Effects of the putative probiotics Bacillus licheniformis, Bacillus pumilus, and Bacillus subtilis on white leg shrimp, Litopenaeus vannamei, immune response, gut histology, water quality, and growth performance.

Background: A commercially significant species in the aquaculture sector globally, particularly in Egypt, is Litopenaeus vannamei. Aim: The experiment's objective was to ascertain how Sanolife PRO-F impacted the growth, water quality, immunological response, and intestinal morphometry of L. vannamei. Methods: In the current investigation, which lasted 12 weeks, Sanolife PRO-F was administered to shrimp post-larvae at diet doses of 0 (control), 1 (group one), 2 (group two), and 3 (group three) g/kg diet, respectively. Each experimental group had three repetitions. Results: In the current study, shrimp fed on probiotic-treated diets showed a considerable improvement in growth performance measures and survival rate, and the nonspecific immune response was also enhanced. Shrimp fed probiotic diets had longer and more intestinal villi overall. Shrimp fed on the G2 and G3 diets showed no appreciable differences in growth or intestinal morphology. With the G2 and G3 diet, the water had lower concentrations of nitrite and ammonia. Conclusion: The study's findings indicate that Sanolife PRO-F treatment at 2-3 g/kg feed promotes the growth of shrimp, immunological response, gut health and function, and water quality.

New Apigenin Glycoside, Polyphenolic Constituents, Anti-inflammatory and Hepatoprotective Activities of Gaillardia grandiflora and Gaillardia pulchella Aerial Parts.

BACKGROUND: Gaillardia grandiflora Hort. ex Van Houte and Gaillardia pulchella Foug are flowering plants widely cultivated in Egypt for their ornamental value. Previous reports demonstrated that sesquiterpene derivatives represent the major compounds in both species. Moreover, only few flavones were identified from genus Gaillardia and few studies on the cytotoxicity of G. pulchella were found. AIM OF THE STUDY: Investigation of the phenolic constituents of the aerial parts of both species and evaluation of their anti-inflammatory and hepatoprotective activities. MATERIALS AND METHODS: The 80% aqueous methanol extracts (AME) were prepared for both plants and evaluated for their biological activities. Phytochemical investigation of both extracts resulted in isolation of twelve compounds, which have been identified on the basis of ultraviolet, 1D and 2D nuclear magnetic resonance spectroscopy and negative ESI-MS. RESULTS: The new 8-hydroxyapigenin 6-O-ß-D-apiofuranosyl-(1'''→6'')-C-ß-D-4C1-glucopyranoside was isolated from G. grandiflora for the first time in nature, along with schaftoside, luteolin 6-C-ß-D-4C1-glucopyranoside 8-methyl ether, apigenin 6-C-ß-D-4C1-glucopyranoside 8-methyl ether, isoorientin, isovitexin, 6-methoxyluteolin and hispidulin, as well as vicenin-2, vitexin, luteolin and apigenin, which were isolated from G. pulchella together with 6-methoxyluteolin. Furthermore, the AME of both species were found to be nontoxic to mice and exhibited significant anti-inflammatory and hepatoprotective activities in dose dependent manner. CONCLUSION: Current results shed light on the phenolic constituents of G. grandiflora and G. pulchella aerial parts and the safety of the AME of both species, in addition to their significant anti-inflammatory and hepatoprotective activities. Both plant species may be promising candidates for natural anti-inflammatory and hepatoprotective drugs. SUMMARY: Phytochemical investigation of Gaillardia grandiflora and Gaillardia pulchella 80% aqueous methanol extracts of the aerial parts led to the isolation of twelve compoundsThe new compound 8-hydroxyapigenin 6-O-ß-D-apiofuranosyl-(1''''→6'')-C-ß-D-4C1-glucopyranoside was isolated from G. grandiflora for the first time in natureSchaftoside, luteolin 6-C-ß-D-4C1-glucopyranoside 8-methyl ether, apigenin 6-C-ß-D-4C1-glucopyranoside 8-methyl ether, isoorientin, isovitexin, 6-methoxyluteolin and hispidulin were isolated from G. grandifloraVicenin-2, vitexin, luteolin, apigenin and 6-methoxyluteolin were isolated from G. pulchellaThe extracts of both species were nontoxic to mice up to 5 g/kg body weightBoth extracts exhibited significant anti-inflammatory and hepatoprotective activities in dose dependent manner Abbreviations used: ALP: Alkaline phosphatase; ALT: Alanine aminotransferase; AME: The 80% aqueous methanol extract of G. grandiflora or G. pulchella aerial parts; AST: Aspartate aminotransferase; br d: Broad doublet; Comp-PC: Comparative paper chromatography; d: Doublet; 2D-PC: Two-dimensional paper chromatography; DMSO-d6: Deuterated dimethyl sulfoxide; G.: Gaillardia; GPx: Glutathione peroxidase; GRd: Glutathione reductase; GSH: glutathione; GST: Glutathione-S-transferase; J: Nuclear spin-spin coupling constant; m: Multiplet; [M-H]-: Molecular ion peak; MDA: Malondialdehyde; m/z: Mass/charge ratio; NO: Nitric oxide; p: Probability; PC: Paper chromatography; Rf: Retention flow; rpm: Rotation per minute; s: Singlet; SDE: The ethanol extract of Scoparia dulcis; SE: Standard error; SOD: Superoxide dismutase; TMS: Tetramethylsilane; λmax: Maximum fluorescence emission wavelength.

