical structuration with a little variation from the EPS-NR. The EPS-NR showed antibacterial activity with the maximum inhibition against S. aureus ATCC 25923. Furthermore, all the EPSs revealed a proinflammatory action through dose-dependent increment of expression of proinflammatory cytokine mRNAs, IL-6, IL-1ß and TNFα.

A lysine-based 2:1-[α/aza]-pseudopeptide series used as additives in polymeric membranes for CO2 capture: synthesis, structural studies, and application.

The current study presents for the first time the synthesis of a new 2:1-[α/aza]-pseudopeptide series possessing charged amino acids (i.e., lysine) and aims at studying the influences of chirality, backbone length, and the nature of the lysine side chains on the conformation of the 2:1-[α/aza]-oligomers in solution using NMR, FTIR spectroscopy and molecular dynamic calculations. The spectroscopic results emphasized the conservation of the ß-turn conformation adopted by the trimers regardless of the chirality which demonstrated a noticeable effect on the conformation of homochiral hexamer (8c) compared with the hetero-analogue (8d). The molecular dynamic calculations predicted that the chirality and the side chain of the lysine residues caused a little distortion from the classical ß-turn conformation in the case of short trimer sequences (7c and 7d), while the chirality and the backbone length exerted more distortion on the ß-turn adopted by the longer hexamer sequences (8c and 8d). The large disturbance in hexamers from classical ß-turn was attributed to increasing the flexibility and the possibility of molecules to adopt a more energetically favorable conformation stabilized by non-classical ß-turn intramolecular hydrogen bonds. Thus, alternating d- and l-lysine amino acids in the 2:1-[α/aza]-hexamer (8d) decreases the high steric hindrance between the lysine side chains, as in the homo analogue (8c), and the distortion is less recognized. Finally, short sequences of aza-pseudopeptides containing lysine residues improve CO2 separation when used as additives in Pebax® 1074 membranes. The best membrane performances were obtained with a pseudopeptidic dimer as an additive (6b'; deprotected lysine side chain), with an increase in both ideal selectivity α CO2/N2 (from 42.8 to 47.6) and CO2 permeability (from 132 to 148 Barrer) compared to the virgin Pebax® 1074 membrane.

Spatial distribution, sources and risk assessment of polycyclic aromatic hydrocarbons in the surficial sediments of the Egyptian Mediterranean coast.

The Egyptian Mediterranean coast (EMC) receives a considerable quantity of polycyclic aromatic hydrocarbons (PAHs). PAHs from EMC sediments were assessed to understand the effects of marine and riverine currents on their distribution. The concentrations of total PAHs ranged between 13,156-34,852 ng/g dw. PAH levels have increased even in areas far from the shoreline under the influence of riverine inputs from the Nile River; this is attributed to the tidally induced riverine freshwater re-suspension of surface sediments in the shallow near-shore section and re-precipitation in the fare stations. PAH levels generally increase as one moves from the western to the eastern part of the studied area, owing to the effect of the marine current. Diagnostic ratios pointed toward different pyrogenic sources. SQGs were used to assess the probability of observing adverse biological effects in benthic organisms in sediment samples. The toxic and mutagenic equivalent quotient for carcinogenic PAHs was extremely high.

Prevalence and risk assessment of microplastics in the Nile Delta estuaries: "The Plastic Nile" revisited.

Recent research is directed toward studying plastic pollution in rivers, and estuaries due to the importance of freshwater bodies in all aspects of life. The river deltas and estuaries are interesting for studying the flux of plastics into the oceans. The Nile River has been identified as a hot spot of plastic litter flux in the eastern Mediterranean basin. In addition, it was nicknamed "Plastic Nile", yet this major river is largely unexplored with a lack of field measurements and adequate surveys. The current study was based on bridging this scientific gap. Three trips were conducted, covering 30 km in the Rosetta branch and 23 km in the Damietta branch, during the high water level in summer 2021, and 10 km off the inlet of Lake Burullus, in spring 2021. Microplastics in surface water ranged from 761 ± 319 to 1718 ± 1008 MPs/m3, and from 167 ± 137 to 1630 ± 1303 MPs/kg of dry sediments. Land use/ land cover mapping using Sentinel-2 images showed several sources of pollution that contribute to plastic contamination in the study area. Thermal analysis indicated seven plastic polymers; including, PE, PP, PET, PEVA, and PTFE, using discarded plastic products as reference materials. Microplastics were composed of colored and glossy fragments of sizes <500 µm, originating from land-based sources. Pollution load, polymer risk assessment, and ecological risk indices were calculated. Based on field observations macro-plastics were retained within the extensive network of infrastructure and dam systems. 80-106 billion MPs/year were estimated to flux from the Nile estuaries into the Mediterranean Sea. The current situation urges the development of binding plans to reduce plastic waste in the Nile Delta, as well as setting environmental monitoring points along the Deltaic coast.

