FACULTY OF LIFE SCIENCES

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Alchornea laxiflora AQUEOUS LEAF EXTRACT: ITS ALLELOPATHIC POTENTIAL AND BIOACTIVE AGENTS
(UNIVERSITY OF ILORIN, 2017-12) OSINEYE, SUAIB OLASUPO
The dangers posed by synthetic herbicides necessitated the need to search for eco-friendly alternatives in some plants by exploring their allelopathic property. Allelopathic plants have the ability to inhibit the growth and survival of surrounding plants. The aim of this experiment was therefore to evaluate the allelopathic potential of Alchornealaxiflora and to isolate the bioactive agents responsible. The objectives of the study were to: (i) screen Alchornealaxiflora aqueous leaf extract for the presence of secondary metabolites; (ii) examine the growth inhibitory, germination inhibitory and genotoxic effects of the extract; (iii) determine the effects of the extract on some photosynthetic and lipid synthesis parameters; (iv) determine the effects of the extract on oxidative stress parameters; (v) determine the effect of the extract on some osmotic stress indicators; (vi) carry out a bioactivity-guided isolation and identification of the bioactive agents; (vii) conduct an autodock analysis using enzymes. The extract was first subjected to Alliumcepa test to evaluate the growth-inhibitory and genotoxic effects (reduction of mitotic index, chromosomal aberration), then tomato seedlings were used to further evaluate its germination inhibitory and other phytotoxic effects (photosynthesis inhibition, lipid synthesis inhibition, oxidative stress induction, osmotic stress induction)with atrazine as standard. Thereafter, it was subjected to standard chromatographic techniques involving bioactivity-guided fractionations, thin layer chromatography and high performance liquid chromatography as well as isolate the bioactive agents responsible for the allelopathic property. Finally, the isolated compounds were subjected to molecular docking techniques using enzymes (Ferredoxin NADP-reductase and Acetyl CoA carboxylase and alpha amylase) to further confirm the bioactive agents. Data were subjected to Analysis of Variance Test at p˂0.05. The findings of the studywere that the extract: i. contained a number of secondary metabolites including alkaloids, phenol, phlobatannins, terpenes, tannin andflavonoids. ii. caused significant reductions (p˂0.05) in growth, germination,and abnormalities in chromosomal activities in tomato seedlingsandAlliumcepa root cells. iii. caused significant reductions (p˂0.05) inChlorophyll contents and activities of Ferredoxin NADP-reductase and Acetyl CoA carboxylase enzymes of tomato seedlings. iv. caused significant increases (p˂0.05) in Malondialdehyde concentrations and decreasesin activities of antioxidant enzymes (Superoxide dismutase, Catalase andPeroxidase). v. caused significant increases (p˂0.05) in protein, proline and total soluble sugarconcentrations. vi. contained Rutin and Quercetrin as its Allelochemicals. vii. contained Rutin and Quercetrin which interacted favourably with the amino acid residues on Alpha amylase, Acetyl-coA carboxylase and Ferredoxin NADP-reductase. The study concluded that Rutin and Quercetrin are partly responsible for the allelopathic activity exhibited by Alchornealaxifloraaqueous leaf extract. It is therefore recommendedthat Alchornealaxifloramay be used as a source of templates for the development of bio-herbicides.
ANTIMALARIAL ACTIVITIES OF SOLVENT EXTRACTS AND FRACTIONS FROM Adansonia digitata STEM BARK IN Plasmodium berghei-INFECTED MICE
(UNIVERSITY OF ILORIN, 2017-12) ADEOYE, AKINWUNMI OLUWASEUN
Malaria is caused by a protozoan of the genus Plasmodium and it is an infectious disease of humans and other animals. In the process of studying human malaria parasites, rodent parasites are recognized as valuable model parasites for the investigation of parasite-host interactions, biology of malaria parasites, drug testing and vaccine development. Increase in the resistance of malaria parasite to synthetic drugs has led to the continuous search for alternative treatment approach from plant origin. Majority of these plants have biologically active polyphenol components and phytochemicals which have protective and therapeutic properties. These phytochemicals are found as secondary plant metabolites that are explored for their bioactivities with application in medicine. Adansonia digitata L. Baobab (Bombacaceae) is rich in these secondary metabolites and is used for the treatment of malaria in Nigeria. The aim of this research was to investigate the possible anti-malarial potential of aqueous and methanolic extracts of A. digitata stem bark and its fraction on Plasmodium berghei infected mice. Aqueous and methanolic (AEAD and MEAD) extract was obtained from Adansonia digitata stem bark utilizing solvent extraction and the fractions was obtained through solvent partitioning. The free radical scavenging activity of the extract in vitro was evaluated. The extract was also evaluated for its antimalarial activity using curative test in mice. Severity of malaria was evaluated by determining the serum and tissue alkaline phosphatase (ALP), aspartate amino transferase (AST) activity, alanine amino tranferase (ALT), the serum C-reactive protein (CRP) and tumor necrosis factor-alpha (TNF-α). Mechanisms of action of the extract were investigated by measuring the degree of tissue peroxidation and tissue antioxidant status, mitochondrial swelling and mitochondria ATPase activity. The efficacy of Adansonia digitata stem bark extract in offering protection against experimental malaria was evaluated and its remediation effect when administered after established infection was also examined. The extract fraction was evaluated for its activity against inhibition of β-hematin formation. The results revealed that Adansonia digitata stem bark extracts exhibited DPPH radical scavenging activity, ferric reducing antioxidant power, metal chelating activity and hydroxyl radical scavenging activity comparable (p<0.05) with those of standard antioxidants. The HPLC-DAD analysis showed significant levels of flavonoids and phenolic acids in the extracts and fractions of Adansonia digitata stem bark. The results revealed that the aqueous and methanolic 21 extracts were active against P berghei NK65 in vivo and were not cytotoxic at both 200 and 400 mg/kg body weight. Methanolic extract exhibited higher antimalarial activity causing 70.15%, 90.18% and 100% inhibition in parasite growth at the dose of 400 mg/kg body weight on day 3, 5 and 7 post-inoculation respectively. The extracts showed a significant dose dependent increase in packed cell volume (PCV), at the two doses when administered after established infection compared to control. Moreover, ethylacetate extract fraction exhibited considerable antiplasmodial activity against inhibition of β-hematin formation (IC50 < 50 μg/ml). However, the ability of the extract fraction to inhibit β-hematin formation was significantly lower (P < 0.05) than that of chloroquine and artesunate. The extracts significantly reduced (P < 0.05) malondialdehyde concentrations in all the tissues investigated compared to the infected untreated mice on day 5 and 7 post-inoculation. Administration of the extract after established infections significantly increased (P < 0.05) glutathione concentrations and activities of superoxide dismutase and catalase in a dose-dependent manner compared to the untreated control on day 5 and 7 post-inoculation. The extracts significantly decreased serum creatinine concentrations but significantly increased (P < 0.05) serum urea and uric acid concentrations. Administration of the extract reduced serum concentrations of sodium ion, potassium ion, calcium ion and chloride ion compared to control but significantly increased serum concentrations of magnesium ion when compared to control. A significant increase in the serum concentrations of CRP and TNF-α of the control animals was observed when compared to the baseline (uninfected mice) and the group treated with extract. Significant relationship exists between serum CRP and TNF-α concentration and percentage parasitemia in the control group. Administration of the extracts lowered the CRP and TNF concentrations. Alkaline phosphatase (ALP) activity of the control group increase significantly (P < 0.05) in all the tissues investigated compared to the group that received the extract. However, a significant reduction in ALT and AST activity of the control group were observed compared to the group that received the extract. Ethylacetate fraction of A. digitata stem bark and the phenolics, quercetin and apigenin induced the opening of mitochondrial membrane permeability transition pore in a concentration dependent manner. The ethylacetate extract fraction also increased (P < 0.05) significantly the mitochondrial ATPase activity in a concentration dependent manner. 22 The results suggest that Adansonia digitata protects against Plasmodium berghei induced-malaria, and that administration of the extract after established infection reduced malaria progression. A significant relationship was obtained between antioxidant activity and phenolic content indicating that phenolic compounds contribute significantly to antioxidant and antimalarial activity of the plant. Bioactive compounds from Adansonia digitata influence mitochondria membrane permeability transition by inducing cell death through mitochondria-mediated pathway of apoptosis. Opening of MMPT pore and induction of apoptotic process could likely be one of the mechanisms of action of antimalarial compound.
PHYTOAVAILABILITY, DYNAMICS AND PARTITIONING OF SOME HEAVY METALS IN VEGETABLE FARMSOILS IN ILORIN METROPOLIS.
