Biochemical responses in mice induced by saxitoxins extracted from the cockles Acanthocardia tuberculatum.

Harmful algae blooms have increased in frequency and geographic range in recent decades, and they produce toxins strains such as saxitoxins (STXs). they block voltage-gated sodium channels and can lead to several poisonings and the death of organisms that pose a significant risk to public and environmental health. The study of STXs toxicity has been carried out but little is known about the response of antioxidant enzymes activities to STXs in mice. The purpose of this study was to evaluate biochemical responses and oxidative stress induced by STXs extracted from Acanthocardia tuberculatum. To this end, daily, mice were treated orally for 7 days with sublethal concentrations (10 mg/100 g mouse). The animal's liver was assessed using biomarkers such as activities of catalase (CAT), thiobarbituric acid reactive substances (TBARS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and succinate dehydrogenase (SDH). In the blood, plasmatic markers were analysed as glutamic oxalic transaminase (GOT), glutamic pyruvic transaminase (GPT), creatinine phosphokinase (CPK), lactate dehydrogenase (LDH), urea and creatinine. Globally, test toxicity test showed a significant decrease in the weight at 10 mg /100 g mouse, and the results showed an increase of GPT, GOT, CPK, LDH, CAT and TBARS activities and the inhibitory effect of GAPDH activities but creatinine, urea and SDH activities showed no significative difference from the control. We concluded that STXs induce oxidative stress breaking in mice the balance of the defence system and causing oxidations reactions. Moreover, STXs affect energy metabolism in mice, however, renal function in mice is not affected by exposure to STXs.

Methyl Red Decolorization Efficiency of a Korea Strain of Aspergillus sp. Immobilized into Different Polymeric Matrices.

Intensive research studies have revealed that fungal decolorization of dye wastewater is a promising replacement for the current process of dye wastewater decolorization. The authors isolated an Aspergillus sp. from the effluent of a textile industry area in Korea and assessed the effects of a variety of operational parameters on the decolorization of methyl red (MR) by this strain of Aspergillus sp. This Aspergillus sp. was then immobilized by entrapment in several polymeric matrices and the effects of operational conditions on MR decolorization were investigated again. The optimal decolorization activity of this Aspergillus sp. was observed in 1% glucose at a temperature of 37 °C and pH of 6.0. Furthermore, stable decolorization efficiency was observed when fungal biomass was immobilized into alginate gel during repeated batch experiment. These results suggest that the Aspergillus sp. isolated in Korea could be used to treat industrial wastewaters containing MR dye.

Biogenic nanopalladium based remediation of chlorinated hydrocarbons in marine environments.

Biogenic catalysts have been studied over the last 10 years in freshwater and soil environments, but neither their formation nor their application has been explored in marine ecosystems. The objective of this study was to develop a biogenic nanopalladium-based remediation method for reducing chlorinated hydrocarbons from marine environments by employing indigenous marine bacteria. Thirty facultative aerobic marine strains were isolated from two contaminated sites, the Lagoon of Mar Chica, Morocco, and Priolo Gargallo Syracuse, Italy. Eight strains showed concurrent palladium precipitation and biohydrogen production. X-ray diffraction and thin section transmission electron microscopy analysis indicated the presence of metallic Pd nanoparticles of various sizes (5-20 nm) formed either in the cytoplasm, in the periplasmic space, or extracellularly. These biogenic catalysts were used to dechlorinate trichloroethylene in simulated marine environments. Complete dehalogenation of 20 mg L(-1) trichloroethylene was achieved within 1 h using 50 mg L(-1) biogenic nanopalladium. These biogenic nanoparticles are promising developments for future marine bioremediation applications.

Heavy metals and antibiotics resistance of bacteria isolated from Marchica lagoon: biodegradation of anthracene on submerged aerated fixed bed reactor.

Heavy metals resistant and polyaromatic hydrocarbon (PAH)-degrading bacteria isolated from Marchica lagoon. Six isolates, Pseudomonas putida, Orchobactrum antropi, Staphylococus epidermidis, Brevundimonas diminuta, Serratia ficaria and Bacillus anthracis were characterized on the basis of biochemical and 16S rDNA. The strains that showed high resistance to heavy metals were also studied for their antibiotics resistance and growth kinetics. The minimal inhibitory concentrations (MIC) of metals at variant concentrations (10-38 mM) were determined in liquid and solid medium. Strains were showed an extreme resistance against metals with the MIC values of Cu (9 mM), Cd (6.25 mM), Cr (2.5 mM), Ag (0,625), and Hg (0,156). Furthermore, growth rates were decreased in the presence of metals (compared to the control). The anthracene elimination from synthetic wastewater was determined in a submerged aerobic fixed-film reactor. Optimal conditions for bacterium growth and biodegradation of anthracene are determined as: temperature of 37 °C, pH 7, and initial anthracene concentration of 20 mg/l. It emerged that at anthracene concentrations 5-40 mg/l, COD removal efficiency were 84.62%; 90.62%; 91.36% and 71.4% respectively.

