Determination of Multiclass Antibiotics in Fish Muscle Using a QuEChERS-UHPLC-MS/MS Method.

The surging global demand for fish has increased aquaculture practices, where antibiotics have become indispensable to prevent diseases. However, the passive incorporation of these compounds into the diet may have adverse effects on human health. In this work, the QuEChERS method combined with ultra-high-performance liquid chromatography tandem mass spectrometry was applied for the determination of 10 multiclass antibiotics (5 quinolones, 2 sulfonamides, 2 diaminopyrimidines, and 1 macrolide) in muscle tissue of farmed fish (European sea bass and gilt-head sea bream). The applied method demonstrated acceptable recovery values, mostly between 70 and 120%, with limits of quantification of the method meeting the established EU maximum residue limits. The analysis of twenty fish samples in duplicate revealed that most antibiotics were not present, with the only exception of oxolinic acid and tilmicosin in European sea bass, which were below the limit of quantification of the method.

Treatment of textile wastewater using monolayered ultrafiltation ceramic membrane fabricated from natural kaolin clay.

Fabrication, characterization and application of ceramic membrane developed from Tunisian natural kaolin clay for textile wastewater treatment are presented in this study. The morphology and properties of the resulting membrane sintered at 1000°C for 3 h were then determined by Scanning Electron Microscopy (SEM), mechanical and chemical resistance and water permeability. Separation performance of the membrane was evaluated during the treatment of textile wastewater. SEM images reveal the homogeneous surface of the membrane. The membrane displayed good chemical and mechanical resistances as well. Its permeability was of 21.2 L.h -1.m-2.bar-1, indicating that separation performance could occur in the domain of Ultrafiltration (UF). Performances of the membrane during the treatment of raw and biologically pretreated textile effluents are promising in terms of the removal of colour (99% for the raw effluent and 100% for the biologically pretreated effluent), chemical oxygen demand (COD) (80% for the raw effluent and 93% for the biologically pretreated effluent) and turbidity (98% for the raw effluent and 100% for the biologically pretreated effluent).

Overexpression of a Wheat Aquaporin Gene, TdPIP2;1, Enhances Salt and Drought Tolerance in Transgenic Durum Wheat cv. Maali.

In this study, we generated transgenic durum wheat cv. Maali overexpressing the wheat plasma membrane aquaporin TdPIP2;1 gene under the control of PrTdPIP2;1 promoter or under the constitutive PrCaMV35S promoter. Histochemical analysis of the fusion PrTdPIP2;1::TdPIP2;1::GusA in wheat plants showed that the ß-glucuronidase (GUS) activity was detected in the leaves, stems and roots of stably transformed wheat T3 plants. Our results showed that transgenic wheat lines overexpressing the TdPIP2;1 gene exhibited improved germination rates and biomass production and retained low Na+ and high K+ concentrations in their shoots under high salt and osmotic stress conditions. In a long-term study under greenhouse conditions on salt or drought stress, transgenic TdPIP2;1 lines produced filled grains, whereas wild-type (WT) plants either died at the vegetative stage under salt stress or showed drastically reduced grain filling under drought stress. Performing real time RT-PCR experiments on wheat plants transformed with the fusion PrTdPIP2;1::GusA, we showed an increase in the accumulation of GusA transcripts in the roots of plants challenged with salt and drought stress. Study of the antioxidant defence system in transgenic wheat TdPIP2;1 lines showed that these lines induced the antioxidative enzymes Catalase (CAT) and Superoxide dismutase (SOD) activities more efficiently than the WT plants, which is associated with lower malondialdehyde and hydrogen peroxide contents. Taken together, these results indicate the high potential of the TdPIP2;1 gene for reducing water evaporation from leaves (water loss) in response to water deficit through the lowering of transpiration per unit leaf area (stomatal conductance) and engineering effective drought and salt tolerance in transgenic TdPIP2;1 lines.

Clay-Na as a sorbent for the extraction of anti-inflammatory compounds in water samples.

In this work, clay-Na particles are used as the adsorbent for the solid-phase extraction of acidic compounds. The novel sorbent under study is based on high-specific surface area, cation-exchange capacity designed specifically to offer ion-exchange properties with the goal being to selectively extract a group of acidic compounds. The effects of the extraction parameters including extraction elution solvent, sample volume and pH. In optimum conditions, the repeatability for one fiber (n = 3), expressed as % relative standard deviation, was between 0.3 and 4.3% for the acid compounds. The detection limits for the studied acidic compounds were between 0.1-0.6 µg/L. The developed method offers the advantages of being simple to use and having a low cost of equipment.

