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Drought stress adversely affects plant health and productivity. Recently, drought-resistant bacterial isolates are used to combat drought resistance in crops. In this in vitro study, 20 bacterial isolates were isolated from harsh soil; their drought tolerance was evaluated using four concentrations of polyethylene glycol (PEG) 6000. The two most efficient isolates (DS4 and DS9) were selected and identified using 16S rRNA genetic sequencing. They were registered in the NCBI database and deposited under accession numbers MW916285 and MW916307 for Bacillus cereus (DS4) and Bacillus albus (DS9), respectively. These isolates were screened for plant growth-promoting properties compared to non-stressed conditions. Biochemical parameters; Proline, salicylic acid, gibberellic acid (GA), indole acetic acid (IAA), antioxidant activity, and antioxidant enzymes were measured under the same conditions, and in vitro seed germination was tested under stress conditions and inoculation with selected isolates. The results showed that under the harsh conditions of PEG6000, DS4 produced the highest amount of IAA of 1.61 µg/ml, followed by DS9 with 0.9 µg/ml. The highest amount of GA (49.95 µg/ml) was produced by DS9. On the other hand, the highest amount of siderophore was produced from DS4 isolate followed by DS9. Additionally, DS4 isolate recorded the highest exopolysaccharide (EPS) content of 3.4 mg/ml under PEG (-1.2 MPa) followed by DS9. The antioxidant activity increased in PEG concentrations depending manner, and the activity of the antioxidant enzymes increased, as catalase (CAT) recorded the highest activity in DS4 with an amount of 1.095 mg/ml. additionally, an increase in biofilm formation was observed under drought conditions. The isolated mixture protected the plant from the harmful effects of drought and showed an increase in the measured variables. Under unstressed conditions, the highest rates of emulsification index (EI 24%) were obtained for DS4 and DS9, at 14.92 and 11.54, respectively, and decreased under stress. The highest values of germination, total seedling length, and vigor index were obtained upon inoculation with the combination of two strains, and were 100%, 4.10 cm, and 410, respectively. Therefore, two strains combination is an effective vaccine capable of developing and improving drought tolerance in dryland plants.

The rapid spread of late wilt disease among maize cultivations has resulted in serious economic losses in many countries. Harpophora maydis is the main cause of this destructive vascular disease. Here we evaluate the fungicidal activity of chitosan and nano-chitosan against six aggressive isolates of H. maydis collected from different Egyptian governorates. Pathogenicity tests for these isolates show that the highest disease severity was found for the Giza isolate. The isolates were tested for their response to the fungicide Permis, chitosan, and nano-chitosan treatments in vitro and in vivo. Nano-chitosan treatments fully inhibited the radial growth of H. maydis isolates at concentrations of 5 and 10 mM, compared to the full control growth (9 cm in diameter). On the other hand, in vitro, in vivo, and molecular diagnosis results showed high antifungal activity of chitosan and nano-chitosan compared to the Permis fungicide. Chitosan at the nano and normal scales proved a potent ability to enhance plant resistance in response to H. maydis. Disease severity (DS%) was extremely decreased among the tested cultivars by using nano-chitosan; the highest percentage was obtained on Giza 178 cv, where the DS% was 21.7% compared to 42.3% for the control. Meanwhile, the lowest percentage was obtained on Giza 180 cv with DS% 31.2 and the control with 41.3%. The plants treated with nano-chitosan showed the highest growth parameters for all cultivars. Such natural treatments could reduce the impact on the environment as they are non-pollutant natural compounds, protect the plants by reducing fungal activity, and induce plant resistance.

