The endocannabinoid system in inflammatory bowel system
- Authors: Ababio, Frank James Kweku
- Date: 2014
- Subjects: Inflammatory bowel diseases , Gastrointestinal system , Gastrointestinal system -- Diseases
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10346 , http://hdl.handle.net/10948/d1020338
- Description: Crohn’s disease (CD) and ulcerative colitis (UC) constitute the two major forms of inflammatory bowel disease (IBD), which are disorders of chronic inflammation in the gastrointestinal tract that are associated with significant morbidity and socioeconomic burden. IBD patients with long-standing intestinal inflammation are more prone to developing colorectal cancer (CRC). Until now, none of the existing IBD treatments is able to heal the mucosal ulcerations satisfactorily. The endocannabinoid system (ECS), which comprises of endogenous cannabinoid ligands, their receptors, and metabolic enzymes, has been implicated in gut homeostasis, visceral sensation, inflammation and gastrointestinal motility. Available studies in rodent models of IBD suggest that enhancing the ECS tone may reduce inflammation and improve mucosal integrity. This evidence indicates that the components of the ECS seem well positioned to exert a protective role in IBD and also to offer a great opportunity for therapeutic exploitation. Despite the role of the ECS in the gut, the presence and function of the components of the ECS is not well characterised in human IBD. The primary aim of the study was to investigate the state of the major components of the ECS in human IBD and to establish whether IBD is associated with any changes of the components of the ECS. Cannabinoid CB1 and CB2 receptors, enzymes for endocannabinoid biosynthesis PLC, “LRAT”, NAPE-PLD and DAGL, and endocannabinoid metabolic enzymes FAAH and MAGL were analysed from colonic tissue samples of CD, UC and control patients by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) to determine the relative mRNA expression of the above genes. The RT-qPCR analysis showed that the mRNA expression of PLC, LRAT, and NAPE-PLD were unchanged in both CD and UC, whiles DAGL mRNA was decreased in UC but was unchanged in CD. The endocannabinoid degradation enzymes, FAAH mRNA expression was also unchanged in CD but decreased in UC, whereas the mRNA expression of MAGL was significantly decreased in both CD and UC. NAPE-PLD/FAAH and DAGL/MAGL ratios, an estimation of the balance of AEA and 2-AG levels, showed that AEA and 2-AG levels could be increased and unchanged, respectively, in IBD. The mRNA expression of CB1 was significantly decreased in CD and UC whilst CB2 mRNA expression was unchanged in both forms of IBD. The study demonstrated that the components of the ECS which were investigated were present in colonic tissues of both IBD patients and healthy individuals, but they appear to be off balance in CD and UC patients. The decreased CB1 receptors in IBD patients could be an important modifier in the disease and could also provide a possible pathoaetiological mechanism linking IBD and CRC. Although these findings look promising, more studies with larger sample size are required to characterise the components of the ECS in human IBD.
