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Item Analysis of aquatic insects’ communities of Awba reservoir and its physico-chemical properties(Maxwell Scientific Organization, 2011-06) Popoola, K. O. K.; Otalekor, AThis study was conducted to assess the Awba reservoir insects’ communities and the health status through the determination of insects’ abundance, composition, distribution and water qualities parameters. Water samples and insects were collected bi-weekly from August through December, 2009. Insects were sampled using standard entomological methods, while water samples was analyzed using standard Winkler’s titrimetric and APHA methods to determine the chemical properties. Water analyses and insects’ identifications were conducted in the laboratory in Department of Zoology, University of Ibadan, Oyo State. The results show that only DO and phosphate-phosphorus had significant difference (p<0.05). A total of 1,154 insects were recorded, Chironomidae and Culicidae were most abundance. The chemical properties and the distinct taxa found in the water suggest that the water body is polluted and may be dangerous to the health of people around the reservoir.Item ANTI-MOSQUITO PROPERTIES OF METHANOL EXTRACT OF Clerodendrum polycephalum Baker(2018-02) ADEWOYIN, F.BABSTRACT Mosquitoes are vectors of various diseases of public health importance and their control has been premised on the use of synthetic insecticides. However, these insecticides are laden with problems of high cost, environmental hazards and development of resistance in many species of mosquitoes. Alternatives to synthetic substances are the natural products. Plant species, Clerodendrum inerme, have been found to inhibit the growth of Aedes aegypti and Culex quinquefasciatus larvae. However, there is paucity of information on the anti-mosquito properties of the Nigerian species, C. polycephalum Baker. Therefore, the aim of the study was to investigate the anti-mosquito properties of the methanol extract of the leaves of C. polycephalum. Fresh leaves (1.4 kg) of C. polycephalum were harvested and brought to the laboratory. The leaves were air-dried, powdered, extracted with methanol and concentrated to dryness using rotary evaporator. The crude methanol extract was subjected to phytochemical analysis according to standard method and separated into n-hexane, dichloromethane, ethyl acetate and ethanol fractions using vacuum liquid chromatography. The most active fraction (ethanol), was subjected to column chromatography. Gas Chromatography - Mass Spectroscopy (GC-MS) was used to analyse the subfraction which was ultra-violet active. The crude methanol extract and fractions (250–8000 ppm) were tested for larvicidal activity. The methanol extract was further tested for anti-oviposition, effects on growth and development on Aedes aegypti, Anopheles gambiae and Culex quinquefasciatus mosquitoes together with corresponding positive (lambda-cyhalothrin) and negative controls (dimethylsulfoxide) using World Health Organisation procedures. Toxic effects, determined by mortality, on representative non-target organisms was evaluated on adult Biomphalaria glabrata (snail) and tadpoles of Bufo regularis. Ten each of the organisms were used per concentration. Data were analysed using descriptive statistics and student’s t-test at α0.05. Tannins, flavonoids, saponins and alkaloids were identified from the crude extract. Seventeen compounds were detected from ethanol subfraction, with quantities varying from 0.0 to 23.7%. Prominent compounds include 4-((1E)-3-Hydroxy-1-propenyl)-2-methoxyphenol (7.5%), 2-Hydroxy-5-methylbenzaldehyde (6.1%) and Ethyl iso-allocholate (23.7%). Larvicidal activities of crude methanol extract and fractions were in the order ethanol (100.0%) > methanol (96.6%) > n-hexane (7.5%) > DCM/ethyl-acetate (0.0%) at 2000ppm after 24h. Susceptibilities of mosquito larvae to methanol extract were also in the order A. aegypti > A. gambiae > C. quinquefasciatus. Mortality of the larvae was significantly different from the positive control. Anti-oviposition index increased (46.4-89.9) with extract concentrations. Morphological deformities of larvae and pupae were observed at the higher doses of 2000 ppm (A. aegypti), 5000 ppm (A. gambiae) and 7000 ppm (C. quinquefasciatus). Mosquito adult emergence inhibitions (24.0-100.0%) were recorded at 1100–5600 ppm concentration levels. The crude extract had no effect on the tadpoles at 250–1000 ppm, but the mortality of the snail at 250–2000ppm, varied from 10.0–95.0%. Clerodendrum polycephalum leaves contained biological components capable of inhibiting the survival and development of mosquitoes and could be exploited in the control of mosquitoes.Item CYTOGENETIC AND SYSTEMIC TOXICITY INDUCED BY SILVER AND COPPER(II) OXIDE NANOPARTICLES AND THEIR MIXTURE IN THE SOMATIC CELLS OF THREE EUKARYOTIC ORGANISMS(2019-06) OGUNSUYI, O.ISilver (Ag) and copper(II) oxide (CuO) nanoparticles are used in personal care products because of their antimicrobial properties. Their continual release into the environment may enhance genotoxic effects in the ecosystem, a condition widely reported from in vitro studies. However, in vivo, there is insufficient information on DNA and systemic damage, as well as the effect of the mixture of these nanoparticles in aquatic and terrestrial biota. This study was designed to investigate the genetic and systemic toxicity of Ag and CuO nanoparticles, singly and combined in somatic cells of three eukaryotic organisms and their mechanism of DNA damage. The selected eukaryotic organisms were onion (Allium cepa Linnaeus), mud catfish (Clarias gariepinus Burchell) and mice (Mus musculus Linnaeus). Cytogenotoxicity of Ag, CuO and their mixture (1:1) was investigated at different concentrations using the A. cepa chromosome aberration assay (0, 5-80 mg/L; n=64), micronucleus assessment in peripheral blood of juvenile catfish (0, 6.25-100 mg/L; n=80) and bone marrow of male mice (0, 18.75-300 mg/kg; n=64). Haematological parameters [haemoglobin concentration, Packed Cell Volume (PCV), Red Blood Cell (RBC) and White Blood Cell (WBC) counts] were assessed in catfish and mice. The histopathology of their liver and fish