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Theses and dissertations (Applied Sciences)

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    Efficacy and mechanisms of antiretroviral drugs removal by algaefrom wastewater treatment plants
    (2024-05) Reddy, Karen; Bux, Faizal; Kuttan Pillai, Sheena Kumari; Renuka, Nirmal; Moodley, Brenda
    The presence, risks, and fate of pharmaceutical pollutants in the environment have raised concerns worldwide. South Africa, with the largest population consuming antiretroviral (ARV) drugs in Africa, faces challenges in efficiently removing these compounds from water bodies. This study's primary focus was to investigate the efficiency and mechanisms of nevirapine (NVP) removal by algae isolated from wastewater treatment processes. It included the isolation and screening of algal strains from wastewater treatment plants for their potential to remove ARV drugs, optimizing culture conditions to enhance removal efficiency, determining the potential mechanisms employed by selected algal strains for NVP remediation, and assessing the associated metabolic responses of algal cells to NVP using gene expression and metabolomics analyses. Eleven green indigenous fresh water microalgal isolates were screened from wastewater treatment plants (WWTPs) in KwaZulu-Natal, resulting in the selection of two strains, Coelastrella tenuitheca and Tetradesmus obliquus, based on their growth rates, biomass productivity and toxicity tolerance. In the ecotoxicity study, the calculated IC50 values of NVP (0–100 mg L−1) on selected algal strains after 96 h of exposure were 23.45 mg L−1 (C. tenuitheca) and 18.20 mg L−1 (T. obliquus), which far exceeds the concentration of NVP found in wastewater. Hence, T. obliquus and C. tenuitheca was selected for further NVP remediation studies using different cultivation conditions. A concentration range of 0-4000 ng L-1 of NVP was tested to assess the potential for NVP removal by both microalgae (autotrophic cultivation). Lower concentrations of NVP (up to 200 ng L−1) have shown to have a positive impact on microalgae growth. Specifically, in T. obliquus, the highest dry cell weight of 941.27 mg L−1 was obtained when exposed to a NVP concentration of 50 ng L−1. Both microalgae showed varying removal efficiencies (19.53–74.56%) when exposed to different NVP concentrations. During the late log phase on day 8, T. obliquus achieved the highest NVP removal efficiency, removing 74.56% of the NVP, while C. tenuitheca achieved a removal rate of 48% at an NVP concentration of 50 ng L−1. The photosynthetic efficiency (Fv/Fm and rETR) of both microalgal species was found to be unaffected by environmental concentrations of NVP (up to 4000 ng L−1) during the mid-log phase of growth. Furthermore, the scanning electron microscopy (SEM) analysis demonstrated that both algal species produced distinct ridges on their cell surfaces after NVP uptake. Additional evaluations were conducted on the microalga, T. obliquus, for the removal of NVP at 4000 ng L-1, as well as their cellular response (expression of antioxidant enzymes and metabolomics) and biomass production under different cultivation modes (autotrophic, heterotrophic, and mixotrophic). The highest NVP removal efficiency was observed under mixotrophic (80.13%) growth on day 8, whilst heterotrophic and autotrophic cultivation modes removed 70.30% and 64.40%, respectively. Mass balance calculations showed that the primary removal mechanism was identified as biodegradation, with a relatively low contribution from bioadsorption (2.39-3.36%) and bioaccumulation (0.55- 0.87%). Fourier-transform infrared (FTIR) spectroscopy results of harvested microalgal cells displayed bands in the region of 950-1000 cm-1, indicating the presence of aromatic C-H rings found in NVP. Additionally, 6 possible biotransformation products of NVP were identified by untargeted liquid chromatography-time of flight mass spectrometry. Additionally, under autotrophic conditions, the gene expression analysis revealed heightened activities of superoxide dismutase (sod1), glutathione peroxidase (gpx1) and catalase (cat2) in T. obliquus. The upregulation of antioxidant genes enhances the organism's ability to defend against oxidative stress induced by NVP. The expression levels of antioxidant genes were significantly reduced during heterotrophic and mixotrophic growth, suggesting microalgae can overcome oxidative stress with glucose supplementation. To further investigate the cellular level response of microalgal cells to NVP, metabolomic analysis was carried to out to identify and quantify key algal metabolites during mixotrophic cultivation. The increase in activity of the fatty acid biosynthesis pathway and carbohydrate synthesis was observed by T. obliquus in the presence of NVP under mixotrophic growth conditions. The findings from this study emphasize the significant potential of microalgae in the field of ARV drug remediation.
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    Isolation and characterization of prebiotic oligosaccharides from algal extracts and their effect on gut microflora
    (2016) Hadebe, Nontando; Odhav, Bharti
    Prebiotics are defined as non-digestible oligosaccharides (NDOs) or polysaccharides (NDPs), which promote the growth of beneficial lactic acid bacteria in the colon. Algae are rich in polysaccharides and can be exploited as prebiotics for functional food ingredients to improve human and animal health. Currently, inulin is the most widely used ingredient in the prebiotics market, which is produced from live plants and requires expensive production processing. There is a vast repository of marine life with algae as a major source of nutrients. Therefore, this study provides an alternative source for prebiotic production and examines marine and freshwater algae that promote the growth of two strains of Lactobacillus delbrueckii subs. (Lactobacillus lactis and Lactobacillus bulgaricus) and one strain of Bifidobacterium spp. (Bifidobacterium longum). Monosaccharides of the oligosaccharide fraction of marine and freshwater algal extracts were investigated with the use of thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) after acidic hydrolysis of cell matrix polysaccharides. A total of fifty-five marine and freshwater aqueous algal extracts were assessed for their effect on the growth of L. lactis, B. longum and L. bulgaricus over a 96 hour period. Relative to the negative control, 34.5% algal extracts showed improved growth on one or more probiotic bacteria. The optimum time for maximum bacterial growth was noted at 48 h for all the tested aqueous algal extracts. Five marine and freshwater algal cultures (Spirulina platensis, Chlorococcum spp., Dunaliella salina, Scenedesmus magnus, Chlorella spp. and algal extract no. 48) from various aquatic environments in Kwa-Zulu Natal showed the best growth dynamics and demonstrated the greatest potential as sources of biomass for prebiotic production. These algal extracts were able to significantly increase the growth of at least one of the three probiotic bacteria (p < 0.05). Aqueous algal extract from S. platensis was regarded as the best algal source for prebiotics as it demonstrated a greater stimulatory effect on the growth of all three probiotic bacteria (L. lactis, B. longum and L. bulgaricus) compared to tested aqueous algal extracts and the inulin used as a positive control. The results obtained by HPLC for characterization confirmed TLC data, as xylose and galactose were detected by both chromatograms. These data indicated that xylose and galactose were present in aqueous algal extracts from S. magnus and S. platensis and galactose in aqueous algal extract no. 48. Xylose was most abundant in aqueous algal extracts from S. platensis (3mg/ml) and S. magnus (2.3mg/ml). In conclusion aqueous algal extracts from S. platensis, Chlorococcum, D. salina, S. magnus, Chlorella and algal extract no. 48 are potential sources for prebiotic production. Spirulina platensis extract was regarded as the best algal source. Xyose and galactose characterized by HPLC in algal extracts make up oligosaccharides that function as prebiotic compounds for stimulation of probiotic bacteria. There is a great scope for successful production of prebiotics from algal sources in South Africa.