- Full Text:
- Date Issued: 2014
- Authors: Ababio, Frank James Kweku
- Date: 2014
- Subjects: Inflammatory bowel diseases , Gastrointestinal system , Gastrointestinal system -- Diseases
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10346 , http://hdl.handle.net/10948/d1020338
- Description: Crohn’s disease (CD) and ulcerative colitis (UC) constitute the two major forms of inflammatory bowel disease (IBD), which are disorders of chronic inflammation in the gastrointestinal tract that are associated with significant morbidity and socioeconomic burden. IBD patients with long-standing intestinal inflammation are more prone to developing colorectal cancer (CRC). Until now, none of the existing IBD treatments is able to heal the mucosal ulcerations satisfactorily. The endocannabinoid system (ECS), which comprises of endogenous cannabinoid ligands, their receptors, and metabolic enzymes, has been implicated in gut homeostasis, visceral sensation, inflammation and gastrointestinal motility. Available studies in rodent models of IBD suggest that enhancing the ECS tone may reduce inflammation and improve mucosal integrity. This evidence indicates that the components of the ECS seem well positioned to exert a protective role in IBD and also to offer a great opportunity for therapeutic exploitation. Despite the role of the ECS in the gut, the presence and function of the components of the ECS is not well characterised in human IBD. The primary aim of the study was to investigate the state of the major components of the ECS in human IBD and to establish whether IBD is associated with any changes of the components of the ECS. Cannabinoid CB1 and CB2 receptors, enzymes for endocannabinoid biosynthesis PLC, “LRAT”, NAPE-PLD and DAGL, and endocannabinoid metabolic enzymes FAAH and MAGL were analysed from colonic tissue samples of CD, UC and control patients by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) to determine the relative mRNA expression of the above genes. The RT-qPCR analysis showed that the mRNA expression of PLC, LRAT, and NAPE-PLD were unchanged in both CD and UC, whiles DAGL mRNA was decreased in UC but was unchanged in CD. The endocannabinoid degradation enzymes, FAAH mRNA expression was also unchanged in CD but decreased in UC, whereas the mRNA expression of MAGL was significantly decreased in both CD and UC. NAPE-PLD/FAAH and DAGL/MAGL ratios, an estimation of the balance of AEA and 2-AG levels, showed that AEA and 2-AG levels could be increased and unchanged, respectively, in IBD. The mRNA expression of CB1 was significantly decreased in CD and UC whilst CB2 mRNA expression was unchanged in both forms of IBD. The study demonstrated that the components of the ECS which were investigated were present in colonic tissues of both IBD patients and healthy individuals, but they appear to be off balance in CD and UC patients. The decreased CB1 receptors in IBD patients could be an important modifier in the disease and could also provide a possible pathoaetiological mechanism linking IBD and CRC. Although these findings look promising, more studies with larger sample size are required to characterise the components of the ECS in human IBD.
- Full Text:
- Date Issued: 2014
Prevalence and seasonal changes of gastro-intestinal parasites of ovine on three different veld types in communal farming areas of the Eastern Cape Province, South Africa
- Jansen, Mlungisi Selby https://orcid.org/0000-0002-6735-1054
- Authors: Jansen, Mlungisi Selby https://orcid.org/0000-0002-6735-1054
- Date: 2023
- Subjects: Gastrointestinal system , Parasites , Traditional farming
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10353/28233 , vital:73956
- Description: The current study aimed to investigate the prevalence and seasonal changes of gastro-intestinal parasites GIP of ovine grazed don three different veld types in the communal farming areas of the Eastern Cape Province. A total of 1242 sheep faecal samples were collected monthly between January 2012 and December 2015 to investigate the prevalence and season occurrence of internal parasites. From September to November 2018, a semi-structured questionnaire survey was conducted in three agro-ecological zones, humid Wartburg, semi-humid Allen waters, and arid region Cradock commonages to investigate farmers’ knowledge of the prevalence and occurrence of GIP in sheep. The prevalence of gastro-intestinal parasites was significantly higher in the humid zone roundworms 81 percent and coccidia 37 percent, followed by the semi-humid zone roundworms 75 percent and coccidia 22 percent and the arid zone was the lowest roundworms 71 percent