gill was done using standard protocols. Mechanism of DNA damage was investigated by analysing hepatic oxidative stress biomarkers [Superoxide Dismutase (SOD), reduced Glutathione and Malondialdehyde] in both catfish and mice. Interaction Factor (IF) of the mixture was calculated according to standard method. Data were analysed using descriptive statistics and ANOVA at α0.05. In A. cepa, there was a concentration-dependent increase in the percentage frequency of dividing cells with Ag (1.3-1.6 fold); and decrease with CuO (1.1-16.8 fold) as well as mixture (1.5-2.7 fold). The frequency of aberrant chromosomes significantly increased only with Ag (3.3-8.7 fold) and mixture (1.5-4.6 fold) compared with control. Micronuclei induction with Ag, CuO and their mixture significantly increased in catfish (1.1-1.9, 1.4-2.2 and 1.6-2.9 fold), and mice (1.0-2.9, 1.1-4.8 and 1.5-3.1 fold), respectively. Haemoglobin concentration, PCV, RBC and WBC significantly decreased only in both nanoparticles and their mixture for catfish. Gill lamella hyperplasia and hepatocellular necrosis were observed in catfish and mice respectively. In catfish, there were significant alterations in SOD activities (1.1-2.2 fold increase with Ag and CuO; and 1.6-2.0 fold decrease with mixture). Alongside, reduced Glutathione and Malondialdehyde levels (1.1-1.8; and 1.1-2.4 fold increase with Ag and CuO, respectively; and 1.1-2.8 fold decrease with mixture) were altered. In mice, there were significant alterations in SOD activities (1.1-1.6 fold decrease with Ag and CuO; and 1.3-1.6 fold increase with mixture), Malondialdehyde (1.1-1.5 fold increase with Ag and mixture; and 1.1-2.0 fold decrease with CuO) and reduced Glutathione levels (1.1-1.2 fold increase with Ag and decrease with CuO). The IF showed that interaction between Ag and CuO was antagonistic for cytogenotoxicity and oxidative stress. Silver, copper(II) oxide and their mixture induced genomic disruption in the three organisms with systemic anomalies in Clarias gariepinus and Mus musculus. Oxidative stress in the exposed cells was responsible for the observed DNA damage.Item EARTHWORM DIVERSITY AND ABUNDANCE IN SELECTED DUMPSITES IN IBADAN, AND TOXICITY OF CONTAMINATED WATER ON ASSOCIATED FAUNA(2016-08) ADEWOYIN, O.AHuge quantities of wastes which are indiscriminately disposed into uncontrolled dumpsites and flowing waters around residential areas are generated from Ibadan, metropolis. These may adversely affect soil, surface- and ground-water qualities and the fauna. Limited information exists on toxic effects that wastes from these non- designated sites have on associated fauna. Earthworms are one of the dominant fauna in soils and are pollution bioindicators. Therefore this study was designed to investigate physico-chemical parameters, earthworm diversity and abundance, and acute toxicity of contaminated-water on some fauna in selected dumpsites in Ibadan. One-hundred and twenty water samples from streams (Irefin, Gege, Gbagi, Odinjo, and Omi-Adio) receiving domestic wastes; 60-composite soil samples each from dumpsites (Ojokondo, Olodo, Moniya, Idi-Ope and Oremeji) and stream banks; 12- composite soil samples from a control site at University of Ibadan (UI); 72 groundwater samples from wells around the dumpsites and UI were purposively collected once every two months from March 2008 to February 2010. Earthworms from top-soil were sampled using 0.25x0.25 m2 quadrat, handpicked, identified and density determined following standard procedures. Earthworm species’ diversity and evenness were determined using Shannon-Wiener diversity index and Shannon’s evenness, respectively. Water samples were analysed for physico-chemical parameters [including Dissolved-Oxygen (DO), Biochemical-Oxygen-Demand (BOD), and Chemical-Oxygen-Demand (COD)] and soil samples including lead and zinc using standard methods and results were compared with NESREA standards. The 48-hour-LC50 of stream-samples were determined for Cloeon perkinsi (mayfly) larvae (CP); while 96-hour-LC50 for Rana temporaria tadpoles, Clarias gariepinus frys (CG1) and fingerlings (CG2) were determined using Probit method. Data were analysed using descriptive statistics and ANOVA at p=0.05. Three earthworm species (Eudrilus euginiae, Dichogaster modigliani and Hyperiodrilus africanus) were identified at UI, streams and dumpsites. Highest density of Eudrilus euginiae (30.0±13.7/m2) was recorded at UI and this was significantly different from other sites, while lowest (6.4±2.2/m2) was recorded at Ojokondo. Highest densities of Hyperiodrilus africanus (32.7±14.6/m2) and Dichogaster modigliani (40.0±0.0/m2) were recorded at UI and Gbagi, respectively; while lowest value of 4.0±0.0/m2 was recorded at Irefin and Gege. Eudrilus euginiae and Dichogaster modigliani were not found at Gege and Idi-Ope, respectively, while Hyperiodrilus africanus was not found at Odinjo, Moniya and Olodo. Earthworm diversity (0.5) was highest in UI and lowest (0.3) at Gege. Earthworms were most evenly distributed at Odinjo (2.4) and least at Ojokondo (0.8). The DO (mg/L) for streams ranged from (0.4±0.4) to (2.6±0.6) and were lower than permissible limit for aquatic life. Lead (260.6±77.7mg/kg, 269.6±46.4mg/kg) and zinc (456.9±69.9mg/kg, 1685.1±420.3mg/kg) values in Gege and Ojokondo soils, respectively were significantly higher than NESREA limits. In all groundwater samples, BOD (46.7±21.0mg/L) and COD (154.0±7.0mg/L) exceeded NESREA limits. The 48-hour- LC50 of stream-water to CP (Irefin, 12.7%; Gege, 8.6%), and 96-hour-LC50 to CGI (Gege, 0.8%; Gbagi, 2.8%) and CG2 (Gege, 3.3%; Irefin, 0.6%) indicated high toxicity of the sites. The low earthworm abundance, poor physico-chemical qualities and high toxicity of the study sites revealed that the soil, streams and groundwater were polluted in Ibadan. Consequently, there is need for adequate management and disposal of solid- wastes to prevent further environmental contaminationItem GENETIC AND SYSTEMIC TOXICITY INDUCED BY TITANIUM DIOXIDE AND ZINC OXIDE NANOPARTICLES AND THEIR MIXTURE IN SOMATIC AND GERM CELLS OF MICE(2018-11) FADOJU, O.MTitanium dioxide (TiO2) and zinc oxide (ZnO) nanoparticles are components of personal care products whose