Phytochemical profiling, cytotoxic, anti-migration, and anti-angiogenic potential of phenolic-rich fraction from Peganum harmala: in vitro and in ovo studies.

Peganum harmala has been extensively employed in Algerian traditional medicine practices. This study aimed to explore the impact of n-butanol (n-BuOH) extract sourced from Peganum harmala seeds on cell proliferation, cell migration, and angiogenesis inhibition. Cytotoxic potential of n-BuOH extract was evaluated using MTT (3-(4,5-dimethylthiazol-2-yl) 2,5 diphenyltetrazolium bromide) assay against human breast adenocarcinoma MCF-7 cells, cell migration was determined using scratch assay, and anti-angiogenic effect was evaluated through macroscopic and histological examinations conducted on chick embryo chorioallantoic membrane. Additionally, this research estimated the phytochemical profile of n-BuOH extract. Fifteen phenolic compounds were identified using Ultra-performance liquid chromatography UPLC-ESI-MS-MS analysis. In addition, the n-BuOH extract of P. harmala exhibited potent antioxidant and free radical scavenging properties. The n-BuOH extract showed potent cytotoxicity against MCF-7 cell with an IC50 value of 8.68 ± 1.58 µg/mL. Furthermore, n-BuOH extract significantly reduced migration. A strong anti-angiogenic activity was observed in the groups treated with n-BuOH extract in comparison to the negative control. Histological analysis confirmed the anti-angiogenic effect of the n-BuOH extract. This activity is probably a result of the synergistic effects produced by different polyphenolic classes.

Screening of Wild Lactic Acid Bacteria from Algerian Traditional Cheeses and Goat Butter to Develop a New Probiotic Starter Culture.

Twenty-five lactic acid bacterial (LAB) strains have been isolated from traditional goat butter and three types of cheese (dry Klila, frech Klila, and Bouhezza) and evaluated for technological abilities, probiotic properties, and potentials as starter cultures. The twenty-five LAB strains comprised eight strains belonging to Lactobacillus, four strains belonging to Lactococcus, eleven strains belonging to Enterococcus, and two strains belonging to Leuconostoc. A non-hierarchical cluster analysis was performed in order to select the performing strains. After carrying out the preliminary phenotypic characterizations and the probiotic potential, three strains designated as BM10, B15, and C30 belonging to the genus Lactobacillus and Enterococcus with good tolerance to acidity were selected. The strains showed a significant resistance to 0.5% bile salts and 0.4% phenol. Hemolytic activity was not detected; in addition, good hydrophobicity and autoaggregation was obtained. A significant antimicrobial activity was exhibited by all selected strains against Listeria innocua. Genotypic identification by 16S rRNA allowed the identification of B15, BM10, and C30 as Lactobacillus plantarum, Lactobacillus casei, and Enterococcus durans, respectively. The results of the current study suggest that the strains isolated from Algerian fermented dairy products have high potential as probiotic starter cultures in the goat butter and cheese industry.

Biodegradation of chlorpyrifos by a Weissella confusa strain and evaluation of some probiotic traits.

Pesticides play an important role in agriculture; however, their excessive use causes several problems such as pollution of ecosystems and risks to human health. The presence of microorganisms able to degrade these pollutants can reduce their negative effect. The objective of this study was to test the capacity of Weissella confusa Lb.Con to tolerate or to degrade the chlorpyrifos pesticide. The results showed the capacity of the strain to tolerate a concentration of 200 µg/ml of chlorpyrifos. The strain Lb.Con has a remarkable capacity to grow in glucose-free MRS medium which contains different concentrations of chlorpyrifos. HPLC analysis showed that this strain was able to remove about 25% of chlorpyrifos. The evaluation of some probiotic properties showed that the strain Lb.Con had a remarkable resistance to the gastrointestinal conditions and a good antibacterial activity towards the pathogenic bacteria. The probiotic potential was evaluated to verify the possible use of W. confusa Lb.Con to detoxify harmful chlorpyrifos contained in food.

Removal of diclofenac by a local bacterial consortium: UHPLC-ESI-MS/MS analysis of metabolites and ecotoxicity assessment.

