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Aflatoxin exposure in Southwestern Uganda and the ameliorative effects of selected medicinal plants on induced aflatoxicosis in wistar rats

(Makerere University, 2023-11) Murokore, Biryomumaisho Justus

Introduction: Over 5.5 billion people, worldwide are chronically exposed to aflatoxins. This contributes to 40% of the African disease burden. Aflatoxins are mycotoxins produced by Aspergillus flavus and A. parasiticus, A. nomius, and A. tamarii, which are fungal species. Aflatoxin B1 (AFB1) is the most prevalent and carcinogenic among them. Since prevention of their production is not practical, we ingest them making toxicity control the easiest option. Consumption of high phenolic medicinal plant ameliorates the toxicological effects of aflatoxins. Objectives: The goal of the study was to assess dietary aflatoxin exposure in selected foodstuffs in Southwestern Uganda and the ameliorative effects of selected medicinal plants on aflatoxicosis in wistar rats. Specifically, the study set out to: (i) determine the aflatoxin susceptible food consumption frequency, prevalence and levels of aflatoxins in household foodstuffs in Southwestern Uganda; (ii) determine the phenolic, and flavonoid contents and antioxidant capacity of different selected medicinal plants (iii) assess the ameliorative effects of the selected medicinal plants on aflatoxin induced changes in liver, kidney and immune functions. Methods: Flours of Arachis hypogaea (groundnuts), Zea mays (maize), Eleusine coracana (millet), and Sorghum bicolor (sorghum), the most commonly aflatoxin contaminated foods, were systematically sampled, following a consumption frequency questionnaire. Samples were analysed for aflatoxin content using competitive ELISA. Ocimum gratissimum, Allium sativum, Cymbopogon citratus, Zingiber officinale, and primary grades and green tea, Camellia sinensis, were analysed for phenolics and flavonoids, following Folin-ciocalteu and Aluminium chloride methods respectively. Their antioxidant capacity was determined following DPPH and FRAP methods. Ninety-six male wistar rats were grouped into 16 groups of 6 and treated for 30 days as follows; 1. Control, 2. Olive oil. 3. AFB1 Low-dose (100µg/kg), 10. AFB1 High-dose (300µg/kgAFB1). 4-9. AFB1 Low-dose + 2% extracts, 11-16. AFB1 High-dose + 2% extract. Liver and kidney functions, histopathology, and immune function indicators were evaluated using standard methods. Results: The highest aflatoxin prevalence, levels and consumption frequency were observed in maize and groundnuts (Tables 4.1.2 and 4.1.1, respectively). Indoor dried/processed medicinal plants yielded higher total phenolic content (TPC) than fresh ones. In addition, 40 minutes extracts of medicinal plants yielded higher TPC. Green tea yielded the highest DPPH (93.82%) and FRAP (39.04µg AAE/mL), and tea harvested from Buganda (4,371 µgQE/g) the highest total flavonoid content (TFC). Green and black tea, C. sinensis, O. gratissimum, and C. citratus reduced the damaging effects of aflatoxin toxicity in liver functions, histopathology and blood and immunity parameters. Conclusion: Groundnut and maize were the main foods through which aflatoxin exposure occurs. For maximum TPC, it is better to use processed medicinal plants and longer extraction periods. Green and black tea, C. sinensis, O. gratissimum, and C. citratus have ameliorative effects in aflatoxicosis. Community awareness in aflatoxin contamination avenues and health effects, and encouragement to regularly consume high phenolic medicinal plants is urgently needed.
Amphibian diversity patterns in the Namulonge agricultural landscape

(Makerere University, 2017) Akoth, Sisiria

Uganda’s agricultural landscapes are patterns of cultivated areas and semi-natural elements. Although comprising only a small fraction of the total area, semi-natural elements provide habitats for most of the landscape biodiversity. Agricultural intensification has increasingly destroyed or fragmented semi-natural elements resulting into species decline. Insights into the effects of landscape structure on species’ distributions within and among semi-natural habitats are needed to effectively conserve biodiversity in agricultural landscapes. The influence of land use practices on amphibian species diversity was investigated in areas of intensive and subsistence agriculture in Namulonge for 12 weeks. Amphibians were sampled using Visual Encounter Surveys while environmental parameters were measured using digital multimetre. Altogether, 19 amphibian species were recorded in the Namulonge agricultural landscape. The Mann-Whitney U test showed no significant difference in species diversity between the land use areas. Characteristics of the landscape were used to predict patterns of amphibian species distribution in canonical correspondence analysis (CCA). The model identified plant materials, water and ambient temperature as the most important variables in predicting species distribution. Areas with relatively intact natural vegetation cover supported a few more species than completely converted areas. Such pockets of vegetation provide microhabitat conditions suitable for amphibian species occurrence. The foregoing observation suggests that there is value at landscape-level to preserve natural/semi-natural pockets of vegetation to enhance biodiversity in any agricultural environment.
Analgesic, anti-inflammatory and oxytocic activities of Dracaena steudneri Engl. stem bark aqueous extract in Wistar rats

(Academic journals, 2022) Katuura, Esther ; Tenywa, Mercy Gladys ; Agaba, Amon Ganafa ; Ajayi, Clement Olusoji ; Tolo, Casim Umba

Dracaena steudneri Engl. (Dracaenaceae) is a medicinal plant that is used in labour by traditional birth attendants to induce labour, relieve pain and treat various diseases; albeit no documentation. This study determined the analgesic, anti-inflammatory and oxytocic effects of the D. steudneri stem bark aqueous extract. D. steudneri stem bark was collected, authenticated, oven-dried at 45ºC for 48 h, pulverized and extracted using the decoction method. The analgesic effect of the aqueous extract was determined using the acetic acid writhing test, anti-inflammatory activity was determined using paw oedema method while oxytocic activity was determined using abortion and other standard test procedures and were tested at 12.1 24.1 and 48.2 mg/kg. The results of the analgesic study showed that the extract reduced the number of abdominal writhing between 53.51 and 92.40% at 6.02 and 48.2 mg/kg dose, respectively while the standard drug showed a reduction of 36.58% at 10 mg/kg. The anti-inflammatory results showed percentage inhibition of 18.6% (2 h), 22% (2 h) and 18.9% (6 h) at 12.1, 24.1 and 48.2 mg/kg, respectively. The in vivo oxytocic activity showed average time taken for the rats to deliver was between 20.63 h at lowest dose (12.1 mg/kg) and 8.25 h at highest dose (48.0 mg/kg) while Oxytocin the reference drug exhibited, 22 h as the average time taken for delivery.
Analysis of microbial quality of common bean (Phaseolus vulgaris l.) seeds from five bean producing districts in Uganda