continuous release into the environment may enhance co-exposure, with potential risks to the ecosystem. In vitro studies have shown their potential to induce genetic damage. However, there is dearth of information on in vivo induction of DNA and systemic damage, alongside their interactive effects. This study was designed to investigate genetic and systemic toxicity and mechanism of DNA damage by TiO2 and ZnO nanoparticles and their mixture in mice. Male Swiss mice (=24.0±2.0g; n=80; 6-8 weeks old) were intraperitoneally exposed to distilled water (Control) and 9.4, 18.8, 37.5, 75.0 and 150.0 mg/kg concentrations of each of the nanoparticles and their mixture (1:1) for 5 days (5 mice/group) to assess micronucleus induction and cytomorphological abnormalities in the bone marrow of mice. Haematological parameters [Haemoglobin, Packed Cell Volume (PCV), Red Blood Cell (RBC) and White Blood Cell (WBC) counts] were assessed following standard procedures. Mechanism of DNA damage was evaluated by oxidative stress [Superoxide dismutase (SOD), reduced Glutathione and Malondialdehyde in the liver and kidney] parameters following standard methods. Sperm count, motility, abnormalities and concentrations of Luteinizing Hormone (LH), Follicle Stimulating Hormone (FSH) and Testosterone were evaluated in another group of mice (=30.0±2.0g; n=80; 11-15 weeks old), intraperitoneally exposed with the same nanoparticle concentrations (5 mice/group) at 35-day exposure. Liver, kidney and testis were sectioned for histopathological analysis. The Interaction Factor (IF) of nanoparticle mixture was calculated according to standard method. Data were analysed using descriptive statistics and ANOVA at α0.05. The nanoparticles and mixture induced micronuclei, but significant only for TiO2 (16.8±2.1-53.3±18.5) compared with the control (3.7±0.9). Blebbed, target, hyperchromic and hypochromic erythrocytes were the observed cytomorphological anomalies. The mixture exerted a significant reduction only in the WBC count. In the liver, there was a significant decrease in SOD (unit/mg protein) activities (1.3-1.5; 1.4-2.0; and 1.2-1.6 fold for TiO2, ZnO and mixture, respectively), with increase in Malondialdehyde (nmol/mg protein) levels (1.1-1.7; 1.2-1.8; and 1.7-1.7 fold for TiO2, ZnO and mixture, respectively). In the kidney, there were significant alterations in SOD: 1.2-1.3; and 1.1-1.4 fold decrease for TiO2 and ZnO, respectively; and 1.3-2.0 fold increase for the mixture. While Malondialdehyde levels increased (1.2-1.4; 1.4-1.6; and 1.7-1.9 fold for TiO2, ZnO and mixture, respectively). Both organs showed alterations in reduced Glutathione levels (1.0-1.5 fold decrease for TiO2; 1.0-1.1 fold increase for ZnO and mixture) indicating systemic toxicity. A significant decrease in sperm count and motility; and increase in abnormalities (1.3-8.0; 1.2-2.6; 4.6-12.1 fold for TiO2, ZnO and mixture, respectively), with a concomitant decrease in the serum level of LH and increase in FSH and Testosterone were observed. Hepatocellular and spermatogenic cell necrosis and degeneration of tubular epithelial cells were observed. The IF indicated synergism. Titanium dioxide and zinc oxide nanoparticles and their mixture induced genomic and systemic damage in somatic and germ cells of mice; with the mixture synergistically evoking the highest toxic response. Oxidative stress might be one of the mechanisms of cytogenotoxicity.Item GENETIC AND SYSTEMIC TOXICITY INDUCED BY TITANIUM DIOXIDE AND ZINC OXIDE NANOPARTICLES AND THEIR MIXTURE IN SOMATIC AND GERM CELLS OF MICE(2018-11) FADOJU, O.MTitanium dioxide (TiO2) and zinc oxide (ZnO) nanoparticles are components of personal care products whose continuous release into the environment may enhance co-exposure, with potential risks to the ecosystem. In vitro studies have shown their potential to induce genetic damage. However, there is dearth of information on in vivo induction of DNA and systemic damage, alongside their interactive effects. This study was designed to investigate genetic and systemic toxicity and mechanism of DNA damage by TiO2 and ZnO nanoparticles and their mixture in mice. Male Swiss mice (=24.0±2.0g; n=80; 6-8 weeks old) were intraperitoneally exposed to distilled water (Control) and 9.4, 18.8, 37.5, 75.0 and 150.0 mg/kg concentrations of each of the nanoparticles and their mixture (1:1) for 5 days (5 mice/group) to assess micronucleus induction and cytomorphological abnormalities in the bone marrow of mice. Haematological parameters [Haemoglobin, Packed Cell Volume (PCV), Red Blood Cell (RBC) and White Blood Cell (WBC) counts] were assessed following standard procedures. Mechanism of DNA damage was evaluated by oxidative stress [Superoxide dismutase (SOD), reduced Glutathione and Malondialdehyde in the liver and kidney] parameters following standard methods. Sperm count, motility, abnormalities and concentrations of Luteinizing Hormone (LH), Follicle Stimulating Hormone (FSH) and Testosterone were evaluated in another group of mice (=30.0±2.0g; n=80; 11-15 weeks old), intraperitoneally exposed with the same nanoparticle concentrations (5 mice/group) at 35-day exposure. Liver, kidney and testis were sectioned for histopathological analysis. The Interaction Factor (IF) of nanoparticle mixture was calculated according to standard method. Data were analysed using descriptive statistics and ANOVA at α0.05. The nanoparticles and mixture induced micronuclei, but significant only for TiO2 (16.8±2.1-53.3±18.5) compared with the control (3.7±0.9). Blebbed, target, hyperchromic and hypochromic erythrocytes were the observed cytomorphological anomalies. The mixture exerted a significant reduction only in the WBC count. In the liver, there was a significant decrease in SOD (unit/mg protein) activities (1.3-1.5; 1.4-2.0; and 1.2-1.6 fold for TiO2, ZnO and mixture, respectively), with increase in Malondialdehyde (nmol/mg protein) levels (1.1-1.7; 1.2-1.8; and 1.7-1.7 fold for TiO2, ZnO and mixture, respectively). In the kidney, there were significant alterations in SOD: 1.2-1.3; and 1.1-1.4 fold decrease for TiO2 and ZnO, respectively; and 1.3-2.0 fold increase for the mixture. While Malondialdehyde levels increased (1.2-1.4; 1.4-1.6; and 