Several potential genotoxins found in water samples arise from anthropogenic activities. Acid mine effluent resulting from coal mining poses serious environment concerns all over the world. The use of toxicity tests to evaluate the quality of streams add value by providing site-specific toxicological data. Treatment systems such as the use of natural wetlands (passive) or conventional physical and chemical pH-neutralised processes (active) are employed mainly to meet certain water quality guidelines. Nonetheless, potential genotoxins or residues remain which influence the quality of discharged effluents. The objective of this study was to evaluate the genotoxic potential of acid mine drainage (AMD) released into a natural stream following treatment by passive and active methods. This study aimed to identify the extent of AMD mutagenicity and genotoxicity to African Vero monkey kidney cell line and a fish gill cell line (RTgill-W1) using two assays, the Ames test, and the comet assay, as a rapid and effective screening tool. The Ames test performed without metabolic activation using Salmonella typhimurium TA98 and TA100 strains showed no indication of mutagenicity in the water samples tested. Differing results were however obtained for the comet assay using the African Vero monkey kidney cell line and a fish gill cell line (RTgill-W1), which revealed DNA fragmentation and variations in morphologies indicative of genotoxicity in the water samples following the two treatment processes. A significant reduction in DNA damage was observed in water samples following active treatment of the AMD, evidenced by reduced damage frequency and a lowered comet score. This bioassay confirms the urgency of integrating high-throughput screening in aquatic toxicity assessment at genetic levels, giving further evidence that in-vitro bioassays can be incorporated for use in short-term genotoxicity assays. The result suggests that the comet assay proved sensitive at detecting genotoxicity, supporting the integration of this into environmental monitoring frameworks targeted at AMD-contaminated sites.

Hot water and hydroethanolic (70:30) extracts were prepared from 15 plant species, which were investigated to discover eco-friendly and less expensive tick control methods as an alternative to synthetic acaricides. A contact bioassay was used to determine the acaricidal activity of these extracts against the cattle tick, Rhipicephalus turanicus (Acari: Ixodidae) at a concentration of 20% (200 mg/mL). The hydroethanolic extracts had better activity than the hot water extracts against R. turanicus. The hydroethanolic extract from Tabernaemontana elegans (leaves) had the best mortality (87.0%). This was followed by Calpurnia aurea (stems) with a mortality of 75.0%, Schkuhria pinnata (whole plant) with a mortality of 67.0% and Aloe rupestris (leaves) with a mortality of 66.6%. The toxicity of the plant extracts was also investigated and it was found that most of the hydroethanolic and hot water extracts were either safe or very safe on human Vero kidney and liver HepG2 cells. From this study, it was evident that botanicals have the potential to be developed as environmentally benign natural acaricides against R. turanicus.

ABSTRACT Four compounds including beauvericin, parahydroxybenzaldehyde, indole-3-carboxylic acid and quinizarin were isolated from endophytic fungus Epicoccum nigrum and their cytotoxicity, antibacterial and antioxidant activity were evaluated. Beauvericin had remarkable activity against two Gram-negative strains (Bacillus cereus and Salmonella typhimurium) with respective MIC values of 3.12 and 6.25 µg/ml. All the compounds had weak cytotoxic effect on both normal and tumor cells. LC50 values ranged from 40.42 to 86.56 µg/ml, 31.87 to 86.57 µg/ml and 21.59 to 67.27 µg/ml on Vero cells, THP-1 and RAW 264.7 respectively. The present study showed that these compounds could be developed for the formulation of antioxidant-rich therapeutic diets and as a therapeutic agent against bacterial infections.

Reactive oxygen species (ROS) production and resultant oxidative stress (OS) has been implicated as a pathway of toxicity in animal species exposed to pollutants. The gills of aquatic animals and the liver and kidneys of mammalian species are specific cellular sites of toxicity. Oxidative effects of acid mine drainage effluent (following passive and active treatment) impacting a natural stream were assessed using selected cell lines. Levels of pollutants such as heavy metals in acid mine drainage (AMD) effluent can be quantified following treatment, but it is unknown whether this is associated with equivalent reduction in toxicity. ROS production by AMD untreated (U) and after treatment (T) was quantified in a fish gill cell line (RTgill-W1) and in two mammalian cell lines (C3A human liver and Vero monkey kidney). ROS production was determined using the oxidant sensitive fluorogenic probe, 2', 7'-dichlorofluorescein diacetate (DCFH-DA) following exposure to U and T, AMD water. Treatment of AMD water caused reduction in levels of Al, Zn, Fe, Si and Mn while levels of Cr, Cu, Ar and Hg remained unchanged. A dose-dependent increase in ROS production was observed for U and T. ROS formation decreased from 14% to 4.5%, 16.4% to 7.2% and 25.3% to 17.7% in the RTgill-W1, C3A, and Vero cell lines exposed to 100% AMD water, U and T. The presence of Mn and/or other ions in treated water and subsequent ROS formation indicates that water could still be toxic to cells and requires further processing. The DCFH-DA assay in several cell lines can be used to rapidly bio-monitor quality of AMD water related to formation of ROS and subsequent cellular effects. However, cut-off levels for cellular toxicity must be established to ensure safety of this water for aquatic animals and for animal and human consumption.

BACKGROUND: Limonoids are highly oxygenated compounds with a prototypical structure. Their occurrence in the plant kingdom is mainly confined to plant families of Meliaceae and Rutaceae. Owing to their wide range of pharmacological and therapeutic properties, this study was aimed at investigating the potential nitric oxide (NO) and acetylcholinesterase (AChE) inhibitory activity and the cytotoxicity of three limonoids: trichilia lactone D5 (1), rohituka 3 (2) and dregeanin DM4 (3), isolated from Trichilia welwitschii C.DC RESULTS: Results indicated that the three limonoids had low cytotoxicity towards Vero cells with LC50 values ranging from 89.17 to 75.82 (μg/L. Compounds (2) and (3) had lower cytotoxicity compared to puromycin and doxorubicin used as reference cytotoxic compounds. Compound (1) (LC50 of 23.55 (μg/L) had good antiproliferative activity against RAW 264.7 cancer cells. At the lowest concentration tested (0.5 μg/mL), compound (2) and (3) released the lowest amount of nitric oxide (2.97 and 2.93 μM, respectively). The three limonoids had anti-AChE activity with IC50 values ranged of 19.13 (μg/L for (1), 34.15 (μg/L for (2) and 45.66 (μg/L for (3), compared to galantamine (IC50 of 8.22 ( g/mL) used as positive control CONCLUSION: The limonoid compounds studied in this work inhibited nitric oxide production in LPS-stimulated macrophages and had anti-AChE activity. Trichilia lactone D5 had potential antiproliferative activity against RAW 264.7 cancer cells. The limonoids had low cytotoxicity towards Vero cells lines. This study provided further examples of the importance of limonoids compounds as potential AChE inhibitors and anti-inflammatory agents targeting the inhibition of NO production