Spectroscopic identification of new ellagitannins and a trigalloyl-glucosylkaempferol from an extract of Euphorbia cotinifolia L. with antitumour and antioxidant activity.

From an extract of leaves and small branches of Euphorbia cotinifolia L., 17 polyphenols were isolated including two new ellagitannins and a trigalloyl-glucosylkaempferol. Based on extensive spectral data (UV, ESI-MS, 1H NMR, DEPT and 1D/2D NMR) and chemical studies, their structures were characterized as 1-O-galloyl-3,6-hexahydroxydiphenoyl-D-B1,4-glucopyranose (5), 1-O-galloyl-3,6-valoneoyl-D-B1,4-glucopyranose (6), and kaempferol 3-O-(2",3",6"-tri-O-galloyl)-beta-D-glucopyranoside (13). Biological evaluation indicated that the 80% aqueous methanol extract (AME), chloroform extract (CE), and some pure compounds have potent scavenging activity in the DPPH assay with SC50 values lower than that of ascorbic acid, especially 5, 7-9, and a mixture of hyperin 6"-gallate (11) and isoquercitrin 6"-gallate (12). Moreover, using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) cell viability assay, 6 and 8 exhibited the highest inhibition of human hepatocellular carcinoma cells (Hep-G2), while AME, CE, 5, 7, 9, and the mixture of 11 and 12 were found to be moderate growth inhibitors according to their IC50 values. In addition, AME, 5, and 8 exhibited significant antiproliferative activity against colon carcinoma cells (HCT-116); however, CE and the other examined compounds displayed moderate to low antitumour activity against HCT-116 cells.

Polyphenolic profile and bioactivity study of Oenothera speciosa Nutt. aerial parts.

Two new flavonol glycosides, myricetin 4'-O-alpha-L-rhamnopyranoside (1) and quercetin 3'-O-alpha-L-rhamnopyranoside (2), together with a novel biflavonol compound, speciin (3), as well as eleven phenolic metabolites, namely myricitrin (4), europetin 3-O-alpha-L-(1)C(4)-rhamnopyranoside (5), quercitrin (6), hyperin (7), rhamnetin 3-O-beta-galacto-pyranoside (8), caffeic acid (9), caffeic acid methyl ester (10), chlorogenic acid (11), chlorogenic acid methyl ester (12), gallic acid (13) and gallic acid methyl ester (14), were identified from the 80 % methanol extract of the aerial parts (leaves and stems) of Oenothera speciosa Nutt. (Onagraceae). In addition myricetin (15), quercetin (16) and ellagic acid (17) were identified from the chloroform extract. The structures were established depending on their chemical and physical analyses (UV, HR-ESIMS, 1D and 2D NMR). It was found that 80 % aqueous methanol extract of O. speciosa is non-toxic to mice up to 5 g kg(-1)b wt. The investigated extract exhibited significant antihyperglycaemic and anti-inflammatory activities in a dose dependant manner. Also, the 80 % methanol extract, myricitrin(4) and hyperin(7) showed potent antioxidant activity in vitro using 1,1-diphenyl 2-picryl hydrazyl (DPPH) radical assay.

Antidiabetic and antioxidant activities of major flavonoids of Cynanchum acutum L. (Asclepiadaceae) growing in Egypt.