(UNIVERSITY OF ILORIN, 2018) BEN-UWABOR, PATIENCE OLAYINKA
Vegetables are food materials that provide vitamins, minerals and fibres. These plants are raised during dry season (irrigated) and rainy season (rain fed). Anthropogenic activities involved in the raising of vegetables have been of great concern. There is little or no available information on the safety and qualities of irrigated vegetables. The objectives of this study were to: (i) determine the total heavy metal (HM) load in some farm soils and water; (ii) determine the phytoavailable heavy metals in soil; (iii) determine the heavy metal fractionation; (iv) determine available heavy metals in the vegetables; (v) investigate the heavy metal transfer and pathway in the vegetables; and (vi) evaluate the effects of the agronomic activity on the nutritional value of vegetable accessions. The HM contents of the soil and water were determined using Atomic Absorption Spectrophotometer. Five vegetable accessions: Amaranthus hybridus (NG/AA/03/11/010, NG/AO/11/08/039 and NGBO125) and Corchorous olitorius (NG/OA/Jun/09/002, NG/OA/ 04/010) obtained from National Centre for Genetic Resources and BioTechnology, Ibadan were planted in different soils and irrigated with water collected from the different sites. The HM and the proximate analysis of vegetables were carried out. Data generated were subjected to Analysis of Variance and the means separated by Duncan Multiple Range Test set at 5% level of significance. The findings of the study were that the: i. range of heavy metal content of soils (mg/kg) and water (mg/l) respectively, were: 0.21 4.67 and 0.00-2.75 (Cd); 1.40–4.79 and 2.41-30.08 (Cu); 1.29-11.67 and 5.25-82.00 (Pb) in soils (mg/kg) and 0.00 0.07 and 0.00 0.07 (Cd); 0.00 0.34 and 0.52 1.68 (Cu); 0.04-0.63 and 0.21-0.54 ( Pb) for dry and rainy season respectively; ii. phytoavailable HM range in soils (mg/kg) were: 0.11-0.67 and 0.00-0.13 (Cd), 0.17- 3.86 and 1.80-9.25 (Cu) and 0.50-5.83 and 0.276.25 (Pb) for dry and rainy, respectively; iii. range of soils HM fractionation were: 0.00-1.67 and 0.00-1.17 (Cd); 0.21-6.67 and 0.00-11.67 (Cu) and 0.00-7.33 and 0.00-18.33 (Pb). iv. available HM in the accessions were: 0.00-1.67 and 0.00-1.17 (Cd); 0.17-6.97; 0.00-23.00 (Cu); 0.00-6.67 and 0.00-18.33 (Pb) for dry and rainy season, respectively. However, most accessions accumulated Cd and Pb above the European Union (EU) and WHO/FAO standards for vegetables; v. five vegetable accessions were good accumulators of Cd, Cu and Pb with bioaccumula tion coefficient of accessions greater than 0.5; and vi. accessions had 6.18-10.14% (H2O); 1.21-5.56% (ash), 3.19-5.38% (crude fat and oil); 6.26-21.55% (crude protein); 1.07-2.58% (crude fibre) and 62.71-76.83% (crude protein) and 6.53-9.44% (H2O); 1.06-7.14% (ash); 2.84-5.83% (crude fat and oil); 7.02 23.50% (crude protein); 0.93 4.60% (crude fibre); 58.19 75.18% (carbohydrate) in dry and rainy season, respectively. However, the soils and water used made Cd, Cu and Pb more significantly (p<0.05) available to the vegetables. The study concluded that soil and water used for raising vegetables are sources of HM pollution which can lead to health hazard. Farmers should therefore, be advised and encouraged to use treated wastewater for irrigation.
INFLUENCE OF SELECTED BOTANICAL GARDEN STRUCTURE,PLANNING AND DESIGN ON ECOSYSTEM SERVICES AND ATMOSPHERIC AIR QUALITY
(UNIVERSITY OF ILORIN, 2018) ALABI, JOHN OMONIYI
Pollution due to smoke, dust and other pollutants posed serious challenges to atmospheric air and human health. The study was to investigate the influence of selected botanical garden structures (BGS), planning and design on Ecosystem services (ESS) and atmospheric air quality (AAQ) in Lagos, Ibadan, Akure, Ilorin, Kano and Jos universities botanical gardens (BG) in Nigeria. The objectives of the study were to: (i) identify and determine landscape status (LS) of tree species (TS); (ii) identify tree crown characteristic indices (TCCI) for shade and shelter; (iii) determine TS air pollution tolerance index (APTI); (iv) determine level of air quality index (AQI); and (v) design BGS prototype with TS that can enhance provision of ESS. Study areas were divided into several plots using transects lines for TS and lichens (L); samples were collected from each plot, frequency values (FV) for TS and L were used to determine the LS. Cluster analysis (CA) of the TS and tree crown characteristics indices (TCCI) were performed, by assigned codes 1 – 6 to the taxonomic character (TC) in ascending order according to the observation and measurement. Ascorbic acid, relative water content, pH and total chlorophyll were evaluated from fresh leaves; values obtained were used to calculate APTI. Pollutants and metrological parameters were determined using standard methods and weather equipment; validated with LaMotte air-sampler for inside-outside BGs AQI during dry (DS) and rainy seasons (RS), obtained values were used for AQI and correlation analysis. Layout plan (LP) was used to design a complete BGS, planting and leaf morphology (LM) plans, based on computer- aided architectural design (CAAD). The study revealed that: i. TS of 180 belonging to 25 families (F) were represented in the study areas, LS showed abundant (16F) and rare (9F). FV ranged from 9.71 to 0.94 TS and 8.95 to 0.01 L of good and moderate level of AAQ. Moraceae has the highest numbers of TS (19) accounting for 10.56% LS of TS; ii. CA of TS in the 6BGs gave percentage clusters of 63%, outliers 37% and TCCI ranged from 3 to 7 and 1 to 2.67 for shade and shelter respectively; iii. APTI for Mangifera indica has high value of tolerance 29.46 in all the BGs, while sensitive species Ficus bengamina has lowest value 5.91; iv. DS and RS AQI ranged from of 34.49 to 40.47 and 31.36 to 37.52 (inside BGs) and 84.60 to 96.38 and 74.38 to 84.82 (outside BGs). However, significant difference p≤0.05 existed among the BGs, pollutants and AQI; v. BGS prototype designed by CAAD ranged from LP, building 9825mm by 19870mm, planting and LM plans for TS family of Apocynaceae, Moraceae, Araucariaceae and Mimosaceae as sensitive species and Anacardaceae, Meliaceae, Annonaceae and Mytaceae as tolerant species. The study concluded that, TS families of Apocynaceae, Anacardiaceae, Meliaceae, Moraceae, Mimosaceae, Myrtaceae and Araucariaceae out of 25F were good candidates for ESS such as: lichens occurrence, shading, pollutant removal and AQI improvement by BGS. The BGS is therefore, recommended for provision of ESS to the society.
IN VITRO AND IN VIVO ANTIMALARIAL, ANTIOXIDANT AND TOXICOLOGICAL EFFECTS OF METHYL GALLATE AND PALMATINE COMBINATION
(UNIVERSITY OF ILORIN, 2018-02) ADEGUNLOYE, ADEGBENRO PETER
Resistance of malaria parasite to conventional drugs and the high cost of potent ones have necessitated the search for new drugs. Methyl gallate and palmatine are compounds with known in vitro activities against multidrug-resistant strains of Plasmodium falciparum, but without in vivo activities, which are associated with plants from which they have been isolated. This, therefore, necessitates the evaluation of antimalarial activities of combination of isolated compounds. Thus, the objectives of this study were to evaluate: (i) in vitro antimalarial activity of methyl gallate and palmatine combination (MGPAL); (ii) interaction of methyl gallate and palmatine with selected P. falciparum proteins in silico; (iii) antimalarial activity of MGPAL in P. berghei-infected mice; (iv) antimalarial activity of MGPAL when co-administered with a bio-enhancer in P. berghei-infected mice; (v) antioxidant activities of MGPAL in vitro and in P. berghei-infected mice when co-administered with a bio-enhancer; and (vi) in vivo toxicity of MGPAL when co-administered with a bio-enhancer using selected organ function indices. In vitro antimalarial activity of methyl gallate and palmatine combination was determined using inhibition of β-hematin formation assay. Suppressive and curative antimalarial activities of the individual drugs and their combination, with or without piperine (bio-enhancer), were evaluated using P. berghei NK65-infected mice. Antioxidant activities of MGPAL in vitro and in P. berghei-infected mice when co-administered with piperine were evaluated. Toxicological effects of MGPAL when co-administered with piperine were also evaluated in rats. Data were analyzed using Analysis of Variance at P<0.05. The findings of this study were that: i. MGPAL inhibited β-hematin formation in vitro (IC50 - 0.73 µg/mL, indicating antimalarial activity), with the compounds exhibiting synergistic interaction when combined in ratio 3:2; ii. Methyl gallate and palmatine exhibited higher affinity for plasmepsins I, II and III than for other proteins studied in silico; iii. MGPAL in ratio 3:2 (MGPAL3:2), in the absence of piperine, exhibited no in vivo antimalarial activity, causing less than 30% reduction in parasitemia; iv. MGPAL3:2, in the presence of piperine, caused more than 30% reduction in parasitemia (especially at 12.5 and 25 mg/kg body weight in the suppressive and curative antimalarial tests) in P. berghei NK65-infected mice mainly on days 6 and 8 post-inoculation; v. MGPAL3:2 demonstrated DPPH, nitric oxide, hydroxyl radical scavenging activities and induced antioxidant defense system in murine malaria model in the presence of piperine; and vi. MGPAL3:2, in the presence of piperine, significantly increased (p<0.05) plasma conjugated bilirubin concentration (at the highest dose), HDL-cholesterol concentration (at doses higher than 12.5 mg/kg body weight) and liver aspartate aminotransferase activity (at doses higher than 25 mg/kg body weight) but significantly reduced (p<0.05) atherogenic index and Low density lipoprotein-cholesterol concentration (at doses higher than 6.25 mg/kg body weight) and plasma Ca2+ concentration (at the highest dose) compared to controls. The study concluded that, in the presence of piperine, MGPAL3:2 exhibited antimalarial activity, ameliorated ROS-mediated secondary complications of malaria and adversely affected some liver and kidney functions, especially at higher doses. Further studies on the mechanisms of action of the compound combination should be carried out.