and coccidia 14 percent. Roundworms had significantly higher counts P 0.05 mostly in hot-wet seasons of the year spring and summer and low during dry cold months of the year autumn and winter across all veld types. The humid zone had significantly higher counts P 0.05 in the seasonal occurrence of roundworms and coccidia, followed by humid zone, and very low counts were encountered in the arid zone during the study period. Seasonal occurrence of roundworm species was significant across all veld types, 64 percent of farmers were males and 36 percent were females. The study reveals that helminths and coccidia are major causative agents causing parasitic infections in livestock production, and therefore, good animal health practices management practices including proper hygiene should be followed to prevent parasitic infection in small ruminants. , Thesis (MSc) -- Faculty of Science and Agriculture, 2023
- Full Text:
- Date Issued: 2023
- Authors: Jansen, Mlungisi Selby https://orcid.org/0000-0002-6735-1054
- Date: 2023
- Subjects: Gastrointestinal system , Parasites , Traditional farming
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10353/28233 , vital:73956
- Description: The current study aimed to investigate the prevalence and seasonal changes of gastro-intestinal parasites GIP of ovine grazed don three different veld types in the communal farming areas of the Eastern Cape Province. A total of 1242 sheep faecal samples were collected monthly between January 2012 and December 2015 to investigate the prevalence and season occurrence of internal parasites. From September to November 2018, a semi-structured questionnaire survey was conducted in three agro-ecological zones, humid Wartburg, semi-humid Allen waters, and arid region Cradock commonages to investigate farmers’ knowledge of the prevalence and occurrence of GIP in sheep. The prevalence of gastro-intestinal parasites was significantly higher in the humid zone roundworms 81 percent and coccidia 37 percent, followed by the semi-humid zone roundworms 75 percent and coccidia 22 percent and the arid zone was the lowest roundworms 71 percent and coccidia 14 percent. Roundworms had significantly higher counts P 0.05 mostly in hot-wet seasons of the year spring and summer and low during dry cold months of the year autumn and winter across all veld types. The humid zone had significantly higher counts P 0.05 in the seasonal occurrence of roundworms and coccidia, followed by humid zone, and very low counts were encountered in the arid zone during the study period. Seasonal occurrence of roundworm species was significant across all veld types, 64 percent of farmers were males and 36 percent were females. The study reveals that helminths and coccidia are major causative agents causing parasitic infections in livestock production, and therefore, good animal health practices management practices including proper hygiene should be followed to prevent parasitic infection in small ruminants. , Thesis (MSc) -- Faculty of Science and Agriculture, 2023
- Full Text:
- Date Issued: 2023
Biological activities and mechanisms of action of two ethnobotanically selected South African medicinal plants on some bacteria associated with gastrointestinal infections
- Olajuyigbe, Olufunmiso Olusola https://orcid.org/0000-0002-7889-0416
- Authors: Olajuyigbe, Olufunmiso Olusola https://orcid.org/0000-0002-7889-0416
- Date: 2012-08
- Subjects: Medicinal plants , Herbs -- Therapeutic use , Gastrointestinal system
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10353/25439 , vital:64249
- Description: In this study, 36 plant species representing 24 families were found to be commonly used for the treatment of a variety of gastrointestinal disorders in Eastern Cape, South Africa. The family Fabaceae had the highest number of species. Out of these, 47.06percent were used in the treatment of dysentery alone while 46.15percent were used in the treatment of diarrhoea. Acacia mearnsii De Wild and Ziziphus mucronata subsp. mucronata Willd were selected for this research because they are extensively used in folkloric medicine in South Africa and there was lack of scientific reports that documented their biological activities. The phytochemical screening, antioxidant activities, in vitro antimicrobial activities, cytotoxicity, the synergistic potentials and mechanisms of actions of these plants were investigated. The phytochemical screening and the antioxidant activities of the two species showed that the quantity of the phenolic compounds, flavonoids and proanthocyanidins detected differ significantly in the various extracts. Of the aqueous, acetone, ethanolic