Solid-phase extraction of drugs with cuttlefish bone powder as a sorbent.

We investigated cuttlefish bone powder for the solid-phase extraction of naproxen, ibuprofen, and carbamazepine. The basic principles controlling the extraction are presented to aid in the choice of the nature and quantity of the extracting phase according to the sample matrix and the solute properties, based on the mechanisms of phase retention. Their retention mechanism is based on hydrogen bonding and electrostatic interactions. The results show a significant recovery rate for the three drugs, selectivity, and low cost. The method has successfully reduced the amount of tested pharmaceuticals with recoveries >87% at pH 4.

Removal of two anionic reactive textile dyes by adsorption into MgAl-layered double hydroxide in aqueous solutions.

Textile dyes pose a significant challenge for water pollution due to the poor degradability of their complex aromatic structures (e.g., RR-120 and RBB-150). In order to minimize the harmful effects of RR-120 and RBB-150, the capacity of MgAl-layered double hydroxide for removing of these contaminants was studied herein. Batch adsorption experiments were conducted to investigate the effect of various operating parameters, such as solution pH, contact time, dye concentration, and temperature in order to provide optimal conditions for removal. Structural and morphological analyses were used to highlight the assembly and/or interaction LDH-dye. The state of equilibrium of RR-120 and RBB-150 adsorption was pH- and temperature-dependent and followed the pseudo-second-order rate model. Also, the equilibrium adsorption data of both dyes were found to adopt the Langmuir type isotherm model, which assumes a monolayer arrangement in LDH-dye. Furthermore, the effects of four major coexisting and competing mono- and divalent interlayer anions, such as NO3-, Cl-, CO32-, and SO42-, on the uptakes of RR-120 and RBB-150 were studied and the results showed that NO3- anions had insignificant effect on the uptakes of RR-120 and RBB-150 by MgAl. An equivalent study on the presence of both dyes in competitive trial adsorption/desorption from binary aqueous solution was investigated. And finally, the reuse operation of recovered material after dye adsorption was tested in up to 5 cycles of recyclability.

Extraction of clarithromycin and atenolol by cuttlefish bone powder.

Pharmaceutical products including analgesics, antibiotics, and antiepileptics are often present in wastewater treatment effluents, lakes, and rivers and have been found in waterways of many countries. The adsorption of two common pharmaceuticals present in surface waters, clarithromycin (CLR) and atenolol (ATN), onto cuttlefish bone powder treated with HCl has been investigated. Adsorption experiments were carried out at different pH conditions and the experimental equilibrium data were fitted to the Langmuir and Freundlich. For the same experimental conditions (pH = 7), the estimated adsorption capacities are from 34.5 mg g-1 for CLR and 39.5 mg g-1 for ATN. The influence of adsorption pH was established for each compound. The negative values of ΔG at different temperatures for all samples indicate the spontaneous nature of the sorption process.

Evaluation of Antioxidant, Anticholinesterase, and Antidiabetic Potential of Dry Leaves and Stems in Tamarix aphylla Growing Wild in Tunisia.

Tamarix aphylla (L.) Karst. has a wide geographic distribution and was employed in traditional medicine as astringent, anti-rheumatic and to treat fever. T. aphylla leaves and stems extracts were studied from both chemical and biological points of view to assess the antidiabetic, anticholinesterase and antioxidant potential of this species. The HPLC/Diode Array Detector (DAD) analysis showed the presence of 14 phenolic compounds (gallic, caffeic, p-coumaric, ferulic and ellagic acids, kaempferol, quercetin, quercetin 3-O-galactoside and six flavonol derivatives). This is the first study reporting a comparative study of the biological activities of different extracts from T. aphylla. High activities were obtained against DPPH radical, superoxide anion radical (O2∙-) and nitric oxide radical (• NO) in a concentration-dependent manner, the most active extracts being the polar ones. T. aphylla also showed moderate protective effects against acetylcholinesterase, but no effects were observed against butyrylcholinesterase. Against α-glucosidase the MeOH extracts displayed IC50 values from 8.41 to 24.81 µg/ml.

Simultaneous detection of antibiotics and other drug residues in the dissolved and particulate phases of water by an off-line SPE combined with on-line SPE-LC-MS/MS: Method development and application.