ASSESSMENT OF STORAGE METHODS ON THE SHELF LIFE, NUTRITIONAL COMPOSITION AND FUNGAL SPOILAGE OF TOMATO (SOLANUM LYCOPERSICUM L.) FRUITS
(UNIVERSITY OF ILORIN, 2018-02) GARUBA, TAOFEEQ
Tomato (Solanum lycopersicum L.) fruit is an important component of daily diet. The fruit is perishable and commonly attacked by fungi during storage. Hence, it becomes imperative to enhance its shelf life and minimize its spoilage. The objectives of this study were to evaluate the efficacy of storage structures and botanicals in prolonging the shelf life of four varieties of tomato fruits under different storage conditions, determine the effects of storage conditions on nutritional composition of four varieties of tomato fruits, evaluate the efficacy of botanicals on fungal load during storage, isolate and identify fungi associated with deteriorated tomato fruits using both morphological and molecular tools and determine the degree of virulence of fungal isolates and their biomass in different carbon and nitrogen-rich media. Four varieties of tomato were used in this study: two local varieties (Hausa and Yoruba varieties) and two improved varieties (Tropimech and Roma VF varieties). Storage structures used were plastic crate, raffia basket and pot in pot refrigerator, while botanicals (wood ash of Vitellaria paradoxa, sawdust of Khaya ivonresis and Oryza sativa straw) were preservatives. Each botanical and sampled fruits from each variety were mixed in ratio 1:2 and stored accordingly. The shelf life was studied and lycopene content, proximate and mineral composition and fungal load were determined. The isolated fungi were identified using macromorphological and micromorphological features. Internal Transcribed Spacer (ITS) regions of the ribosomal DNA (rDNA) of fungi was amplified and sequenced. Pathogenicity test and frequency of occurrence for each isolate were carried out. Biomass of the isolates in response to carbon and nitrogen sources were determined. All data were analyzed using a statistical software called Statistical Package for Social Science (SPSS), version 16.00. One-way Analysis of Variance (ANOVA) was used to determine the differences within the variety. means were separated using Dunca 7 Multiple Range Test (DMRT). Univariate analysis of variation (under General Linear Model) was used to determine the interactions among the fixed factors (variety, storage and botanicals). Statistical software Origin 7.0 was used to plot the line graphs as well as bar charts. The findings from the study revealed that the shelf life of all the varieties was prolonged in pot in pot refrigerator for up to 20 days and storage structures had no significant effect on the loss of firmness at p≤0.05. Proximate analysis showed that moisture was highest (95.92%), followed by carbohydrate (9.04%) in all varieties irrespective of storage structure and botanicals. Mineral composition of all stored tomato fruits was significantly influenced by interactions between variety, storage and botanical at p≤0.05. Sawdust had higher antifungal potential than rice straw and ash by reducing the fungal loads in the tomato fruits. Aspergillus japonicus, Rhizopus oryzae, Curvularia geniculata, Fusarium proliferatum and Fusarium oxysporum were associated with spoilt tomato fruits of all the varieties. Sequencing of the ITS regions of rDNA of the isolates confirmed their identities. All the fungal isolates were pathogenic to different degrees with the local tomato varieties more susceptible than improved varieties. Curvularia geniculata occurred less frequently than other isolates. Biomass of each isolate was dependent on carbon and nitrogen sources in the media. The study concluded that pot in pot refrigerator was the only suitable structure in elongating the shelf life of tomato fruits. Sawdust was very efficacious to reduce incidence of fungal spoilage in tomato fruits, followed by rice straw. Pot in pot refrigerator and sawdust are recommended to minimize postharvest losses of tomato fruits.
ABORTIFACIENT ACTIVITIES AND BIOCHEMICAL ASSESSMENT OF ETHANOLIC EXTRACT OF Citrullus colocynthis PODS IN PREGNANT FEMALE WISTAR RATS
(UNIVERSITY OF ILORIN, 2018-04) ADETUNJI, JULIANA BUNMI
The use of abortifacient herbs is now gaining acceptance over abortifacient drugs due to adverse effects and high cost. Citrullus colocynthis pods (Bitter Apple in English), have been claimed to be used as an abortifacient in Nigeria without scientific evidence to substantiate it. This study was therefore carried out to evaluate the abortifacient activities of ethanolic extract of Citrullus colocynthis pods (EECCP) on pregnant female Wistar rats. The specific objectives were to: (i) qualitatively and quantitatively determine the secondary metabolites and mineral elements in EECCP; (ii) evaluate the abortifacient activity of EECCP on the pregnant female Wistar rats; and (iii) assess the biochemical changes on the pregnant Wistar rats after administration of EECCP. The secondary metabolites in EECCP were screened using standard methods. Sixty pregnant female rats (165.27 ± 7.13g) were randomized into six groups (A-F). Group A (control) received orally 0.5 ml of olive oil, Group B received 1.84mg/kg body weight (bwt) of Mifepristone (reference drug) while those in Groups C, D, E and F received orally 100, 200, 400 and 800 mg/kg bwt of EECCP once daily for 9 days post pregnancy after which abortifacient parameters, biochemical changes, chromosomal aberration, and histological evaluation were done.Data were subjected to analysis of variance and Duncan Multiple Range Test at p˂0.05. The results revealed that: i. EECCP contained alkaloids, saponins, tannins, phlobatannins, glycosides, phenolics, steroids, terpenes, flavonoids, iron, sodium, copper, chromium, potassium, manganese, zinc, nickel, magnesium and calcium with alkaloids (5.2mg/ml) and calcium (27.17mg/l) being the highest; ii. the number of life foetuses, survival ratio, corpora lutea and implantation index were significantly reduced (p<0.05) by EECCP whereas the number of dead foetus, rats that aborted, resorption index, pre- and post-implantation increased significantly; iii. EECCP significantly reduced (p< 0.05) the levels of progesterone, oestrogen, prolactin, luteinizing hormone and follicle stimulating hormone; iv. the decrease in liver and kidney aminotransferases activity by EECCP was accompanied by corresponding increase in the serum enzymes; v. EECCP significantly reduced (p<0.05) albumin, total bilirubin, uric acid, total protein, urea, creatinine, glucose, phosphorus, high density lipoprotein cholesterol in the liver, kidney and serum whereas chloride, total cholesterol, low density lipoprotein cholesterol and atherogenic index increased significantly; vi. EECCP produced increase (p<0.05) in metaphase chromosomal aberration and decreased the mitotic index; vii. EECCP significantly increased malondialdehyde concentrations but reduced significantly (p<0.05) the catalase and superoxide dismutase activities; viii. EECCP significantly reduced the white blood cells, platelet count, mean corpuscular haemoglobin and mean corpuscular volume; and ix. there were degeneration of corpus luteum, loss of both the theca external layers and oocytes in the ovary, erosion of endometrium, rupture of blood vessels, shrinkage of nucleus and sinusoidal spaces in the liver as well as vacuolation of the parenchyma in the kidney. The study concluded that EECCP possesses abortifacient activities at the least toxic dose of 400mg/kg body weight. Therefore, EECCP should be further explored for development of abortifacient drug.