1.7-1.9 fold for TiO2, ZnO and mixture, respectively). Both organs showed alterations in reduced Glutathione levels (1.0-1.5 fold decrease for TiO2; 1.0-1.1 fold increase for ZnO and mixture) indicating systemic toxicity. A significant decrease in sperm count and motility; and increase in abnormalities (1.3-8.0; 1.2-2.6; 4.6-12.1 fold for TiO2, ZnO and mixture, respectively), with a concomitant decrease in the serum level of LH and increase in FSH and Testosterone were observed. Hepatocellular and spermatogenic cell necrosis and degeneration of tubular epithelial cells were observed. The IF indicated synergism. Titanium dioxide and zinc oxide nanoparticles and their mixture induced genomic and systemic damage in somatic and germ cells of mice; with the mixture synergistically evoking the highest toxic response. Oxidative stress might be one of the mechanisms of cytogenotoxicity.Item GENETIC POLYMORPHISMS ASSOCIATED WITH HYPERTENSION IN THE ETHNIC POPULATIONS OF CALABAR AND UYO, NIGERIA(2012-08) KOOFFREH, M.EHypertension is a public health challenge due to its high prevalence, and is a major risk factor for cardiovascular diseases. Hypertension is a complex disease resulting from an interaction of genes and environmental factors. Inconsistent association between polymorphisms of the renin angiotensin aldosterone, the atrial natriuretic peptide systems and hypertension has been reported among various ethnic groups, but not for the Efiks and Ibibios in south-south Nigeria. This study was designed to determine the frequency of gene polymorphisms of these two systems and their association with hypertension in Calabar and Uyo, Nigeria. A population-based case control design was used. A total of 1224 participants, 612 each of patients and controls were randomly recruited from hypertension clinics and the general population. Genotyping of the M235T allele of the angiotensinogen, Insertion/Deletion allele (I/D) of the angiotensinogen converting enzyme, A1166C allele of the angiotensin II type I receptor and C664G allele of the atrial natriuretic peptide genes to identify variants was performed using polymerase chain reaction and restriction enzyme digestion. The Hardy-Weinberg equation was used to calculate the allele and genotype frequencies. Plasma angiotensinogen levels were measured by Enzyme Linked Immunosorbent Assay. Hypertensinogenic factors such as age, familial history, physical exercise and drinking were assessed using questionnaires. Descriptive statistics, chi-square, multiple regression analysis and odds ratio were used to analyze the data. The frequency of the genotypes M235M, M235T, T235T of the M235T allele for the Efiks were 0.4, 7.7, 92 % in patients and 0, 6, 94 % in controls; for the Ibibios were 0.5, 1.2, 87 % in patients and 0, 7, 93 % in controls. The I/D genotypes II, ID, DD frequencies for the Efiks were 11, 44, 46 % in patients and 16, 45, 39 % in controls; for the Ibibios were 11, 40, 49 % in patients and 13, 49, 38 % in controls. The frequency of the A1166C carriers was 1 % while 99 % of the study population had the wild type A1166A genotype for the A1166C allele. Only the CC genotype was observed for the C664G allele. These frequencies did not conform to the Hardy-Weinberg assumptions. There were no significant differences between the genotype frequencies of patients and controls. Plasma angiotensinogen values were significantly higher in the patients with M235T allele than in the controls. Age was a positive predictor for systolic blood pressure (SBP, r = 0.60) in patients and diastolic blood pressure (DBP, r = 0.56) in controls. Other hypertensinogenic variables were not predictors for SBP and DBP in the population (p < 0.05). The Insertion/Deletion allele was a risk factor for hypertension, (O.R = 1.15). A high frequency was observed for the M235T allele and the Insertion/Deletion allele, which was associated with an increased risk for hypertension. The lack of association between the alleles of the M235T, A1166C and the C664G and hypertension suggests that other loci or environmental factors are involved in the disease outcome.Item HEAVY METALS ACCUMULATI ON IN SOIL, WATER, MAIZE AND CHICKEN AT ORI-ILE BATTERY WASTE DUMPSITE, OLODO, IBADAN, NIGERIA(2016-06) ONONUGA, A . OBattery waste consists of toxic Heavy Metals (HMs) and the Ori-Ile battery waste dumpsite, Olodo, Ibadan has elicited public health concerns. Available literature revealed that maize absorbs and accumulates toxic HMs from polluted soils of irrigated farmlands. But, there is limited information on its accumulation in chicks fed with maize-based feeds at Olodo, where maize is predominantly grown for poultry feed and human consumption. This study was designed to investigate accumulation of some HMs in soil, water, maize and chicks from the vicinity of Ori-Ile battery waste dumpsite, Olodo, Ibadan, Nigeria. An auto-battery Waste Dumpsite (WD), Ori-Ile, Olodo was purposively selected for the study. One hundred and samples l six topsoithirty were purposively collected (every two months, 2009) July to2008March from waste dumpsite and along North, South, East and West (N,S,E,W) directions at 5 m intervals from the edge of WD. Thirty two groundwater samples were collected 25 m away from . directionsW E,S,N, along WD Control soil and water samples were collected from Moor Plantation (MP), Ibadan. Soil and groundwater samples were analysed for HMs. Maize was planted in Direction with Highest HMs Concentration (DHHMC) for three months. The maize-parts weregrains) leaves, stems, (roots, harvested and analysed for HMs. Broiler feed protein) crude 19.7%-(18.8 was formulated from part of harvested-grains using standard method. Thirty broilers (day-old) were obtained from a farm, acclimatised for two-weeks on commercial feed and subdivided into two equal groups. The broilers were then fed on Formulated Feed from Harvested Grains (FFHG) and Formulated Feed from Control Grains (FFCG) for additional six weeks. Five chicks from each broiler group were sacrificed at four, six and eight-week old to determine lead, cadmium and iron accumulation in plasma, skin, liver and femur. Similar organs from Free-Range Chicks around WD (FRCMP) MP and (FRCWD) were also