(Makerere University, 2022-04-25) Isout, Sylas

Being a basic and an affordable source of protein, common bean (Phaseolus vulgaris L.) is grown widely in Uganda and the world at large. The quality of bean seeds has gained little attention yet remains a significant aspect of the value chain of production and consumption in Uganda. The aim of this study was to assess the microbial quality of common beans from five bean producing districts of Uganda. One hundred bean samples were collected from farmers and traders purposively selected from five bean producing districts of Uganda (Arua, Oyam, Hoima, Nakaseke, and Sironko districts). Three replicates of twenty samples of bean seeds randomly obtained from each district were subjected to microbial quality analysis using whole seed and agar plate standard protocols under laboratory conditions. The modified blotter method was used to test germinability and health of bean seeds in the laboratory. Standard seedling symptom tests were conducted in screen house experiments using a completely randomized design to assess germination and health of bean seedlings. Seed health tests revealed that 85% of the bean samples were contaminated with seed borne fungi. Five seed borne fungal genera were isolated from the seed samples, with Aspergillus (42.8 %) and Fusarium (37.1 %) being the most common fungi followed by Penicillium (18.9 %) while Colletotrichum (12.2 %) and Rhizopus (6.2%) were the least occurring fungi. Xanthomonas sp. was the only bacteria isolated from 15% of the samples tested. In the laboratory, bean seeds exhibited relatively poor germination (58.2%), with 30.9% of seeds having fungal outgrowths. While in the screen house, only 63.2% of seeds germinated, with 23.9% of ungerminated seeds being rotten. Disease assessment in the screen house revealed that, 25.1% of seedlings were diseased with various symptoms. Six seed borne diseases were recorded including; Fusarium wilt, Angular leaf spot (ALS), Ascochyta blight, Anthracnose, Common Bacterial Blight (CBB) and Bean common mosaic virus (BCMV). Determinately, the beans available in Uganda are of low/poor quality. Poor quality seeds contaminated with seed borne pathogens may result in poor germination rates, poor health of seedlings and reduced crop stand and yield. Therefore, improving access to clean and healthy seed for farmers is inevitable. Farmers are urged to embrace good bean production practices especially disease management in the field and post-harvest handling practices in order to improve production of safe and quality beans.
Anti-infective activity, phytochemistry and cytotoxicity of Dalbergia Melanoxylon Guill. & Perr and Ormocarpum Sennoides ssp. Zanzibaricum Brenan & J.B. Gillett

(Makerere University, 2025-03-26) Chalo, Duncan Mutiso

Infectious diseases account for 90% of the disease burden worldwide with a significant rampage being felt in developing countries. This is coupled by the widespread antibiotic resistance to available conventional drugs. Infectious agents have been reported to raise the risk of human cancer which is a major cause of mortality worldwide. The aim of this study was to investigate Dalbergia melanoxylon Guill. & Perr and Ormocarpum sennoides subsp. zanzibaricum Brenan & J.B. Gillett as potential anti-infective and anticancer agents. Extraction of root bark of D. melanoxylon was done using CH2Cl2/MeOH (1:1) while O. sennoides subsp. zanzibaricum was extracted using 80% methanol and partionioned to obtain n-hexane, ethylacetate, n-butanol and water extracts. The structures and absolute configuration of the isolated compounds were elucidated by Mass spectrometry, Nuclear magnetic resonance and Electronic circular dichroism spectroscopy. The antibacterial activity of the crude extract and the isolated compounds against Bacillus subtilis and Aliivibrio fischeri was determined in a turbidimetric assay. The pure compounds were subjected to a panel of human pathogenic bacteria (Enterococcus faecalis, Escherichia coli, Mycobacterium vaccae, Staphylococcus aureus, methicillin resistant Staphylococcus aureus and Pseudomonas aeruginosa using agar diffusion assay. The crude extracts were tested for antifungal activity against Phytophthora infestans, Botrytis cinerea, and Septoria tritici using turbidimetric assay. The pure compounds were subjected to further antifungal testing against human pathogenic fungi (Sporoidiobolus salmonicolor, Candida albicans and Penicillium chrysogenum) using agar diffusion assay. The crude extracts of O. sennoides subsp. zanzibaricum and D. melanoxylon (500 μg/ml) were tested for anthelmintic activity using Caenorhabditis elegans. The cytotoxicity (CC50) of the crude extracts against the human prostate cancer (PC-3) and colon adenocarcinoma (HT-29) was performed using colorimetric cell viability assays. The cytotoxicity assay against HeLa cells was performed on the most promising active pure compound using the methylene blue assay.Phytochemical investigation of root and root bark of O. sennoides subsp. zanzibaricum and D. melanoxylon resulted in the isolation of twenty compounds (1-20), of which seven are new. The root bark of D. melanoxylon yielded six previously undescribed prenylated isoflavanones (1-6), alongside isoflavanoids (7-10), neoflavones (11-13) and alkyl hydroxycinnamates (14). A new biflavonoid, trime-chamaejasmin (15) alongside two known biflavonoids (16-17), bi-4-phenyldihydrocoumarin (18), isoflavan (19), triterpenoid (20) were also isolated from the roots of O. sennoides subsp. zanzibaricum. The crude extracts at 50 and 500 μg/mL exhibited promising antibacterial activity against Bacillus subtilis. Sophoraisoflavanone A (10) caused inhibition against both B. subtilis and Aliivibrio fischeri assays at 99% at 100 µM. Kenusanone H (7) and (3R)-tomentosanol B (9) exhibited promising antibacterial activity against MRSA and M. vaccae showing minimum inhibitory concentrations between 0.8 and 6.2 µg/mL. Both crude extracts showed promising antifungal activity against all phytopathogens at 125 µg/mL. Kenusanone H (7) at 42 µg/mL showed very promising activity against S. tritici (74%) and B. cinerea (99%). At low 0.05 μg/mL, both crude extracts showed no cytotoxic effect against PC-3 and HT-29 cells. Kenusanone H (7) was highly cytotoxic against HeLa cells with CC50 of 1.8 ± 1.4 µg/mL (4.2 µM). The crude extracts of both O. sennoides subsp. zanzibaricum and D. melanoxylon are not toxic in general but show selective antibacterial and antifungal activities. Kenusanone H (7) proved to be a lead drug candidate as an antibacterial, antifungal and cytotoxic compound.
The Anti-mycobacterial activity and safety profile of selected Crinum species in Northwestern Uganda