EFFECTS OF CHRONIC ADMINISTRATION OF NIFEDIPINE ON THE REPRODUCTIVE SYSTEM OF WISTAR RATS
(UNIVERSITY OF ILORIN, 2018-05) ABIOYE, ADESOLA IDOWU ABDUR – RAHEEM
Nifedipine, one of the antihypertensive drugs is associated with various side effects on organs of the body but there is paucity of information on its effects on female reproductive system of Wistar rats. The study evaluated the effects of Nifedipine on the female reproductive system. The objectives of the study were to: (i) investigate the effect of Nifedipine on the oestrous cycle of the non – pregnant Wistar rats; (ii) determine the effect of Nifedipine on the serum levels of oestradiol, progesterone, and gonadotrophins in non - pregnant and pregnant Wistar rats; (iii) investigate the effect of Nifedipine on the foetuses at term; and (iv) demonstrate any histoarchitectural changes in the ovary of the treated animals. The study was carried out in two experimental stages: in the first stage, eighteen cycling non-pregnant Wistar rats were used. They were divided into three groups with each group comprising six rats. Rats in group I were administered 10 ml/kg body weight of distilled water (control), group II was administered aqueous solution of Nifedipine 1.5mg/kg body weight, while rats in group III were administered 3 mg/kg body weight. In the second stage, eighteen pregnant Wistar rats were used. They were grouped and treated as the cycling non-pregnant Wistar rats. Doses of distilled water and the drug solutions were delivered orogastrically once daily for 21 days. Vaginal smears cytology was done daily throughout the 21 days in the non-pregnant rats and the cell types recorded. All the medications were administered through the orogastric route. At the end of the experiment, the rats were euthanized using 20 mg/kg body weight of ketamine hydrochloride. Whole body perfusion fixation was done using 4% paraformaldehyde. Blood specimens were taken for evaluation of sex hormones. The ovaries were removed and fixed in 4% paraformaldehyde solution before tissue processing. In the pregnant groups, the pup-containing uteri were similarly harvested for morphological evaluation. Data were analysed using Student’s t-test at 0.05 level of significance. The findings of the study were that: (i) there was total disruption of the oestrous cycles in Nifedipine treated in both the low and high-doses compared with the controls; (ii) the serum levels of oestradiol was 41.0 ± 6.14 pgm/ml for the control, 75.83 ± 12.61 pgm/ml for non-pregnant and 2036.0 ± 50.57 pgm/ml for the pregnant rats; (iii) luteinizing hormone concentration was 13.6 ± 5.51 mIU for non-pregnant, 5.9 ± 0.64 mIU for the pregnant and 9.48 ± 1.45 mIU for the control; (iv) the serum levels of progesterone was 26.2 ± 2.33 ng/ml in non-pregnant rats, 98.4 ± 1.47 ng/ml in pregnant and 24.2 ± 0.96 ng/ml in the control; (v) there were no obvious congenital malformation affecting the foetuses; and (vi) there was loss of cortico-medullary demarcation in the treated groups. The study concluded that there were structural ovarian changes associated with use of Nifedipine in Wistar rats. The study recommended that Nifedipine should be avoided in control especially in non-pregnant individuals.
Larvicidal Properties, Toxicological Effects and Characterisation of Bioactive Principle(s) of Garcinia kola Seeds Fractions on Clarias gariepinus
(UNIVERSITY OF ILORIN, 2018-05) ASOGWA, TOBECHUKWU NNAEMEKA
Malaria is a protozoan disease transmitted by the bite of an infected female Anopheles mosquito. Globally, there is a marked increase in resistance to well-known antimalaria drugs. The increasing drug resistance of the parasite and insecticide resistance of the vector dims the possibility of eradicating malaria in the tropics. Vector control are of the most successful method for reducing incidences of mosquito-borne diseases. Garcinia kola seeds (GKS) are used in Opi Nsukka for water guard to prevent Mosquito larvae in water. No literature has reported on the larvicidal property of GKS fractions. Therefore, this study was to investigate the larvicidal property, toxicological effect, and characterization of bioactive principles of GKS fractions with the view of providing a more cost effective method of malaria eradication. The specific objectives were to: (i) determine the presence of secondary metabolites and fractionate the aqueous and methanolic GKS. (ii) determine the larvicidal property of the aqueous and methanolic GKS fractions on mosquito larvae. (iii) identify the active components in the fractions with the highest larvicidal property (iv) characterize the bioactive principle(s) responsible for the larvicidal activity (v) propose a probable mechanism of action of the bioactive agent(s) (vi) determine the effect of GKS extracts on the Liver, brain and kidney function parameters, of Clarias gariepinus(vii) determine the binding conformations of the components of the extracts with mosquito larva acetylcholine esterase and Na+/K+-ATPase. Screening of the extracts for secondary metabolites and fractionation were done using standard methods. The larvicidal property of the GKS fractions on mosquito larvae were done using standard methods. Furthermore, the effects of the extracts on the rate of inhibition of mosquito larva acetylcholine esterase and Na+/K+-ATPase were done using standard methods. The components of the extracts were analysed by GC-MS and HPLC-DAD and in silico docking simulations were done on mosquito larva acetylcholine esterase and Na+/K+-ATPase. The toxicological effect on Clarias gariepinuswere done using standard methods. The data were subjected to analysis of variance and Duncan multiple range test and statistical significance was set at p˂0.05. The results revealed that the aqueous and methanolic extracts of GKS contains alkaloids, tannins, phenolics, glycosides, saponin, flavonoids, steroids and HPLC-DAD revealed the presence of Apiginin, Ellagic acid and Luteolin. The larvicidal activities of 5th aqueous and 4th methanolic extracts of GKS fractions (96.92±0.42, and 89.72±0.03 respectively) were higher than the standard larvicide fenthione (81.27±0.24). Mosquito Larva Na+/K+-ATPase and AChE activity by GKS fractions were suppressed in dose dependent manner. In silico screening of GKS fractions suggested that Apiginin, Ellagic acid and Luteolin are potent inhibitor of Mosquito LarvaAcetylcholinesterase and Na+/K+-ATPase. The administration of GKS extracts had no toxic effect on the liver, brain and kidney function parameters, of Clarias gariepinus taking the water The study concluded that Apiginin, Ellagic acid and Luteolin effectively inhibited Na+/K+ ATPase and Acetylcholinesterase and thereby increasing the cholinergic response in the mosquito larva. The increase in cholinergic response may cause uncontrollable muscle contraction in the larva and its death
Anti-diabetic Activity of Aqueous Extract of Chrysophyllum albidum Stem Bark, its Fractions and Toxicity in Alloxan-induced Diabetic Rats
(UNIVERSITY OF ILORIN, 2018-06) YUSUF, BABALOLA OLA
Diabetes mellitus is a chronic metabolic disease that is caused by a relative lack of insulin and/or reduced insulin activity resulting in hyperglycaemia. The rise in poverty level coupled with side effects of available anti-diabetic drugs necessitated the evaluation of a medicinal plant, Chrysophyllum albidum, claimed to be used in the management of diabetes. The objectives of the study were to: (i) evaluate the anti-diabetic activity of aqueous extract of C. albidum stem bark (AECASB); (ii) carry out solvent partitioning of AECASB and evaluate their anti-diabetic activity; (iii) carry out bioactivity-guided chromatographic fractionation (CF) of the most effective solvent-partitioned fraction (SPF) of AECASB and evaluate their anti-diabetic activity; (iv) identify the bioactive principles in the most effective CF of AECASB; (v) carry out safety evaluation of the antidiabetic principle(s)-rich CF of AECASB and (vi) propose the mechanism of action of the antidiabetic principle(s)-rich CF of AECASB. A total of 340 female rats (180.80 ± 8.50g) were assigned into group A (received 1.0 ml of distilled water) while the diabetic rats {induced by intraperitoneal administration of 120 mg/kg body weight (bw) of alloxan} in groups B, C, D, E, F, G were orally administered distilled water, glibenclamide (reference antidiabetic drug), 25, 50, 100 and 200 mg/kg bw of AECASB respectively, once daily for 14 days. Crude, SPFs (ethyl acetate, residue, n-butanol)andCF (1-10) were also screened. Biochemical assessments and histological examinations were also carried out. Data were analysed with Analysis of Variance and Tukey’s post-hoc test at p < 0.05. The findings in this study revealed that: i. AECASB reversed the alloxan treatment related increases in fasting blood glucose (FBG), serum urea, creatinine, hepatic glucose (HG), amylase activity and related decreases in albumin, glycogen, erythrocytic indices, glucokinase (GK) and glucokinase-6-phosphate dehydrogenase (G6PDH) activities. ii. AECASB also restored the deranged histoarchitecture of the pancreas caused by alloxan; iii. ethyl acetate-partitioned fraction reduced (p<0.05) FBG, HG, α-amylase activity, total cholesterol, triglycerides, malondialdehyde (MDA), ameliorated the obliteration observed in pancreas due to alloxan treatment while glycogen, high density lipoprotein (HDL), G6PDH, SOD and catalase activities increased significantly (p<0.05); iv. Chromatographic fractions F5 and F7 decreased (p<0.05) FBG, HG, organ body-weight ratio, glycosylated haemoglobin, C-reactive protein, glucose-6-phosphatase and fructose-1,6-bisphosphatase activities, TC, triglycerides, atherogenic index, MDA while BW, serum insulin, glycogen, HDL, GK, G6PDH, catalase, SOD activities significantly (p<0.05) increased; v. procyanidin,stigmasterol, catechin, epicatechin, epigallocatechin,hexadecane,eleagnine, octadecanoic acid and eicosane were detected in F7; vi. CFs 5 and 7 did not alter urea, creatinine, albumin, bilirubin, alkaline phosphatase, alanine aminotransferase and aspartate aminotransferase activities; vii. catechin, stigmasterol and procyanidin B5 are the likely bioactive principles in CF7; and viii. mode of anti-diabetic action of AECASB and F7 may be via regenerating the pancreas and enhancing the activities of glucose metabolising and antioxidant enzymes. The study concluded that CF7 produced the most profound anti-diabetic activity and is relatively safe. AECASB can be explored as lead drug for the management of diabetes.