analysed for metals. Histopathological analysis of chicks’ liver and kidney were done using standard procedure. The HMs in all samples were determined using atomic absorption spectrophotometry. Contamination, Bio-concentration and Bio-accumulation Factors BaF) and BcF (CF, were determined for soil, maize and chicks respectively using standard methods. Data were analysed using descriptive statistics, ANOVA and T-test at p=0.05. iii The HMs concentration (mg/kg) from the WD was 123.1,±258.4 :Cd 1436.7,±4273.8:Pb 791.5±7910.0:Fe while that for 94.8,±274.3 :Cd 1107.9,±4693.8 :Pb :North 740.0;±8346.7 :Fe South: 867.0,±4353.3:Pb 603.5;±8189.6:Fe 71.4,±255.2:Cd 65.6,±248.2:Cd 832.9,±4351.3 :Pb :East 639.5; ±8130.0:Fe West: 1020.8,±4698.3 , 676.8±7851.3 86.9,±278.4 respectively. These were significantly higher than control 353.9)±976.3 1.2,±2.2 39.8,±(157.0 and NESREA limits 50).:Cd 164,:(Pb Soil CF values were greater than 6 indicating severe contamination. The HMs concentrations in groundwater 0.002, ±0.003:Cd 0.015,±0.017:(Pb mg/L) 0.015±0.033:Fe were significantly higher than control but less than NESREA limits 1.0). :Fe 0.005,:Cd 0.01,:(Pb Lead and cadmium in DHHMC maize-parts were significantly higher than control. Roots had concentration of . mg/L 0.19±2.84:Cd and 1.98±40.95:Pb In all maize-parts, BcF of HMS was < 1. Four-week old FFHG broilers’ liver had highest lead, cadmium and iron 0.003,±0.011 0.002,±(0.014 ly) respective mg/L 28.023±302.01 where four- weeks>six-weeks>eight-weeks. In FFHG and FRCWD chicks, lead and cadmium were significantly higher than FFCG and FRCMP. Lead and cadmium BaF values for all chicks were < 1 while iron was > 1. In FRCWD, lead iron and 0.015)±(0.068 8.48)±(298.0 were highest in the liver, while 0.002) ±(0.013 cadmium was highest in the skin. Necrosis, severe diffused hepatic degeneration and interstitial haemorrhages were observed in FFHG and FRCWD chicks. High accumulation of heavy metals found in the soils of Ori-Ile battery waste dumpsite, Olodo, Ibadan bio-accumulated in maize-roots and in chicken organs.Item HOST CYTOKINE GENE POLYMORPHISMS AND PARASITE GENETIC VARIABILITY IN DETERMINING THE DISEASE OUTCOME OF Plasmodium falciparum INFECTION(2012-09) OYEDEJI, S.IMalaria, the world’s most important tropical parasitic disease is caused by Plasmodium species. Infection with Plasmodium falciparum can result in one of three possible disease outcomes: Asymptomatic (AS), Uncomplicated Malaria (UM) or Severe Malaria (SM). Information on host genetic factors and parasite genetic diversity can improve understanding of the disease pathogenesis. In this study, the genetic diversity of P. falciparum isolates as well as polymorphisms in host cytokine genes was investigated in relation to the outcome of P. falciparum infection. Four hundred and thirty-seven children recruited from the Specialist Hospital, Lafia, Nasarawa State were assigned into UM or SM based on malaria severity, determined by clinical and laboratory diagnoses. Asymptomatic children recruited from primary schools within the study location constituted the control group (AS). Plasmodium falciparum infection was confirmed by PCR-based assay of SSUrRNA genes. Genetic diversity of P. falciparum was analyzed by genotyping the polymorphic domains of the Merozoite Surface Protein 2 (MSP-2). Host cytokine genes investigated included Interleukin-18 (IL-18), IL-18 receptor alpha (IL-18Rα) and Tumour Necrosis Factor alpha (TNF-α). Sequencing of MSP-2 gene and of the pro-inflammatory cytokines was carried out using ABI PRISM® 3100. Sequences were analyzed using the BioEdit Sequence Alignment software. Genotype and allelic frequencies were analyzed by Chi-square test. The level of significance was set at P=0.05. All participants had P. falciparum infection. Polyclonality was significantly higher in the AS (61%) and UM (60%) groups compared with the SM (34%) group. Mean multiple infections was 2.1 ±1.0 in AS, 2.0 ±1.0 in UM and 1.3 ±0.6 in SM. A total of 32, 35 and 28 distinct MSP-2 alleles were found in the AS, UM and SM groups respectively. Frequency of the 3D7 allele type was significantly higher in UM (51%) and SM (54%) groups compared to the AS group (38%). Sequence analysis of the central variable region of the MSP-2 gene showed that the FC27-type sequence was characterised by two unique subtypes and hybrid sharing sequence with the two subtypes. The 3D7-type sequence was characterised by three subtypes of repetitive domains: a GSA-rich repeat unit, a TPA repeat motif and a poly-Threonine stretch. Three single nucleotide polymorphisms (SNPs): -656G/T, -607C/A and -137G/C were identified at the promoter region of IL-18 gene. The -656G/T and -607C/A SNPs were found to be in complete linkage disequilibrium. The genotype frequency of -607AA was significantly higher in the AS group compared to SM cases (χ2=4.26, P<0.05). Likewise, four SNPs were identified at the promoter and Exon 1 of the IL-18Rα: -661T/C, -175G/A, -93C/T and Ex1 +21C/G but none was associated with disease outcome based on statistical level of significance. Exons 2 to 11 of IL-18Rα gene were relatively conserved. Furthermore, two SNPs: -308G/A and -238G/A were identified at the promoter region of TNF-α but none was associated with disease outcome. Plasmodium falciparum was found to be genetically heterogeneous. Higher carriage of Plasmodium falciparum 3D7 alleles indicates higher risk of developing symptomatic malaria. There was association between IL18 -607AA genotype and asymptomatic infection, probably indicating a protective role.Item HOST GENETICS AND EXPOSURE IN SUSCEPTIBILITY TO SOIL-TRANSMITTED HELMINTH INFECTIONS AMONG HOUSEHOLDS IN IGBO-ORA, NIGERIA(2019-03) OLUWATOBA, O.AHOST GENETICS AND EXPOSURE IN SUSCEPTIBILITY TO SOIL-TRANSMITTED HELMINTH INFECTIONS AMONG HOUSEHOLDS IN IGBO-ORA, NIGERIAItem MOLECULAR ANALYSIS OF MECHANISMS AND IDENTIFICATION OF FACTORS OF PYRETHROID RESISTANCE IN ANOPHELES GAMBIAE SENSU LATO IN SOUTHWESTERN NIGERIA AND SOUTHERN BENIN REPUBLIC(2011) DJOUAKA-FOLEFACK, J.RThe development of resistance to insecticides by Anopheles mosquitoes continues to threaten the success of malaria control programmes in West Africa. Local data on mechanisms and factors causing