and methanolic extracts of A. mearnsii, the ethanolic extract had the highest flavonoids while the acetone extract had the highest phenolic contents. The proanthocyanidins were highest in the methanol extract while aqueous extracts had the least phytochemicals. Aqueous extract showed the least ferric reducing power but methanol extract indicated the highest reducing power. The reducing power of the extracts was lower than those obtained from the reference standard such as butylated hydroxytoluene (BHT), rutin and ascorbic acid. 2,2’-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) diammonium salt showed that ethanol extract exhibited the highest antioxidant activity at the highest concentration tested. Also, 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay indicated that ethanol extract had the highest radical scavenging activity at the lowest concentration and the activities of all the extracts decreased with increase in their concentrations. In Z. mucronata subsp. mucronata, the phenolics were significantly higher than the flavonoids and proanthocyanidin contents in all the extracts investigated. The ethanol extract had the highest antioxidant activity, followed by the acetone extract while the aqueous extract was the least active. Reacting with ABTS, the 50percent inhibitory concentrations (IC50) were (0.0429 ± 0.04 mg/ml) for aqueous, (0.0317 ± 0.04 mg/ml) for acetone and (0.0306 ± 0.04 mg/ml) for ethanol extracts while they inhibited DPPH radical with 50percent inhibitory concentration (IC50) values of 0.0646 ± 0.02 mg/ml (aqueous), 0.0482 ± 0.02 mg/ml (acetone) and 0.0422 ± 0.03 mg/ml (ethanol). The investigation showed that a positive linear correlation existed between the total phenolic content and antioxidant activity of the extracts and that these plants have strong antioxidant property and free radical scavenging capability. The in vitro antibacterial activities of Acacia mearnsii and Z. mucronata subsp. mucronata showed that their minimum inhibitory concentrations ranged between 0.039 mg/ml and 1.25 mg/ml. With the exception of acetone extract of A. mearnsii having MICs greater than 1.0 mg/ml for Enterococcus faecalis ATCC 29212 and Bacillus subtilis KZN, all other isolates had MICs less than 0.7 mg/ml. In all the bacteria treated with Z. mucronata subsp. mucronata extracts, Enterobacter cloacae ATCC 13047 had MIC greater than 1 mg/ml in methanol extract, Enterococcus faecalis ATCC 29212 and Staphylococcus aureus ATCC 6538 had MICs greater than 1 mg/ml in acetone extract while all other isolates were highly susceptible to the different extracts of Z. mucronata subsp. mucronata and had MICs less than 0.7 mg/ml. While aqueous extract was as active as the alcoholic extracts in A. mearnsii, that of Z. mucronata had no effect. The ethanol extracts exhibited the highest degree of antibacterial activity in both plants. This study, also, showed that the antifungal activity of A. mearnsii ranging 0.3125 – 5.0 mg/ml was higher than those of the different extracts of Z. mucronata subsp. mucronata ranging 1.25 – 10.0 mg/ml. It is evident from the results of the brine shrimp lethality assay that the crude extracts of A. mearnsii with the LC50 equaled 112.36 µg/ml and having the highest levels of toxicity (100percent) death at 500 μg/ml was non toxic (LC50 > 100 μg/ml) while the LC50 for Z. mucronata subsp. mucronata equaled 90.27 µg/ml indicated a low level of toxicity. The effects of combining the crude extracts of these plants with eight antibiotics were investigated by means of checkerboard and agar diffusion methods. On using the methanol extract of A. mearnsii, the agar diffusion assay showed that extract-kanamycin combination had zones of inhibition ≥ 20 ± 1.0 mm in all the bacteria tested (100percent), followed by extract chloramphenicol (90percent) > extract-ciprofloxacin = extract-tetracycline (70percent) > extract amoxicillin (60percent) > extract-nalidixic acid (50percent) > extract-erythromycin (40percent) > extract metronidazole (20percent). The checkerboard showed synergistic interaction (61.25percent), additivity/indifference (23.75percent) and antagonistic (15percent) effects. I, therefore, concluded that the antibacterial potentials of the antibiotics were improved and combining natural products with antibiotic could be a potential source of resistance-modifying agents useful against multi-drug resistant bacteria. The influences of these extracts on the ultrastructures, elemental components, protein and lipid leakages of five different bacteria were determined as the possible mechanisms of action of the extracts investigated. The