(Global Scientific Journals, 2021) Katuura, Esther ; Waako, Paul ; Chandia, Milton

Crinum species is a source of many bioactive molecules with both antimicrobial and anti-tumor properties. Chloroform, methanol and aqueous extracts of Crinum scabrum and Crinum macowanii bulbs and leaves were investigated for their anti-mycobacterial activity against Mycobacteria tuberculosis using agar well diffusion, and broth dilution methods. Rifampicin streptomycin resistant (R.S), pan African sensitive (H37Rv) and wild type (Sou 14827) strains of the bacteria were used. The investigation aimed to determine the anti-mycobacterial activity and safety profile of the crude extracts of Crinum scabrum and Crinum macowanii. The extracts had antimycobacterial activity that ranged between 1-5µg/ml. Their patterns of inhibition varied with the plant extract, solvent used for extraction and the organisms tested. Different concentrations of methanol extract were compared with similar concentrations of chloroform and aqueous extracts for their maximum zones of inhibition. All the extracts of Crinum macowanii were found inactive against Mycobacteria tuberculosis. Methanol leaf extract of Crinum scabrum was the most active, with minimum inhibitory concentration (MIC) less than1µg/ml and minimum bactericidal concentration (MBC) of 1µg/ml. acute toxicity test in mice for Crinum scabrum leaf was found to have LD50 greater than 2000mg/kg. In conclusion, the leaf of Crinum scabrum is a natural source of new anti-mycobacterial compound that is tolerable and effective in treatment of tuberculosis. The study recommends herbalists to use the leaves instead of bulb of C. scabrum Alani Davis for treating TB patients. In future, a comparative study should be done on activity of crude methanol leaf extract of C. scabrum and combination of the drugs used to treat multidrug resistant MTB. The active principles in the leaves of C. scabrum Alani Davis should also be isolated, identified and tested on strains of Mycobacteria tuberculosis resistant to at least two or three drugs
Antiplasmodial activities of extracts and pure compounds from Securidaca Longipedunclata Fresen (Polygalaceae)

(Makerere University, 2023-01-30) Ochora, Douglas Ongeri

Introduction: Malaria is the most significant parasitic disease globally especially in Sub-Saharan Africa. The disease kills a child every two minutes. This is coupled with increased malaria parasites resistance to the available drugs. The challenges of parasites resistance call for search of novel antimalarial compounds from plants. This led to the determination of antiplasmodial activities of extracts and pure compounds from Securidaca longipedunculata Fresen (Polygalaceae). Cytotoxicity activities of extracts and pure compounds was also determined. Methodology: The roots extracts were subjected to a combination of chromatographic separations. Structures of isolated compounds were elucidated by mass spectrometry and Nuclear Magnetic Resonance spectroscopy. Crude roots, stems and leaves extracts, pure compounds and drug-extract combinations were each tested for immediate ex vivo and in vitro antiplasmodial activities using SYBR Green I method against W2, D6, 3D7 and DD2 reference strains of Plasmodium falciparum. Cytotoxicity assays were also done using the antiproliferative assay using drug-sensitive and multidrug-resistant cancer cell lines. Results: Roots extracts yielded a novel benzophenone 2,3,4,5-tetramethoxybenzophenone (37), alongside seven known compounds. Both methanol (MeOH) and ethyl acetate (EtOAc) roots extracts were active with IC50 values of less than 5 ng/mL against the D6 and 3D7 strains of P. falciparum. The MeOH extract showed good antiplasmodial activity against field isolates with IC50 value of 9.8±1.3 ng/mL and IC50 value of 1.4±0.07 ng/mL against W2 strain. Compounds 41 and 44 exhibited in vitro activity with IC50 value of 19.7 μM and 14.5 μM respectively and compounds 37 and 38 displayed ex vivo antiplasmodial activities with IC50 value of 28.8 μM and 18.6 μM, respectively all against D6 strain. Synergism was displayed across all fixed doses of artemether-roots extract, lumefantrine-stems extract, and lumefantrine-leaves extract combinations against D6, DD2, and W2 strains of P. falciparum, respectively. The artemether-roots extract combinations displayed the highest synergism across all fixed doses with an average FIC50 value of 0.403±0.068 against D6 strain. For cytotoxicity assay, all the extracts (apart from MeOH leaves extract) and pure compounds were not toxic to the normal BEAS and LO2 cell-lines. Conclusion: The observed synergism and high antiplasmodial activities of the species indicates its potential use as an alternative antimalarial drug in controlling the occurrence of malaria resistance.
Aspects of bats ecology in Kaptum cave in Mt. Elgon, Eastern Uganda

(Makerere university, 2019-10-14) Matovu, Benard

Few studies have addressed the ecology of cave bats in Uganda. This study aimed at assessing the diversity; roosting and feeding ecology of micro chiropterans as well as aspects of human disturbance in Kaptum cave of Mt Elgon Eastern Uganda. Data collection was conducted during two field visits in July/August 2016 and October/November 2016 survey periods. Mist nets and a harp trap were used to capture bats for examination and identification of species present. Infrared trail trap cameras were used to augment the monitoring of roosting habits and activity patterns of the bats in the cave; as well as record instances of human disturbance in the cave. A portable weather station (Novalyx WS-18) was used to record the microclimatic conditions in the different sections of the cave to evaluate their influence on choice of roost. Kaptum cave contains three main chambers each dominated by different genera or species of insectivorous bats. These chambers vary in structural complexity and size. The cave contained six species of insectivorous bats namely; Nycteris thebaica, Rhinolophus landeri, R. hildebrandtii, Hipposideros caffer, H. ruber, and Myotis bocagii. These generally roosted and segregated themselves in the three different cave chambers and hence referred to as: “Nycteris”, “Rhinolophus” and “Hipposideros” chambers. The bats exhibited similar and slight variations activity and roosting patterns. From evidence of insect remains in the roost, the diet of the bats in Kaptum Cave consisted of eight insect orders (Lepidoptera, Coleoptera, Orthoptera, Dictyoptera, Hymenoptera, Isopteran, Hemiptera, and Odonata) with the order Lepidoptera constituting the bulk of insects preyed upon. The specific prey items of each bat species could not be established since most insect remains were recovered from a section the cave we referred to as the “Nycteris corner” dominated Nycteris thebaica, where most of the bats fed from. The Nycteris corner was however, frequented by the other species (Rhinolophids and Hipposiderids) at any time in a 24-hr period. Temperature and relative humidity varied significantly in all the cave chambers, and these had a big influence on segregation of bats in the cave. There were several encounters and signs of human disturbance and use of the cave such as: cave mass tourism, extraction of volcanic crystal stones, making fireplaces, and collection bat guano. Continued human presence in the cave could have serious implications for roost stability; but also, it could predispose humans to potentially harmful aerosols and pathogens associated with bats and bat guano
Assemblages of Hawkmoths (Sphingidae: Lepidoptera) in Kalinzu Forest South-western Uganda