resistance in the region are scanty. This study was designed to investigate the environmental factors and mechanisms implicated in resistance to pyrethroids by Anopheles gambiae in southwestern Nigeria and southern Benin Republic. Larvae of Anopheles mosquito were collected in 2007 from 19 localities in the six states of southwestern Nigeria and 18 localities in the seven divisions of southern Benin and reared to adults. These were identified morphologically and with Polymerase Chain Reactions (PCR). They were also bioassayed for susceptibility to pyrethroids. Molecular characterisation of pyrethroid resistant phenotypes was carried out using PCR and microarray analyses of the expressed genes. Dissolved Oxygen (DO) and pH were determined using a digital multipurpose meter while physical appearances of breeding sites were assessed visually. Xenobiotic factors such as Spilled Engine Oil (SEO) and agricultural pesticides that might contribute to the emergence of resistance in Anopheles populations were examined through bioassay. Associations between pyrethroid resistance with environmental factors and molecular profiles of Anopheles were evaluated using Chi square. A. gambiae complex genotyped in Nigeria comprised of 73.6 % A. arabiensis and 26.3 % A. gambiae sensu stricto; while those genotyped in Benin were 92.9 % A. gambiae s.s. and 7.0 % A. melas. Pyrethroid resistance in Nigeria and Benin were recorded in 68.4 % and 94.4 % of the localities examined respectively. Breeding sites contaminated with SEO (B-SEO) or Pesticide Residues (B-PR) had low DO (B-SEO = 13.4 ± 1.5 mg/l, B-PR=12.2 ± 1.7 mg/l), the Non-contaminated Breeding sites (B-NC) had higher levels of DO (B-NC=33.1 ± 2.3) and mainly produced pyrethroid-susceptible Anopheles (p<0.05). Significant variations in pH were not recorded. Differences in habitation by resistant-Anopheles in breeding sites contaminated with SEO or pesticide residues were observed. A. gambiae found around the two agricultural sites (Houeyiho, Benin and Ajibode, Nigeria) exposed to synthetic pesticides showed significant levels of pyrethroid resistance with mortality rates of 70.0 % and 89.7% respectively. A. gambiae larvae survived at SEO concentrations below 11.8x10-3 μL/cm2. Ninety six percent of larval mortality resulted from direct cuticle contact with SEO whereas only four percentage mortality was from larval suffocation. A cross resistance phenomenon was recorded with SEO and pyrethroids. A. gambiae showed the presence of elevated frequencies of knock down resistance West (kdr-W) mutations in Benin samples (kdr-W ranged from 0.6 to 0.9) and absence of kdr-W in Nigeria samples. Two detoxification genes (CYP6P3 and CYP6M2) were up-regulated in resistant-Anopheles. Additional detoxification genes specific to agricultural and SEO sites were also over-expressed in the resistant populations. There was an association between residual synthetic pesticides, spilled engine oil and emergence of pyrethroid-resistance in A. gambiae in Nigeria and Benin Republic. The diversified profile of identified metabolic genes reflected the influence of a range of xenobiotics on selection of resistance in mosquitoes.Item MOLECULAR MECHANISMS OF MULTIPLE INSECTICIDE RESISTANCE IN Anopheles funestus sensu stricto AT AKAKA-REMO, SOUTHWESTERN NIGERIA(2019-03) ATOYEBI, S.MThe use of insecticides against malaria vectors has been a good approach to control malaria, but its efficacy is threatened by mosquito resistance to the lethal doses. In Nigeria, Anopheles funestus is gradually becoming an important malaria vector, especially in rural areas. However, despite recurring cases of Insecticide Resistance (IR) in some Anopheles species, there is a dearth of information on the molecular mechanisms of IR in An. funestus. Therefore, this study was designed to investigate the molecular mechanisms of IR in An. funestus at Akaka-Remo, Southwestern Nigeria. Early morning collections of adult mosquitoes were conducted from October 2014 to April 2015 in 30 rooms (25 houses) at Akaka-Remo (population density = 5,585). The mosquitoes were identified with standard keys, but only An. funestus (F0) was identified to sub-species level by Polymerase Chain Reaction (PCR). The F0 was analysed for Plasmodium Infection Rate (PIR) by TaqMan real-time PCR, and screened for Knockdown resistance (Kdr) mutations using next-generation sequencing. The gene sequences generated were analysed with bioinformatics tools. The F0 was further subjected to forced-egg laying technique to generate the F1. The mortality rate of 2-5 day old F1 exposed to permethrin- (0.75%), deltamethrin- (0.05%), DDT- (4%), dieldrin- (4%) and bendiocarb (0.1%)-treated papers was determined using WHO standard procedures. The contribution of detoxifying enzymes to IR was assessed by first exposing the F1 to three standard synergists: Piperonyl Butoxide (PB:4%), S,S,S-Tributyl Phosphorotrithioate (STP:0.25%) and Diethyl Maleate (DM:8%); and to permethrin and DDT, respectively. Microarray and real-time PCR were used to identify differentially expressed genes in permethrin and DDT-resistant populations. The frequency of resistant and susceptible alleles in DDT (L119F-GSTe2) and dieldrin (A296S-RDL) resistance markers was determined in the F0 and F1 genotype using TaqMan real-time PCR. Data were analysed using descriptive statistics and Student’s t-test at 0.05. A total of 375 mosquitoes were collected and identified as An. funestus (83.8%), An. gambiae (6.9%), Culex species (5.6%), Aedes species (1.3%) and Mansonia species (2.4%). All Anopheles funestus were further identified as An. funestus sensu stricto, with a PIR of 8.0%; and no Kdr mutations. Permethrin, deltamethrin, DDT, dieldrin, and bendiocarb induced 68.0, 87.0, 10.0, 8.0 and 84.0 % mortalities, respectively in the F1, indicating resistance. Exposure of the F1 to the three synergists with permethrin induced 100% mortality each; while PB, STP and DM with DDT, respectively induced 30.0, 81.8 and 71.4% mortalities. These suggest that detoxifying enzymes contributed to IR in the mosquitoes. Overexpressed genes in the resistant populations were GSTe2, GSTd3, GSTd1-5, GSTU2, CYP6P2, CYP6P9a, CYP6P4a and CYP9K1. The frequency of