Diclofenac (DCF) belongs to the class of nonsteroidal anti-inflammatory drugs, which is one of the most consumed by population and detected in raw sewage. Several studies have reported variable removal rates by biodegradation of diclofenac in wastewater treatment plants (WWTPs). This study deals with the evaluation of the biodegradation of DCF by a bacterial consortium (obtained from pure cultures of Enterobacter hormaechei D15 and Enterobacter cloacea D16), which were isolated from household compost and Algerian WWTP, respectively, as sole carbon source and by co-metabolism, using glucose as carbon source. A 98% removal rate of DCF was observed when it is used as the sole carbon source, whilst only 44% of DCF was removed in co-metabolic conditions. Two metabolites were identified using ultra-high-performance liquid chromatography coupled to electrospray injection tandem mass spectrometry analysis (UHPLC-ESI-MS/MS); one of them was identified as 4'-hydroxy-DCF, and the second metabolite was suspected to be a nitro derivative of DCF, according to comparison with the literature. Biodegradation of DCF by this bacterial consortium generates relatively safe final by-products.

Effect of probiotic supplementation on oxidative stress markers in rats with diclofenac-induced hepatotoxicity.

In this study, we have investigated the effect of an antioxidant probiotic pretreatment toward an overdose of diclofenac in rats (100 mg/kg bw). Rats were treated daily with the probiotic Streptococcus salivarius St.sa (109 CFU) during seven successive days and then received a single treatment with diclofenac overdose in distilled water. Liver transaminases (alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase), histology, glutathione (GSH) and malondialdehyde (MDA) level were investigated. In addition, both antioxidant enzyme activity and its mRNA gene expression were studied to evaluate diclofenac hepatotoxicity. The results indicated that probiotic pretreatment reduced diclofenac-induced hepatotoxicity through enhancement of the studied hepatic markers and regulation of antioxidant enzyme expression and activity. These findings indicate that the probiotic pretreatment protects rat liver against the oxidative stress induced by diclofenac overdose.

Assessment of Streptococcus salivarius sp thermophiles Antioxidant Efficiency and its Role in Reducing Paracetamol Hepatotoxicity.

BACKGROUND: Probiotics have attracted a great attention aiming to develop natural non-toxic antioxidants, because of their role in decreasing the risk of reactive oxygen species [ROS] accumulation. OBJECTIVES: The purpose of this study was to assess the antioxidant activity of a probiotic Streptococcus salivarius ssp thermophillus [St.sa] and to evaluate its protective effect against the oxidative stress induced by a toxic dose of paracetamol in Wistar rats. MATERIALS AND METHODS: Several assays were used to investigate the in vitro antioxidant capacity of the strain. To evaluate the protective effect against oxidative stress induced by paracetamol in liver, hepatic marker enzymes, the antioxidant enzyme activities, malondialdehyde [MDA] and glutathione [GSH] content in liver tissues were investigated. RESULTS: The strain has shown a considerable ability to scavenge DPPH free radical [89.43%],a good resistance to hydroxyl radicals [47%], a considerable ability to chelate iron ions [33.21%] and a good inhibitory effect against plasma lipid peroxidation [54.36%]. Significant changes in liver function tests, antioxidant enzyme activities, MDA and GSH levels in paracetamol treated group were obtained compared to control group. Pretreatment with probiotic removed significantly the inhibition of antioxidant enzymes and suppressed MDA increase and GSH depletion. The analysis of the level of mRNA expression of antioxidant enzymes showed no significant differences in the expression of the enzymes in treated or non-treated groups. CONCLUSION: This finding emphasizes the protective role of probiotics against ROS generated during the treatment with paracetamol.

Metabolic and Co-Metabolic Transformation of Diclofenac by Enterobacter hormaechei D15 Isolated from Activated Sludge.

The presence of non-steroidal anti-inflammatory drugs, such as diclofenac (DCF), in the environment, is an emerging problem due to their harmful effects on non-target organisms, even at low concentrations. We studied the biodegradation of DCF by the strain D15 of Enterobacter hormaechei. The strain was isolated from an activated sludge, and identified as E. hormaechei based on its physiological characteristics and its 16 S RNA sequence. Using HPTLC and GC-MS methods, we demonstrated that this strain metabolized DCF at an elimination rate of 52.8%. In the presence of an external carbon source (glucose), the elimination rate increased to approximately 82%. GC-MS analysis detected and identified one metabolite as 1-(2,6-dichlorophenyl)-1,3-dihydro-2H-indol-2-one; it was produced as a consequence of dehydration and lactam formation reactions.

Biological Removal of the Mixed Pharmaceuticals: Diclofenac, Ibuprofen, and Sulfamethoxazole Using a Bacterial Consortium.