(Makerere University., 2023-02-01) Kasangaki, Dismas

Intensification in anthropogenic activities has significantly lowered habitat quality, accelerated habitat loss and habitat isolation within forest ecosystems. However, there is less documentation on recovery of communities against reference landscapes in afrotropical forests. This study investigated the diversity and faunal composition of sphingid moths (Lepidoptera: Sphingidae) in old-growth and regenerating forest in Kalinzu Central Forest Reserve, South-western Uganda. The goal of the study was to generate information on how habitat recovery impacts insect fauna. The study was guided by the following questions: 1) how do sphingidae moth diversity patterns in regenerating forest compare with those in old-growth forest? 2) How does hawkmoth species diversity correlate with habitat vegetation variables? and 3) what phylogenetic relationships exist among hawkmoths in Kalinzu Forest? Sphingid moths were sampled between July 2019 and February 2020, using a combination of two 20W Ultraviolet light traps (FL20S-BLB and FL20-BL), operated on a battery. Habitat vegetation variables measured or estimated included: basal area of trees, understorey cover, tree density, understorey density and understorey complexity. Phylogenetic relationships among hawkmoths based on mt(CO1) gene were inferred using Maximum Likelihood method and General Time Reversible model in MEGA X. A total of 378 hawkmoth individuals belonging to 22 species were collected. Individual-based extrapolation curves did not reach asymptote implying that more species would be recorded with additional sampling effort. The overlap of curves at 95% CI indicated that richness between forest types did not differ significantly. Fisher’s α diversity was not significantly different between forest types (t(2.2) = 1.786, p > 0.05), also agreeing with very low beta diversity (βChao-Jaccard = 0.0499). Species dominance did not differ significantly between old-growth and regenerating forest (t(3.438) = -0.96). The highest proportion of species in both forest types were closed forest species. Analysis of similarity revealed no significant difference in hawkmoth community composition between forest types (R = -0.241, p > 0.05). Negative value of ANOSIM statistic signifies greater variability within a particular forest portion, than between forests. The genetic distances based on mt(CO1) between any two species of Hawkmoths ranged between 10% and 22%. The findings of this study add to the evidence that species richness does not always decrease with habitat modification. As a way of conserving biodiversity, natural regeneration of formerly degraded habitats should be considered.
Assessing the impact of biological control in the Rivers Kagera, Mara, Pangani and Sigi.

(Makerere University, 2007-07) Katagira, F.

Water hyacinth Eichhornia crassipes (Martius) Solms-Laubach, is considered to be the worst aquatic weed worldwide. Biological control using Neochetina weevils was initiated in Tanzania in1995; however this control mechanism was successful in Lake Victoria where water hyacinth was reduced by 80%, but failed in some water bodies (Rajabu et al., 2000; LVEMP, 2001). Studies were conducted during 2003 to 2005 to assess the impact of biological control in the rivers Kagera, Mara, Pangani and Sigi and to evaluate potential effect of nitrogen, phosphorus and silt on weevil establishment in greenhouse conditions at Kyaka near Kagera river in Kagera region, Tanzania. The objectives were achieved by studying the following aspects: (i) The status of water hyacinth and Neochetina weevils in fresh water rivers (Kagera, Mara, Pangani and Sigi rivers) of Tanzania; (ii) Effect of nitrogen and phosphorus on weevil establishment and damage on water hyacinth; (iii) Effect of silt on establishment and performance of Neochetina weevils on water hyacinth; and (iv) Effect of water quality on weevil establishment and damage on water hyacinth. The study on rivers involved sampling of water hyacinth and determination of population densities and growth. Weevil densities, stages of development and feeding scars were also determined. Results showed that Kagera river had the highest water hyacinth densities (44.2 plants m-2) followed by Mara (32.8 plants m-2), Pangani (27.2 plants m-2 ) and least in Sigi (22.8 plants m-2). Plant height was highest in Mara (51.04 cm), followed by Kagera, Pangani and the least height in Sigi (28.6 cm). Other plant parameters namely, leaf length and width and root length followed the same trend. Kagera river had the highest number of ramets per plant (1.13) followed by Mara, Sigi and Pangani (0.69). The number of adult weevils per plant was highest in Sigi (1.93) and Pangani (1.78) and lowest in Kagera and Mara (0). The number of weevil feeding scars per plant was highest in Pangani (22.1) and Sigi (17.81). Overall Mara (1.4) and Kagera (0.21) had the lowest. Water hyacinth growth suppression and population reduction was only evident in Pangani and Sigi rivers and was associated with abundance and feeding by weevils. On the other hand, the weed continued to proliferate in Kagera and Mara rivers in Lake Victoria basin despite monthly weevil releases. The effect of N and P on water hyacinth growth and Neochetina weevil establishment and efficacy was conducted under greenhouse conditions. It consisted of 0 (control), 50 mg N L-l , 100 mg N L-l, 42.5 mg P L-l and 85 mg P L-l and their combinations. Weevil damage was suppressed at higher N (100 mg N L-l) but not high P application rates. Plants fed with high levels of N had low counts of adult weevils and pupae. On the other hand, growth of water hyacinth was limited in low N by 40%, yet weevil numbers, eggs, larvae and pupae per plant as well as weevil damage, increased at low N. Similarly, the number of adult weevil feeding scars per plant was higher (P<0.01) in lower N than at high levels. However, P concentration had no significant effect on number of weevil feeding scars per plant. Total plant N and P varied among nutrient application rates and N uptake depended on P availability; and plant N content increased with increase in P application rate. There was a significant positive correlation (P<0.001) between number of eggs and larvae per plant. The number and length of roots per plant increased with P application while application of N resulted to significant reduction in number and length of roots per plant. Water quality in the Kagera, Mara, Pangani and Sigi rivers differed. The highest total nitrogen was found in Kagera (0.99 mg N L-l) while the lowest was in Sigi (0.12 mg N L-l). Similary, total phosphorus was highest in Kagera (0.36 mg P L-l), while the lowest was in Sigi (0.12 mg P L-l). Likewise, electrical conductivity was highest in Kagera (969.8 µscm-l) follwed by Mara while Pangani and Sigi had the lowest indicating pollution loads in Kagera and Mara rivers. Water temperature was high in Pangani (30.39 oC) and Sigi (29.4 oC), while Mara had (26.2 oC) and Kagera had the lowest (26.23 oC). Air temperatures and relative humidity followed the same trend. It was concluded that high N and P levels might be partly responsible for poor weevil damage on water hyacinth in eutrophic habitat such as in Kagera and Mara rivers. On addition, the low temperatures and relative humidity may have contributed by limiting the weevil activities. The study on the effect of silt on the water hyacinth and weevil development and efficacy consisted of control (no silt applied) and silt at levels 17 kg, 51 kg and 85 kg in 50 litres of water applied in order to achieve distances of 20, 10 and 0 cm between the root tips and silt topmost level/surface. Silt and water for the experiments was from Lake Victoria and Kagera river. There was reduced weevil reproduction in treatments with silt as evidenced by low numbers of eggs, larvae, pupae and adult weevils. In contrast water hyacinth plant populations, leaf length and number of ramets were significantly enhanced (P<0.05) by silt application. Water hyacinth grown in water and silt from Kagera river, had higher weevil feeding scars per plant were 32.2 at low silt (17 kg) and 8.7 at 85 kg silt level indicating reduced weevil damage at high silt. A significant and positive relationship (r = 0.881, P < 0.01) was obtained between water hyacinth leaf length and lamina width. Silt levels appear to directly affect different life stages of the weevil, thereby reducing the ability of the insects to multiply to populations that can suppress growth of water hyacinth. Sediment loads in Kagera and Mara rivers may have contributed to poor establishment of Neochetina weevils and favoured water hyacinth proliferation.
Assessment of the distribution and conservation status of non-haplochromis fish species in Uganda.

