The effect of in vitro digestion on selected biological activities of Hypoxis sobolifera corms
- Authors: Van Rooyen, Anzel
- Date: 2013
- Subjects: Potatoes -- Therapeutic use , Medicinal plants
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10344 , http://hdl.handle.net/10948/d1020058
- Description: In South Africa part of the cultural and religious beliefs of the African people is the use of traditional remedies to treat diseases. These remedies are obtained from medicinal plants (Steenkamp, 2003). One of the most frequently traded plants in the Eastern Cape is Hypoxis, commonly known as Afrika patat, or African potato. South African traditional healers instruct patients to brew the fresh Hypoxis corm as a tea and then ingest it (Steenkamp, 2006a). This prompted an investigation into the digestive stability of a traditionally prepared Hypoxis extract. The H. sobolifera extracts were digested using a simulated gastric/small intestinal digestion and their biological activity determined. The hot water H. sobolifera extract before digestion only showed cytotoxic activity against cancer cell lines at very high concentrations which are not likely to be achieved under normal ingestion circumstances. In Chang liver cells on the other hand, chronic exposure to the hot water H. sobolifera extract increased glucose uptake in amounts similar to that of metformin. On the negative side, the glucose utilization stimulation was lost due to the simulated digestion process. The significant inhibition of AGEs by hot water H. sobolifera extract (IC50 of 6.3 Ig/ml) is a very encouraging result as treatment in the management of diabetes. This activity was only slightly reduced by the in vitro digestion process. Also observed was enzyme inhibition activity by traditionally prepared H. sobolifera, with ∝-amylase being inhibited (IC50 of approximately 250 Ig/ml) and therefore preventing or limiting starch breakdown. From the DPPH results it was clear that H. sobolifera, even when digested, is a potent anti-oxidant (IC50 of 134.4 Ig/ml when undigested compared to 162.9 when digested with β-glucosidase added to stomach digestive step). HPLC and TLC experiments revealed that rooperol which has previously been thought to be the compound responsible for the anti-oxidant activity in Hypoxis extracts, was absent from the traditional extract of H. sobolifera and therefore cannot be the sole compound exhibiting anti-oxidant activity; other compounds such as phenolics may be contributing. The phenolic and flavonoid content results revealed very highconcentrations of these compounds in the traditionally prepared H. sobolifera extract. These compounds may therefore play major roles in all of the biological activities observed from treatment with Hypoxis spp. The ROS results yielded interesting and promising results. Using standard or traditionally prepared H. sobolifera extracts, activation of differentiated U937 cells with PMA was greatly enhanced by cotreatment with the extracts, while extracts on their own did not cause significant activation. Future studies should investigate this property of the extracts as a promising immune boosterThe HPLC results showed that hypoxoside was undetectable in the hot water traditional extract and the TLC anti-oxidant experiment proved that rooperol is not present in the hot water traditional extract after treatment with β-glucosidase. This indicates that neither one of the Hypoxis compounds previously believed to be responsible for the biological activities observed are present in the extract when prepared the traditional way. Therefore, the biological activities observed in this study can be attributed to other phytochemical compounds.