scanning electron microscopy indicated varied ultrastructural changes in the morphology of bacterial cells treated with the extracts. The X-ray microanalysis showed significant differences between the elemental contents of extract-treated and untreated bacteria while lipids and proteins were leaked to a great extent from the extract-treated bacterial strains in comparison with the untreated ones. The possible mechanisms of action of the extracts may include inhibition of a significant step in peptidoglycan assembly, inhibition of metabolic processes, disruption of cell wall and cell membranes resulting in the efflux of lipid and protein in all the bacteria tested. The possible mechanism of action involved in the lipid and protein leakages in the bacterial cells could be attributed to lipid peroxidation and protein oxidation owing to the antioxidant activities of the extracts that were active beyond the protective levels. I concluded that the morphological changes and the observed leakages showed rapid killing, significant membrane depolarization resulting in leakages and efflux of disintegrated cellular materials. In general, this study has justified the ethnotherapeutic importance of A. mearnsii and Z. mucronata subsp. mucronata in the treatment of microbial infections by indicating the possible mechanisms of action of the crude extracts on the tested bacteria. , Thesis (PhD) -- Faculty of Science and Agriculture, 2012
- Full Text:
- Date Issued: 2012-08
- Authors: Olajuyigbe, Olufunmiso Olusola https://orcid.org/0000-0002-7889-0416
- Date: 2012-08
- Subjects: Medicinal plants , Herbs -- Therapeutic use , Gastrointestinal system
- Language: English
- Type: Doctoral theses , text
- Identifier: http://hdl.handle.net/10353/25439 , vital:64249
- Description: In this study, 36 plant species representing 24 families were found to be commonly used for the treatment of a variety of gastrointestinal disorders in Eastern Cape, South Africa. The family Fabaceae had the highest number of species. Out of these, 47.06percent were used in the treatment of dysentery alone while 46.15percent were used in the treatment of diarrhoea. Acacia mearnsii De Wild and Ziziphus mucronata subsp. mucronata Willd were selected for this research because they are extensively used in folkloric medicine in South Africa and there was lack of scientific reports that documented their biological activities. The phytochemical screening, antioxidant activities, in vitro antimicrobial activities, cytotoxicity, the synergistic potentials and mechanisms of actions of these plants were investigated. The phytochemical screening and the antioxidant activities of the two species showed that the quantity of the phenolic compounds, flavonoids and proanthocyanidins detected differ significantly in the various extracts. Of the aqueous, acetone, ethanolic and methanolic extracts of A. mearnsii, the ethanolic extract had the highest flavonoids while the acetone extract had the highest phenolic contents. The proanthocyanidins were highest in the methanol extract while aqueous extracts had the least phytochemicals. Aqueous extract showed the least ferric reducing power but methanol extract indicated the highest reducing power. The reducing power of the extracts was lower than those obtained from the reference standard such as butylated hydroxytoluene (BHT), rutin and ascorbic acid. 2,2’-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) diammonium salt showed that ethanol extract exhibited the highest antioxidant activity at the highest concentration tested. Also, 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay indicated that ethanol extract had the highest radical scavenging activity at the lowest concentration and the activities of all the extracts decreased with increase in their concentrations. In Z. mucronata subsp. mucronata, the phenolics were significantly higher than the flavonoids and proanthocyanidin contents in all the extracts investigated. The ethanol extract had the highest antioxidant activity, followed by the acetone extract while the aqueous extract was the least active. Reacting with ABTS, the 50percent inhibitory concentrations (IC50) were (0.0429 ± 0.04 mg/ml) for aqueous, (0.0317 ± 0.04 mg/ml) for acetone and (0.0306 ± 0.04 mg/ml) for ethanol extracts while they inhibited DPPH radical with 50percent inhibitory concentration (IC50) values of 0.0646 ± 0.02 mg/ml (aqueous), 0.0482 ± 0.02 mg/ml (acetone) and 0.0422 ± 0.03 mg/ml (ethanol). The investigation showed that a positive linear correlation existed between the total phenolic content and antioxidant activity of the extracts and that these