Isolation, characterization, and antimicrobial activity of communic acid from Juniperus phoenicea.

OBJECTIVES: A mixture of Z and E communic acid is isolated for the first time from the cones of Juniperus phoenicea. Its biological activity was studied. METHODS: The plant material was extracted in a Soxhlet apparatus with n-hexane, the resulting extract was subjected to column chromatography (CC) on silica gel. The structure elucidation of the constituents of the isolated fraction was identified by comparison of its spectroscopic properties 1H and 13C NMR data with those reported in the literature. The antimicrobial assay of hexanic extract and isolated compounds was carried out by the disc diffusion and micro-dilution methods. RESULTS: A mixture of two diterpene acids isomers was isolated, with a high yield (68%). Their chemical structures were confirmed after comparing their spectral data with published reports. These natural products exhibited a significant antibacterial and antifungal activity against the tested strains. Indeed, for Bacillus cereus, Staphylococcus aureus, and Pseudomonas aeruginosa, the inhibition zone diameters (36-37 mm) was better than penicillin, novobiocin, and amoxicillin. For Candida albicans activity, it show that the mixture possess an activity similar to that of Metrazol. Against Escherichia coli, the inhibitory activity was found less than Amoxicillin. This is the first report of isolation of communic acid from J. phoenicea. CONCLUSIONS: These results showed that the cones of J. phoenicea were an important source of communic acid, and its hexanic extract had the greatest potential antibacterial activity against both Gram-negative and Gram-positive bacteria and C. albicans.

Prediction of Anti-COVID 19 Therapeutic Power of Medicinal Moroccan Plants Using Molecular Docking.

The emerging pathogen SARS-CoV2 causing coronavirus disease 2019 (COVID-19) is a global public health challenge. To the present day, COVID-19 had affected more than 40 million people worldwide. The exploration and the development of new bioactive compounds with cost-effective and specific anti-COVID 19 therapeutic power is the prime focus of the current medical research. Thus, the exploitation of the molecular docking technique has become essential in the discovery and development of new drugs, to better understand drug-target interactions in their original environment. This work consists of studying the binding affinity and the type of interactions, through molecular docking, between 54 compounds from Moroccan medicinal plants, dextran sulfate and heparin (compounds not derived from medicinal plants), and 3CLpro-SARS-CoV-2, ACE2, and the post fusion core of 2019-nCoV S2 subunit. The PDB files of the target proteins and prepared herbal compounds (ligands) were subjected for docking to AutoDock Vina using UCSF Chimera, which provides a list of potential complexes based on the criteria of form complementarity of the natural compound with their binding affinities. The results of molecular docking revealed that Taxol, Rutin, Genkwanine, and Luteolin-glucoside have a high affinity with ACE2 and 3CLpro. Therefore, these natural compounds can have 2 effects at once, inhibiting 3CLpro and preventing recognition between the virus and ACE2. These compounds may have a potential therapeutic effect against SARS-CoV2, and therefore natural anti-COVID-19 compounds.

Molecular characterization of mutations associated with resistance to second line drugs in Mycobacterium tuberculosis patients from Casablanca, Morocco.

The emergence and spread of extensively drug-resistant tuberculosis (XDR-TB) is a serious threat to global health. Therefore, its rapid diagnosis is crucial. The present study aimed to characterize mutations conferring resistance to second line drugs (SLDs) within multidrug Mycobacterium tuberculosis (MDR-MTB) isolates and to estimate the occurrence of XDR-TB in Casablanca, Morocco. A panel of 200 MDR-TB isolates was collected at the Pasteur Institute between 2015-2018. Samples were subjected to drug susceptibility testing to Ofloxacin (OFX), Kanamycin (KAN) and Amikacin (AMK). The mutational status of gyrA, gyrB, rrs, tlyA and eis was assessed by sequencing these target genes. Drug susceptibility testing for SLDs showed that among the 200 MDR strains, 20% were resistant to OFX, 2.5% to KAN and 1.5% to AMK. Overall, 14.5% of MDR strains harbored mutations in gyrA, gyrB, rrs and tlyA genes. From the 40 OFXR isolates, 67.5% had mutations in QRDR of gyrA and gyrB genes, the most frequent one being Ala90Val in gyrA gene. Of note, none of the isolates harbored simultaneously mutations in gyrA and gyrB genes. In eight out of the 200 MDR-TB isolates resistant either to KAN or AMK, only 25% had A1401G or Lys89Glu change in rrs and tlyA genes respectively. This study is very informative and provides data on the alarming rate of fluoroquinolone resistance which warrants the need to implement appropriate drug regimens to prevent the emergence and spread of more severe forms of Mycobacterium tuberculosis drug resistance.