- Full Text:
- Date Issued: 2013
- Authors: Van Rooyen, Anzel
- Date: 2013
- Subjects: Potatoes -- Therapeutic use , Medicinal plants
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10344 , http://hdl.handle.net/10948/d1020058
- Description: In South Africa part of the cultural and religious beliefs of the African people is the use of traditional remedies to treat diseases. These remedies are obtained from medicinal plants (Steenkamp, 2003). One of the most frequently traded plants in the Eastern Cape is Hypoxis, commonly known as Afrika patat, or African potato. South African traditional healers instruct patients to brew the fresh Hypoxis corm as a tea and then ingest it (Steenkamp, 2006a). This prompted an investigation into the digestive stability of a traditionally prepared Hypoxis extract. The H. sobolifera extracts were digested using a simulated gastric/small intestinal digestion and their biological activity determined. The hot water H. sobolifera extract before digestion only showed cytotoxic activity against cancer cell lines at very high concentrations which are not likely to be achieved under normal ingestion circumstances. In Chang liver cells on the other hand, chronic exposure to the hot water H. sobolifera extract increased glucose uptake in amounts similar to that of metformin. On the negative side, the glucose utilization stimulation was lost due to the simulated digestion process. The significant inhibition of AGEs by hot water H. sobolifera extract (IC50 of 6.3 Ig/ml) is a very encouraging result as treatment in the management of diabetes. This activity was only slightly reduced by the in vitro digestion process. Also observed was enzyme inhibition activity by traditionally prepared H. sobolifera, with ∝-amylase being inhibited (IC50 of approximately 250 Ig/ml) and therefore preventing or limiting starch breakdown. From the DPPH results it was clear that H. sobolifera, even when digested, is a potent anti-oxidant (IC50 of 134.4 Ig/ml when undigested compared to 162.9 when digested with β-glucosidase added to stomach digestive step). HPLC and TLC experiments revealed that rooperol which has previously been thought to be the compound responsible for the anti-oxidant activity in Hypoxis extracts, was absent from the traditional extract of H. sobolifera and therefore cannot be the sole compound exhibiting anti-oxidant activity; other compounds such as phenolics may be contributing. The phenolic and flavonoid content results revealed very highconcentrations of these compounds in the traditionally prepared H. sobolifera extract. These compounds may therefore play major roles in all of the biological activities observed from treatment with Hypoxis spp. The ROS results yielded interesting and promising results. Using standard or traditionally prepared H. sobolifera extracts, activation of differentiated U937 cells with PMA was greatly enhanced by cotreatment with the extracts, while extracts on their own did not cause significant activation. Future studies should investigate this property of the extracts as a promising immune boosterThe HPLC results showed that hypoxoside was undetectable in the hot water traditional extract and the TLC anti-oxidant experiment proved that rooperol is not present in the hot water traditional extract after treatment with β-glucosidase. This indicates that neither one of the Hypoxis compounds previously believed to be responsible for the biological activities observed are present in the extract when prepared the traditional way. Therefore, the biological activities observed in this study can be attributed to other phytochemical compounds.
- Full Text:
- Date Issued: 2013
The medicinal plant Sutherlandia Frutescens regulates gene expression to reverse insulin resistace in rats
- Authors: Fortuin, Melissa
- Date: 2013
- Subjects: Insulin resistance , Medicinal plants , Genetic regulation , Insulin resistance -- Animal models
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10349 , http://hdl.handle.net/10948/d1020823
- Description: Obesity can lead to Type 2 Diabetes, both conditions increase in association with physical inactivity and high-energy diets, resulting in elevated blood glucose, decreased insulin sensitivity and increased insulin resistance. Sutherlandia frutescens (S.frutescens), an anti-diabetic plant, reverses and prevents insulin resistance in a rat model and human cell culture model. Gene expression analysis in hepatocyte cultures, identified genes down regulated in insulin resistance and up regulated by S.frutescens. These included genes encoding vesicle transporter proteins, hypothesised to be linked to hepatic lipid accumulation and lipid droplet formation during insulin resistance. The aim of this study was to investigate critical genes involved in lipid droplet formation, vesicle assembly and transport in high fat diet (HFD)-induced insulin resistant rat liver tissue during the development of insulin resistance and the reversal of these changes by S.frutescens. Rats were fed a low fat diet (LFD) or HFD supplemented with S.frutescens for 2, 4 and 8 weeks. Rats fed a HFD for 12 weeks developed insulin resistance, confirmed by plasma glucose and insulin levels (compared to normal controls). Groups of these rats were gavaged with S. frutescens (50mg/kg BW), Metformin (13mg/kg BW) or water for a further 4 weeks and starved for 12 hours, anaesthetized and blood removed by heart puncture. Liver was stored in RNA-Later™ for qRT-PCR and snap-frozen in liquid nitrogen for western blotting and confocal microscopy analysis. Changes in expression of vesicle transporter genes VAMP3 and NSF were analysed by qRT-PCR and changes in the protein expression by western blotting analysis. Proteins were localised within the liver by confocal immunohistochemistry using ZEN lite™ software. Statistical analysis was performed using One-Way ANOVA and unpaired t-test. mRNA gene expression of vesicle transport components VAMP3, NSF and SNAP25 showed relatively moderate changes with considerable individual variation within control or experimental groups. Uncorrelated changes in mRNA and protein products were found and may be due to differential regulation by siRNA. Proteins also showed altered staining patterns in high fat diet rats that reverted towards normal on S. frutescens treatment, potentially reflecting functional changes associated with transport of lipid-filled vesicles.