Due to their widespread use in human and animal healthcare, antibiotics and other drug residues are ubiquitous in the aquatic environment. Given their potential impacts on ecosystem functioning and public health, the quantification of environmental drug residues has become a necessity. Various analysis techniques have been found to be suitable for reliable detection of such compounds. However, quantification can be difficult because these compounds are present at trace or ultra-trace levels. Consequently, the accuracy of environmental analyses depends on both the efficiency and the robustness of the extraction and quantification method. In this work, an off-line solid-phase extraction (SPE) combined with on-line SPE-LC-MS/MS was applied to the simultaneous extraction and quantification of 26 pharmaceutical products, including 18 antibiotics, dissolved in a water phase. Optimal conditions were determined and then applied to assess the contamination level of the targeted drug residues in water collected from four sites in Northern France: a river, the input and output of an aerated lagoon, and a wastewater treatment plant. Drug residues associated with suspended solid matter (SSM) were also quantified in this work using pressurized liquid extraction (PLE) combined with an on-line SPE-LC-MS/MS system in order to complete an assessment of the degree of total background pollution.

Enolizable Carbonyls and N,O-Acetals: A Rational Approach for Room-Temperature Lewis Superacid-Catalyzed Direct α-Amidoalkylation of Ketones and Aldehydes.

An efficient catalytic room-temperature direct α-amidoalkylation of carbonyl donors, that is, ketones and aldehydes with unbiased N,O-acetals, is described. Sn(NTf2 )4 is an optimal catalyst to promote this challenging transformation at low loading and the reaction shows promising scope. A comprehensive and rational evaluation of this reaction has led to the establishment of an empirical scale of nucleophilic reactivity for a broad set of ketones that should be helpful in the synthetic design and development of carbonyl α-functionalization methods.

Sequential Friedel-Crafts-Type α-Amidoalkylation/Intramolecular Hydroarylation: Distinct Advantage of Combined Tf2NH/Cationic LAu(I) as a Consecutive or Binary Bicatalytic System.

The combined use of Tf2NH and L(Au)(+)X(-) as a dual or binary catalytic system clearly improves the efficiency and enlarges the scope of the tandem intermolecular Friedel-Crafts α-amidoalkylation/intramolecular hydroarylation sequence, compared to an "all gold" multicatalysis approach.

Tissue and cellular localization of tannins in Tunisian dates (Phoenix dactylifera L.) by light and transmission electron microscopy.

A histological approach including light microscopy and transmission electron microscopy (TEM) was used to provide accurate information on the localization of condensed tannins in the edible tissues and in the stone of date fruits (Phoenix dactylifera L.). Light microscopy was carried out on fresh tissues after staining by 4-dimethylaminocinnamaldehyde (DMACA) for a specific detection of condensed tannins. Thus, whether under light microscopy or transmission electron microscopy (TEM), results showed that tannins are not located in the epidermis but more deeply in the mesocarp in the vacuole of very large cells. Regarding the stones, tannins are found in a specific cell layer located at 50 µm from the sclereid cells of the testa.

Functional analysis of the durum wheat gene TdPIP2;1 and its promoter region in response to abiotic stress in rice.

In a previous work, we demonstrated that expression of TdPIP2;1 in Xenopus oocytes resulted in an increase in Pf compared to water injected oocytes. Phenotypic analyses of transgenic tobacco plants expressing TdPIP2;1 generated a tolerance phenotype towards drought and salinity stresses. To elucidate its stress tolerance mechanism at the transcriptional level, we isolated and characterized the promoter region of the TdPIP2;1 gene. A 1060-bp genomic fragment upstream of the TdPIP2;1 translated sequence has been isolated, cloned, and designated as the proTdPIP2;1 promoter. Sequence analysis of proTdPIP2;1 revealed the presence of cis regulatory elements which could be required for abiotic stress responsiveness, for tissue-specific and vascular expression. The proTdPIP2;1 promoter was fused to the ß-glucuronidase (gusA) gene and the resulting construct was transferred into rice (cv. Nipponbare). Histochemical analysis of proTdPIP2;1::Gus in rice plants revealed that the GUS activity was observed in leaves, stems and roots of stably transformed rice T3 plants. Histological sections prepared revealed accumulation of GUS products in phloem, xylem and in some cells adjacent to xylem. The transcripts were up-regulated by dehydration. Transgenic rice plants overexpressing proTdPIP2;1 in fusion with TdPIP2;1, showed enhanced drought tolerance, while wild type plants were more sensitive and exhibited symptoms of wilting and chlorosis. These findings suggest that expression of the TdPIP2;1 gene regulated by its own promoter achieves enhanced drought tolerance in rice.