Microplastics contamination in commercial fish from Alexandria City, the Mediterranean Coast of Egypt.

Plastic waste is a major threat to various marine ecosystems. Due to being semi-enclosed basin with dense population, the Mediterranean Sea has been considered as a hot spot for plastic pollution. Alexandria was listed as one of the major cities contributing to plastic waste in the eastern Mediterranean basin. Accordingly, microplastics (MPs) abundance and composition were investigated in the digestive tracts of commercial fish species from two major fishing areas in Alexandria; Abu-Qir Bay and Eastern Harbor, which are affected by plastic pollution. The incidence and average densities of MPs were 91.8 ± 8.4% and 11.7 ± 9.5 items fish-1, similar to highly polluted regions in the southeastern Mediterranean Sea. The average MPs concentration was significantly higher in Sparus aurata (38.3 ± 28.4 items fish-1) than all species, except for Siganus rivulatus and Boops boops. Polyethylene and poly ethylene-vinyl acetate were the dominant polymers in the fish digestive tracts. Four types of plastic polymers were recorded in Siganus rivulatus and five in Parupeneus macronemus. The dominance of glossy fragments (sizes <500 µm) in Abu-Qir Bay indicated land-based source of pollution from domestic, agricultural, and industrial wastes. The dominance of larger plastic filaments and colored fragments in the Eastern Harbor suggested secondary MPs, originating from the fragmentation of larger plastic items of sea-based sources, such as fishing and recreational activities.

Horizontal and vertical segregation of polycyclic aromatic hydrocarbons in the Egyptian Mediterranean coast.

Egyptian Mediterranean coast receives significant amounts of polycyclic aromatic hydrocarbons (PAHs) from industrial exhausts, riverine inputs, maritime shipping and fishers, and oil and natural gas production and exploration. The present study considers the first exhaustive assessment for the dissolved PAHs along the Egyptian Mediterranean coast (Alexandria to Manzallah) to monitor their spatial distribution and investigate the effect of the marine currents and the role of microorganisms in their distribution. Surface water levels ranged between 124.97 and 301.02 ng L-1 with an average 223.68 ± 41.11 ng L-1. The distribution increases from west to east based on the water circulation in the Mediterranean Sea. The levels in near shore stations were lower than those of middle and onshore stations. The intensive existence of micro-organisms near shore stations consumes great part of PAHs, while this bio-remediation process decreases gradually away from the shoreline leaving relative high concentrations of dissolved PAHs in the middle and onshore stations. Middle and deep-water levels ranged between 312.75 and 1042.95 ng L-1 with an average 633.47 ± 225.53 ng L-1. Deeper waters showed higher PAHs concentrations where the average concentrations of 50 m stations (868.12 ± 138.35 ng L-1) ˃ 30 m stations (629.49 ± 143.85 ng L-1) ˃ 10 m stations (402.79 ± 59.46 ng L-1). The wind-induced waves re-suspend rich PAHs sediment particles to increase its concentration in the water column. Carcinogenic toxic equivalent quotient (TEQ) for total detected PAHs in the middle and deep water represented more than double (75.46 ng TEQ L-1) the value in the surface water (34.76 ng TEQ L-1). The diagnostic ratios and principal component analysis indicated mainly pyrogenic origin in surface, middle, and deep waters.

Production and Characterization of Exopolysaccharide From Newly Isolated Marine Probiotic Lactiplantibacillus plantarum EI6 With in vitro Wound Healing Activity.

Because of its safety, biological activities, and unique properties, exopolysaccharide (EPS) from lactic acid bacteria (LAB) has been developed as a potential biopolymer. A few studies have investigated the EPS produced by marine LAB. This study reports the wound healing activity of an EPS produced by a marine isolate identified as Lactiplantibacillus plantarum EI6, in addition to assessing L. plantarum EI6's probiotic properties. EI6 demonstrated promising antimicrobial activity against different pathogenic bacteria, as well as the ability to withstand stomach pH 3, tolerate 0.3% bile salt concentration, and exhibit no signs of hemolysis. Furthermore, EI6 was able to produce 270 mg/L of EPS upon growth for 48 h at 37°C in an MRS medium enriched with 1.0% of sucrose. The chemical features of the novel EI6-EPS were investigated: the UV-vis estimated a high carbohydrate content of ~91.5%, and the FTIR emphasized its polysaccharide nature by the characteristic hydroxyl, amide I, II, & III, and glycosidic linkage regions. The GC-MS and NMR analyses revealed the existence of five monosaccharides, namely, rhamnose, galactose, mannose, glucose, and arabinose, existing mainly in the pyranose form and linked together by α- and ß-glycosidic linkages. EI6-EPS was found to be safe (IC50 > 100 µg/ml) and induced human skin fibroblasts (HSF) proliferation and migration. These findings imply that EI6 can be used as a safe source of bioactive polymer in wound care.