- Full Text:
- Date Issued: 2013
- Authors: Fortuin, Melissa
- Date: 2013
- Subjects: Insulin resistance , Medicinal plants , Genetic regulation , Insulin resistance -- Animal models
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10349 , http://hdl.handle.net/10948/d1020823
- Description: Obesity can lead to Type 2 Diabetes, both conditions increase in association with physical inactivity and high-energy diets, resulting in elevated blood glucose, decreased insulin sensitivity and increased insulin resistance. Sutherlandia frutescens (S.frutescens), an anti-diabetic plant, reverses and prevents insulin resistance in a rat model and human cell culture model. Gene expression analysis in hepatocyte cultures, identified genes down regulated in insulin resistance and up regulated by S.frutescens. These included genes encoding vesicle transporter proteins, hypothesised to be linked to hepatic lipid accumulation and lipid droplet formation during insulin resistance. The aim of this study was to investigate critical genes involved in lipid droplet formation, vesicle assembly and transport in high fat diet (HFD)-induced insulin resistant rat liver tissue during the development of insulin resistance and the reversal of these changes by S.frutescens. Rats were fed a low fat diet (LFD) or HFD supplemented with S.frutescens for 2, 4 and 8 weeks. Rats fed a HFD for 12 weeks developed insulin resistance, confirmed by plasma glucose and insulin levels (compared to normal controls). Groups of these rats were gavaged with S. frutescens (50mg/kg BW), Metformin (13mg/kg BW) or water for a further 4 weeks and starved for 12 hours, anaesthetized and blood removed by heart puncture. Liver was stored in RNA-Later™ for qRT-PCR and snap-frozen in liquid nitrogen for western blotting and confocal microscopy analysis. Changes in expression of vesicle transporter genes VAMP3 and NSF were analysed by qRT-PCR and changes in the protein expression by western blotting analysis. Proteins were localised within the liver by confocal immunohistochemistry using ZEN lite™ software. Statistical analysis was performed using One-Way ANOVA and unpaired t-test. mRNA gene expression of vesicle transport components VAMP3, NSF and SNAP25 showed relatively moderate changes with considerable individual variation within control or experimental groups. Uncorrelated changes in mRNA and protein products were found and may be due to differential regulation by siRNA. Proteins also showed altered staining patterns in high fat diet rats that reverted towards normal on S. frutescens treatment, potentially reflecting functional changes associated with transport of lipid-filled vesicles.