Comparative study of antioxidant power, polyphenols, flavonoids and betacyanins of peel and pulp of three Tunisian Opuntia forms.

The antioxidant activity and the chemical composition of methanol extracts from peel and pulp belonging to two species of Tunisian prickly pears Opuntia ficus indica (spiny and thornless forms) and Opuntia stricta has been studied. The antioxidant capacity was measured by DPPH radical scavenging activity. The Total Phenolic Compound (TPC) and the total flavonoid content were determined by the Folin-Ciocalteu method and colorimetric method, respectively. The phenolic compounds were identified and quantified by High Performance Liquid Chromatography (HPLC) coupled with an electrospray ionization mass spectrometry (ESI-MS). The results showed that O. stricta fruits present the best antioxidant activities than the two forms of O. ficus indica while the TPC was more important in O. ficus indica than in the O. stricta fruits. The peels have higher flavonoids than pulps and the thornless has more flavonoid than the spiny. The RP-HPLC and ESI-MS analysis detected two classes of phenolic compounds and betalain pigments. Isorhamnetin derivatives are the dominant flavonol glycoside identified in O. ficus indica (spiny: 65.25 µg g(-1); thornless: 77.03 µg g(-1)) and O. stricta peels (19.22 µg g(-1)).

Cloning and heterologous expression of a thermostable pectate lyase from Penicillium occitanis in Escherichia coli.

The entire pectate lyase cDNA (Pel1) of Penicillium occitanis was cloned from a cDNA bank and sequenced. The ORF exhibited a great homology to Penicillium marneffei and conservation of all features of fungal pectate lyases such as the barrel structure with "eight right-handed parallel ß-helix" architecture. The structure modeling also showed the interesting resemblance with thermostable pectate lyases since several specific residues were also shared by Pel1 and these thermostable enzymes. Having shown that the enzyme retains its activity after endoH-mediated deglycosylation, we investigated its expression in Escherichia coli BL21 using the pET28-a vector. This expression was shown to be optimum when cells were induced at room temperature in 2YT medium rather than at 37 °C and LB medium. In such conditions, the recombinant protein was apparently produced more in soluble form than as inclusion bodies. The effect of NaCl concentration was investigated during the binding and elution steps of recombinant His-tagged enzyme on MagneHis Ni-particles. The purified enzyme was shown to retain its thermo-activity as well as a great tolerance to high concentration of NaCl and imidazole.

A modified cyclen azaxanthone ligand as a new fluorescent probe for Zn2+.

A new cyclen derivative L, bearing a methyl-chromeno-pyridinylidene hydrazone moiety, was synthesized and studied in MeOH, as potential fluorescent "OFF-on-ON" sensors for Zn(ii). Photophysical properties of this ligand being PET regulated, L was only weakly emissive in the absence of metal ions (OFF). L fluorescence was increased modestly upon addition of one equivalent of Zn(II), and further increased upon addition of a second equivalent. Therefore, Zn : L behaved as a highly sensitive ON sensor for zinc. This efficiency was correlated to Zn(II) coordination via the hydrazone moiety of the fluorophore, producing an efficient CHelation-Enhanced Fluorescence (CHEF) effect. A complementary theoretical study carried out with DFT calculations further elucidated the optical properties.

Detailed polyphenol and tannin composition and its variability in Tunisian dates (Phoenix dactylifera L.) at different maturity stages.

The polyphenol profile of two Tunisian varieties of dates including flavanols, flavonols, flavones, and hydroxycinnamates was characterized. Three tissue zones (flesh, peel, and stone) and three maturity stages were considered. Phenolic compounds were analyzed using reversed phase high-performance liquid chromatography coupled to UV-visible and electrospray mass spectrometry. Procyanidin oligomers and polymers were characterized and quantified using phloroglucinolysis prior to HPLC analysis. Procyanidin polymers based on (-)-epicatechin structure were by far the most concentrated polyphenols in ripe dates, accounting for 95% of total polyphenols with an average concentration of 14 g/kg in the fresh edible parts of the fruit. Interestingly, procyanidins were also highly concentrated in the stones. The concentration and average degree of polymerization (DPn) of the procyanidins decreased according to maturity. Other phenolics, including caffeoylshikimic acid hexoside, caffeoyl-sinapoyl monohexoside and dihexoside, and acetylated flavonols, were tentatively identified for the first time in the fruit.

