Programme: Biotechnology

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    SINGLE NUCLEOTIDE POLYMORPHISMS OF Pfdhfr RESISTANCE GENE AMONG SYMPTOMATIC PATIENTS’ ISOLATES FROM SELECTED HOSPITALS IN IFO LGA, OGUN STATE
    (Covenant University Ota, 2025-10) SULE, Queen Elizabeth; Covenant University Dissertation
    Malaria remains a primary universal health concern, particularly in endemic areas where drug resistance poses a serious threat to the effectiveness of key treatment and prevention strategies. Sulfadoxine-pyrimethamine (SP), commonly used for malaria prophylaxis, is increasingly compromised by resistance associated with mutations in the Plasmodium falciparum dihydrofolate reductase (Pfdhfr) gene. This study aimed to assess the prevalence of P. falciparum infection and identify the single-nucleotide polymorphisms (SNPs) in the Pfdhfr resistance gene among symptomatic patients in Ifo Local Government Area, Ogun State, Nigeria. Five hundred patients with severe P. falciparum infection were recruited, and demographic data were recorded. Blood samples were analysed for P. falciparum stages and parasitemia levels using microscopy. DNA was extracted from samples with high parasitemia and genotyped for Pfdhfr mutations using PCR, followed by visualisation on 1% agarose gel electrophoresis. Microscopy confirmed P. falciparum malaria in 300 patients (60%). A significantly higher prevalence (71.05%) was recorded in the 0–4 years’ age group, while males accounted for 64.31% of cases (p < 0.05). Parasitemia levels (greater than 200 parasites/100 μL) were more pronounced in males than in females, and were highest among individuals aged 0–4 years. Among the 10.67% Pfdhfr genotypes identified, males exhibited a higher frequency (6.0%) than females. The overall prevalence of pfdhfr SNPs in N51I, C59R, S108, and I64L was (96%), (96%), (100%), and (0%), respectively. tripple mutant halotype (N51I+ C59R+S108), prevance was 92%. Males have a higher mutation rate (60%) than females (40%). The overall prevalence of pfdhfr SNPs in N51I, C59R, S108, and I164L was (96%), (96%), (100%), and (0%), respectively. tripple mutant halotype (N51I+ C59R+S108), prevance was 92%. Males have a higher mutation rate (60%) than females (40%). Also, individuals aged 0-4 years (20%) and 15-20 years (20%) show higher SNPs than the other age groups. The study highlights a high prevalence of P. falciparum and emerging Pfdhfr resistance mutations, emphasising the need for continuous surveillance and targeted interventions in malaria-endemic regions, such as Ifo LGA, Nigeria.
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    EVALUATION OF EFFECTS OF Trichoderma asperellum STRAINS IN BIOCONTROL OF CHARCOAL-ROT CAUSING Macrophomina phaseolina (Tassi) Goidanich IN COWPEA
    (Covenant University Ota, 2025-09) OYESOLA, Olushola Luke; Covenant University Thesis
    Macrophomina phaseolina, a soil-borne fungal pathogen, is responsible for loss in cowpea at both seedling and adult stages. Synthetic fungicides, although proven effective in M. phaseolina control, lead to severe cowpea poisoning and pose significant health risks to humans and animals. Trichoderma are efficient bioagents for managing plant pathogens, representing a promising strategy for managing M. phaseolina. This study evaluated the antifungal efficacy of some Trichoderma asperellum strains against M. phaseolina. The Trichoderma isolates obtained from soil were macroscopically, microscopically and genotypically identified based on the Internal Transcribed Spacer genes. Bioactive compounds were isolated from selected Trichoderma species and evaluated using Gas Chromatography-Mass Spectrometry (GC-MS). The inhibitory potential of the suspensions of selected Trichoderma species against M. phaseolina was tested singly in vitro and in combination in vivo. The in vivo experiments were carried out in the screenhouse across pre-planting, post-germination and seed treatment for eighty days, and plant growth indicators such as plant height, stem girth and leaf number were assessed at ten-day intervals. Disease incidence and severity were also determined following standard methods. After the termination of the in vivo experiment, yield assessment was carried out to determine the cowpea pod number, pod weight, and seed number, as well as the fresh and dry weights of leaves, stems and roots. A histopathological assessment