Seven flavonoids were isolated from the butanol fraction of the methanolic extract of the aerial parts of Cynanchum acutum L. (Asclepiadaceae). All of which have been isolated for the first time from the genus Cynanchum. Their structures were established as quercetin 3-O-beta-galacturonopyranoside (1), quercetin 7-O-beta-glucopyranoside (2), tamarixtin 3-O-beta-galacturonopyranoside (3), kaempferol 3-O-beta-galacturonopyranoside (4), 8-hydroxyquercetin 3-O-beta-galactopyranoside (5), tamarixtin 3-O-alpha-rhamnopyranoside (6), and tamarixtin 7-O-alpha-arabinopyranoside (7) on the basis of their chromatographic properties, chemical and spectroscopic data. The major isolated flavonoids 1, 2 and 3 were found to exhibit significant antioxidant and antidiabetic activities (by measuring blood glucose and insulin levels). This is the first report about the antioxidant and antidiabetic activities of compounds 1-3.

An acylated flavonol glycoside and hydrolysable tannins from Callistemon lanceolatus flowers and leaves.

INTRODUCTION: Callistemon lanceolatus DC. (Myrtaceae) is a plant rich in polyphenols, and is used as anticough, antibronchitis and insecticide in folk medicine. Because of the biological importance of plant polyphenols, particularly tannins, a phytochemical study was of interest to investigate the constitutive poyphenols in the extracts of flowers and leaves. OBJECTIVE: To avoid time-consuming methodology for isolation of a complex mixture of known metabolites, HPLC-ESI/MS was employed for fast picking up of the new compounds followed by identification of the structures with UV and one- and two-dimensional NMR. METHODOLOGY: Flowers and leaves were separately extracted with hot aqueous methanol under reflux (70 degrees C). Pre-isolation of the total extracts was achieved through column chromatographic fractionation on polyamide with water-methanol for gradient elution. The main fractions were purified using repeated column chromatography on cellulose and/or Sephadex LH-20 with suitable eluents. HPLC-ESI/MS analyses were carried out in the single ion monitoring (SIM) and negative ion modes. The pure compounds in methanol-water (1:1) were analysed by direct infusion ESI/MS. Final structure elucidation was obtained by one- and two-dimensional NMR. RESULTS: Two new metabolites namely quercetin 3-O-beta-D-glucuronopyranoside n-butyl ester (1) and n-butylgallate 4-O-(2',6'-di-O-galloyl)-beta-D-glucopyranoside (4) along with nine known ones were identified from the aqueous methanol extracts of flowers and leaves. CONCLUSION: The study has shown that Callistemon lanceolatus is rich in polyphenols. HPLC-ESI/MS may be used, in negative ion mode, as an efficient and rapid analytical tool for investigating complex plant extracts.

Isoflavonoid glycosides and rotenoids from Pongamia pinnata leaves.

Chromatographic separation of a 70% aqueous methanol extract (AME) of Pongamia pinnata (Linn.) Pierre (Leguminosae) leaves has led to the isolation of two new isoflavonoid diglycosides, 4'-O-methyl-genistein 7-O-beta-D-rutinoside (2) and 2',5'-dimethoxy-genistein 7-O-beta-D-apiofuranosyl-(1"'-->6")-O-beta-D-glucopyranoside (6), and a new rotenoid, 12a-hydroxy-alpha-toxicarol (5), together with nine known metabolites, vecinin-2 (1), kaempferol 3-O-beta-D-rutinoside (3), rutin (4), vitexin (7), isoquercitrin (8), kaempferol 3-O-beta-D-glucopyranoside (9), 11,12a-dihydroxy-munduserone (10), kaempferol (11), and quercetin (12). Their structures were elucidated on the basis of chemical and spectroscopic analyses.

Anticancer and antioxidant tannins from Pimenta dioica leaves.

Two galloylglucosides, 6-hydroxy-eugenol 4-O-(6'-O-galloyl)-beta-D-4C1-glucopyranoside (4) and 3-(4-hydroxy-3-methoxyphenyl)-propane-1,2-diol-2-O-(2',6'-di-O-galloyl)-beta-D -4C1-glucopyranoside (7), and two C-glycosidic tannins, vascalaginone (10) and grandininol (14), together with fourteen known metabolites, gallic acid (1), methyl gallate (2), nilocitin (3), 1-O-galloyl-4,6-(S)-hexahydroxydiphenoyl-(alpha/beta)-D-glucopyranose (5), 4,6-(S)-hexahydroxydiphenoyl-(alpha/beta)-D-glucopyranose (6), 3,4,6-valoneoyl-(alpha/beta)-D-glucopyranose (8), pedunculagin (9), casuariin (11), castalagin (12), vascalagin (13), casuarinin (15), grandinin (16), methyl-flavogallonate (17) and ellagic acid (18), were identified from the leaves of Pimenta dioica (Merr.) L. (Myrtaceae) on the basis of their chemical and physicochemical analysis (UV, HRESI-MS, 1D and 2D NMR). It was found that 9 is the most cytotoxic compound against solid tumour cancer cells, the most potent scavenger against the artificial radical DPPH and physiological radicals including ROO*, OH*, and O2-*, and strongly inhibited the NO generation and induced the proliferation of T-lymphocytes and macrophages. On the other hand, 3 was the strongest NO inhibitor and 16 the highest stimulator for the proliferation of T-lymphocytes, while 10 was the most active inducer of macrophage proliferation.

