The antifungal activity of an aqueous Tulbaghia violacea plant extract against Aspergillus flavus
- Authors: Belewa, Xoliswa Vuyokazi
- Date: 2015
- Subjects: Medicinal plants , Antifungal agents , Fungi -- Biotechnology
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10948/5858 , vital:21001
- Description: Phytochemical analysis of both HEA1 and the crude plant extract showed the presence of phenolics, tannins and saponins. Saponins were the predominant secondary metabolites and were mostly abundant in the plant extract and to a lesser extent in the active compound. Steroidal saponins, tannins and phenolics were also detected in the plant extract, but only the phenolics were detected in the active compound. The results of the phytochemical analysis showed that those compounds that were not present in the active compound could be removed from the crude extract during the TLC purification process. Investigation on the mechanism of action of the crude plant extract on the sterol production by A. flavus showed that the plant extract affected ergosterol biosynthesis by causing an accumulation of oxidosqualene in the ergosterol biosynthetic pathway resulting in a decline in ergosterol production. An oscillatory response in lanosterol production was observed in the presence of the plant extract, which may be an adaptation mechanism of A. flavus to unfavourable conditions and compensation for the loss of enzyme activity which may have occurred as a result of the accumulation of oxidosqualene. The antifungal activity of the plant extract on ergosterol production by A. flavus may also be due to saponins which target the cell membrane and ergosterol production in fungi. The effect of the plant extract on the fungal cell wall of A. flavus also showed that the plant extract caused a decline in β-(1, 3) glucan production by inhibiting β-glucan synthase. The plant extract also affected the chitin synthesis pathway of A. flavus, by causing a decline in chitin production, which was due to the inhibition of chitin synthase. Investigation of chitinase production using 4MU substrates showed that the plant extract caused an accumulation of chitobioses, by activating chitobiosidases and endochitinases. A decline in N-acetylglucosaminidase activity in the presence of the plant extract was observed and this prevented the formation of N-acetylglucosamine. The accumulation of chitobiosidase and endochitinase may be as a result of autolysis that may be triggered by A. flavus as a survival mechanism in the presence of the plant extract and as a compensatory mechanism for the loss of β-glucans and chitin. The antifungal effect of the plant extract on various components of the cell wall of A. flavus, makes T. violacea aqueous plant extract an ideal chemotherapeutic agent against both human and plant pathogens of Aspergillus. The broad spectrum of antifungal activity of T. violacea against A. flavus also eliminates any chances of the fungus developing resistance towards it and would make it a candidate for use as a potential antifungal agent. Further identification and possible chemical synthesis is needed to shed light on the safety and efficacy of the active compound for further development as a chemotherapeutic agent.
- Full Text:
- Date Issued: 2015
- Authors: Belewa, Xoliswa Vuyokazi
- Date: 2015
- Subjects: Medicinal plants , Antifungal agents , Fungi -- Biotechnology
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10948/5858 , vital:21001
- Description: Phytochemical analysis of both HEA1 and the crude plant extract showed the presence of phenolics, tannins and saponins. Saponins were the predominant secondary metabolites and were mostly abundant in the plant extract and to a lesser extent in the active compound. Steroidal saponins, tannins and phenolics were also detected in the plant extract, but only the phenolics were detected in the active compound. The results of the phytochemical analysis showed that those compounds that were not present in the active compound could be removed from the crude extract during the TLC purification process. Investigation on the mechanism of action of the crude plant extract on the sterol production by A. flavus showed that the plant extract affected ergosterol biosynthesis by causing an accumulation of oxidosqualene in the ergosterol biosynthetic pathway resulting in a decline in ergosterol production. An oscillatory response in lanosterol production was observed in the presence of the plant extract, which may be an adaptation mechanism of A. flavus to unfavourable conditions and compensation for the loss of enzyme activity which may have occurred as a result of the accumulation of oxidosqualene. The antifungal activity of the plant extract on ergosterol production by A. flavus may also be due to saponins which target the cell membrane and ergosterol production in fungi. The effect of the plant extract on the fungal cell wall of A. flavus also showed that the plant extract caused a decline in β-(1, 3) glucan production by inhibiting β-glucan synthase. The plant extract also affected the chitin synthesis pathway of A. flavus, by causing a decline in chitin production, which was due to the inhibition of chitin synthase. Investigation of chitinase production using 4MU substrates showed that the plant extract caused an accumulation of chitobioses, by activating chitobiosidases and endochitinases. A decline in N-acetylglucosaminidase activity in the presence of the plant extract was observed and this prevented the formation of N-acetylglucosamine. The accumulation of chitobiosidase and endochitinase may be as a result of autolysis that may be triggered by A. flavus as a survival mechanism in the presence of the plant extract and as a compensatory mechanism for the loss of β-glucans and chitin. The antifungal effect of the plant extract on various components of the cell wall of A. flavus, makes T. violacea aqueous plant extract an ideal chemotherapeutic agent against both human and plant pathogens of Aspergillus. The broad spectrum of antifungal activity of T. violacea against A. flavus also eliminates any chances of the fungus developing resistance towards it and would make it a candidate for use as a potential antifungal agent. Further identification and possible chemical synthesis is needed to shed light on the safety and efficacy of the active compound for further development as a chemotherapeutic agent.