(Makerere University., 2023-09-12) Akoth, Dorothy

Assessing the conservation status of species according to their extinction risk is a major step in conservation planning and resource prioritization. To date, most of the assessments have been conducted on global level; yet, most conservation efforts are best conducted at national or local levels. Also, the global assessments often do not fit individual countries whose biodiversity may have unique threats that are specific and distinct from those at the global level. Elsewhere, this problem has been addressed by developing national red lists; in Uganda, this approach is also taking root, except that it has only focused on terrestrial biota, with limited attention to freshwater biodiversity. To address this gap, this study used species occurrence data, integrated with expert knowledge and literature, to revise the list and distribution of non-Haplochromis fish species native to Uganda, determine the national range of these fishes relative to the global range, and assess the extinction risk of these fishes at national level. The study showed that at least 110 non-Haplochromis fish species, belonging to 11 orders, 21 families, and 48 genera exist in Uganda. Species diversity was higher in large water bodies than small waterbodies, which is expected due to diverse habitats in the former, although it could also be attributed to sampling bias with more studies focusing on large waterbodies than small water bodies. The geographical range for 51 fish species (46% of the total number of species) was greater than 50% of the global range, suggesting that they are geographically restricted and are at a higher extinction risk. In terms of conservation status, 31 species (28%) were threatened with extinction at the national level compared to 10 species (9%) which were listed as threatened at global level as of 2021. This variation between national and global assessments suggests that the conservation status for fish species can differ, irrespective of geographical restrictedness of the species. Elsewhere, studies have shown that this scenario is likely to be abated by ensuring flow of information and data between the two assessment frameworks. The data and information from this study can, therefore, be used to improve the global assessments. In addition, this study highlights areas where surveys should be enhanced to improve our knowledge of the distribution and occurrence of the non-Haplochromis fish species. For the ease of future assessments, it is important that baseline data collection is prioritized in areas or basins where there are no occurrence records or where species are listed as Data Deficient.
Barriers to implementation of Uganda’s national industrial policy: A case study of the Steel Mills Industry

(Makerere University, 2014-12) Lubwama, Festo

This research is a descriptive study of Uganda’s national industrial policy implementation state. The study was conducted in the Steel mills industry with a major aim of assessing the process of implementing Uganda’s industrial policy and the challenges this process is facing to devise means for better policy implementation. Other objectives included; analysis of the policy making and implementation strategies for industrialisation, determining progress of implementation and impact of the policy in the Steel mills industry, determining challenges to implementing the National Industrial Policy (NIP), and then making recommendations for better industrial policy implementation in Uganda. The research involved interpretation of both qualitative and quantitative data collected from the policy makers, implementers, the Steel mills industry, and independent industrial experts/associations. The research found that the National Industrial Policy and its implementation strategy are competent and able to guide Uganda’s industrialisation if well followed. However, the key stakeholders expressed scepticism about whether they are being and/or shall be implemented. Some of the challenges facing Uganda’s industry should not be existing if the policy was being implemented. This indicates the little impact the policy is having in the manufacturing sector. The major challenges to policy implementation included the lack of a clear industrialisation strategy and lack of commitment to implement the NIP characterised by a poor implementation infrastructure with no National Council/Body to oversee implementation of the policy. The research concluded that though the policy might be good and a timely intervention, it still requires the government to fully commit to creating an environment that supports/facilitates implementation of the policy especially providing resources, legal support and the other supportive policies. The industry should also come together and form associations and other innovative solutions to solve their problems given government’s inefficiency. The government should, however, put in place a Statutory Council to oversee the entire implementation of the NIP.
Bio-efficacy of insecticides used for vector control in Busia and Tororo districts, Eastern Uganda