Trastuzumab (Trz) is a humanized monoclonal antibody targeting epidermal growth factor receptor 2 (HER2; ErbB2). The combined administration of Trz and doxorubicin (DOX) has shown potent anti-cancer efficacy; however, this regimen may be accompanied by severe cardiac toxicity. Mesenchymal stem cells (MSCs)-derived exosomes are nanosized vesicles that play a crucial role in cell–cell communication and have shown efficacy in the treatment of various diseases. In this study, we aim to investigate the cardioprotective effects of MSCs-derived exosomes in a DOX/Trz- mediated cardiotoxicity model, and the possible mechanisms underlying these effects are elucidated. Forty-nine male rats were randomly assigned into four groups: Group I (control); Group II (Dox/Trz); Group III (protective group); and Group IV (curative group). Cardiac hemodynamic parameters, serum markers of cardiac injury, oxidative stress indices, and cardiac histopathology were investigated. Further, transcript profile of specific cardiac tissue injury markers, apoptotic markers, and fibrotic markers were analyzed using qRT-PCR, while the protein expressions of pAkt/Akt, pERK/ERK, pJNK/JNK, pJNK/JNK, and pSTAT3/STAT3 were evaluated by ELISA. Additionally, cardiac mirR-21 and miR-26a were assessed. A combined administration of DOX/Trz disrupted redox and Ca2+ homeostasis in cardiac tissue induced myocardial fibrosis and myofibril loss and triggered cardiac DNA damage and apoptosis. This cardiotoxicity was accompanied by decreased NRG-1 mRNA expression, HER2 protein expression, and suppressed AKT and ERK phosphorylation, while triggering JNK phosphorylation. Histological and ultra-structural examination of cardiac specimens revealed features typical of cardiac tissue injury. Moreover, a significant decline in cardiac function was observed through biochemical testing of serum cardiac markers and echocardiography. In contrast, the intraperitoneal administration of MSCs-derived exosomes alleviated cardiac injury in both protective and curative protocols; however, superior effects were observed in the protective protocol. The results of the current study indicate the ability of MSCs-derived exosomes to protect from and attenuate DOX/Trz-induced cardiotoxicity. The NRG-1/HER2, MAPK, PI3K/AKT, PJNK/JNK, and PSTAT/STAT signaling pathways play roles in mediating these effects.

The rapid spread of late wilt disease among maize cultivations has resulted in serious economic losses in many countries.

Background. Liver transplantation remains the only viable therapy for liver failure but has a severely restricted utility. Here, we aimed to decellularize rat livers to form acellular 3D bio-scaffolds suitable for seeding with induced pluripotent cells (iPSCs) as a tool to investigate the role of Wnt/β-catenin signaling in liver development and generation. Methods. Dissected rat livers were randomly divided into three groups: I (control), II (decellularized scaffolds) and III (recellularized scaffolds). Liver decellularization was established via an adapted perfusion procedure and assessed through the measurement of extracellular matrix (ECM) proteins and DNA content. Liver recellularization was assessed through histological examination and measurement of transcript levels of Wnt/β-catenin pathway, hepatogenesis, liver-specific microRNAs and growth factors essential for liver development. Adult rat liver decellularization was confirmed by the maintenance of ECM proteins and persistence of growth factors essential for liver regeneration. Results. iPSCs seeded rat decellularized livers displayed upregulated transcript expression of Wnt/β-catenin pathway-related, growth factors, and liver specification genes. Further, recellularized livers displayed restored liver-specific functions including albumin secretion and urea synthesis. Conclusion. This establishes proof-of-principle for the generation of three-dimensional liver organ scaffolds as grafts and functional re-establishment.

With growing concern about including unconventional dietary protein sources in poultry diets to substitute the protein sources that are essential for human consumption such as soybean meal, Azolla leaf meal (ALM) has grown in popularity. In our prior experiment, ALM was used at inclusion rates of 5 and 10%. Five per cent inclusion of ALM increased broiler chicken growth performance, the concentration of cecal propionic acid, and activation of skeletal muscle p70S6 Kinase1 (p70S6K1) without having detrimental effects on the meat quality. Those results prompted us to further evaluate the effect of the same inclusion rates of ALM on phase feeding and intestine and liver health of the broiler chicks. The current study hypothesis is that dietary ALM positively affects phase feeding, intestinal morphology and p70S6K1 activation, cecal microbial gene expression, and improves the liver energy status. For this, we enrolled 135 one-day-old broiler chicks and collected growth performance data (starter, grower, and finisher stages) and samples of the gastrointestinal tract to analyse the morphology of the villi, immune-related organs, mucin, and abundance of intestinal p70S6K1. Cecal bacterial species were analysed using qPCR and liver samples were collected to analyse adenosine monophosphate (AMP) and ATP content and selected oxidative stress biomarkers. ALM increased BW and feed intake during the starter and grower phases but did not affect the feed conversion ratio. Liver oxidative stress and AMP: ATP ratio increased in chickens fed on a diet containing 10% ALM (AZ10; P