resistant alleles in L119F-GSTe2 and A296S-RDL was significantly high: F0=77.0%, F1=80.0% and F0=76.0%, F1=90.0%, respectively compared to the susceptible alleles: F0=23.0%, F1=20.0% and F0=24.0%, F1=10.0%, respectively. Overexpression of detoxifying genes and the high frequency of resistant-associated mutations were responsible for multiple insecticide resistance in the Anopheles funestus at Akaka-Remo.Item MOLECULAR PREDICTIVE BIOMARKERS FOR ORAL SQUAMOUS CELL CARCINOMA IN HUMANS IN IBADAN, NIGERIA(2021-08) ONYEGBULA, K.CSquamous Cell Carcinoma is the most prevalent malignant tumour of the oral cavity in humans. Late hospital presentation and diagnosis often result in high mortality, recurrence and metastatic rates. Prognosis is poor with a low 5-year survival rate. There is a possibility that molecular events underlie the aetiology of oral squamous cell carcinoma (OSCC) and could prove useful in predicting OSCC-susceptible individuals, but this information is largely lacking in Nigerian cohorts. Thus, this study was designed to identify molecular predictive diagnostic biomarkers for OSCC from patients in Ibadan. Using a retrospective-prospective study design, a total of 100 (58 males, 42 females) histologically-classified OSCC cases were identified from 1527 tumour cases recorded at the University College Hospital, Ibadan, Nigeria, between January 2004 and December 2015. Patients’ demographic variables were extracted. Archived formalin-fixed, paraffin-embedded tissue samples were retrieved and processed immunohistochemically for Epithelial Membrane Antigen (EMA) and cytokeratin protein expression. The DNA from samples was also profiled for aberrant CpG island methylation and genotypes of rs7528484 polymorphism in RUNX3 gene by methylation-specific and restriction fragment length polymorphism-PCR, respectively. Data were analysed using descriptive statistics, while association between patients’ demographic variables, aberrant CpG island methylation and rs7528484 polymorphism in RUNX3 were assessed by Pearson’s χ2 test at P≤0.05, Monte-Carlo exact test and Odds Ratios (OR) at Confidence Interval (CI) of 95%, respectively. The prevalence of OSCC was 6.5%. The moderately differentiated class was the most prevalent (65.0%), with a general prevalence peak at the seventh decade age group and the palate being the most affected location. EMA was expressed by the well and moderately differentiated classes, while cytokeratin was expressed by the well, moderately and poorly differentiated classes. RUNX3 promoter hypermethylation was detected in 45.0% of OSCC, suggesting that aberrant CpG island promoter hypermethylation in RUNX3 was prevalent in the disease. The rs7528484 polymorphism in RUNX3 was also detected with a genotype distribution of 52.7% (39) homozygote normal (CC), 28.4% (21) heterozygote mutant (CT), 18.9% (14) homozygote mutant (TT), and a C>T allelic ratio of 0.67:0.33. There was significant association between aberrant CpG island promoter hypermethylation in RUNX3 and tumour location (P<0.05). Genotypes of rs7528484 polymorphism in RUNX3 and their alleles were significantly associated with both male and female gender (P<0.05) and histologic class (P<0.05). Mutant genotypes (CT) and (TT) showed odds of predicting OSCC (OR 0.28, 95% CI: 0.1889 - 0.3711) and (OR 0.18, 95% CI: 0.1118 - 0.2482), respectively. Mutant allele (T) showed odds of predicting OSCC (OR 0.66, 95% CI: 0.52 - 0.80). Aberrant CpG island promoter hypermethylation of RUNX3 in combination with tumour location and rs7528484 polymorphism in RUNX3 in combination with gender served as epigenetic and genetic predictors, respectively for oral squamous cell carcinoma, while epithelial membrane antigen expression pattern served as an immunohistochemical predictor for oral squamous cell carcinoma.Item PHYSICO-CHEMICAL PARAMETERS, PLANKTON, MACROZOOBENTHOS AND FISH FAUNA OF IBUYA RIVER, SEPETERI, SOUTH-WESTERN NIGERIA(2016-09) AKPONINE, J.AIbuya River runs across the Old Oyo National Park, a wildlife and recreational park. There is paucity of information on the limnology of the river which will provide information on the ecological status relevant for sustainable management. Therefore, this study was carried out to investigate the physico-chemical parameters, diversity and abundance of plankton, macrozoobenthos and fish fauna of Ibuya River. Surface water (72), plankton (72) and macrozoobenthic (72) samples were collected monthly from September 2012 to February 2014 at randomly selected stations (i-iv) along the river. Water temperature, pH and Dissolved Oxygen (DO) were measured in situ, while hardness, turbidity, phosphate (PO₄3˗), sulphate (SO₄2-), heavy metals including cadmium (Cd), Iron (Fe) and lead (Pb), were determined in the laboratory according to APHA method. Plankton samples were collected with plankton net (mesh size, 55 µm), identified and counted microscopically. Macrozoobenthos were collected with Van-Veen grab (0.086 m2), identified and counted macroscopically. Fish samples were collected with gill net (mesh size 45 mm), identified and counted. All identifications including pollution indicator species were done using standard identification keys. Species diversity was determined with Shannon-Wiener index HI. Descriptive statistics, student t-test, one-way ANOVA, Pearson correlation coefficient and Principal Component Analysis (PCA) were used for analysis of data at α0.05. Water temperature was 24.8±0.2 °C; pH, 7.60±0.04; DO, 4.4±0.2 mg/L; hardness, 39.6±2.1 mg/L CaCO3; turbidity, 19.4±0.9 FTU; Cd, 0.2±0.1 mg/L; Pb, 0.7±0.1 mg/L; Fe, 2.1±0.2 mg/L; PO₄3˗, 24.3±2.8 mg/L; and SO₄2-, 31.8±3.1 mg/L. Turbidity had significant spatial and seasonal variation at p<0.001 while pH and SO₄2- had significant spatial and seasonal variation at p<0.04 and p<0.002, respectively. Cadmium, Pb, and PO₄3˗ exceeded the NESREA permissible limits for surface water (0.003, 0.01 and 3.5 mg/L, respectively). Turbidity correlated significantly with Fe (r=0.6) and SO₄2- (r=0.7). The PCA revealed high positive loading for water temperature (0.7 °C), hardness (0.6 mg/L CaCO3) and turbidity (0.6 FTU). Forty-five species of phytoplankton belonging to four Classes: Bacillariophyceae (25 