Isolation, Purification and Characterization of Cellulase Produced by Aspergillus niger Cultured on Arachis hypogaea Shells
(UNIVERSITY OF ILORIN, 2018-06) SULYMAN, Abdulhakeem Olarewaju
The substantial amounts of waste materials such as shells, tatters, trunks, peels and seeds are produced as a result of agricultural practice. Large quantities of these agrowastes obtained are from heavy consumptions of agricultural products. These wastes particularly groundnut shells are abundant because once the nuts have been removed, the shells are always discarded. Accumulation of these shells constitutes what we called “wastes” and consequently lead to environmental pollution. Transformation of these wastes to expedient products will not only combat environmental pollution arising from unnecessary discarding of shell but also boast the economy of our country. The uses of different agricultural wastes such as corn cob, rice bran, bagasses, wheat bran, banana trunk for production and characterization of cellulase have been studied in the past. Therefore, this study addressed the possible use of Arachis hypogaea shells as a substrate for cellulase production from Aspergillus niger. The A. hypogaea shells were dried and subjected to acid and alkali pretreatment. Proximate analysis was carried out on the treated and untreated substrates. The remains of pretreated substrates were then used as substrates in a shake-flask containing enriched media preparation and A. niger was inoculated. Fermentations were carried out in flasks containing the enriched media inoculated with A. niger for 168 hours. Optimization parameters for maximum cellulase production were determined by varying the fermentation conditions. The crude cellulase was precipitated at 80% ammonium sulphate saturation, followed by dialysis and gel filtration using Sephadex G-100 as stationary phase in chromatographic column. The molecular weight was estimated using sodium dodecyl sulphate polyacrylamide gel electrophoresis. Cellulase activity was assayed by determining the concentration of glucose produced from the hydrolysis of carboxymethylcellulose catalysed by cellulase. The effects of pH and temperature on the activity of purified cellulase were determined. The kinetic parameters (Km and Vmax) were also evaluated. The effects of some cations (Na+, K+, Mg2+, Cu2+, Zn2+, Ca2+, Fe2+, Mn2+, Co2+) and anions (CO32-, Cl-, and SO42-) on the activity of purified cellulase were investigated. Also, the effects of some surfactants (DMSO, Triton X, Tween 20 and Mercaptoethanol) on the activity of purified cellulase as well as detergent compatibility of purified cellulase were determined. The results obtained from proximate analysis of pretreated A. hypogaea shells showed that crude fiber content, alkaline pretreated substrates had the highest percentage of crude fibre with 87.0% compared to acid treated and untreated with 85.5 and 82.0% respectively. There was a significant increase (p ˂ 0.05) in the content of crude protein following the treatment of A. hypogaea shell with alkali compared to acid treated and untreated shells. The alkaline pretreated substrates had the highest percentage of cellulose with 2.2 folds increase when compared to untreated substrate. Also, the percentage of cellulose in the acid treated substrate was higher with a 1.7-fold increased, compared to untreated A. hypogaea. This study also revealed that the: optimal production of crude cellulase was achieved at incubation period of 120 hours, pH of 4, temperature of 40 oC, and inoculum size of 13 cfu/ml; enzyme was purified to 68.12 fold with a yield and specific activity of 3.87% and 484.3 U/mg respectively; Vmax for the cellulase was 9.26 U/ml while the Km was 0.23 mg/ml; molecular weight of the cellulase was 13.5 kDa; optimum pH and temperature for the cellulase activity were 4 and 40 oC respectively; CO32-, Cl- and SO42- decreased the activity of cellulase. Na+ activated the cellulase activity at 1 to 5mM while K+ did not affect the cellulase activity; Mg2+ competitively inhibited cellulase activity while Zn2+, Cu2+, Ca2+ and Fe2+ non-competitively inhibited the activity of the enzyme. Mn2+ and Co2+ enhanced the cellulase activity; and cellulase activity was inhibited by dimethyl sulfoxide, triton X, tween 20 and mercaptoethanol. The study concluded that A. hypogaea shells can be used as a source of carbon by A. niger for the production of cellulase. Therefore, the purified cellulase produced may be explored for the various industrial applications of the enzyme.
Antidiarrhoeal Principle Rich-FractioninMangifera indica Leaves and its Mechanisms of Action in Castor Oil-induced Diarrhoeal Rats
(UNIVERSITY OF ILORIN, 2018-07) SALIMON, SAOBAN SUNKANMI
Diarrhoea is an alteration in bowel movement characterized by increased water content and frequency of stooling. The aqueous extract of Mangifera indica leaves have been validated for their efficacy as an antidiarrhoeal agent. However, the probable mechanism(s) of action remain unknown. The objectives of the study were to: (i) partition the aqueous extract using solvent-solvent extraction; (ii) determine the antidiarrhoeal activity of the solvent-solvent and column chromatographic fractions in female Wistar rats; (iii) establish the probable mechanism of action of the sub-fractions; (iv) evaluate the toxicity of sub-fraction B (SFB) on selected organs; and (v) identify the probable bioactive constituents of SFB. The aqueous extract of M. indica leaves was successively fractionated into ethyl-acetate fraction (EAF), n-butanol fraction (NBF) and residual fraction (RF). The EAF, NBF and RF were evaporated to dryness at 40℃. They were reconstituted and administered to screen for antidiarrhoeal activity using castor oil-induced diarrhoea; enteropooling; and gastrointestinal motility models. Each of the sub-fractions of the RF were also screened for their antidiarrhoeal activity and evaluated for probable mechanism(s) of action. The SFB was also evaluated for its effects on the liver, kidney and small intestine. Data were analysed using One-way Analysis of Variance and Tukey’s Post-hoc test at p < 0.05. The findings revealed that the: i. RF at 14.09 and 56.35 mg/kg body weight (bw) significantly prolonged the onset time of diarrhoea, decreased the total number of faeces, wet faeces, water content and increased the inhibition of defecation by 94.29 and 96.34% respectively; ii. mass and volume of intestinal content significantly decreased at 14.09 mg/kg bw of the RF with an inhibition of 45.20% on intraluminal fluid accumulation; iii. distance travelled by charcoal meal was shortest in diarrhoeal rats treated with 14.09 mg/kg bw of the RF with 39.79% inhibition of peristalsis; iv. the antidiarrhoeal index (ADI) was highest at 14.09 mg/kg bw of the RF (55.19%). At 56.35 mg/kg bw, the ADI (49.87%) compared favourably with that of the reference drug (48.50%); v. highest inhibition of defecation was exhibited by sub-fraction B at 6.47 mg/kg bw while the onset of diarrhoea was most prolonged by sub-fraction A; vi. antienteropooling and antimotility activities were highest in the sub-fraction A and B treated rats, respectively; vii. activities of Na-K ATPase, alkaline phosphatase and the concentrations of glucose, Na+ and K+ were increased by SFB while the levels of Cl , protein carbonyl and nitrite were decreased; viii. levels of acetylcholinesterase, glutathione, superoxide dismutase and intestinal protein were all increased by SFB; ix. sub-fraction B at 12.94 mg/kg bw exhibited structural and functional toxicity on the liver kidney and small intestine; and x. mangiferin and catechin were identified as the probable bioactive constituents of SBF. The study concluded that the antidiarrhoeal principles were present in the RF while the established mechanisms included antisecretory, anti-inflammatory, antioxidant and modulation of intestinal permeability. The antidiarrhoeal principle rich-SBF may therefore be explored as a lead for drug discovery in the treatment of diarrhoea.