AIMS Cyanobacteria are immense sources of several pharmacological active compounds such as flavonoids and carotenoids with anti-inflammatory and antioxidant activity. The potential therapeutic effect of two novel cyanobacterial isolates, Cronbergia siamensis (KY296358.1) and Sphaerospermopsis aphanizomenoides (KU212886.1), against hydrogen peroxide (H2 O2 )-induced oxidative stress damage in the rat model was determined in this study. METHODS AND RESULTS In vitro antioxidant activity of the two studied isolates was evaluated by radical scavenging assay and ferric reducing power. The possible prophylactic activity of S. aphanizomenoides (KU212886.1) against H2 O2 -induced oxidative stress in the rat model was assessed in vivo. Serum alanine transaminase and aspartate transaminase were measured for the liver functions in redox rats. Liver malondialdehyde (MDA), glutathione, oxidized glutathione, nitric oxide, superoxide dismutase (SOD) and catalase (CAT) were assessed as oxidative stress markers. The effect of S. aphanizomenoides on the transcripts level of superoxide dismutase (Mn-SOD) and catalase (CAT) genes in the rat's liver tissues was measured using qRT-PCR. Oral administration of S. aphanizomenoides extract in low and high doses (100, 200 mg kg-1 b.w) resulted in significant improvement in biochemical parameters of liver functions and oxidative stress markers. Also, the endogenous antioxidant defence enzymes and the expression of their related genes (Mn/SOD, CAT) were upregulated. Immunohistochemistry of Caspase-3, an apoptotic marker, showed potent amelioration in the liver tissues. CONCLUSIONS The novel isolate S. aphanizomenoides proved in vitro and in vivo antioxidant activity against redox rat model. SIGNIFICANCE AND IMPACT OF THE STUDY This isolate provides a new source of pharmacological compounds with great importance in pharmacological and medical fields.

Background Very small embryonic-like stem cells (VSELs) are a rare population within the ovarian epithelial surface. They contribute to postnatal oogenesis as they have the ability to generate immature oocytes and resist the chemotherapy. These cells express markers of pluripotent embryonic and primordial germ cells. Objective We aimed to explore the capability of VSELs in restoring the postnatal oogenesis of chemo-ablated rat ovaries treated with bone marrow-derived mesenchymal stem cells (BM-MSCs) combined with pregnant mare serum gonadotropin (PMSG). Methods Female albino rats were randomly assigned across five groups: I (control), II (chemo-ablation), III (chemo-ablation + PMSG), IV (chemo-ablation + MSCs), and V (chemo-ablation + PMSG + MSCs). Postnatal oogenesis was assessed through measurement of OCT4, OCT4A, Scp3, Mvh, Nobox, Dazl4, Nanog, Sca-1, FSHr, STRA8, Bax, miR143, and miR376a transcript levels using qRT-PCR. Expression of selected key proteins were established as further confirmation of transcript expression changes. Histopathological examination and ovarian hormonal assessment were determined. Results Group V displayed significant upregulation of all measured genes when compared with group II, III or IV. Protein expression confirmed the changes in transcript levels as group V displayed the highest average density in all targeted proteins. These results were confirmed histologically by the presence of cuboidal germinal epithelium, numerous primordial, unilaminar, and mature Graafian follicles in group V. Conclusion VSELs can restore the postnatal oogenesis in chemo-ablated ovaries treated by BM-MSCs combined with PMSG.

Salmonella spp is the main cause of foodborne salmonellosis that is considered a public health threat all over the world. The robust usage of antibiotics in Egyptian poultry farms resulted in increasing the prevalence of multi-drug resistant Salmonella enterica. In this study, the authors identify five multi-drug resistant Salmonella enterica serovars. Multidrug-resistant characteristics of these isolates were detected. The taxonomic evidence of these isolates was investigated based on 16S rRNA gene sequence and pairwise sequence alignment between the isolates' sequence and the nearest sequences in the database. In silico restriction maps and phylogenetic trees were also constructed. The obtained sequences were deposited in the database under accession numbers MW311328.1, MW311371.1, MN820824.1, MN822653.1 and MW310702.1 for Salmonella enterica subsp. enterica serovar Enteritidis strains EG.SmE1, EG.SmE2 and Salmonella enterica subsp. enterica serovar Typhimurium strains EG.SmT1, EG.SmT2, EG.SmT3, respectively. The five Salmonella isolates in this study showed multi-drug resistant characteristics. Salmonella Typhimurium isolate EG.SmT3 revealed resistance to more than five antibiotics. Results of pairwise sequence alignment, restriction maps and phylogenetic tree confirmed the close relationship between S. enteritidis isolates (EG.SmE1, EG. SmE2) and S. typhimurium isolates (EG.SmT1, EG.SmT2). However, the other S. typhimurium isolate (EG.SmT3) revealed the lowest identity ratio (98.6%) with the nearest sequence. The relative divergence of this isolate could be attributed to proposed mutations as a result of the vigorous use of antibiotics in Egyptian poultry farms.