- Full Text:
- Date Issued: 2013
In vitro cytotoxic effects of selected Nigerian medicinal plant extracts on cancer cell lines
- Authors: Baatjies, Lucinda
- Date: 2012
- Subjects: Cancer -- Treatment , Cancer cells , Medicinal plants , Plant extracts , Traditional medicine , Public health
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10316 , http://hdl.handle.net/10948/d1008191 , Cancer -- Treatment , Cancer cells , Medicinal plants , Plant extracts , Traditional medicine , Public health
- Description: Cancer is a disease that imposes a heavy burden on public health and poses a challenge to science. The World Health Organization estimates that 80 percent of people in developing countries of the world rely on traditional medicine for their primary health needs, and about 85 percent of traditional medicine involves the use of plant extracts. This is particularly true in Africa where a large percentage of the population depends upon medicinal plants for health care. Therefore, detailed screening and evaluation of bioactive substances for chemotherapeutic purposes of African plants are urgently warranted. Furthermore, this will serve to validate the efficacy and safety of African traditional medicine. The current study investigated the in vitro cytotoxic effects of 17 ethanolic extracts of the following 16 plants used in traditional anticancer medicine in Nigeria: Sapium ellipticum leaves, Sapium ellipticum stembark, Combretum paniculatum, Celosia trigyna, Pupalia lappacea, Justica extensa, Hedranthera barteri leaves, Alternanthera sessilis, Ethulia conyzoides leaves, Lannea nigritana stembark, Combretum zenkeri root, Combretum molle leaves, Adenanthera parvoniana, Lannea acida, Cyathula achyranthoides, Drymaria cordata, Cyathula prostrata, against HeLa cancer cells. Five of the most promising extracts (Sapium ellipticum leaves, Combretum paniculatum, Celosia trigyna, Drymaria cordata, Cyathula prostrata) were selected for further screening against HT29 and MCF-7 cancer cells. Of the five, the first two were investigated further based on their activities in the screening phase. The S. ellipticum leaf extract yielded IC50 values of 88.60 ± 0.03 and 93.03 ± 0.03 μg/ml against HeLa and MCF-7, respectively. The toxicity was also evaluated on normal cells and an IC50 of 77.66 μg/ml was obtained for peripheral blood mononuclear cells (PBMCs). The IC50 values for proliferating and confluent Chang liver cells were both >125 μg/ml. These results suggest that the extract may be selective for specific cell types. Bio-assay guided fractionation of the S. ellipticum ethanolic extract yielded two active fractions; chloroform and ethyl acetate. Two compounds isolated from the chloroform extract were screened against the three cancer cell lines and found to be inactive. Three compounds were isolated from the ethyl acetate fraction and revealed IC50 values < 62.5 and < 31 μg/ml against MCF-7. Unfortunately these two compounds soon lost activity before any further work could be done on them and work was continued with the crude extract.
- Full Text:
- Date Issued: 2012
- Authors: Baatjies, Lucinda
- Date: 2012
- Subjects: Cancer -- Treatment , Cancer cells , Medicinal plants , Plant extracts , Traditional medicine , Public health
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10316 , http://hdl.handle.net/10948/d1008191 , Cancer -- Treatment , Cancer cells , Medicinal plants , Plant extracts , Traditional medicine , Public health
- Description: Cancer is a disease that imposes a heavy burden on public health and poses a challenge to science. The World Health Organization estimates that 80 percent of people in developing countries of the world rely on traditional medicine for their primary health needs, and about 85 percent of traditional medicine involves the use of plant extracts. This is particularly true in Africa where a large percentage of the population depends upon medicinal plants for health care. Therefore, detailed screening and evaluation of bioactive substances for chemotherapeutic purposes of African plants are urgently warranted. Furthermore, this will serve to validate the efficacy and safety of African traditional medicine. The current study investigated the in vitro cytotoxic effects of 17 ethanolic extracts of the following 16 plants used in traditional anticancer medicine in Nigeria: Sapium ellipticum leaves, Sapium ellipticum stembark, Combretum paniculatum, Celosia trigyna, Pupalia lappacea, Justica extensa, Hedranthera barteri leaves, Alternanthera sessilis, Ethulia conyzoides leaves, Lannea nigritana stembark, Combretum zenkeri root, Combretum molle leaves, Adenanthera parvoniana, Lannea acida, Cyathula achyranthoides, Drymaria cordata, Cyathula prostrata, against HeLa cancer cells. Five of the most promising extracts (Sapium ellipticum leaves, Combretum paniculatum, Celosia trigyna, Drymaria cordata, Cyathula prostrata) were selected for further screening against HT29 and MCF-7 cancer cells. Of the five, the first two were investigated further based on their activities in the screening phase. The S. ellipticum leaf extract yielded IC50 values of 88.60 ± 0.03 and 93.03 ± 0.03 μg/ml against HeLa and MCF-7, respectively. The toxicity was also evaluated on normal cells and an IC50 of 77.66 μg/ml was obtained for peripheral blood mononuclear cells (PBMCs). The IC50 values for proliferating and confluent Chang liver cells were both >125 μg/ml. These results suggest that the extract may be selective for specific cell types. Bio-assay guided fractionation of the S. ellipticum ethanolic extract yielded two active fractions; chloroform and ethyl acetate. Two compounds isolated from the chloroform extract were screened against the three cancer cell lines and found to be inactive. Three compounds were isolated from the ethyl acetate fraction and revealed IC50 values < 62.5 and < 31 μg/ml against MCF-7. Unfortunately these two compounds soon lost activity before any further work could be done on them and work was continued with the crude extract.