plants have strong antioxidant property and free radical scavenging capability. The in vitro antibacterial activities of Acacia mearnsii and Z. mucronata subsp. mucronata showed that their minimum inhibitory concentrations ranged between 0.039 mg/ml and 1.25 mg/ml. With the exception of acetone extract of A. mearnsii having MICs greater than 1.0 mg/ml for Enterococcus faecalis ATCC 29212 and Bacillus subtilis KZN, all other isolates had MICs less than 0.7 mg/ml. In all the bacteria treated with Z. mucronata subsp. mucronata extracts, Enterobacter cloacae ATCC 13047 had MIC greater than 1 mg/ml in methanol extract, Enterococcus faecalis ATCC 29212 and Staphylococcus aureus ATCC 6538 had MICs greater than 1 mg/ml in acetone extract while all other isolates were highly susceptible to the different extracts of Z. mucronata subsp. mucronata and had MICs less than 0.7 mg/ml. While aqueous extract was as active as the alcoholic extracts in A. mearnsii, that of Z. mucronata had no effect. The ethanol extracts exhibited the highest degree of antibacterial activity in both plants. This study, also, showed that the antifungal activity of A. mearnsii ranging 0.3125 – 5.0 mg/ml was higher than those of the different extracts of Z. mucronata subsp. mucronata ranging 1.25 – 10.0 mg/ml. It is evident from the results of the brine shrimp lethality assay that the crude extracts of A. mearnsii with the LC50 equaled 112.36 µg/ml and having the highest levels of toxicity (100percent) death at 500 μg/ml was non toxic (LC50 > 100 μg/ml) while the LC50 for Z. mucronata subsp. mucronata equaled 90.27 µg/ml indicated a low level of toxicity. The effects of combining the crude extracts of these plants with eight antibiotics were investigated by means of checkerboard and agar diffusion methods. On using the methanol extract of A. mearnsii, the agar diffusion assay showed that extract-kanamycin combination had zones of inhibition ≥ 20 ± 1.0 mm in all the bacteria tested (100percent), followed by extract chloramphenicol (90percent) > extract-ciprofloxacin = extract-tetracycline (70percent) > extract amoxicillin (60percent) > extract-nalidixic acid (50percent) > extract-erythromycin (40percent) > extract metronidazole (20percent). The checkerboard showed synergistic interaction (61.25percent), additivity/indifference (23.75percent) and antagonistic (15percent) effects. I, therefore, concluded that the antibacterial potentials of the antibiotics were improved and combining natural products with antibiotic could be a potential source of resistance-modifying agents useful against multi-drug resistant bacteria. The influences of these extracts on the ultrastructures, elemental components, protein and lipid leakages of five different bacteria were determined as the possible mechanisms of action of the extracts investigated. The scanning electron microscopy indicated varied ultrastructural changes in the morphology of bacterial cells treated with the extracts. The X-ray microanalysis showed significant differences between the elemental contents of extract-treated and untreated bacteria while lipids and proteins were leaked to a great extent from the extract-treated bacterial strains in comparison with the untreated ones. The possible mechanisms of action of the extracts may include inhibition of a significant step in peptidoglycan assembly, inhibition of metabolic processes, disruption of cell wall and cell membranes resulting in the efflux of lipid and protein in all the bacteria tested. The possible mechanism of action involved in the lipid and protein leakages in the bacterial cells could be attributed to lipid peroxidation and protein oxidation owing to the antioxidant activities of the extracts that were active beyond the protective levels. I concluded that the morphological changes and the observed leakages showed rapid killing, significant membrane depolarization resulting in leakages and efflux of disintegrated cellular materials. In general, this study has justified the ethnotherapeutic importance of A. mearnsii and Z. mucronata subsp. mucronata in the treatment of microbial infections by indicating the possible mechanisms of action of the crude extracts on the tested bacteria. , Thesis (PhD) -- Faculty of Science and Agriculture, 2012
- Full Text:
- Date Issued: 2012-08
An investigation of short-chain fatty acid profiles and influential gastrointenstinal microbiota associated with irritable bowel syndrome
- Authors: Theunissen, Reza
- Date: 2013
- Subjects: Fatty acids in human nutrition , Gastrointestinal system
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10352 , http://hdl.handle.net/10948/d1020943