Indigenous chicken breeds in developing countries have diverse benefits to rural economy as a source of high-quality animal protein. However, there are few reports on the evaluation of economic traits in Egyptian indigenous breeds. Hence, this study aimed to investigate growth performance, carcass characteristics, body measurements and meat quality traits in two indigenous breeds of chickens (Benha line and Golden Montazah) versus Rhode Island Red as a reference worldwide breed. Besides, a time series expression profile of somatotropic axis genes including GH and IGF-1 and their plasma level concentrations were investigated. Benha line chickens (BL) revealed the highest improved estimates of growth performance, carcass characteristics and meat quality traits. In the same manner, it displayed the highest levels of hepatic GH and IGF-1 and muscle IGF-1 gene expression compared to Rhode Island Red (RIR) and Golden Montazah (GM) chickens. Accordingly, BL exhibited the highest levels of plasma IGF-1 and the lowest levels of plasma GH. This result suggests the direct association between growth performance, carcass characteristics and levels of IGF-1 gene expression in the selected chicken breeds. BL is a superior Egyptian genotype with candidate productive traits and competing characteristics, it could be used widely as a proven ancestor of commercial hybrid breeds.

Background The use of zinc oxide in the form of nanoparticles (ZnO-NPs) is of great benefit due to its potent effectiveness and higher bioavailability compared to zinc oxide. This study aimed to investigate the impact of dietary inclusion of different doses of ZnO-NPs on selected serum biomarkers, lipid peroxidation and tissue gene expression of antioxidant enzymes and cytokines in Japanese quail. Eighty Japanese quails (Coturnix japonica) (45 days old) were randomly divided into four groups (20 birds for each) with 4 replicates (5 birds each). Birds in the first group were fed a basal diet alone and served as a control (C). Birds in groups 2–4 were fed the basal diet supplemented with ZnO-NPs at doses of 15 mg/kg, 30 mg/kg and 60 mg/kg for a period of 60 days. At the end of the experiment, all birds were sacrificed to collect blood in a plain vacutainer, whereas liver and brain tissues were stored frozen at -80 °C. The obtained sera were used for the analysis of selected biochemical parameters, whereas tissue homogenates were used for the estimation of zinc, oxidative stress biomarkers and gene expression of selected antioxidant enzymes and cytokines. Results ZnO-NPs (30 and 60 mg/kg) induced a significant decrease in serum triacylglycerol (TAG) compared to the control. ZnO-NPs did not affect the activities of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), total protein, albumin, globulin and tissue zinc concentrations but reduced the malondialdehyde (MDA) levels compared to the control. The liver retained a higher zinc concentration than that of brain tissue. In a dose-dependent manner, ZnO-NPs upregulated the mRNA levels of antioxidant enzymes (superoxide dismutase: SOD1; catalase: CAT; glutathione peroxidase-1: GPX 1) and pro-inflammatory cytokines (interferon α: IFN-α; interleukin 6: IL-6) in liver and brain tissues. Conclusion The current study suggests the inclusion of ZnO-NPs, particularly 60 mg/kg, in the diet of Japanese quails to improve antioxidant and immune status.