Comparative Study of Antioxidant Power, Polyphenols, Flavonoids and Betacyanins of the Peel and Pulp of Three Tunisian Opuntia Forms.

The antioxidant activity and the chemical composition of methanol extracts from peel and pulp belonging to two species of Tunisian prickly pears Opuntia ficus indica (spiny and thornless forms) and Opuntia stricta have been studied. The antioxidant capacity was measured by DPPH radical scavenging activity. The total phenolic compound (TPC) and the total flavonoid content were determined by the Folin-Ciocalteu method and colorimetric method, respectively. The phenolic compounds were identified and quantified by high-performance liquid chromatography (HPLC) coupled with an electrospray ionization mass spectrometry (ESI-MS). The results showed that O. stricta fruits present the best antioxidant activities than the two forms of O. ficus indica, while the TPC was more important in O. ficus indica than in the O. stricta fruits. The peels have higher flavonoids than pulp, and the thornless variety has more flavonoid than the spiny. The RP-HPLC and ESI-MS analysis detected two classes of phenolic compounds and betalain pigments. Isorhamnetin derivatives are the dominant flavonol glycoside identified in O. ficus indica (spiny: 65.25 µg·g(-1); thornless: 77.03 µg·g(-1)) and O. stricta peels (19.22 µg·g(-1)).

A new approach for the assessment of groundwater quality and its suitability for irrigation: a case study of the Korba Coastal Aquifer (Tunisia, Africa).

Groundwater is the main source of water in Mediterranean, water-scarce, semiarid regions of Tunisia, Africa. In this study of the Korba coastal aquifer, 17 water wells were studied to assess their suitability for irrigation and drinking purposes. Assessment parameters include pH, salinity, specific ion toxicity, sodium adsorption ratio, nutrients, trace metals pollutants, and fecal indicators and pathogens. Results indicate that salinity of groundwater varied between 0.36 dS/m and 17.4 dS/m; in addition, its degree of restriction is defined as "none", "slight to moderate", and "severe" for 18, 23, and 59% of the studied wells, respectively. To control salts brought in by irrigation waters, the question arises as to how much water should be used to reach crop and soil requirements. To answer this question, a new approach that calculates the optimum amount of irrigation water considering the electrical conductivity of well water (ECw), field crops, and the semiarid meteorological local conditions for evapotranspiration and rainfall is developed. This is applied to the authors' case study area; barley and lettuce were selected among the commonly grown crops because they are high- and low-salinity tolerant, respectively. Leaching requirements were found to be independent of the crop selected, and depend only on the season, that is, 250 to 260 mm/month in the driest season, with a minimum of 47 mm/month though all seasons. A high bacteriological contamination appears in almost all samples. However, if disinfected and corrected for pH, all the well waters can be used for animal farming (including livestock and poultry), although only 29% could be used for human consumption.

Supercritical SC-CO(2) and Soxhlet n-Hexane Extract of Tunisian Opuntia ficus indica Seeds and Fatty Acids Analysis.

The fatty acids profiles of Tunisian Opuntia ficus indica seeds (spiny and thornless form) were investigated. Results of supercritical carbon dioxide (SC-CO(2)) and soxhlet n-hexane extract were compared. Quantitatively, the better yield was obtained through soxhlet n-hexane: 10.32% (spiny) and 8.91% (thornless) against 3.4% (spiny) and 1.94% (thornless) by SC-CO(2) extract (T = 40°C, P = 180 bar, time = 135 mn, CO(2) flow rate = 15 mL·s(-1)). Qualitatively, the main fatty acids components were the same for the two types of extraction. Linoleic acid was the major compound, SC-CO(2): 57.60% (spiny), 59.98% (thornless), soxhlet n-hexane: 57.54% (spiny), 60.66% (thornless), followed by oleic acid, SC-CO(2): 22.31% (spiny), 22.40% (thornless), soxhlet n-hexane: 25.28% (spiny), 20.58% (thornless) and palmitic acid, SC-CO(2): 14.3% (spiny), 12.92% (thornless), soxhlet n-hexane: 11.33% (spiny), 13.08% (thornless). The SC-CO(2) profiles fatty acids showed a richness with other minority compounds such as C(20:1), C(20:2), and C(22).The seeds oil was highly unsaturated (US = 4.44-5.25), and the rising temperatures donot affect the selectivity of fatty acids extract by SC-CO2: US = 4.44 (T = 40°C) and 4.13 (T = 70°C).