Characterization and distribution of plastic particles along Alexandria beaches, Mediterranean Coast of Egypt, using microscopy and thermal analysis techniques.

Microplastics (MPs) contamination has become a global concern with potential impacts on the marine environment. Alexandria is the second-largest city in Egypt and a significant contributor of plastic litter inputs into the Eastern Mediterranean Sea. The current study provides an in-depth analysis of the plastic particles accumulated along Alexandria beaches. Types, composition, and potential sources of MPs were investigated using microscopy and thermal analysis. A mean value of 389.1 ± 285.9 items kg-1 dry weight was detected in the shore sediments similar to other records from the Eastern Mediterranean region. An average of 457.4 ± 281.8 items m-3 was recorded in the surface water, which was the highest recorded MPs density in onshore waters of the Mediterranean region. Thermogravimetric analysis (TGA) showed that plastics made up 0.5% - 72% of the materials extracted from the sediment samples, and 0.58% - 20.6% from the water samples. Differential scanning calorimetry (DSC) identified ten semi-crystalline polymers. Low-density polyethylene (LDPE) and polyethylene vinyl acetate (PEVA) were the common polymers. The single-use plastic bags and detergents were the land-based sources of marine plastic litter. The sea-based sources included antifouling paints, maintenance of ships, and abandoned fishing gears. Proper management plans of domestic waste input, polluter-pay strategy, and education programs aiming at the Fishermen and how plastic pollution would impact their livelihood are urgently needed.

Considerable Production of Ulvan from Ulva lactuca with Special Emphasis on Its Antimicrobial and Anti-fouling Properties.

In the current study, a significant amount of ulvan was extracted from Ulva lactuca collected from Alexandria coastline, Egypt, using a simple extraction method. According to the chemical analysis, the obtained polysaccharide content is estimated to be 36.50 g/100 g with a high sulfate content of 19.72%. Physio-chemically, the FTIR analysis confirmed the presence of sulfated groups attached to the carbohydrate backbone. The GC-MS results revealed the presence of various monosaccharides with relative abundances in the order: fucopyranose (22.09%) > L-rhamnose (18.17%) > L-fucose (17.46%) > rhamnopyranose (14.29%) > mannopyranose (8.59%) > α-D-glactopyranose (7.64%) > galactopyranose (6.14%) > ß-arabinopyranose (5.62%). In addition, the SEM-EDX depicted an amorphous architecture with a majority wt% for the elements of C, O, and S. The partially purified ulvan demonstrated potent antimicrobial activity against some fish and human pathogenic microbes. The inhibition zone diameter ranged from 11 to 18 mm. On the other hand, the prepared ulvan-chitosan hydrogel significantly improved the antimicrobial activity as the inhibition zone diameter ranged from 12 to 20. Moreover, when compared to the controls, the extracted ulvan demonstrated anti-fouling properties and successfully disrupted the biofilm formed on a glass slide submerged in seawater.

Structural assessment of the bioplastic (poly-3-hydroxybutyrate) produced by Bacillus flexus Azu-A2 through cheese whey valorization.

The demand for the production of biodegradable plastics has significantly increased. Bioplastics have become an essential alternative to the threats of the daily consumable plastics, sourced from fossil fuels, to the environment. Polyhydroxyalkonates (PHAs) are a ubiquitous group of bioderived and biodegradable plastics, however their production is limited by the costs associated mainly with the carbon sources. Herein, this study aims to reduce the PHAs production cost by using a by-product from the dairy industry, i.e., cheese whey (CW), as a sole carbon source. The developed process recruits an aquatic isolate, Bacillus flexus Azu-A2, and is optimized via studying various parameters using the shaking flasks technique. The results showed that the maximum PHA production (0.95 g L-1) and PHA content (20.96%, w/w), were obtained after incubation period 72 h at 45 °C, 100 rpm agitation rate, 50% CWS concentration, pH 8.5, and 1.0 g L-1 ammonium chloride. Physiochemically, Fourier transform infrared spectroscopy (FTIR), gas chromatography-mass spectroscopy (GC-MS), nuclear magnetic resonance (NMR), and energy-dispersive X-ray (EDX) techniques, emphasized the type of the extracted PHA as polyhydroxybutyrate (PHB). The thermal properties of PHB were measured using differential scanning calorimetry (DSC), recording melting transition temperature (Tm) at 170.96 °C. Furthermore, a scanning electron microscope (SEM) visualized a homogenous microporous structure for the thin PHB biofilm. In essence, this study highlights the ability of Bacillus flexus Azu-A2 to produce a good yield of highly purified PHB at reduced production cost from dairy CW. Consequently, the current study paves the way for an improved whey management strategy.