(Makerere University, 2022-02-14) Chemutai, Faith

High malaria transmission rates in Sub-Saharan Africa (SSA) are attributed to the continuous presence of effective and competent Plasmodium vectors, Anopheles gambiae complex and the Anopheles funestus group. These vectors play a key role in transmitting the most dangerous malaria parasite species Plasmodium falciparum. The key vector control interventions in Africa including Uganda, rely heavily on utilisation of long-lasting insecticidal nets (LLINs) and insecticide-based indoor residual spraying (IRS). This study assessed the bio-efficacy of LLINs and IRS as vector control interventions used in Busia and Tororo districts. Samples of Anopheles mosquito larvae were collected from various breeding grounds before rearing them in an insectary. Additionally, a nine-year period malaria cases data set (2012-2020) was obtained from the health management database to depict malaria trends. Emerged adults were dissected under a stereoscopic microscope and identified using standardized morphological keys after respective bioassays on used nets and sprayed walls. A One-Way Analysis of Variance (ANOVA) was used to test for significant differences in the mean composition of mosquito species, mean variations, mortalities by net and wall type. Results showed that mean mortalities of Anopheles mosquitoes to used brands of nets from Busia (9.86±11.35) and Tororo (9.64 ±11.12) varied insignificantly (t=0.119, p=0.906). A disaggregated analysis for each net used revealed that, the DAWA plus 2.0 registered a highest mean mortality of mosquitos, followed by PermaNet 3.0, Olyset and PermaNet 2.0 respectively. The average number of Anopheles mosquitoes that died after 2 weeks (6.86±4.07) and 1 month of chemical spray (6.69±4.01) varied insignificantly (t=0.175, P=0.861). Meanwhile, disaggregated results by time of spray (T0 and T1) and exposure periods, showed that plastered painted wall, had higher mortalities followed by Brick Plain and Mud/Wattle wall substrates, respectively. Considering the malaria cases registered on annual basis, Mann- Kendal test revealed a drastic decline of malaria cases over the nine year period (2012 -2020) in Tororo district with Sen’s slope of -22, while for Busia district it revealed an increase of malaria cases (Sen’s slope +28). Equally, these trends reveal varied spatial patterns over the two districts. Both the districts revealed a high prevalence of malaria between May-November in respect to bimodal rainfall pattern, matching to the perennial transmission setting of Uganda. This study has shown that vector behaviour, biology and physiology need consistent monitoring and surveillance for further entomological characterisation.
Bio-physical suitability of saline crater lakes of Western Uganda for artemia production

( 2019) Nkambo, Mujibu

Despite Artemia being the most widely used live feed for successful larval weaning and rearing, unreliable quality, not being readily available and expensive especially to small farmers hinders its use. This study explored production of Artemia using waters from saline crater lakes of western Uganda. Local Artemia production will contribute towards reducing the low survivals in hatcheries more especially for African catfish (Clarias gariepinus) by providing affordable Artemia of reliable quality and quantity to the hatchery operators. Limno- biophysical chemistry was used to identify the best suited lakes for bio-manipulation to produce Artemia. Limno-biophysical chemical findings indicated the studied lakes ranged between 0.2±0.0m and 2.3±0.3m (depth), 0.0±0.0mgl-1 and 205.0±15.3mgl-1 for salinity, 27.9±0.3°C and 34.4±2.4°C for temperature, 18.6±0.1 µscm-1 and 106.3±3.5 µscm-1 for conductivity, 1.7±0.4 mgl-1 and 6.0±1.0mgl-1 for Dissolved Oxygen and 9.6±0.1 and 11.5±1.0 for pH, with no occurrence of local Artemia resources. Other than Lake Bagusa where Anabaena circinalis was found to dominate the algal biomass, Spirurina platensis was the dominant algal biomass in majority of these lakes. Lake Kikorongo had the highest zooplankton diversity, having Brachionus calyciflorus as the most abundant (50 to 100 individuals /litre). Hatchability experiments using five commercial Artemia strains: Artemia franciscana from Great Salt Lake (GSL), A. franciscana from salt ponds in Vinh Chau (VC), Chinese strain (Chinese), TUZ Parthenogetic Artemia from Kazakhstan (TUZ) and Parthenogenetic Artemia strain from Siberia (PAS) using waters from crater lakes indicated Artemia franciscana (VC) to have significantly higher hatching percentage of 93.9±29.4 and 79.8±15.2 for lake Bagusa, 89.3±6.9 and 92.7±3.6 for lake Katwe, 83.4±10.7 and 83.5±9.1 for lake Maseche in the dry and wet season respectively (P <0.05). Bucket culture experiments of the best performing commercial Artemia strains from the hatchability experiments indicated survival and mean total length of GSL Artemia nauplii cultured at salinities of 40 and 60g/l to be significantly higher than that cultured at salinities of 80 and 100g/l. VC Artemia nauplii cultured at salinity of 40g/l was found to have significantly higher growth rate (length increment) than those at salinities of 60g/l, 80g/l and 100g/l (Asymptotic sig (2-sided tested) ≤ 0.000). Pond culture experiments showed that the highest mean total lengths of GSL and VC Artemia nauplii were 921.0µm and 923.0±91.9µm respectively when cultured at salinities of 60g/l for 10days. No Artemia nauplii were found to survival beyond day 10 of culture at both salinities of 60 and 80g/l. The mean survival of both GSL and VC Artemia nauplii did not significantly vary across the treatments (60 and 80g/l) after the same culture period in the salt ponds (Asymptotic sig (2-sided tested) ≥ 0.200). Although more research is needed to establish the cause for the high mortalities after 8 and 10days of culturing in buckets and ponds respectively, this results show that it possible to culture Artemia using waters from alkaline saline crater once these waters are bio-manipulated to favorable conditions for Artemia production.
Biochemical characterisation of water from River Ntabago and implication on water quality and safety