Artesunate-Procyanidin Hybrid Compound: Synthesis, In vitro and In vivo Antimalarial, Antioxidant and Toxicological Effects
(UNIVERSITY OF ILORIN, 2018-07) TIJJANI, HABIBU
The goal of global eradication of malaria has remained elusive due to the rapid spread of drug resistant parasites, necessitating urgent development of novel antimalarials. Therefore, the aim of this study was to synthesize a hybrid compound of artesunate and procyanidin and evaluate its antimalarial, antioxidant and toxicological effects using various models. The objectives were to: (i) synthesize and chracterise a hybrid compound from artesunate and procyanidin; (ii) evaluate the in vitro antiplasmodial activity and cytotoxicity of the compound; (iii) evaluate its mechanisms of action; (iv) evaluate its suppressive and curative antimalarial activities in Plasmodium berghei NK65-infected mice; (v) evaluate the protein-ligand interactions; (vi) evaluate the in vitro and in vivo antioxidant activities of the compound; and (viii) evaluate its toxic effects. Artesunate-procyanidin hybrid compound was synthesized using standard chemical protocols and the structure was elucidated using Nuclear Magnetic Resonance and Infra-Red spectroscopy. The compound was evaluated for its activity against Plasmodium falciparum W2 strain in vitro and P. berghei NK65 in vivo. Inhibition of β-hematin formation, red blood cell lysis and cytotoxicity against Buffalo Green Monkey (BGM) kidney cell line of the compound were evaluated. The compound was evaluated for antioxidant activity using in vitro and in vivo models and for toxic effects using selected organ function indices of mice. Protein-ligand interactions were evaluated using molecular docking. Data were analyzed using Analysis of Variance at P<0.05. The findings of this study were that the hybrid compound: i. was synthesized as a brownish crystal with two pharmacopores linked together by ester bond; ii. was active against P. falciparum W2 (IC50: 0.0335 µg/ml), though its activity was lower than that of artesunate, but higher than that of procyanidin and it was less toxic to BGM cell line (MLD50: 155 µg/ml); iii. caused higher inhibition of β-haematin formation in vitro (IC50: 42.46 µg/ml) compared to chloroquine (IC50: 55.53 µg/ml), though was lower than that of artesunate (IC50: 17.79 µg/ml) and did not cause red blood cell membrane perturbation; iv. exhibited higher suppressive activity (ED50: <5 mg/kg) than artesunate on day 4 post-inoculation and curative activity (ED50: <5 mg/kg) which favourably compared with artesunate on day 6 post-inoculation against P. berghei NK65 in mice; v. exhibited higher binding affinity for P. falciparum Lactate Dehydrogenase (-9.6 kcal/mol) compared to artesunate (-6.2 kcal/mol) and procyanidin (-8.0 kcal/mol); vi. exhibited higher ferric ion reducing power and DPPH scavenging activity (IC50: 19.68 µg/ml) compared to butylated hydroxytoluene in vitro and caused significant increase (P< 0.05) in glutathione peroxidase, and glutathione-S-transferase activities in RBC,heart, liver, kidney, and brain of P. berghei-infected mice compared to untreated infected controls; and vii. had no significant effect (P>0.05) on atherogenic index and plasma concentrations of sodium, calcium, creatinine, urea and bilirubin but significantly reduced (P< 0.05) brain Na+,K+Adenosine triphosphatase activity in mice at all doses administered compared to controls. The study concluded that hybrid compound exhibited antimalarial activity (which favourably compared with artesunate) and antioxidant activity in vivo. Artesunate-procyanidin hybrid compound may be explored as an alternative therapy for malaria.
RESPONSES OF SELECTED FRUIT VEGETABLES TO INTERCROPPING WITH JUVENILE OIL PALM AND WEED DIVERSITY
(UNIVERSITY OF ILORIN, 2018-08) OLUWATOBI, AYODELE SAMUEL
The wide spacing used for oil palm plantation has led to serious weed infestation from transplanting to canopy closure stages when the oil palm becomes productive. The study was conducted to investigate the responses of some fruit vegetables to intercropping with juvenile oil palm of two years old. The objectives of the study were to: (i) evaluate the growth and yield of the fruit vegetables to three spacing regimes in juvenile oil palm/fruit vegetable intercrop; (ii) determine the effects of intercropping on growth of the juvenile oil palms; (iii) determine the biological and economic efficiencies of oil palm/fruit vegetable intercrop; (iv) investigate weed species diversity in the experimental plots; and (v) establish the effect of the intercrops on weed smothering efficiency, density and biomass. Four fruit vegetables: (i) tomato (accessions NGB 01665 and NG/AA/SEP/09/053); (ii) pepper (NGB 01312 and NGB 01641); (iii) okra (NGB 01184); and (iv) egg plant (NGB 01737), were intercropped within alley of juvenile oil palms (240 m2)during rainy season of 2016 and 2017 at oil palm plantation in Ala, Akure-North Local Government Area, Ondo State. Each vegetable was intercropped at a distance of 1, 2 or 3 m from the juvenile oil palm and control plot (non-intercropped) at 1 X 1 m spacing. Intercropped and non-intercropped juvenile oil palm growths were compared. Weed species distribution and diversity were evaluated. Data were subjected to analysis of variance using Statistical Package for Social Sciences and means were separated using Duncan Multiple Range Test at P<0.05. The findings of the study were that: i. tomato (NGB 01665) and okra (NGB 01184) intercropped recorded highest yield at 3 m from the juvenile oil palm (3.893 and 0.321 tonha-1 respectively); and tomato (NG/AA/SEP/09/053), pepper (NGB 01312 and NGB 01641) and eggplant (NGB 01737) intercropped at 3 m from the juvenile oil palms were higher in yield (3.704, 0.614, 0.570 and 2.063 tonha-1, respectively) than other spacing regimes and control; ii. juvenile oil palms did not show any adverse effects when intercropped with the fruit vegetables during second (2016) and third year (2017) of establishment (canopy spread, 218.20 cm and 433.80 cm, respectively); iii. pepper (NGB 01641)-, okra (NGB 01184)- and eggplant (NGB 01737)-intercrops recorded land equivalency ratio of 1.404, 1.447 and 1.097 respectively, while tomato (NGB 01665 and NG/AA/SEP/09/053) recorded intercropping advantages at 3 m from juvenile oil palm; iv. twenty-eight and 20 different weed species were found at 3 weeks after intercrop and harvesting of the fruit vegetables, respectively. Members of Asteraceae and Poaceae were the most abundant weeds at 3 weeks after intercrop (17.86%) and after harvesting (20.0%), respectively. Pepper control plot had the highest weed species diversity; and v. tomato (NG/AA/SEP/09/053) and eggplant (NGB 01737) exhibited greatest smothering effect on weeds at 3 m from juvenile oil palm (46.64% and 58.414%, respectively). The study concluded that tomato (NG/AA/SEP/09/053) and eggplant (NGB 01737) have highest yield and weed smothering effect respectively. It is therefore recommended that they should be intercropped within the alley of juvenile oil palm.
IMPACTS OF HEAVY METALS IN DUMPSITES ON SOIL AND VEGETATION IN SELECTED LOCATIONS IN KWARA STATE
(UNIVERSITY OF ILORIN, 2018-08) OLUKOYEJO, HANNAH OLUWAKEMI
Indiscriminate dumping of refuse is one of the major sources of pollution to the environment. Clean up of these wastes has led to appearance of dumpsites which has become preferable farmlands. This study was carried out to investigate the impacts of heavy metals (HMs) in dumpsites on soil and vegetation of selected locations and the possibility of reducing the availability of heavy metals (HMs) to the plants. The objectives of the study were to: (i) assess the heavy metal concentrations in the soil and plants from the dumpsites; (ii) develop methods for production of biochar from the waste; (iii) assess the effects of biochar on the yields of Solanum lycopersicon, Amaranthus esculentus, Corchorus olitorious,Abelmoschus esculentum and Tithonia diversifolia; and (iv) assess the mobility potential of the biochar on the HMs in selected plants. Plants and soil samples were collected at 10 km intervals in Oko-Olowo, Offa, Omu-Aran (Urban), and Odo-Ore, Ipee and Aran-Orin (rural). The samples were digested using Aqua regia method after which Cd, Pb, Cr, Zn, Fe and Cu concentrations were determined with Atomic Absorption Spectrophotometer (AAS). Composite dumpsite soil was used to raise the plants and its HMscontent determined by AAS. Biochar was prepared by slow pyrolysis of maize cobs (MB). Scanning Electron Microscopy, Energy Dispersive X-ray and Fourier Transform Infraredwere used to identify the properties of the MB.Data were analysed with Analysis of Variance and Duncan Multiple Range Test at p <05. The findings of the study were that: i. Pb(0.00 – 75.00), Cd(0.00 – 4.00), Zn(4.50 – 1290), Ni(0.00 – 19.5), Cu (2.5 – 225) and Fe (1,390 – 20,850) mg/kg were present in the soils; ii. Pb (0.00 – 7.00), Cd(0.00 – 0.50), Zn( 16.00 – 310.00), Ni(0.00 – 5.50), Cu(1.00 –9.50) and Fe(195.00 – 4,950.00) mg/kg were found in the plants; iii.MB had well-defined pore structure and contained C, O, Si, K and Mg; v. MB increased the yieldsof S. lycopersicon(50%), C. olitorious (6.3%), Abelmoschus esculentum (40.1%) and T. diversifolia (7.5%) but reduced the yield of Amaranthus esculentus (50%); vi. there was significant reduction (p≤0.05) in the HMs content of the selected plants withMB. Abelmoschus esculentum, C. olitorious, Amaranthus esculentus, T. diversifolia and S. lycoperscon on dumpsite soil contained 2.78. 2.23, 2.29, 5.20, 3.50 mg/kg of Pb while those with biochar had 2.36, 0.90, 2.35, 3.67, 2.40 and 2.30 mg/kg, respectively; and vii.Abelmoschus esculentum accumulated Ni, Cu and Zn but excluded Cd, Pb and Fe, Amaranthus esculentus accumulated Cd and Zn but excluded Pb, Ni, Cu and Fe,T.diversifolia,accumulated Cd, Ni and Zn, C. olitorious accumulated Cd and Pb whileS. lycopersiconexcluded all the investigated HMs. The study concluded that indiscriminate dumping of waste contributed to the HMs load of dumpsite soil and vegetation.It is recommended that remediation process be put in place to reduce the HMs load in order to avert the health hazards that may result in humans that consume the vegetables.