Ionophoric properties of atropine: complexation and transport of Na+, K+, Mg2+ and Ca2+ ions across a liquid membrane.

The activity of atropine on the complexation and transport of Na(+), K(+), Mg(2+) and Ca(2+) ions across a liquid membrane was investigated using a spectrophotometric method. Atropine is a natural drug that blocks muscarinic receptors. It is a competitive antagonist of the action of acetylcholine and other muscarinic agonists. Atropine is shown to extract Na(+), K(+), Mg(2+) and Ca(2+) ions from an aqueous phase into an organic one with a preference for Ca(2+) ions. According to a kinetic study, divalent cations (Mg(2+) and Ca(2+)) are more rapidly transported than monovalent ones (Na(+) and K(+)). In both complexation and transport, the flux of the ions increases with the increase of atropine concentration. Atropine might act on the membrane permeability; its complexation and ionophoric properties shed new lights on its therapeutic properties.

Purification and partial characterization of paralytic shellfish poison-binding protein from Acanthocardia tuberculatum.

A paralytic shellfish poison-binding protein (PSPBP) was purified 16.6-fold from the foot of the Moroccan cockles Acanthocardia tuberculatum. Using affinity chromatography, 2.5mg of PSPBP showing homogeneity on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was obtained from 93 mg of crude extract. The purified PSPBP exhibits a specific activity of about 2.78 mU/mg proteins and has estimated molecular weight of 181 kDa. Observation of a single band equivalent to 88 kDa on SDS-PAGE under reducing conditions suggested it to be a homodimer. The optimal temperature and pH for the purified PSPBP were respectively 30 degrees C and 7.0.

Decolorization of some azo dyes by immobilized Geotrichum sp. biomass in fluidized bed bioreactor.

Geotrichum sp. strain, which is able to decolorize azo dyes enzymatically, was used in this study for decolorization of synthetics solutions contaminated by toxic azo dyes orange G, trypan blue, azorubine, and methyl red. The biomass of Geotrichum sp. was immobilized in calcium alginate and polyacrylamide gels and used for the decolorization of tested azo dyes in fluidized bed bioreactor. The highest specific decolorization rate was obtained when the fungal biomass was entrapped in calcium alginate beads. Immobilized biomass in calcium alginate continuously decolorized azo dyes after eight repeated batch decolorization experiments without significant loss of activity whereas polyacrylamide immobilized biomass retained only 10% of its activity after 4 days of incubation. The effects of some physicochemical parameters such as temperature, pH, and dyes concentration on decolorization performance of isolated fungal strain were also investigated.

A comparative study on biosorption characteristics of certain fungi for bromophenol blue dye.

Laboratory investigations of the potential use of dried biomasses of Rhizopus stolonifer, Fusarium sp., Geotrichum sp., and Aspergillus fumigatus as biosorbents for the removal of bromophenol blue (BPB) dye from aqueous solutions were conducted. Kinetics studies indicated that the BPB dye uptake processes can be well described by the pseudo-second-order model. The fungal biomasses exhibited the highest dye biosorption at pH 2.0. The Langmuir adsorption model appears to fit the dye biosorption better than the Freundlich model, with maximum dye uptake capacities ranging from 526 to 1,111 mg/g, depending on the biomass used.

Comparative study on differential accumulation of PSP toxins between cockle (Acanthocardia tuberculatum) and sweet clam (Callista chione).