( 2011-05) Muhwezi, James

This study analysed the biological and chemical components characterising water from River Ntabago. The composition of the water determines its quality and affects the lives of aquatic organisms and other water consumers. The study was carried out from January to July 2009. Human activities such as brewing, farming, and washing fabrics; and natural processes like erosion, affect the biochemical composition of the water, its quality and people’s health. The water has a bad smell, bitter taste and contains black particles that make it look unpleasant. There are complaints of skin irritations and/or ulcerations when the water is used for bathing and abdominal pains when drunk. The objectives of this study were: to determine the phytoplankton composition of the water; to quantify heavy metals in water: cadmium, chromium, cobalt, copper, iron, lead, manganese, nickel, zinc; and anions: chloride, fluoride, sulphate and sulphide. The study also determined water quality parameters: pH, temperature, specific conductance, dissolved oxygen, coliforms, alkalinity, turbidity, hardness, and nutrients; so as to assess the quality of the water and its safety as a source for domestic use. The biological parameters were determined using microcopy, heavy metals using atomic absorption spectrophotometry, while anions and water quality parameters were analysed using various titration and colorimetric methods. The study revealed the concentration of phytoplaktons to be very high and increasing downstream. These reduced water quality through production of: toxins, irritant lipo-polysaccharides, taste and odour substances; reduction of dissolved oxygen and increasing turbidity and colour of the water. Concentrations of coliform bacteria were fairly high. The study also revealed the existence of cadmium, iron, lead, manganese and nickel above their NEMA and WHO recommended values for potable and drinking water, raising concerns for the health of water consumers. The health effects of metals include: lowering energy levels; damaging the central nervous system and mental function; damaging blood composition, lungs, kidneys, liver and other vital organs. Turbidity and alkalinity were shown to be high, hardness fairly moderate, while dissolved oxygen and nutrients were very low in comparison to their recommended values. The biochemical composition and water quality of the river were functions of growing population pressure, changing settlement and development patterns, siltation due to loss of vegetation cover and land degradation, seasonal variations in rainfall, agricultural runoffs, water-flow fluctuation rates, heavy metal discharges from weathered rocks and wetland drainage. The water of River Ntabago is therefore considered unsuitable for human and livestock consumption because of high phytoplankton and heavy metal concentration, high turbidity, low dissolved oxygen and the presence of coliforms. Hence, there is need for: a watershed management program in the area, more research to be done, treatment of water in order to meet standards for potability, planning some alternative water, promotion public awareness and sustainability issues, proper land utilization and improved agricultural practices; frequent surveys and monitoring programs to ensure the quality of water.
Biochemical responses of Robusta coffee to water deficit conditions

(Makerere University, 2025) Nabwire, Joan

Drought stress poses a major challenge to crop productivity by altering the biochemical composition, antioxidant defenses, and morphological development. This study investigated how the biochemical responses and the antioxidant system in Robusta coffee relate to the morphological traits under water deficit conditions. Rooted cuttings of seven CWD-r Robusta coffee varieties (KR1-KR7) were evaluated under four watering regimes (100%, 75%, 50%, and 25% field capacity, FC) in a split-split plot randomized complete block design over two temperature environments, low (LTS) and high (HTS). Morphological parameters (plant height, leaf blade length, leaf blade width, stem internode length, number of stem nodes, and number of stem internodes) were measured fortnightly for four months. Fourier transform infrared spectroscopy was used to quantify key biomolecules (carbohydrates, lipids, and proteins) from the leaves, while the 2,2-diphenyl-1-picrylhydrazyl assay was used to assess total antioxidant activity and capacity monthly for four months. Findings demonstrated significant interactive effects of temperature, moisture regime, and variety on the biochemical profile, of CWD-r Robusta coffee varieties. However, under severe drought stress, this coordination weakens as antioxidants become decoupled from biochemical maintenance, particularly under high temperatures. KR5 consistently maintained the highest biomolecule accumulation across most regimes, while KR2 and KR4 had lower biomolecule reserves under stress. TAA and TAC were stable across varieties but peaked for variety KR6 under mild (75% FC) and moderate (50% FC) moisture stress, highlighting the role of antioxidants in stress mitigation. Correlation analyses revealed robust positive associations among the spectral regions corresponding to the biomolecules, confirming a coordinated response to drought stress. Correlations between the biochemical markers and morphological parameters (such as plant height, stem node/internode counts, and leaf dimensions) were weak to moderate, suggesting that while metabolic adjustments are integral for stress mitigation, the morphological outcomes are governed by a more complex interplay of factors. A multi-trait genotype-ideotype distance index was applied to rank variety performance under distinct stress scenarios. Under well-watered (100% FC) and mild water stress (75% FC), KR1 emerged as the most promising genotype. Under moderate water deficit (50% FC), KR7 and KR3 were identified as the most suitable varieties in the HTS and LTS, respectively. Under 25% FC, KR1 and KR5 showed the strongest combination of biochemical accumulation and antioxidant defenses in the HTS and LTS respectively securing their ranking as the most water stress resilient varieties.
Biogeographical examination of the butterfly fauna in selected West Albertine Rift Forests

(Makerere University, 2009-09) Kasangaki, P.

This study was conducted with the main aim of evaluating the factors that may influence butterfly species richness and distribution (i.e. forest size, altitude, forest disturbance, average rainfall, temperature and distance from the supposed Pleistocene refugium) in 17 West Albertine rift forests (WARF’s), Uganda. Butterflies which were collected by the Forest Department (now National Forests Authority, NFA) and preserved at the Makerere University Museum were used for this study. Relevant biodiversity reports prepared by the Forest Department were reviewed to gather information on the physical and environmental attributes of the forests. A list of 630 butterfly species was complied for the 17 forests. Cluster, Principal component, Canonical correspondence and regression analyses were done to relate and compare butterfly species richness among the forests in order to establish the relationship between the butterfly species richness and the environmental and physical variables. These analyses helped in identifying the most important variables influencing the butterfly species richness and distribution in the forests. Forests with similar physical and environmental conditions had similar species richness. The environmental and physical variables which seemed to influence the species richness and distribution in the forests were forest size, number of plant species, altitude and levels of forests disturbance. Plant species richness appeared to be the most significant factor accounting for 49% of the butterfly species richness in the forests. Distance from the supposed Pleistocene refugium also seemed to have been responsible for species richness in the forests although not significantly. However, more research/survey should be done to involve population analysis (number of individuals other than only the number of species should be recorded) to ascertain the effect of forest disturbance on the butterfly species richness and population densities.
Characterisation of resistant starch in selected banana cultivars