PHOSPHODIESTERASE 5 ACTIVITY AND ENDOTHELIAL MARKERS AFTER ADMINISTRATION OF Cnestisferruginea AND Fadogiaagrestis EXTRACTS TO PAROXETINE-TREATED RATS
(UNIVERSITY OF ILORIN, 2018-08) MURITALA, HAMDALAT FOLAKE
PDE-5 inhibitors such as sildenafil citrate (Viagra) commonly prescribed for the management of erectile dysfunction (ED) has been reported to have failure rate of 30-50% at the attempt of intercourse. This has thus necessitated the need to source for better alternatives in medicinal plants for managing ED and its associated risk factors like cardiovascular diseases (CVD).Therefore, this study investigated the PDE-5 inhibitory activities and modulatory properties of Cnestisferruginea root, Fadogiaagrestis stem and their combinations on endothelial markers of paroxetine-treated rats. The specific objectives of this study were to: (i) determine the secondary metabolites in the plants’ parts; (ii) partially purify PDE-5 from rat penile and cardiac tissues (iii) carry out PDE-5 inhibitory assays of the extracts in the presence of cyclic guanosine monophosphate (cGMP) and cyclic adenosine monophosphate (cAMP); (iv) determine the effects of paroxetine on selected endothelial, erectile and cardiovascular functional biomarkers in rats; and (v) determine the effects of the extracts on the selected markers of erectile, cardiovascular and endothelial dysfunctions of male rats pre-treated with paroxetine. Forty male Wistar rats (200.32 ± 6.33) were used for the invitro PDE-5 inhibitory assays, after which Michealis-Menten constant (Km) and maximum velocity (Vmax) were derived from the double reciprocal plots. For the invivo study, 210 male Wistar rats (210.43 ± 4.04) g were grouped into two consisting of 40 rats in group A (control) and 170 rats in group B (received 10 mg/kg body weight of paroxetine) for 21 days. Eight animals from groups A and B were sacrificed on days 5,8,15 and 22 of paroxetine treatment. The remaining animals in group B were re-assigned into 5 groups of 16 animals each. 8 rats were sacrificed from each group on days 29 and 36 of experimental period. Biochemical assays were done on the tissues and plasma. Data were subjected to Analysis of Variance and Duncan multiple range Test. Statistical significance was set at p<0.05. The results revealed that: i C. ferruginea root extract contained six secondary metabolites while F. agrestis stem extract contained five secondary metabolites with alkaloids as major component of the plants (49.95 ± 0.25 x 10-2 and 59.15 ± 0.25 x 10-2 mg/g) respectively; ii purification folds of isolated penile PDE-5 was 5.6 while that of cardiac PDE-5 was 7.5 when compared with the crude extracts of PDE-5; iii the extracts at 0.025-0.125 µg/mL competitively inhibited PDE-5 activities in a manner similar to sildenafil; iv paroxetine-treatment related PDE-5 inhibition was reversed by the extracts after post-treatment; v paroxetine-treatment related reduction in nitric oxide concentration was significantly (p>0.05) elevated by the extracts and was sustained post-treatment; vi endothelin-1, cGMP, and testosterone concentrations were significantly (p>0.05) decreased during treatment and post-treatment periods. vii paroxetine related increase in creatine kinase, triacylglycerol, total cholesterol and low density lipoprotein cholesterol were significantly (p>0.05) decreased after administration of extracts. The study concluded that the extracts exerted their aphrodisiac activities via inhibition of penile phosphodiesterase-5 in a similar mechanism as sildenafil. Moreover, the extracts can be used for managing ED and CVD.
Ameliorative Effects of Moringa oleifera Leaf-based diet on Malnutrition- induced Skeletal Muscle Degeneration in Rats
(UNIVERSITY OF ILORIN, 2019-09) LAMBE, Muinat Olayinka
Skeletal muscle degeneration (SMD) is a physiological condition characterized by decrease in muscle mass due to reduced protein synthesis and/or degradation. About 1 in 300 people is seen with a SDM. Chemotherapies generally used in the treatment of diseases like cancer and tuberculosis although expensive and inaccessible also contribute to SMD. Hence, the need to explore cheaper, safer and readily available options in Moringa oleifera leaf-based diet (MoLD). The objectives of the study were to: (i) identify the secondary metabolites in M. oleifera leaf; (ii) determine the proximate composition of MoLD; (iii) confirm the induction of skeletal muscle degeneration; (iv) evaluate the effects of MoLD on markers of muscle degeneration; and (v) evaluate the toxicity of MoLD in Wistar rats. Skeletal muscle degeneration was induced by maintaining adult rats on 4% low protein iso-caloric diets (LPID) for four weeks. The control animals were fed on soy-based meal only while the SMD-induced rats were maintained on soy-based diet supplemented with methyl jasmonate (100 μg/kg) + vitamin E (20 mg/kg) + carnitine (3 mg/kg) (Soy-based), 12.5% MoLD and 25% MoLD. The feeding was done for four weeks after which serum and hind limb skeletal muscle homogenates were prepared for biochemical analyses. Data was analyzed using one-way Analysis of Variance and Tukey’s posthoc test at p ˂ 0.05. The findings of this study were that: i. M. oleifera leaf contained terpenoids (2.91mg/g), tannins (9.2mg/g), flavonoids (12.5mg/g), total phenols (42.34mg/g), alkaloids (30.86μg/g) and saponins (55.22μg/g); ii. the crude protein (13.77%) and carbohydrate (62.89%) contents of 25% MoLD was higher than those of 12.5% MoLD but was similar to the control; iii. MoLD significantly (p ˂ 0.05) increased the activities of serum creatinine kinase (CK), lactate dehydrogenase (LDH), aspartate aminotransferase (AST) and alanine transaminase (ALT) and reduced Ca2+- ATPase activity when compared with the control; iv. activities of serum CK, LDH, AST and ALT were significantly (p ˂ 0.05) reduced by the 12.5% and 25% MoLD whereas these enzymes in animals fed soy-based were not comparable with the control; v. activity of cytosolic and mitochondrial Ca2+- ATPase was significantly increased by all the treatments in the order: Soy-based ˃ 25% MoLD ˃ 12.5% MoLD; vi. the 25% MoLD produced significantly (p ˂ 0.05) higher activities of superoxide dismutase, glutathione-S-transferase, glutathione (reduced) and malondialdehyde when compared with other treatments; vii. mitochondrial permeability and reactive oxygen species was significantly reduced while membrane potential was increased in all the treatment groups with the 25% MoLD producing the most profound change; and viii. activities of serum AST, ALT and alkaline phosphatase (ALP) as well as albumin, urea and creatinine concentrations in the 12.5% and 25% MoLD fed animals were not significantly different from those of the control whereas those of Soy-based were higher than the control values. The study concluded that 25% MoLD ameliorated muscle degeneration in rats by positively modulating calcium homeostasis, oxidative stress, energy depletion and is also safe for consumption. The 25% M. oleifera leaf-based diet may be explored as an alternative therapeutic option for managing muscle degeneration.