was carried out on the lower stems of the cowpea plants to determine the effect of M. phaseolina on the cowpea tissues. Phenotypic, microscopic and genotypic characterisation identified all the Trichoderma isolates as Trichoderma asperellum. Among the Trichoderma species isolated and tested, T. asperellum (Tric13), T. asperellum (Tric4) and T. asperellum (Tric12) exhibited significant inhibitory potential (p<0.05) against M. phaseolina in vitro, with inhibition values of 82.51%, 82.41%, and 81.95%, respectively. The production of volatile organic compounds varied among the Trichoderma species. Specifically, Tric13 yielded terpenoids, ketones, sesquiterpenes, cycloalkanes, and alcohols. In contrast, Tric4 produced fatty acids, aldehydes, alkanes, terpenoids, and aromatic and bicyclic compounds, while Tric12 generated carboxylic acids, terpenoids, phenolics, bicyclic and aromatic compounds. The in vivo results showed that Trt7 (plant height = 49.8000 cm; stem girth = 0.53625 cm; leaf number = 21.028), followed by Trt3 (plant height = 49.8250 cm; stem girth = 0.49986 cm; leaf number = 18.611), recorded higher cowpea biomass across the Trichoderma formulations than the negative control. Zero disease incidence was observed in Trt3 and Trt7 (0%) upon treatment with the Trichoderma formulations. Also, zero disease severity was noted in Trt3 and Trt7 (0%), compared to the negative controls, which displayed 100% incidence and severity. The cowpea yield assessment showed that the highest pod weight (11.3 g) and pod number (8) were recorded in Trt7 (pre-planting), and seed number (32) was recorded in Trt5 (seed treatment). The histopathological assessment of the cowpea showed the efficacy of the treatments in inhibiting the growth of M. phaseolina, minimising its tissue entry, and xylem vessel occlusion. This study established the efficacy of Trichoderma in M. phaseolina biomanagement and the synergistic potential of different Trichoderma asperellum strains in its biocontrol.
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    CHARACTERISATION OF PROSTATE TISSUE AND EXPRESSED PROSTATIC SECRETION IN PATIENTS WITH PROSTATE DISORDERS IN LAGOS NIGERIA
    (Covenant University Ota, 2025-01) SAMUEL ABOSEDE ESTHER; Covenant University Ota
    Prostate cancer (PCa) is the second most prevalent cancer in men, particularly affecting those of Black African descent. Nigeria currently has the fourth highest risk for PCa mortality in the world. The microbiome of the prostate has emerged as a critical factor in understanding the aetiology and progression of prostate diseases, such as prostate cancer (PCa), benign prostatic hyperplasia (BPH), benign stromal hyperplasia (BSH) and prostatitis (PRO). This study was conducted to comparatively characterize the microbiome present in prostate tissue and expressed prostatic secretion (EPS) from patients diagnosed with PCa, BPH, BSH and PRO. A total of 30 study participants comprising of 15 prostate cancer, 10 benign prostatic hyperplasia, 2 benign stromal hyperplasia and 3 prostatitis subjects. Samples were collected from the urology clinic of Lagos State University Teaching Hospital Ikeja and analysed to identify and quantify bacterial species, assessing the diversity and composition of the microbial communities. Subjects without prostate (15) cancer were used as control subjects. By employing cultural and 16SrRNA sequencing techniques, uro-pathogens were isolated from the samples. The antibiotic susceptibility testing was carried out on these isolates. Prostate tissue and EPS samples from BPH patients demonstrated a higher prevalence of bacterial taxa, including Staphylococcus scuri, Bacillus mycoides, Staphylococcus aureus, Proteus vulgaris, Klebsiella pneumoniae, Pseudomonas aeruginosa, Streptococcus pyogene and Bacillus subtilis. Conversely, PCa patients exhibited an increased presence of pathogenic bacteria such as Escherichia coli Klebsiella oxytoca, Pseudomonas fluorescens, Citrobacter freudii, Pseudomonas putida, Staphylococcus condimentii, and Proteus mirabilis, which have been implicated in chronic inflammation and carcinogenesis. A high abundance of Lactobacillus vaginalis, Staphylococcus carnosus and Zymononas mobilis were observed in the prostate tissue. PCa-associated microbiome displayed reduced microbial diversity compared to other prostate disorders, suggesting a possible dysbiosis linked to cancer progression.