The presence of 4-nitrophenol (4-NP) in the wastewater industry causes toxicity and inhibition of the anaerobic degrading bacteria. The anaerobes in the multistage anaerobic reactor were loaded by 30.0 mg/gVS Graphene nanoparticles (MAR-Gn) as an electron acceptor to detoxify wastewater industry. The half maximal inhibitory concentration (IC50) was reduced from 455 ± 22.5 to 135 ± 12.7 μg Gallic acid equivalent/mL at 4-NP loading rate of 47.9 g/m3 d. Furthermore, 4-NP was decreased by a value of 83.7 ± 4.9% in MAR-Gn compared to 65.6 ± 4.8% in control MAR. The 4-aminophenol (4-AP) recovery was accounted for 44.8% in the MAR-Gn at an average oxidation-reduction potential (ORP) of − 167.3 ± 21.2 mV. The remaining portions of 4-NP and 4-AP in the MAR-Gn effluent were efficiently removed by baffled high rate algal pond (BHRAP), resulting in overall removal efficiency of 91.6 ± 6.3 and 92.3 ± 4.6%, respectively. The Methanosaeta (52.9%) and Methanosphaerula (10.9%) were dominant species in MAR-Gn for reduction of 4-NP into 4-AP. Moreover, Chlorophyta cells (Chlorella vulgaris, Scenedesmus obliquus, Scenedesmus quadricauda and Ulothrix subtilissima were abundant in the BHRAP for complete degradation of 4-NP and 4-AP.

Cyanobacteria are immense sources of several pharmacological compounds such as flavonoids and carotenoids with anti-inflammatory and antioxidant activity. Two novel cyanobacterial isolates were isolated and identified by sequencing of 16S rRNA gene. The results revealed that the two isolates are Sphaerospermopsis aphanizomenoides (KU212886.1) and Cronbergia siamensis (KY296358.1). The BLAST alignment showed that the nearest NCBI deposited sequences were Cronbergia siamensis (NR153750.1) to C. siamensis (KY296358.1) and Sphaerospermopsis aphanizomenoides (LN846950.1) to S. aphanizomenoides (KU212886.1) with identity ratio 94% and 93%, respectively. The phylogenetic trees confirmed the same identity ratios on the roots of clades. The two newly identified isolates could be new species with novel and unique characteristics.

Conjugated linoleic acid (CLA) is known for its multiple benefits including improvement of growth, increasing lean mass, and anti-carcinogenic effects. However, when used in long-term supplementations CLA does not improve semen parameters in boar and bull and reduces fertility in Japanese quails. The content of unsaturated fatty acids in dietary lipids plays a significant role in spermatogenesis owning the high proportion of unsaturated fatty acids in plasma membrane of sperms. Whether CLA plays a role in testicular tissue and epididymal fat is still unknown. Therefore, in this study we hypothesize that long-term supplementation of equal proportion of CLA isomer mix (c9,t11-CLA and t10,c12- CLA) in rabbit bucks might alter male reproductive potentials. Twelve V-Line weaned male rabbits were used in 26 weeks trial, rabbits were individually raised and randomly allocated into three dietary groups. Control group (CON) received a basal diet, a group received 0.5% CLA (CLA 0.5%), and a group received 1% CLA (CLA 1%). Rabbits were euthanized at the end of the trial and several parameters were evaluated related to growth, semen quality, and testicular and epididymal tissue histopathology and transcriptome. The long-term supplementation of CLA increased feed intake by 5% and body weight by 2-3%. CLA 1% decreased sperm progressive motility. In testicular tissue L-carnitine and α-tocopherol were decreased by CLA supplementation. In epididymal fat, CLA tended to decrease concentration of polyunsaturated fatty acids, the expression of SCD5 gene was upregulated by CLA 1% and CASP3 gene was upregulated by CLA 0.5%. Transcription of PPARG was downregulated by CLA. Feeding 1% CLA also decreased testicular epithelial thickness. Long-term supplementation of CLA modestly enhanced male rabbit growth, but negatively impacted male reproduction, especially at high dose of CLA.