species), Chlorophyceae (Nine species), Euglenophyceae (Eight species) and Cyanophyceae (Three species) were recorded. Merismopedia punctata (52.1%) dominated the phytoplankton population. Zooplankton from three groups: rotifers (15 species), crustaceans (five species) and insects (one species) were encountered. Mesocyclops leuckarti (11.6%) was the most abundant zooplankton. Diversity index for phytoplankton was highest in station iv; stations ii and iii recorded highest HI for zooplankton. Eight species of macrozoobenthos were recorded with the gastropod, Indoplanorbis exustus (30.9%) dominating and the insect, Chironomus species (11.8%) was the least abundant. Twenty-four fish species were recorded. Family Cichlidae (22.6%) was the most abundant. Pollution indicator species were abundant and included the phytoplankton, Merismopedia punctata (52.1%) and the macrozoobenthos, Melanoides tuberculata (24.7%). This study provided baseline information on the ecological status of Ibuya River. However, the composition and diversity of both plankton and macrozoobenthos could be potentially used as bio-indicators for assessing and monitoring Ibuya River.Item PREVALENCE AND INTENSITY OF NEMATODE PARASITES OF Poecilia reticulata PETERS (1859) IN FOUR WASTEWATER DRAINS OF LAGOS STATE, NIGERIA(2013-02) AKINWALE, M.M.APoecilia reticulata (guppy) a common ornamental tropical fish is found in many wastewater drains in Nigeria. Guppies feed on copepods which are intermediate hosts of some nematode parasites of culturable fish species. The restriction on the importation of ornamental fishes into Nigeria has enhanced the demand for local species, usually sourced from the wild. There is dearth of information on the parasites of ornamental fishes in Nigeria. This study was aimed at determining prevalence and mean intensity of the nematode parasites of P. reticulata from four waste water drains in Lagos State.Item SUSCEPTIBILITY AND DIAGNOSTIC PROTEOMIC BIOMARKERS FOR URINARY SCHISTOSOMIASIS AND ASSOCIATED BLADDER PATHOLOGIES AMONG ADULTS IN EGGUA, OGUN STATE, NIGERIA(2017-07) ONILE, S.OThe failure to elicit an adequate immune response to the adult Schistosoma haematobium worm, and continuous strong inflammatory responses to the eggs have been the main causes of bladder pathology in chronic schistosomiasis. The identification of bladder pathology-associated biomarkers is necessary to enable early detection of the disease in a non-invasive manner. The aim of this study was to identify candidate-biomarkers for susceptibility and diagnosis of schistosomiasis and schistosomiasis-associated bladder pathologies in adults. A total of 371 respondents, comprising 130 males and 241 females from Eggua, Ogun State were randomly recruited into a cross sectional study from August 2012 to May 2014. Semi-structured pretested questionnaires were administered to obtain information from consenting respondent. They were screened for S. haematobium ova and bladder pathologies by microscopy and ultrasonography, respectively. Host susceptibility to bladder pathologies and schistosomiasis was determined by Polymerase Chain Reaction genotyping of glutathione-S-transferase (GSTT1 and GSTM1) genes, and Interleukin (IL4 and IL13) genes, respectively. Label-free quantification mass spectrometry-based proteomics approach was used to identify protein biomarkers in the urine. Samples were categorised as Schistosomiasis, Bladder Pathology (BP), Pathology and Schistosomiasis (PS). No Pathology and Schistosomiasis (NPS) served as controls. Descriptive statistics, odds ratios (OR) and Chi-square test were used at α0.05 to determine association between schistosomiasis and bladder pathologies. False Discovery Rate (FDR) analysis was also used to determine significant biomarkers. The mean age of respondents was 48.6 ±0.6 years. The prevalence of schistosomiasis was in 42 (11.4 %) males and 66 (17.9 %) females. Majority (74.1%) had light mean intensity of infection (33.3±0.04 eggs/10mL urine). Bladder pathologies included abnormal bladder wall thickness (29.0%), abnormal bladder shape (7.1%), bladder masses (3.1%) and bladder calcification (2.2%). There was a significant association between urinary schistosomiasis and BP. Respondents with GSTM1 and GSTT1 polymorphisms expressed elevated risks of BP (OR = 4.3, 95% CI 2.0 - 9.2 and OR = 4.2, 95% CI 1.5 – 12.0, respectively); with the PS having more GST polymorphisms than BP. Polymorphisms in IL 4-590 and IL 13-1055 were observed in 24.1% and 9.3% schistosomiasis cases, respectively. The IL 13-1055 polymorphisms did not indicate susceptibility to schistosomiasis in males (OR 0.7, 95% CI 0.3-2.1) but a slight risk was found in females (OR 1.1, 95% CI 0.7-1.7). A total of 1306 proteins and 8752 unique peptides were observed (FDR = 0.01). Human host (54) and parasite-derived (36) potential biomarkers were found for schistosomiasis and associated pathologies. These included new potential biomarkers in schistosomiasis (Sialidase-1, Growth factor 15, Programmed cell death 1 ligand-2) and PS (Arylsulfatase A and Phosphatidylethanolamine-binding protein 4). Candidate proteins were identified for the generation of new diagnostic markers for chronic urinary schistosomiasis and its bladder pathologies.Item Ultraviolet radiation of schistosoma mansoni. II. post-hatching radiation effect on some aspects of miracidia behaviour and infectivity(2000) Hassan, A. A.; Oyerinde, J. P. O.Batches of Schistosoma mansorti miracidia were irradiated with ultraviolet light for varying time intervals to determine the influence of radiation on the transmission potential of radiated miracidia. There was a decrease in the survival rate of hatched-free swimming miracidia that corresponded with the increasing radiation exposure time. The activity rate of radiated miracidia also decrease with increasing radiation exposure time, showing a 69.3% reduction in the mean rate of movement of miracidia irradiated for 30 minutes and 40.4% reduction in the miracidia exposed for 0.5 minutes. The infectivity rates of the free-swimming miracidia exposed to 5.0,15.0 and 30.0 minutes uv radiation were significantly different (P<0.001, dα=4.171,5.07,5.227) compared with the non- irradiated miracidium