- Full Text:
- Date Issued: 2015
The effect of tulbaghia violacea plant extract on the growth of aspergillus species
- Authors: Belewa, Xoliswa Vuyokazi
- Date: 2009
- Subjects: Plant products , Plant extracts , Traditional medicine
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10315 , http://hdl.handle.net/10948/d1008186 , Plant products , Plant extracts , Traditional medicine
- Description: Traditional medicine has become an important part of healthcare worldwide. It is estimated that about 25 percent of prescribed medicines contain plant products or active compounds derived from plants. In South Africa, traditional medicine forms part of the culture and tradition of most communities. Garlic compounds have been shown to have a variety of antimicrobial properties. Amongst these are antifungal, antibacterial, antiviral and anti protozoal activities. Allicin and its breakdown products have been shown to be the main active compounds which possess these properties. Tulbaghia violacea has been used for the treatment of a variety of illnesses including asthma, fever, oesophageal cancer, constipation and hypertension. This study investigated the antifungal nature of T.violacea on the morphology, spore germination and lipid synthesis of Aspergillus flavus and Aspergillus parasiticus. The results of this study showed that the plant extract inhibited A. flavus growth at a minimal inhibitory concentration of 15mg/ml and was fungicidal at 20mg/ml and above. A. parasiticus was not inhibited at 25mg/ml indicating resistance to the inhibitory component of the plant extract. A measure of metabolic activity using the XTT assay showed reduced metabolic activity in the presence of increasing concentrations of the plant extract. Higher extract concentrations resulted in higher percentage inhibition of fungal growth for both fungal species with up to 98 percent inhibition being observed for the highest extract concentrations for both fungi. Germination was also delayed in the presence of 15mg/ml plant extract concentration by up to 60hr for A. flavus and 48hr for A. parasititcus. The TEM results showed increased thickening of the cell wall with higher extract concentrations. The thickening was greater for A. flavus than for A. parasiticus. Cell wall thickening may be the reason for the delay in germination in both species. Lipid production was reduced in the presence of plant extracts when compared to the control. The plant extracts inhibited triglyceride production at 15mg/ml for both A. flavus and A. parasiticus. The results therefore indicate that T. violacea extracts are antifungal and probably affect germination through interactions with the cell wall. It is possible that the extract affects lipid production in Aspergillus species.
- Full Text:
- Date Issued: 2009
- Authors: Belewa, Xoliswa Vuyokazi
- Date: 2009
- Subjects: Plant products , Plant extracts , Traditional medicine
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10315 , http://hdl.handle.net/10948/d1008186 , Plant products , Plant extracts , Traditional medicine
- Description: Traditional medicine has become an important part of healthcare worldwide. It is estimated that about 25 percent of prescribed medicines contain plant products or active compounds derived from plants. In South Africa, traditional medicine forms part of the culture and tradition of most communities. Garlic compounds have been shown to have a variety of antimicrobial properties. Amongst these are antifungal, antibacterial, antiviral and anti protozoal activities. Allicin and its breakdown products have been shown to be the main active compounds which possess these properties. Tulbaghia violacea has been used for the treatment of a variety of illnesses including asthma, fever, oesophageal cancer, constipation and hypertension. This study investigated the antifungal nature of T.violacea on the morphology, spore germination and lipid synthesis of Aspergillus flavus and Aspergillus parasiticus. The results of this study showed that the plant extract inhibited A. flavus growth at a minimal inhibitory concentration of 15mg/ml and was fungicidal at 20mg/ml and above. A. parasiticus was not inhibited at 25mg/ml indicating resistance to the inhibitory component of the plant extract. A measure of metabolic activity using the XTT assay showed reduced metabolic activity in the presence of increasing concentrations of the plant extract. Higher extract concentrations resulted in higher percentage inhibition of fungal growth for both fungal species with up to 98 percent inhibition being observed for the highest extract concentrations for both fungi. Germination was also delayed in the presence of 15mg/ml plant extract concentration by up to 60hr for A. flavus and 48hr for A. parasititcus. The TEM results showed increased thickening of the cell wall with higher extract concentrations. The thickening was greater for A. flavus than for A. parasiticus. Cell wall thickening may be the reason for the delay in germination in both species. Lipid production was reduced in the presence of plant extracts when compared to the control. The plant extracts inhibited triglyceride production at 15mg/ml for both A. flavus and A. parasiticus. The results therefore indicate that T. violacea extracts are antifungal and probably affect germination through interactions with the cell wall. It is possible that the extract affects lipid production in Aspergillus species.
- Full Text:
- Date Issued: 2009
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