(Makerere University, 2022-12-14) Kajubi, Ali

In the green stage of the banana, starch accumulates most of which is resistant to degradation by amylases. The resistant starch is utilised in industry for development of nutraceuticals and as a functional food for countering the advance of corresponding NCDs and promoting a healthy gut. Depending on the genotype and cultivar, the starch composition and corresponding properties vary significantly across the banana family. The aim of this study was to determine the molecular, physicochemical and functional properties of the resistant starch (RSII) in economically important banana cultivars of Uganda. Banana RS was isolated from 15 different cultivars using a combination of alkaline and enzyme treatment. The resultant powder (RSII) was then assayed for RS content and polymer composition. Granule structure, size and distribution was determined using light and scanning electron microscopy (SEM). The molecular organisation and crystallinity of the RS was deduced using Fourier Transform Infrared Spectroscopy (FTIR) and the corresponding functional properties of the starches were also explored. The RS content significantly varied across the banana cultivars from 29.3±7.9 to 79.27±3.2 g/100g dry weight basis in Ndiizi and Kayinja respectively. The granules isolated RS were smooth, elliptical with ridges and a few irregulars having a diameter of approximately 17 to 36µm in Kibuzi and Kivuvu respectively as analysed by SEM. The results revealed significant differences in microstructure and physicochemical characteristics among the resistant starch samples with amylose contents of the RS ranging from 9.2% in Fhia17 to 22.4% in Kayinja. Generally, all the banana RS exhibited similar chemical structure of the A and B Types but had varying intensity ratios of the crystalline peaks 0.95 to 1.09 in Narita23 and Kayinja respectively. Multivariate analysis revealed strong correlations among the RS properties, prompting a conclusion that the effects of most RS characteristics are strongly interrelated. The RS content, amylose, and granule size contributed more to the grouping of the cultivars than other properties. Group 1 consisted of Kivuvu, Kayinja and Nfuuka that would be suitable for development of high RS, low moisture content products such as nutritional supplements and extruded foods. In addition to FTIR combined with chemometric data analysis presenting a powerful tool for reliable discrimination between banana RS, results of this study are useful in determining the prospective applications of banana resistant starch as a functional ingredient in the food and non-food industries.
Characterizing phenotypic and genotypic resistance to insecticide in Anopheles gambiae s.l. from the West Nile region in Uganda

(Makerere University, 2024) Kiwanuka, Marvious

In sub-Saharan Africa, Anopheles gambiae sensu lato and Anopheles funestus mosquitoes are the major vectors of Plasmodium that cause malaria; a leading cause of mortalities and morbidity in humans. In order to control Anopheles mosquitoes, Uganda over the past decades has been using indoor residual spraying (IRS) and long-lasting insecticidal nets (LLINs) as recommended by the World Health Organization. However, the effectiveness of these methods has been reduced due to several factors, including reported resistance to insecticides and behavioural change among Anopheles mosquitoes. The management of resistance to insecticides is challenged by limited information on the resistance profile of primary malaria vectors in high transmission regions like the West Nile. This study determined the species diversity, susceptibility and genotypic resistance to insecticides among Anopheles gambiae s.l. collected from October 2023 to January 2024 in Maracha and Terego districts. Larval samples from selected aquatic sites were reared to adults in the insectary at the IDRC laboratory in Butabika, Kampala. Morphologically identified adult females of Anopheles gambiae s.l. were exposed to different diagnostic dosages of deltamethrin, alpha-cypermethrin, pirimiphos-methyl, bendiocarb and pyrethroid (deltamethrin and alphacypermethrin) with piperonyl butoxide (PBO) in the WHO-tube at 25°C and 80% humidity. Percentage mortalities after insecticide exposure were determined after 24 hours. Morphologically identified adults were further differentiated into subspecies using multiplex polymerase chain reaction. Resistance markers Vgsc-L995F/L1014F (kdr-West), Vgsc-L995S/L1014S (kdr-East) and Cyp4j5-L43L were characterised using PCR methods in insecticide exposed mosquitoes. Results showed that the districts of Maracha and Terego consisted of two subspecies: Anopheles gambiae s.s and An. arabiensis. Anopheles gambiae s.l showed high levels of resistance to pyrethroids (3134% mortality) which was significantly associated to Vgsc-L995F/L1014F and VgscL995S/L1014S in Terego (p= 0.03) and Maracha (p= 0.04). The species however had moderate resistance to bendiocarb and full susceptibility to pirimiphos-methyl (100% mortality). Resistance to insecticide was not significantly attributed to Cyp4j5-L43L in samples from both districts. PBO partially improved the susceptibility to pyrethroids but was still below the recommended level by WHO. This indicates presence of metabolic resistance in Anopheles gambiae s.l in West Nile region. Understanding mutations that cause metabolic resistance is therefore important in order to develop more effective vector control method.
Coffee phytochemical compounds with potential antioxidant and antifungal properties for cosmeceutical application

(Makerere University, 2024-04) Nankya, Margret

Factory Robusta, Factory Arabica undergrades and NFCT3 abandoned Robusta coffee genotype were examined for biochemical components including phytochemicals, antioxidant activities and anti-fungal properties. The results were used to formulate a coffee cream which was tested for anti-fungal activity. Bioactive compounds were extracted by cold maceration method and the quantitative analysis of phytochemicals was done using UV Vis spectrophotometry and High performance Liquid Chromatography. The antioxidant activity was determined using the 2, 2- diphenyl-1-picrylhydrazyl (DPPH) and Ferric Reducing Antioxidant Power (FRAP) methods whereas the antifungal activity of coffee green bean extracts and the cream were determined using Agar well diffusion method. Quantitative analysis revealed presence of different phytochemicals in varying quantities and these included; Polyphenols, flavonoids, caffeine and chlorogenic acids. Factory Robusta coffee undergrades had a significantly (P≤0.05) higher concentration of all phytochemicals except for flavonoids where NFCT3 had the highest flavonoid content (1.073 %w/w) followed by factory Arabica coffee (0.75 %w/w) and factory Robusta had the lowest content (0.57%w/w). Genotype NFCT3 had the highest antioxidant activity (63.33%) followed by Factory Robusta undergrades (62.44%) while Factory Arabica undergrades had the lowest antioxidant activity (61.67%) when DPPH assay was used. When FRAP assay was used, Factory Robusta undergrades had the highest Ferric Reducing Antioxidant power (0.55nm) followed by NFCT3 genotype (0.53nm) with factory Arabica having the lowest (0.51nm). Amongst the sampled coffee extracts, only Genotype NFCT3 exhibited antifungal activity. Non targeted analysis of coffee extracts using GC MS tandem showed presence of different compounds and eight of these were only present in NFCT 3 which are responsible for its antifungal activities. A cream with NFCT3 extracts was formulated and the findings showed that it inhibited growth of Aspergillus niger strain. In conclusion, results showed presence of various phytochemicals and great antioxidant and antifungal properties exhibited by NFCT3 and gives a scientific basis for their incorporation in cosmeceutical industry.

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