Foodborne salmonellosis is a global threat to public health. In the current study, we describe the isolation and characterization of two broad-spectrum, lytic Salmonella phages: SPHG1 and SPHG3 infecting a multidrug-resistant Salmonella Typhimurium EG.SmT3. Electron microscopy and whole genome analysis identified SPHG1 as a Myovirus, while SPHG3 as a new member of the genus “Kuttervirus” within the family Ackermannviridae. SPHG1 and SPHG3 had a lysis time of 60 min. with burst sizes of 104 and 138 PFU/cell, respectively. The two phages were robust at variable temperatures and pH ranges that match the corresponding values of most of the food storage and processing conditions. A phage cocktail containing the two phages was stable in the tested food articles for up to 48 h. The application of the phage cocktail at MOIs of 1000 or 100 resulted in a significant reduction in the viable count of S. Typhimurium by 4.2 log10/sample in milk, water, and on chicken breast. Additionally, the phage cocktail showed a prospective ability to eradicate and reduce the biofilm that formed by S. Typhimurium EG.SmT3. A phage cocktail of SPHG1 and SPHG3 is considered as a promising candidate as a biocontrol agent against foodborne salmonellosis due to its broad host ranges, highly lytic activities, and the absence of any virulence or lysogeny-related genes in their genomes.

The limited rabbit resources in Egypt are threatened by the danger of extinction, whereas genetic diversity studies of native breeds could play a vital role in conservation and improvement of these breeds. In this study, 3 native rabbit breeds: Gabali (G), Baladi Red (BR) and Baladi Black (BB), in addition to New Zealand White (NZW), were genotyped using 12 microsatellite markers. All the typed microsatellites were polymorphic by average number of alleles 5.25 per locus. Observed and expected heterozygosity per locus averaged 0.62 and 0.68, respectively. The average polymorphic information content was 0.71 and the highest polymorphic information content was recorded in locus SOL33 by 0.85. All the studied loci except SAT7 and SAT2 showed deviation from Hardy-Weinberg equilibrium with significant level. The inbreeding coefficient of the individuals relative to the total population was 0.07. The within-population heterozygote deficit averaged 0.07 and ranged from 0.141 in BR to 0.015 in BB breeds. The highest pairwise differentiation among the populations was recorded between BB and NZW (0.071), while the lowest value was recorded between BR and both of G (0.038) and BB (0.039). The lowest pairwise Nei’s genetic distance was recorded between BR and BB (0.190), while the highest was recorded between NZW and BB breeds (0.409). BR and G populations were clustered together forming an admixed mosaic cluster. BR recorded the highest contribution in the aggregate genetic diversity based on the three prioritisation methods used.

Cyanobacteria are natural enormous sources of various biologically active compounds with great contributions in different industries. This study aimed to introduce molecular and biochemical characterization for four novel cyanobacterial isolates obtained from Egyptian wastewater canals. Besides, In vitro biological activity of these isolates and their potential ability to take up nutrients and heavy metals from wastewater were examined. The obtained accession numbers were KY250420.1, KY321359.1, KY296359.1 and KU373076.1 for Nostoc calcicola, Leptolyngbya sp, Nostoc sp, and Nostoc sp, respectively. The isolate Leptolyngbya sp (KY321359.1) showed the lowest identity (90%) with other deposited sequences in database. While the isolate Nostoc sp (KU373076.1) showed the highest total phenolic content as well as the highest levels of caffeic, ferulic and gallic acids. Consequently, it appeared the highest antioxidant scavenging activity. All cyanobacterial isolates revealed potent ability to take up nutrients and heavy metals from wastewater. Generally, this study provides a taxonomic and molecular evidence for four novel cyanobacterial isolates with antioxidant activity and potent phycoremediation ability.

Cyanobacteria are natural enormous sources of various biologically active compounds with great contributions in different industries. This study aimed to explore and characterize novel cyanobacterial isolates with antioxidant activity and potent phycoremediation ability from Egyptian wastewater canals. The in vitro biological activity of these isolates and their potential ability to take up nutrients and heavy metals from wastewater were examined. The obtained isolates were sequenced and deposited in database under accession numbers, KY250420.1, KY321359.1, KY296359.1 and KU373076.1 for Nostoc calcicola, Leptolyngbya sp., Nostoc sp., and Nostoc sp., respectively. Leptolyngbya sp. (KY321359.1) showed the lowest identity (90%) with the nearest deposited sequence in database. While the isolate Nostoc sp. (KU373076.1) showed the highest total phenolic content as well as the highest levels of caffeic, ferulic and gallic acids. Consequently, it presented the highest antioxidant scavenging activity. All studied isolates revealed potent ability in chelating nutrients and removing heavy metals from wastewater. In conclusion, this study provides a taxonomic, biochemical and molecular evidence of four novel cyanobacterial isolates with antioxidant activity and potential phycoremediation ability.