- Full Text:
- Date Issued: 2012
The anticancer activity of Cyathula prostrata on two malignant cell lines
- Authors: Schnablegger, Gerald
- Date: 2010
- Subjects: Medicinal plants
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10309 , http://hdl.handle.net/10948/1563 , Medicinal plants
- Description: Plants have always been a source of medicine and are still being used by traditional healers in the rural part of Africa, Asia and India to treat a range of illnesses including cancer. The in vitro anticancer activity of an 80 percent ethanol extract of Cyathula prostrata, an annual branching shrub used by traditional healers in Nigeria to treat cancer was investigated. No previous studies have outlined the possible pathways and mechanisms used by cancer cells when treated with C. prostrata. Dose response analysis was performed to determine the effective cytotoxic concentrations of C. prostrata on HeLa (cervical cancer cell line) and U937 (myelo-monocytic cell line). The IC50 values were 100.8 μg/ml and 64.4 μg/ml for HeLa and U937 cells, respectively. All further experiments were performed using 125 μg/ml C. prostrata extract and 50 μM cisplatin as positive control. With the use of the fluorescent DNA binding dye propidium iodide, the induction of tumour cell death by C. prostrata extract has been linked to cell cycle arrest in the G1 phase at 24 and 48 h. In both cell lines, more than 80 percent of the C. prostrata treated cells were found in the G1 phase after 48 hours of treatment. The annexin V-FITC/PI assay revealed an increase in the percentage apoptotic cells from 4.9 percent to 53.1 percent at 24 h and 8.3 percent to 50.3 percent at 48 h. Since apoptosis induction can occur via a number of different pathways, distinct features were used as markers to investigate the mode of action of this C. prostrata extract. Markers such as activated caspase-8, p21 and cyt-c, were investigated with the aid of fluorescently labelled (FITC) antibodies with analysis using flow cytometry. No change in p21 levels was observed in response to treatment with the extract for up to 48 h. Cell cycle arrest in G1 was therefore not induced by this cyclin-CDK inhibitor. Increase in caspase-8 activation was observed in response to treatment with the extract with no cyt-c release from the mitochondria. The lack of cyt-c release was due to no change in mitochondrial membrane potential, which was investigated with the aid of fluorescent mitochondrial dyes and flow cytometric techniques. Caspase-8 activation is unique to the extrinsic apoptotic pathway. The results from this study therefore show that C. prostrata extract induces apoptosis via the extrinsic pathway and that this activation in independent of the mitochondria. The levels of hTERT, the catalytic subunit of telomerase, were investigated as an additional molecular target for C. prostrata. This was also investigated using FITC labelled antibodies and flow cytometry. A decrease in hTERT levels was observed following C. prostrata treatment. The findings from this study suggest that the extract acts through multiple targets, by inducing: cell cycle arrest in the G1 phase through an unknown mechanism; apoptosis through an extrinsic death receptor pathway and replicative senescence through inhibition of telomerase.