At the western Mediterranean coast of Morocco, the cockle (Acanthocardia tuberculatum) contained persistent high levels of paralytic shellfish toxins for several years, while other bivalve molluscs such as sweet clam (Callista chione) from the same vicinity were contaminated seasonally to a much lesser extent. In order to understand the causes of this prolonged contamination, a comparative study on PSP decontamination between sweet clam and cockle was conducted from November 2001 until June 2002. PSP toxicity was analysed by automated pre-column oxidation (Prechromatographic oxidation and LC-FD) in several organs of both species, namely digestive gland, foot, gill, mantle, muscle and siphon for sweet clams. The results showed that cockle sequester PSP toxins preferably in non-visceral organs (Foot, gill and mantle) contrary to sweet clam that sequester them in visceral tissues (digestive gland). The toxin profile of cockle organs indicated dominance of dcSTX, whereas sweet clam tissues contained especially C-toxins. Substantial differences in toxin profile between cockle and sweet clam, from the same area as well as from the composition of PSP toxin producer, Gymnodinium catenatum, confirm the bioconversion of PSP toxins in cockle.

Bacterial population and biodegradation potential in chronically crude oil-contaminated marine sediments are strongly linked to temperature.

Two of the largest crude oil-polluted areas in the world are the semi-enclosed Mediterranean and Red Seas, but the effect of chronic pollution remains incompletely understood on a large scale. We compared the influence of environmental and geographical constraints and anthropogenic forces (hydrocarbon input) on bacterial communities in eight geographically separated oil-polluted sites along the coastlines of the Mediterranean and Red Seas. The differences in community compositions and their biodegradation potential were primarily associated (P < 0.05) with both temperature and chemical diversity. Furthermore, we observed a link between temperature and chemical and biological diversity that was stronger in chronically polluted sites than in pristine ones where accidental oil spills occurred. We propose that low temperature increases bacterial richness while decreasing catabolic diversity and that chronic pollution promotes catabolic diversification. Our results further suggest that the bacterial populations in chronically polluted sites may respond more promptly in degrading petroleum after accidental oil spills.

Biosorption of mercury from aqueous solution by Ulva lactuca biomass.

The mercury biosorption onto non-living protonated biomass of Ulva lactuca, as an alternative method for mercury removal from aqueous solutions, was investigated. Batch equilibrium tests showed that at pH 3.5, 5.5 and 7 the maxima of mercury uptake values, according to Langmuir adsorption isotherm, were 27.24, 84.74 and 149.25 mg/g, respectively. The ability of Ulva lactuca biomass to adsorb mercury in fixed-bed column, was investigated as well. The influence of column bed height, flow rate and effluent initial concentration of metal was studied. The adsorbed metal ions were easily desorbed from the algal biomass with 0.3 N H(2)SO(4) solution. After acid desorption and regeneration with distilled water, the biomass could be reused for other biosorption assays with similar performances.

Bioremediation of Southern Mediterranean oil polluted sites comes of age.

Mediterranean Sea is facing a very high risk of oil pollution due to the high number of oil extractive and refining sites along the basin coasts, and the intense maritime traffic of oil tankers. All the Mediterranean countries have adopted severe regulations for minimizing pollution events and bioremediation feasibility studies for the most urgent polluted sites are undergoing. However, the analysis of the scientific studies applying modern 'meta-omics' technologies that have been performed on marine oil pollution worldwide showed that the Southern Mediterranean side has been neglected by the international research. Most of the studies in the Mediterranean Sea have been done in polluted sites of the Northern side of the basin. Those of the Southern side are poorly studied, despite many of the Southern countries being major oil producers and exporters. The recently EU-funded research project ULIXES has as a major objective to increase the knowledge of the bioremediation potential of sites from the Southern Mediterranean countries. ULIXES is targeting four major polluted sites on the coastlines of Egypt, Jordan, Morocco and Tunisia, including seashore sands, lagoons, and oil refinery polluted sediments. The research is designed to unravel, categorize, catalogue, exploit and manage the diversity and ecology of microorganisms thriving in these polluted sites. Isolation of novel hydrocarbon degrading microbes and a series of state of the art 'meta-omics' technologies are the baseline tools for improving our knowledge on biodegradation capacities mediated by microbes under different environmental settings and for designing novel site-tailored bioremediation approaches. A network of twelve European and Southern Mediterranean partners is cooperating for plugging the existing gap of knowledge for the development of novel bioremediation processes targeting such poorly investigated polluted sites.

The prevalence of resistance-associated mutations to protease and reverse transcriptase inhibitors in treatment-naïve (HIV1)-infected individuals in Casablanca, Morocco.