- Description: Microbiota are present in large numbers and as a diverse population within the gastrointestinal tract. There are approximately 400 different species of microbiota which may be beneficial, harmful or both, but each play an important role in the regulation and modulation of the hosts’ bowel processes (McOrist et al. 2008; Dethlefsen et al. 2008). Many of these colon microbiota allow for saccharolytic fermentation of non-digestible dietary fibres and carbohydrates into by-products and intermediates, followed by a subsequent conversion into short chain fatty acids (SCFAs) (mainly n-butyric acid, propionic acid and acetic acid) each of which play an important role in maintaining colon homeostasis (Topping & Clifton 2001). A balance of ‘good’ microbiota (e.g., Bacteroides spp./ Bifidobacteria spp.) and ‘bad’ microbiota (e.g., Veilonellae) and the optimal production of various SCFAs within the gut could possibly allow for proper functioning of the large intestine and assist in decreasing the onset of various colonic disorders such as Irritable Bowel Syndrome (IBS). The sample group for the study consists of male and female patients, with an average age of 40 to 50 years old, whom of which have been diagnosed with either constipation IBS (C-IBS) or diarrhoea IBS (D-IBS) via the Rome III criteria system for IBS diagnosis. DNA and SCFA extractions were optimised for human stool, colonic fluid and tissue biopsy sample obtained from the aforementioned patients. Optimization steps allowed for starting material with high analysis integrity. Different methods of microbiota analysis, such as ARISA, were investigated; however, real-time qPCR was selected as the best method to identify and quantify specific microbiota. Extracted SCFAs were separated via gas chromatography and identified and quantified via Mass Spectrometry. Significant changes in microbial content and SCFA profiles were found to be associated with healthy and IBS patients. Results obtained would however be influenced by external factors typical of clinical studies of this nature. This study allows for opportunities for future research into understanding IBS.
- Full Text:
- Date Issued: 2013
- Authors: Theunissen, Reza
- Date: 2013
- Subjects: Fatty acids in human nutrition , Gastrointestinal system
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10352 , http://hdl.handle.net/10948/d1020943
- Description: Microbiota are present in large numbers and as a diverse population within the gastrointestinal tract. There are approximately 400 different species of microbiota which may be beneficial, harmful or both, but each play an important role in the regulation and modulation of the hosts’ bowel processes (McOrist et al. 2008; Dethlefsen et al. 2008). Many of these colon microbiota allow for saccharolytic fermentation of non-digestible dietary fibres and carbohydrates into by-products and intermediates, followed by a subsequent conversion into short chain fatty acids (SCFAs) (mainly n-butyric acid, propionic acid and acetic acid) each of which play an important role in maintaining colon homeostasis (Topping & Clifton 2001). A balance of ‘good’ microbiota (e.g., Bacteroides spp./ Bifidobacteria spp.) and ‘bad’ microbiota (e.g., Veilonellae) and the optimal production of various SCFAs within the gut could possibly allow for proper functioning of the large intestine and assist in decreasing the onset of various colonic disorders such as Irritable Bowel Syndrome (IBS). The sample group for the study consists of male and female patients, with an average age of 40 to 50 years old, whom of which have been diagnosed with either constipation IBS (C-IBS) or diarrhoea IBS (D-IBS) via the Rome III criteria system for IBS diagnosis. DNA and SCFA extractions were optimised for human stool, colonic fluid and tissue biopsy sample obtained from the aforementioned patients. Optimization steps allowed for starting material with high analysis integrity. Different methods of microbiota analysis, such as ARISA, were investigated; however, real-time qPCR was selected as the best method to identify and quantify specific microbiota. Extracted SCFAs were separated via gas chromatography and identified and quantified via Mass Spectrometry. Significant changes in microbial content and SCFA profiles were found to be associated with healthy and IBS patients. Results obtained would however be influenced by external factors typical of clinical studies of this nature. This study allows for opportunities for future research into understanding IBS.
- Full Text:
- Date Issued: 2013
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