BIOCHEMICAL STUDY OF ANTIDIARRHOEAL FRACTIONS OF Annona senegalensis ROOT AND STEM BARKS IN CASTOR OIL-INDUCED DIARRHOEAL ALBINO RATS
(UNIVERSITY OF ILORIN, 2020-01) AHMED, MARYAM ABIOLA
Diarrhoea is a gastrointestinal tract disorder characterized by frequent stooling. The use of the aqueous extract of Annona senegalensis root and stem bark for the treatment of diarrhoea has been documented. However, limited information exists on the components, mechanism of action and safety. The aim of the study was to carry out biochemical assessment of the antidiarrhoeal fractions from A. senegalensis root and stem barks in castor oil-induced diarrhoeal rats. The objectives of the study were to: (i) evaluate the antidiarrhoeal activity of solvents extract of A. senegalensis stem and root barks; (ii) fractionate the bioactive extracts; (iii) explore possible mechanism(s) of action of the bioactive fraction(s); (iv) evaluate the toxicity of bioactive sub-fractions; and (v) identify chemical compounds in the bioactive sub-fractions. Aqueous, n-hexane and dichloromethane stem and root bark extracts of A. senegalensis were screened for antidiarrhoeal activity using castor oil-induced diarrhoeal, gastrointestinal tract (GIT) motility inhibition and antienteropooling models. Bioactive aqueous extract was partitioned into hexane, dichloromethane and ethylacetate fractions. Bioactive fractions were fractionated into sub-fractions using column chromatography. The mechanism(s) of action of the bioactive sub-fractions was determined by evaluating their antioxidant properties, intestinal fluid electrolyte concentration, small intestinal Na+- K+ ATPase and cyclooxygenase II activities. Kidney and liver function indices were evaluated after administration of bioactive sub-fractions. Active principles were identified using gas chromatography-mass spectrometry. Data were subjected to analysis of variance and Duncan’s multiple range test at p < 0.05. The findings of this study were that: i. aqueous stem (AS) and dichloromethane root (DR) extracts at 100 mg/kg body weight (bw) significantly decreased the number of wet feaces while GIT motility inhibition and antienteropooling activity were significantly increased by aqueous root (AR) and dichloromethane stem (DS) extracts; ii. inhibition of defecation was highest (85.25%) in the ethylacetate fraction of AS (EFAS), while antienteropooling and GIT motility inhibition was highest in the dichloromethane fraction (DFAR) and ethylacetate fraction (EFAR) of AR respectively; iii. sub-fraction 1 of EFAS (EFAS1) and fraction 2 of DR (DR2) at 25 mg/kg bw exhibited the highest inhibition of defecation; iv. anti-motility activity was highest in sub-fraction 2 of EFAR (EFAR2) at 25 mg/kg bw while antienteropooling activity was highest in fraction 3 of DS (DS3) and sub-fraction 1 of DFAR (DFAR1); v. EFAS1 had the least IC50 for diphenyl-1-picrylhydrazyl, highest hydrogen peroxide scavenging activity, significantly increased Na+-K+ ATPase activity and decreased cyclooxygenase II activity; vi. chloride and Na+ concentration were significantly decreased by EFAR2 and DS3 respectively. vii. alanine aminotransferase and Na+ significantly increased while urea decreased at 200 and 400mg/kg bw of DFAR1, EFAR2, EFAS1, DS3 and DR2; viii. chemical compounds identified were; catechol and ethyl eicosapentanoate in EFAS1; piperidnyloxy, thiazoline and quinazoline in EFAR2; and aldosterone derivatives in DFAR1 and DS3. The study concluded that antidiarrhoeal principles present in DS3, EFAS1, DR2, EFAR2 and DFAR1 exhibited different mechanism of action. Fourteen days administration of these principles was not safe. Identified bioactive principles may be explored for the development of antidiarrhoeal drugs.
Morphological and Molecular Systematics of Freshwater Fish Diversity and Physico-chemical Parameters of Jebba Hydro-electric Power Reservoir, Nigeria
(UNIVERSITY OF ILORIN, 2021-01) OLADIPO, SEGUN OLAYINKA
Fishery activities in Jebba Hydro-electric Power Reservoir (JHPR) contribute to socio-economic development of Jebba town and Nigeria. There is paucity of information on fish biodiversity in JHPR, hence, this study aimed at identifying fish species and their ecology. The objectives of the study were to: (i) investigate the physico-chemical parameters in up- and down-streams of JHPR; (ii) identify fish species, abundance and diversity; (iii) determine the relationship between fish diversity and physico-chemical parameters; (iv) determine phylogenetic relationships of the barcoded fish species; and (v) determine intra- and inter-generic diversities of the barcoded fish species. The study of physico-chemical parameters in relation to fish composition and diversity in JHPR was carried out between January, 2017 and December, 2018. The physico-chemical parameters of water in the reservoir were determined using standard methods. Fishes were collected with the aid of gill and cast nets of different mesh sizes and identified morphologically utilizing morphometric and meristic features; and molecular approach adopting DNA barcoding. Simpson’s, Shannon-Wiener and Margalef’s indices and relationship between the physico-chemical parameters and fish abundance/diversity in the sites/seasons were statistically analysed with correlation matrix and factor analysis in XLstat at p≤0.05. The phylogenetic relationships, intra- and inter-specific divergence of the barcoded fish species were determined applying MEGA 5.0. The findings of the study were that: (i) the range for dissolved oxygen (DO), chemical oxygen demand (COD), turbidity and nitrate concentration in up- and down-streams of JHPR was 2.0-5.5mg/L, 20.50-35.67mg/L, 1.2-375NTU and 0.2-3.4mg/L respectively; (ii) there was a significant (p<0.05) seasonal variation in turbidity with higher mean value of 175.40±34.49NTU observed in rainy season; (iii) eighty-three fish species belonging to forty-three genera were identified morphologically while 170 nucleotide sequences generated confirmed all the species identification except for Chromidotillapia guntheri, Ctenopoma kingsleayae, Malapterurus electricus and Dasyatis garouaensis; (iv) fish species distribution in the family Cichlidae and Mochokidae were more abundant in up- and down-streams of JHPR, but diversity indices showed higher species richness (Margalef=9.73) and diversity (H= 3.95; 1-D = 0.98) at the downstream; (v) positive correlations (≥0.73) between DO, nitrate concentration and fish abundance were obtained for all fish families except Claroteidae, Latiidae, and Tetraodontidae, while turbidity positively correlated (≥0.71) with the abundance of Mormyridae, Gymnarcidae, Characidae, Distichodontidae, Bagridae, Malapteruridae, Dasyatidae, Polypteridae and Schilbedae; (vi) all the barcoded sequences showed levels of genetic distinctiveness among species in the phylogenetic tree and taxonomical hierarchy; and (vii) a range of 0-18.83% and 0.00-0.45% were generated for intra- and inter-generic divergence respectively in the barcoded species. The study concluded that JHPR harboured high diversity of fish species which was influenced by seasonal variations in turbidity, DO, COD and nitrate concentrations. The study recommended that morphological and molecular methods be used for fish species identification, biodiversity monitoring, ecology and management.
Efficiency of Different Methods of Breaking Dormancy and Seedling Development in Parkia biglobosa and Prosopis africana
(UNIVERSITY OF ILORIN, 2021-03) ADEYEMI, CHRISTIANA OREOLUWA
Seed dormancy is an inactive phase of plant life that is greatly influenced by environmental and physiological conditions. High level of dormancy displayed in seeds of Parkia biglobosa (PB) and Prosopis africana (PA) delay seed germination, growth and development of the plants. This study was carried out to assess the efficiency of different methods of breaking dormancy to enhance seed germination and seedling establishment of PB and PA. The objectives of the study were to: (i) determine the proximate composition and secondary metabolites of the collected seeds; (ii) investigate the effect of different methods of breaking dormancy on the germination capacity; (iii) quantify the generated Reactive Oxygen Species (ROS) in the seeds; (iv) evaluate the effect of germination on the enzyme activities; (v) examine the seed coat morphology; and (vi) determine the elemental constituents of the seed coats. Fruits of PB were collected from Jigawa (PBJG), Kwara (PBKW), Kaduna (PBKD) and Katsina (PBKT) while PA were from Niger (PANG), Kwara (PAKW), Kaduna (PAKD) and Kebbi (PAKB) states respectively. Seeds from the fruits were subjected to chemical, mechanical and hormonal treatments to break the dormancy. Chemical scarification was carried out using concentrated HCl, HNO3 and H2SO4 for 5, 10, 15 and 20 minutes each. Mechanical scarification with sandpaper was used to remove part of the seed coat. Seeds were treated with auxin and gibberellin at 150, 200 and 250 mg/l for 12, 24 and 36 hours. Proximate composition, secondary metabolite, ROS levels, α-amylase, protease and lipase activities were evaluated using standard methods. Scanning electron microscope was used to examine the seed coat morphology. Elemental constituents of the seeds were determined using Electron Dispersive X-ray. Data were analysed using Analysis of Variance at p < 0.05. The findings of the study were that: xix i. PBJG had the highest moisture content (9.29%) and crude fat (1.26%), PBKT produced the highest ash content (3.81%). Highest alkaloid (1.02%) and steroid contents (3.80%) were obtained in PBJG and PBKD, respectively; ii. PBJG, PBKW and PBKD had 100% germination capacity (GC) at 15 and 20 minutes H2SO4 scarification, PAKB and PANG had the highest GC at 20 minutes of scarification (93.3%). Only PBKW displayed 100% GC with mechanical scarification. The PBJG and PAKB treated with 250 mg/l of gibberellin at 30 hours produced the highest GC (76.7% and 70.0%), respectively; iii. hydrogen peroxide {32.72 μmol g/dry weight (DW)}, superoxide anion (23.66 μmol g DW-1) and hydroxyl radicals (82.72) were contained in PBJG and PBKD, respectively; iv. activities of α-amylase (0.081μ/mol/min), protease (0.34μ/mol/min) and lipase (0.99×10-5 U/mol) were highest in the germinating seed of PBJG, PBKW and PBJG at 7 days of planting respectively; v. seeds morphology was undulate and thin with raised seed coat sculpture; and vi. calcium (29.6%) was the most abundant elemental constituents of the seed coat followed by potassium (26.2%) and antimony (23.4%). The study concluded that dormancy breaking was most achieved using H2SO4 scarification. The study recommended that H2SO4 scarification be employed in the breaking of seed dormancy in PB and PA.

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