BACKGROUND: The widespread use of antiretroviral agents and the growing occurrence of HIV-1 strains resistant to these drugs have given rise to serious concerns regarding the transmission of resistant viruses to newly infected persons, which may reduce the efficacy of a first-line antiretroviral therapy. METHODOLOGY: RNA was extracted from plasma samples of 98 treatment-naïve individuals with a plasma HIV RNA viral load of at least 1,000 copies/ml. Both protease (pr) and reverse transcriptase (rt) were amplified and sequenced using an automated sequencer. National Agency for AIDS Research (ANRS) and Stanford HIV database algorithms were used for interpretation of resistance data. RESULTS: In the protease segment, various minor mutations were present in the majority of the sequenced samples with high frequencies. Only two major mutations, M46L and V82L, were separately found in three individuals of 71 (4.2%) with one carrying both mutations. In the reverse transcriptase gene, no NNRTIs-associated resistance mutations were detected. Only one patient of 70 (1.4%) carried the F77L mutation that is associated with NRTIs resistance. Genetic subtyping revealed that 74.6% of samples were infected with subtype B, 15.5% with CRF02_AG, 4.2% with CRF01_AE, 1.4% with C, 2.8% with G and 1.4% with subtype F2. CONCLUSIONS: The low prevalence of major mutations associated with resistance to antiretroviral drugs (ARVs) among drug-naïve individuals studied suggests that the routine of drug resistance testing may be unnecessary for all Moroccan individuals newly diagnosed or all patients beginning antiretroviral therapy. Nevertheless, continuous surveillance is required since greater access to antiretroviral drugs is expected in Morocco.

Electron transfer from NADH bound to horse liver alcohol dehydrogenase (NAD+ dependent dehydrogenase): visualisation of the activity in the enzyme crystals and adsorption of formazan derivatives by these crystals.

The crystals of holoenzyme from native and cross-linked alcohol dehydrogenase exhibit electron transfer from NADH to phenazinium methosulfate (PMS), and then to the tetrazolium salt sodium 3,3'-{1-[(phenylamino)carbonyl]-3,4-tetrazolium}-bis(4-methoxy-6-nitro)benzenesulfonate (XXT). The slow dissociation of the cofactor and/or the conformational change associated can now be bypassed. The reduction product, formazan, did not diffuse out of the crystals in buffer and the crystals turned colored. In the presence of dimethyl sulfoxide or dimethoxyethane, the formazan diffused out to the solution. The reaction rates were found to be, respectively, 18% and 15% of the redox reaction rate of ethanol with cinnamaldehyde, close to the activity determined for the enzyme in solution in the presence of dimethoxyethane. The use of system PMS-tetrazolium salt is a useful tool to visualize the activity of dehydrogenases and other electron transferring systems in the crystalline state. The adsorption of formazan by the alcohol dehydrogenase crystals occurs in solution.

Decolorization of azo dyes with Enterobacter agglomerans immobilized in different supports by using fluidized bed bioreactor.

Immobilized cells of Enterobacter agglomerans, able to reduce azo dyes enzymatically, were used as a biocatalyst for the decolorization of synthetic medium containing the toxic azo dye methyl red (MR). This bacterial strain exhibits high ability to completely decolorize 100 mg/L of MR after only 6 h of incubation under aerobic conditions. Cells of E. agglomerans were immobilized in calcium alginate, polyacylamide, cooper beech, and vermiculite, and were used for the decolorization of MR from synthetic water by using a fluidized bed bioreactor. The highest specific decolorization rate was obtained when E. agglomerans was entrapped in calcium alginate beads and was of about 3.04 mg MR/g cell/h with a 50% conversion time ( t(1/2)) of about 1.6 h. Moreover, immobilized cells in calcium alginate continuously decolorized MR even after seven repeated experiments without significant loss of activity, while polyacrylamide-, cooper beech-, and vermiculite-immobilized cells retained only 62, 15, and 13% of their original activity, respectively.

Purification and partial characterization of azoreductase from Enterobacter agglomerans.

Azoreductase, an enzyme catalyzing the reductive cleavage of the azo bond of methyl red (MR) and related dyes, was purified to electrophoretic homogeneity from Enterobacter agglomerans. This bacterial strain, isolated from dye-contaminated sludge, has a higher ability to grow, under aerobic conditions, on culture medium containing 100mg/L of MR. The enzyme was purified approximately 90-fold with 20% yield by ammonium sulfate precipitation, followed by three steps of column chromatography (gel-filtration, anion-exchange, and dye-affinity). The purified enzyme is a monomer with a molecular weight of 28,000 Da. The maximal azoreductase activity was observed at pH 7.0 and at 35 degrees C. This activity was NADH dependent. The K(m) values for both NADH and MR were 58.9 and 29.4 microM, respectively. The maximal velocity (V(max)) was 9.2 micromol of NADH min(-1)mg(-1). The purified enzyme is inhibited by several metal ions including Fe(2+) and Cd(2+).