Synthesis of chitosan-acrylic acid/multiwalled carbon nanotubes composite for theranostic 47Sc separation from neutron irradiated titanium target.

A simple and efficient separation method of carrier-free 47Sc from neutron irradiated titanium target using a novel chitosan-acrylic acid/multiwalled carbon nanotubes (CS-AA/MWCNTs) composite was established. The synthesis of the CS-AA/MWCNTs composite was achieved using gamma radiation-induced template polymerization. The grafting efficiency (GE%) of AA on CS onto the surface of f-MWCNTs reached a maximum of~84% under the optimized conditions (30 wt% CS, 1.0 wt% AA, 0.15 wt% f-MWCNTs, >0.2 wt% N,N'-Methylenebisacrylamide (NMBA), and irradiation dose ~25 kGy). Different analyses (FT-IR, SEM, TGA and DTA) were examined for confirming the structural morphology and mechanical properties of the new synthesized composite. Interestingly, the CS-AA/MWCNTs composite depicted a selective adsorption of Sc(III) rather than Ti(IV) ions at pH 5 with adsorption efficiency of ~93.93%. The ionic exchange separation of no-carrier-added (NCA)47Sc(III) from irradiated TiO2 target on CS-AA/MWCNTs composite packed column efficiently eluted 47Sc(III) by 91 ± 0.8% using 1 M HCl solution. The quality control tests (radionuclidic, radiochemical, and chemical purities) for the eluted 47Sc(III) clarified its high purity and validity for cancer theranostics.

Synthesis of Thermoresponsive Copolymers with Tunable UCST-Type Phase Transition Using Aqueous Photo-RAFT Polymerization.

Currently, the phase transition of aqueous binary systems containing thermoresponsive (co)polymers, and exhibiting upper critical solution temperature (UCST), is exclusively investigated in dilute solutions, which can limit the knowledge of their UCST-type phase transition. Herein, a photo-RAFT polymerization approach, using acrylamide (AAm) and acrylonitrile (AN) as monomer models, is used to prepare well-controlled poly(AAm-co-AN) copolymers "in situ" in highly concentrated dispersions (60 wt%). The impact of the copolymer concentration and the chemical composition (as a variation of AN fraction in the copolymers) on the cloud point temperature (TCP ) are investigated using turbidity measurements. Importantly, the results show that upon increasing the polymer concentration, a sharp increase of TCP up to a maximum point is observed, representing the UCST, before the decrease of TCP at higher polymer concentrations. Finally, a model equation is developed to fit the UCST values of poly(AAm-co-AN), which can be useful to design new poly(AAm-co-AN) copolymers with a desired UCST for a specific application.

Cyclohexamer [-(d-Phe-azaPhe-Ala)2-]: good candidate to formulate supramolecular organogels.

Molecular self-assembly is a fascinating process which has become an area of great interest in supramolecular chemistry, as it leads in certain cases to molecular gels. Organogels formulated from low molecular weight compounds (LMWOGs) have attracted much interest in the past decades due to their applications as new soft materials. Herein, we report on the ability of the cyclic pseudopeptide cyclo-[-(d-Phe-azaPhe-Ala)2-] (2) to self-assemble in some aromatic solvents and to form organogels driven by non-covalent forces, mainly hydrogen bonding and π-stacking interactions. Comprehensive FTIR and NMR studies emphasized that this cyclic aza-peptide adopts a ß-turn conformation at low concentration in toluene, while an equilibrium between the monomeric states (intramolecular forces) and the supramolecular structures (intra- and intermolecular forces) is established at high concentration (gel state). Rheological investigations of the organogels highlight the dependence of their stiffness (up to ∼4 kPa) and sol/gel transition temperatures (up to 100 °C) as a function of the solvent and concentration of gelator used. The formulation of fibrous structures confirmed the phenomenon of self-assembly. Finally, we found that cyclo-[-(d-Phe-azaPhe-Ala)2-] is an effective organogelator for application in phase selective gelation (PSG) of organic solvents from aqueous/organic mixtures with recovery percents up to 96%.