Antioxidant flavonol glycosides from Schinus molle.

Chromatographic separation of aqueous MeOH extract of the leaves of Schinus molle L. has yielded two new acylated quercetin glycosides, named isoquercitrin 6''-O-p-hydroxybenzoate (12) and 2''-O-alpha-L-rhamnopyranosyl-hyperin 6''-O-gallate (13), together with 12 known polyphenolic metabolites for the first time from this species, namely gallic acid (1), methyl gallate (2), chlorogenic acid (3), 2''-alpha-L-rhamnopyranosyl-hyperin (4), quercetin 3-O-beta-D-neohesperidoside (5), miquelianin (6), quercetin 3-O-beta-D-galacturonopyranoside (7), isoquercitrin (8), hyperin (9), isoquercitrin 6''-gallate (10), hyperin 6''-O-gallate (11) and (+)-catechin (14). Their structures were established on the basis of chromatographic properties, chemical, spectroscopic (UV, 1H, 13C NMR) and ESI-MS (positive and negative modes) analyses. Compounds 4-9 and 11 exhibited moderate to strong radical scavenging properties on lipid peroxidation, hydroxyl radical and superoxide anion generations with the highest activities shown by 6 and 7 in comparison with that of quercetin as a positive control in vitro.

Phytochemical constituents and hepatoprotective activity of Viburnum tinus.

From the leaves of Viburnum tinus L. (Adoxaceae) two acylated iridoid glucosides (viburtinoside A and B), a coumarin diglucoside scopoletin 7-O-beta-D-sophoroside and a natural occurred dinicotinic acid ester 2,6-di-C-methyl-nicotinic acid 3,5-diethyl ester were isolated. In addition to these, 10 known compounds were isolated, namely two bidesmosidic saponins, a hexamethoxy-flavone and five flavonol glycosides, as well as suspensolide A and oleanolic acid were isolated for the first time in this genus and species, respectively. The structures were determined mainly by spectroscopic methods (UV, IR, ESI-MS, 1H-, 13C NMR and DEPT). Toxicity of the investigated extract was determined (LD50=500 mg/kg). CCl4-induced hepatotoxicity has been evaluated in terms of the determination of alanine aminotransferase (ALT), aspartate aminotransferase (AST), lipid peroxide and nitric oxide levels in serum and compared using adult male rats weighing 150-180 g. Their highly elevated levels were significantly reduced by treatment with the investigated aqueous methanol extract in dose-dependent manner.

Pharmacologically active ellagitannins from Terminalia myriocarpa.

A new ellagitannin, methyl (S)-flavogallonate (14) along with fourteen known compounds, gallic acid, methyl gallate, ethyl gallate, 2,3-di-O-[( S)-4,5,6,4',5',6'-hexahydroxybiphenyl-2,2'-diyldicarbonyl]-(alpha/beta)-D-glucopyranose (4), vitexin, isovitexin, orientin, iso-orientin, kaempferol 3-O-beta-D-rutinoside, rutin, neosaponarin, ellagic acid, flavogallonic acid (13), and (alpha/beta)-punicalagin (15) have been isolated from the leaves of Terminalia myriocarpa Heurck. Protective effect of the major and structurally related compounds 4, 13, 15 and the new compound 14 against CCl 4 -induced hepatotoxicity has been evaluated and compared, using adult male rats weighing 200-250 g. Serum levels of glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), lipid peroxide and nitric oxide production were significantly increased by administration of CCl 4 to rats and then reduced significantly only by treatment with compounds 4, 14 and 15 in a dose-dependent manner. Comparison of the protective properties of these compounds showed that compound 14 is more potent than compound 15 than 4 and that compound 13 has a non-significant effect at the used two dose levels.