- Full Text:
- Date Issued: 2010
- Authors: Schnablegger, Gerald
- Date: 2010
- Subjects: Medicinal plants
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10309 , http://hdl.handle.net/10948/1563 , Medicinal plants
- Description: Plants have always been a source of medicine and are still being used by traditional healers in the rural part of Africa, Asia and India to treat a range of illnesses including cancer. The in vitro anticancer activity of an 80 percent ethanol extract of Cyathula prostrata, an annual branching shrub used by traditional healers in Nigeria to treat cancer was investigated. No previous studies have outlined the possible pathways and mechanisms used by cancer cells when treated with C. prostrata. Dose response analysis was performed to determine the effective cytotoxic concentrations of C. prostrata on HeLa (cervical cancer cell line) and U937 (myelo-monocytic cell line). The IC50 values were 100.8 μg/ml and 64.4 μg/ml for HeLa and U937 cells, respectively. All further experiments were performed using 125 μg/ml C. prostrata extract and 50 μM cisplatin as positive control. With the use of the fluorescent DNA binding dye propidium iodide, the induction of tumour cell death by C. prostrata extract has been linked to cell cycle arrest in the G1 phase at 24 and 48 h. In both cell lines, more than 80 percent of the C. prostrata treated cells were found in the G1 phase after 48 hours of treatment. The annexin V-FITC/PI assay revealed an increase in the percentage apoptotic cells from 4.9 percent to 53.1 percent at 24 h and 8.3 percent to 50.3 percent at 48 h. Since apoptosis induction can occur via a number of different pathways, distinct features were used as markers to investigate the mode of action of this C. prostrata extract. Markers such as activated caspase-8, p21 and cyt-c, were investigated with the aid of fluorescently labelled (FITC) antibodies with analysis using flow cytometry. No change in p21 levels was observed in response to treatment with the extract for up to 48 h. Cell cycle arrest in G1 was therefore not induced by this cyclin-CDK inhibitor. Increase in caspase-8 activation was observed in response to treatment with the extract with no cyt-c release from the mitochondria. The lack of cyt-c release was due to no change in mitochondrial membrane potential, which was investigated with the aid of fluorescent mitochondrial dyes and flow cytometric techniques. Caspase-8 activation is unique to the extrinsic apoptotic pathway. The results from this study therefore show that C. prostrata extract induces apoptosis via the extrinsic pathway and that this activation in independent of the mitochondria. The levels of hTERT, the catalytic subunit of telomerase, were investigated as an additional molecular target for C. prostrata. This was also investigated using FITC labelled antibodies and flow cytometry. A decrease in hTERT levels was observed following C. prostrata treatment. The findings from this study suggest that the extract acts through multiple targets, by inducing: cell cycle arrest in the G1 phase through an unknown mechanism; apoptosis through an extrinsic death receptor pathway and replicative senescence through inhibition of telomerase.
- Full Text:
- Date Issued: 2010
Pharmacological Evaluation and Medicinal Potential of Vachellia Karroo (Hayne) Banfi & Galasso Pods traditionally used to treat Sexually Transmitted Diseases in the Eastern Cape, South Africa.
- Authors: Maposa, Sandisiwe
- Date: 2019
- Subjects: Medicinal plants
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10353/14583 , vital:40017
- Description: Vachellia karroo is widely used in folk medicine in the Eastern Cape Province South Africa, however, the pods are usually discarded as waste. The current study was conducted to evaluate the phytochemical content and the antioxidant activity of V. karroo pods. The pods were extracted using acetone, distilled water, hexane and methanol. The total phenol, flavonoid, proanthocyanidin, alkaloid and saponin contents of the various extracts were determined spectrometrically and antioxidant activity was evaluated using inhibition of 2,2-diphenyl-1- picrylhydrazyl (DPPH), 2,2´-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid (ABTS), nitric oxide (NO) radicals, while total antioxidant capacity (TAC) was estimated by the phosphomolybdenum assay. V. karro pods had significant amounts of total phenols, flavonoids and proanthocynadin, although, proanthocynadin was not detected in the aqueous and hexane extracts. Methanol and acetone extracts showed higher phenolic, proanthocynadin and flavonoids contents (52.47 ± 6.82 and 29.31 ± 1.49), (334.8±85.1 and 231.22 ±1.80),(288±4.26 & 208.2±17.7) compared to the aqueous and hexane extracts. The alkaloid and saponin contents were 26.67 and 12.85 % respectively. The IC50 values of the methanol extract for DPPH, ABTS and total antioxidant capacity (TAC) were 0.345, 0.017 and 0.116 mg/mL, respectively. Methanol extract of V. karroo pod showed higher polyphenolic content of all the extracts analysed with corresponding strong free radical scavenging potential. These underutilized pods could serve as a new source of antioxidant compounds which could help in combating various ailments. The great antioxidant activity displayed by V. karroo pods extract supports the therapeutic use of this plant in traditional medicine and are attributed to the phytochemical content.The current information suggests that extracts from Vachellia karroo pods might be a ix cheap potential source of natural antioxidants that could be of great importance for the treatment of free radical related diseases. Furthermore, it makes a case for the utilization of the pods instead of discarding them as waste materials. The evaluation of antimicrobial potential of Vachellia karroo extracts was carried out using agar dilution assay against 8 bacterial strains-4 gram-positive [ Enterococcus faecalis (ATCC 29212), Staphylococcus aureus (OK), Bacillus subtilis KZN, Bacillus cereus, and Streptococcus pyogenes] and 4 gram-negative strains[ Vibrio cholera, Klebsiella pneumonia (ATCC 4352), Pseudomonas aeruginosa (ATCC 19582), Salmonella typhi (OK) ]. Six fungal isolates[ Trichophyton mucoides ATCC 201382, Candida albicans (ATCC 10231), Candida glabatra, Penicillium chrysogenum, Aspergillus fumigatus, Penicillum aurantiogriseum] were usedor antifungal assessment. The methanol extract exhibited broad-spectrum activity, with Gram-positive strains being more sensitive than Gram-negative strains. The extracts also showed great inhibition against Candida albicans the fungal isolate responsible for causing candidiasis in women. Since the pods showed promising antimicrobial activity, they could serve as a cheap source for the treatment and management of these sexually transmitted infections. The brine shrimp toxicity test revealed successful hatching of the cysts was in the order: Aqueous extract> methanol extract> hexane extract> acetone extract. The hatching of nauplii was in a concentration dependent fashion, with hatching success decreasing with increase in concentration of extracts. Lethality of extracts determined based on Meyerʼs index of toxicity, showed that the acetone and hexane extracts of V. karroo were moderately toxic. The results of this study indicated that aqueous and methanolic extracts of Vachellia karroo pods were not toxic, therefore supporting its traditional therapeutic usage. This implies that rather than discard x the pods as is waste as it being currently done, Vachellia karroo pods could be salvaged and processed along with the leaves, thus reducing environmental pollution. Proximate parameters (moisture, ash, crude fibre, crude fat, proteins, and carbohydrate) were evaluated using ALASA methods, and elemental analysis by ICP-OES technique. Nutritional analysis showed that V. karroo pods had low content of crude fat and high content of crude fibre, ash, crude protein, and carbohydrate sufficient to meet the recommended dietary allowances. The pods were rich in major minerals Ca, K, P and Mg, with sufficient amount of trace elements Na, Fe, Zn, and Cu. The outcome of this study suggests that Vachellia karroo pods have good nutritional potentials to support or complement the recommended dietary allowance and could be a cheap source of essential nutrients that could help in ameliorating most nutritional challenges as well as contribute remarkably to the amount of nutrient intake in human and animal diets.
