Water toxicity evaluations: Comparing genetically modified bioluminescent bacteria and CHO cells as biomonitoring tools.

The use of water for drinking and agriculture requires knowledge of its toxicity. In this study, we compared the use of genetically modified bioluminescent (GMB) bacteria whose luminescence increases in the presence of toxicants and Chinese Hamster Ovary (CHO) cells for the characterization of the toxicity of water samples collected from a lake and streams, hydroponic and aquaponic farms, and a wastewater treatment plant. GMB bacteria were used to probe genotoxicity, cytotoxicity and reactive oxygen species-induced effects in the whole water samples. Unlike GMB bacteria, the use of CHO cells requires XAD resin-based pre-concentration of toxic material present in water samples for the subsequent cytotoxicity assay. In addition to the examination of the toxicity of the water from the different sources, the GMB bacteria were also used to test the XAD extracts diluted to the concentrations causing 50% growth inhibition of the CHO cells. The two biomonitoring tools provided different results when they were used to test the above-mentioned diluted XAD extracts. A pre-concentration procedure based on adsorption by XAD resins with subsequent elution was not sufficient to represent the material responsible for the toxicity of the whole water samples toward the GMB bacteria. Therefore, the use of XAD resin extracts may lead to major underestimates of the toxicity of water samples. Although the toxicity findings obtained using the GMB bacteria and CHO cells may not correlate with each another, the GMB bacteria assay did provide a mechanism-specific biomonitoring tool to probe the toxicity of water samples without a need for the pre-concentration step.

Synergistic antibacterial effects of polyphenolic compounds from olive mill wastewater.

Polyphenols or phenolic compounds are groups of secondary metabolites widely distributed in plants and found in olive mill wastewater (OMW). Phenolic compounds as well as OMW extracts were evaluated in vitro for their antimicrobial activity against Gram-positive (Streptococcus pyogenes and Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli and Klebsiella pneumoniae). Most of the tested phenols were not effective against the four bacterial strains when tested as single compounds at concentrations of up to 1000 µg mL(-1). Hydroxytyrosol at 400 µg mL(-1) caused complete growth inhibition of the four strains. Gallic acid was effective at 200, and 400 µg mL(-1) against S. aureus, and S. pyogenes, respectively, but not against the gram negative bacteria. An OMW fraction called AntiSolvent was obtained after the addition of ethanol to the crude OMW. HPLC analysis of AntiSolvent fraction revealed that this fraction contains mainly hydroxytyrosol (10.3%), verbascoside (7.4%), and tyrosol (2.6%). The combinations of AntiSolvent/gallic acid were tested using the low minimal inhibitory concentrations which revealed that 50/100-100/100 µg mL(-1) caused complete growth inhibition of the four strains. These results suggest that OMW specific fractions augmented with natural phenolic ingredients may be utilized as a source of bioactive compounds to control pathogenic bacteria.

Direct effects of phenolic compounds on the mammary gland: In vivo and ex vivo evidence.

We assessed the potential of Pistacia lentiscus (lentisk) phenolic compounds to enhance production of milk composition in lactating goats and caprine primary mammary epithelial cells (MEC). Damascus goats were given a lentisk infusion (LI) or fresh water (FW) to drink, in a crossover design. Milk from LI vs. FW goats was 0.43% richer in fat and 30% in omega 3 fatty acids. Lentisk infusion enhanced antioxidant capacity of plasma and milk by 37% and 30% respectively, and induced transcriptional activation of antioxidant genes. To assess the direct effect of polyphenols on milk quality in terms of composition and antioxidant capacity, we used plasma collected from goats fed hay (HP) or browsed on phenolic compounds-rich pasture (primarily lentisk; PP) as a conditioning medium for primary culture of MEC. PP increased 2-fold cellular triglyceride content and 2.4-fold intracellular casein, and increased ATP production and non-mitochondrial oxygen consumption. Taken together, the results imply that lentisk phenolic compounds affect blood, MEC and milk oxidative status, which increase fat production by the mammary gland.

Antioxidant activity of phenolic fractions in olive mill wastewater.

Olive mill wastewater (OMW) contains a substantial amount of valuable antioxidant phenols that can be recovered for industrial application as food additives and pharmaceuticals. The present study was aimed at extracting different phenolic OMW fractions, and determining their antioxidant potential. Five different OMW fractions were obtained using fractionation techniques, their antioxidant potential determined by DPPH, ORAC and a ß-carotene bleaching test. The total phenol level ranged between 115 and 170 mg/l. The phenolic compounds present in individual fractions were identified using the HPLC-PAD method, where the main compounds were hydroxytyrosol, tyrosol, caffeic acid, vanillic acid, verbascoside, oleuropein, ferulic acid, and p-coumaric acid. The five OMW fractions showed different antioxidant levels depending on the test used. DPPH test showed that the fraction of alkyl aromatic alcohols (AAAs) was the best with EC(50) of 20 mg/l and the pure hydroxytyrosol with 2 mg/l. ORAC test showed that AAA and semi hydrolysed total phenol (s-TP) fractions were significantly better than Trolox when compared to 20 mg/l of Trolox.