Background: The presence of pharmaceuticals at low concentrations (ng to µg) in the environment has become a hot spot for researchers in the past decades due to the unknown environmental impact and the possible damages they might have to the plantae and fauna present in the aquatic systems, as well as to the other living organisms. Objectives: The aim of the present investigation was to develop a bacterial consortium isolated from different origins to evaluate the ability of such a consortium to remove a mixture of pharmaceuticals in the batch system at lab scale, as well as assessment of its resistance to the other micropollutants present in the environment. Material and Methods: Using a closed bottle test, biodegradation of the mixed pharmaceuticals including Diclofenac (DCF), Ibuprofen (IBU), and Sulfamethoxazole (SMX) (at a concentration of 3 mg.L-1 of each drug) by the bacterial consortium was investigated. The test was carried out under metabolic (pharmaceutical was used as the sole source of carbon) and co-metabolic condition (in the presence of glucose). Finally, the ability of the bacterial consortium to resist other micropollutants like antibiotics and heavy metals was investigated. Results: Under the metabolic condition, the mixed bacteria (i.e., consortium) were able to metabolize 23.08% and 9.12% of IBU, and DCF at a concentration of 3 mg.L-1 of each drug, respectively. Whereas, in co-metabolic conditions, IBU was eliminated totally, in addition, 56% of the total concentration of DCF was removed, as well. In both metabolic and cometabolic conditions, removal of SMX was not observed. The selected bacteria were able to resist to most of the applied antibiotics and the used heavy metals, except mercury, where only one strain (S4) was resistant to the later heavy metal. Conclusion: Results suggest that the developed consortium might be an excellent candidate for the application in the bioremediation process for treating ecosystems contaminated with the pharmaceutical.

Technological and Probiotic Traits of the Lactobacilli Isolated From Vaginal Tract of the Healthy Women for Probiotic Use.

BACKGROUND: For biotechnological application, selected lactic acid bacteria strains belonging to the genera Lactobacillus (Lb) are proposed as an alternative to the antibiotics for the prevention and treatment of urogenital tract infections. OBJECTIVES: Isolating and selecting vaginal lactobacilli strains for probiotic use based on their technological and probiotic aptitudes. MATERIALS AND METHODS: The vaginal isolates were examined for their essential characteristics as the potential probiotic such as low pH tolerance, bile-salt and simulated human intestinal fluid (SIF) resistance, adhesion to the vaginal epithelial cells (VECs), aggregation and coaggregation, surface hydrophobicity, antimicrobial activity, acid production, antibiotic resistance, and resistance to spermicides. The best strain was identified by PCR. RESULTS: From 70 lactobacilli isolates and according to the 16 rDNA sequences, isolates B6 and B10 showed the closest homology (99%) to the Lb. gasseri and Lb. plantarum respectively. They produced hydrogen peroxide (H2O2), tolerant to acid, bile, simulated human intestinal fluid, present a strong adhesion, highest percentages of aggregation, and antibacterial activity. These strains are resistant to the spermicide and actively acidify the growth medium. CONCLUSIONS: Strains Lb. plantarum B10 and Lb. gasseri B6 have a strong potential probiotic confirming their value as a tool for prevention against urinary and vaginal infections.

Enhanced production of lipase by the thermophilic Geobacillus stearothermophilus strain-5 using statistical experimental designs.

Statistically based experimental designs were applied to optimize the cultural conditions for the production of a glycerol-inducible lipase from the thermophilic Geobacillus stearothermophilus strain-5. The effect of nineteen culture conditions on enzyme production was evaluated using Plackett-Burman factorial design. Tween 80, K(2)HPO(4), glycerol and glucose were the most significant factors in improving enzyme production. The selected parameters were then further investigated using central composite design to define the optimal process conditions. Maximal enzyme activity (578 U/ml) was reached under the following conditions: glycerol, 2.24% (v/v); Tween 80, 0.76% (v/v); glucose, 0.76% (w/v) and K(2)HPO(4), 0.38% (w/v) which is about five folds the activity in basal medium. A verification experiment was carried out to examine model validation and revealed more than 98% validity.

Emulsification properties of biosurfactant produced from Pseudomonas aeruginosa RB 28.

Biosurfactant produced from P. aeruginosa RB 28 was extracted, purified and characterized. Thin layer chromatography results showed that the extract contained two different compounds. The identification of the nature of the two compounds showed that they were glycolipids and rhamnose was the sugar moiety in these glycolipids. It was concluded that these compounds were rhamnolipids. The production of biosurfactant was started at late log phase and reached its maximal level (2.7 g L(-1)) at the stationary phase. Study of some rhamnolipid properties showed that sunflower oil, heptadecane and paraffin were efficiently emulsified and emulsions formed with vegetable oils (olive oil, corn oil and sunflower oil) were more stable than emulsions formed with hydrocarbons.