- Full Text:
- Date Issued: 2019
- Authors: Maposa, Sandisiwe
- Date: 2019
- Subjects: Medicinal plants
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10353/14583 , vital:40017
- Description: Vachellia karroo is widely used in folk medicine in the Eastern Cape Province South Africa, however, the pods are usually discarded as waste. The current study was conducted to evaluate the phytochemical content and the antioxidant activity of V. karroo pods. The pods were extracted using acetone, distilled water, hexane and methanol. The total phenol, flavonoid, proanthocyanidin, alkaloid and saponin contents of the various extracts were determined spectrometrically and antioxidant activity was evaluated using inhibition of 2,2-diphenyl-1- picrylhydrazyl (DPPH), 2,2´-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid (ABTS), nitric oxide (NO) radicals, while total antioxidant capacity (TAC) was estimated by the phosphomolybdenum assay. V. karro pods had significant amounts of total phenols, flavonoids and proanthocynadin, although, proanthocynadin was not detected in the aqueous and hexane extracts. Methanol and acetone extracts showed higher phenolic, proanthocynadin and flavonoids contents (52.47 ± 6.82 and 29.31 ± 1.49), (334.8±85.1 and 231.22 ±1.80),(288±4.26 & 208.2±17.7) compared to the aqueous and hexane extracts. The alkaloid and saponin contents were 26.67 and 12.85 % respectively. The IC50 values of the methanol extract for DPPH, ABTS and total antioxidant capacity (TAC) were 0.345, 0.017 and 0.116 mg/mL, respectively. Methanol extract of V. karroo pod showed higher polyphenolic content of all the extracts analysed with corresponding strong free radical scavenging potential. These underutilized pods could serve as a new source of antioxidant compounds which could help in combating various ailments. The great antioxidant activity displayed by V. karroo pods extract supports the therapeutic use of this plant in traditional medicine and are attributed to the phytochemical content.The current information suggests that extracts from Vachellia karroo pods might be a ix cheap potential source of natural antioxidants that could be of great importance for the treatment of free radical related diseases. Furthermore, it makes a case for the utilization of the pods instead of discarding them as waste materials. The evaluation of antimicrobial potential of Vachellia karroo extracts was carried out using agar dilution assay against 8 bacterial strains-4 gram-positive [ Enterococcus faecalis (ATCC 29212), Staphylococcus aureus (OK), Bacillus subtilis KZN, Bacillus cereus, and Streptococcus pyogenes] and 4 gram-negative strains[ Vibrio cholera, Klebsiella pneumonia (ATCC 4352), Pseudomonas aeruginosa (ATCC 19582), Salmonella typhi (OK) ]. Six fungal isolates[ Trichophyton mucoides ATCC 201382, Candida albicans (ATCC 10231), Candida glabatra, Penicillium chrysogenum, Aspergillus fumigatus, Penicillum aurantiogriseum] were usedor antifungal assessment. The methanol extract exhibited broad-spectrum activity, with Gram-positive strains being more sensitive than Gram-negative strains. The extracts also showed great inhibition against Candida albicans the fungal isolate responsible for causing candidiasis in women. Since the pods showed promising antimicrobial activity, they could serve as a cheap source for the treatment and management of these sexually transmitted infections. The brine shrimp toxicity test revealed successful hatching of the cysts was in the order: Aqueous extract> methanol extract> hexane extract> acetone extract. The hatching of nauplii was in a concentration dependent fashion, with hatching success decreasing with increase in concentration of extracts. Lethality of extracts determined based on Meyerʼs index of toxicity, showed that the acetone and hexane extracts of V. karroo were moderately toxic. The results of this study indicated that aqueous and methanolic extracts of Vachellia karroo pods were not toxic, therefore supporting its traditional therapeutic usage. This implies that rather than discard x the pods as is waste as it being currently done, Vachellia karroo pods could be salvaged and processed along with the leaves, thus reducing environmental pollution. Proximate parameters (moisture, ash, crude fibre, crude fat, proteins, and carbohydrate) were evaluated using ALASA methods, and elemental analysis by ICP-OES technique. Nutritional analysis showed that V. karroo pods had low content of crude fat and high content of crude fibre, ash, crude protein, and carbohydrate sufficient to meet the recommended dietary allowances. The pods were rich in major minerals Ca, K, P and Mg, with sufficient amount of trace elements Na, Fe, Zn, and Cu. The outcome of this study suggests that Vachellia karroo pods have good nutritional potentials to support or complement the recommended dietary allowance and could be a cheap source of essential nutrients that could help in ameliorating most nutritional challenges as well as contribute remarkably to the amount of nutrient intake in human and animal diets.
- Full Text:
- Date Issued: 2019
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