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Abstract The high global bacterial infection burden has created need to investigate the neglected potential drivers of pathogenic bacteria, to inform disease prevention. Kampala is facing a proliferation of herbalists, selling herbal medicine (HM), of largely unregulated microbiological quality. We evaluated the bacterial contamination burden in HM sold in Kampala, to support evidence-based redress. The total viable loads (TVL), total coliform counts (TCC), E. coli counts, and prevalence of selected bacterial strains in 140 HM were examined using conventional culture, following the guidelines of World Health Organization (WHO), and Uganda National Drug Authority (NDA). Data were analyzed using D'Agostino-Pearson test, frequencies, proportions, Chi-square, and Mann–Whitney U test with STATA version-15.0. Fifty (35.7%), fifty-nine (42.1%), and twelve (8.6%) HM were unsafe for human use because they exceeded WHO’s permissible limits for TVL, TCC, and E. coli counts respectively. Solids had significantly higher mean TVL than liquids. Violation of NDA’s guidelines was significantly associated with high TVL. Fifty-nine bacteria, viz., Klebsiella pneumoniae (n = 34; 57.6%), Escherichia. coli (12; 20.3%), Staphylococcus aureus (7; 11.9%), Klebsiella oxytoca (3; 5.1%), Bacillus cereus, Pseudomonas aeruginosa, and Enterobacter spp. (1; 1.7% each), were isolated from 45 (32.1%) samples. These bacteria can cause severe clinical diseases, and promote deterioration of HM potency.

Abstract Background Antimicrobial resistance is swiftly increasing all over the world. In Africa, it manifests more in pathogenic bacteria in form of antibiotic resistance (ABR). On this continent, bacterial contamination of commonly used herbal medicine (HM) is on the increase, but information about antimicrobial resistance in these contaminants is limited due to fragmented studies. Here, we analyzed research that characterized ABR in pathogenic bacteria isolated from HM in Africa since 2000; to generate a comprehensive understanding of the drug-resistant bacterial contamination burden in this region. Methods The study was conducted according to standards of the Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA). We searched for articles from 12 databases. These were: PubMed, Science Direct, Scifinder scholar, Google scholar, HerbMed, Medline, EMBASE, Cochrane Library, International Pharmaceutical Abstracts, Commonwealth Agricultural Bureau Abstracts, African Journal Online, and Biological Abstracts. Prevalence and ABR traits of bacterial isolates, Cochran’s Q test, and the I2 statistic for heterogeneity were evaluated using MedCalcs software. A random-effects model was used to determine the pooled prevalence of ABR traits. The potential sources of heterogeneity were examined through sensitivity analysis, subgroup analysis, and meta-regression at a 95% level of significance. Findings Eighteen studies met our inclusion criteria. The pooled prevalence of bacterial resistance to at least one conventional drug was 86.51% (95% CI = 61.247–99.357%). The studies were highly heterogeneous (I 2 = 99.17%; p

Abstract Background Globally, diarrheal and respiratory diseases are among the main causes of mortality and morbidity. In Uganda, cities are facing proliferation of trade in herbal medicines (HM), including those for diarrhea and/or cough. Information on the economic, and the ethnopharmacological aspects of these HM is scarce, deterring the sector from achieving optimal capacity to support national development. We profiled the anti-diarrhea and/or anti-cough HM, and the basic economic aspects of HM trade in Kampala city, to support ethnopharmacological knowledge conservation and strategic planning. Methods A cross-sectional survey was conducted on 65 herbalists using semi-structured questionnaires. This was supplemented by an observational survey using a high-resolution digital camera. Data were collected following the guidelines for research on HM, established by Uganda National Drug Authority, and World Health organization. Results Eighty-four plant species from 41 families were documented. Fabaceae and Myricaceae had the highest number of species (9, 10.7% each). Citrus limon (L.) Osbeck was the most commonly cited for cough, with a relative frequency of citation (RFC) of 1.00, and its relative medical importance was not significantly different from the other top 5 species except for Azadirachta indica A.Juss (RFC = 0.87). Entada abyssinica A. Rich (RFC = 0.97) was the most cited for diarrhea. Trees (34, 40.5%) were mostly used, and mainly harvested from wild habitats (55.2%) in 20 districts across Uganda. These HM were mainly sold as powders and concoctions, in markets, shops, pharmacies, and roadside or mobile stalls. The highest prices were Uganda Shillings (UGX) 48,000 ($13.15)/kg for Allium sativum L, and UGX 16,000 ($4.38)/kg for C. limon. All participants used HM trade as a sole source of basic needs; majority (60.0%) earned net monthly profit of UGX. 730,000 ($200) ≤ 1,460,000 ($400). The main hindrances to HM trade were the; disruptions caused by the COVID-19 pandemic (n = 65, 100%), and the scarcity of medicinal plants (58, 89.2%). Conclusion There is a rich diversity of medicinal plant species traded in Kampala to treat diarrhea and cough. The HM trade significantly contributes to the livelihoods of the traders in Kampala, as well as the different actors along the HM value chain throughout the country.

Abstract Background Infectious diseases remain a leading cause of mortality and morbidity around the world, and those caused by bacteria are common in the East African region. In this region, trade and consumption of herbal medicine has been expanding in the recent decades. Herbal medicines may be contaminated with pathogenic bacteria; however, there is limited information due to fragmented studies in East Africa. In this meta-analysis, we critically analyzed original research related to the incidence of pathogenic bacterial contaminants of HM in the East African region since 2000. The aim was to create a comprehensive understanding of the extent and dynamics of bacterial contamination in HM, to guide future research and concerted public health protection in the region. Methodology The study was conducted according to the standards of the Preferred Reporting Items for Systematic Reviews and Meta-analyses. We searched and evaluated published articles from eleven electronic databases (Google Scholar, PubMed, HerbMed, MEDLINE, Science Direct, Scifinder Scholar, Cochrane Library, International Pharmaceutical Abstracts, EMBASE, Biological Abstracts and Commonwealth Agricultural Bureau Abstracts). Prevalences of different bacterial species, Cochran’s Q test, and the I 2 statistic for heterogeneity were evaluated using a software called MedCalcs. Random and fixed effects models were used to determine the pooled prevalence of clinically significant bacteria from studies which were included in this meta-analysis. The potential sources of heterogeneity were examined through sensitivity analysis, sub-group analysis, and meta-regression at 95% level of significance. Results Fourteen studies met our inclusion criteria. Overall, the studies were highly heterogeneous (I 2 = 98.48%) and there was no evidence of publication bias. Escherichia coli was the most prevalent contaminant. Salmonella spp. and Shigella spp. were the most frequently reported primary pathogens with pooled prevalence of 10.4% and 6.3%, respectively. Our findings are in tandem with recent systematic reviews conducted in Europe and Asia, but are in discrepancy with the reviews recently conducted in southern Africa. Conclusion and recommendations The East African herbal medicine industry poses considerable health risks to communities through dissemination of clinically significant bacteria. Presence of enteric bacterial contaminants indicates possible fecal pollution of herbal medicine region-wide. Adequate research pertaining to microbial safety of herbal medicine in the East African countries remains highly desired. The latter will enable establishment of strong, region-wide herbal safety mechanisms in order to support comprehensive public health protection in East Africa.

Abstract Background In Uganda, medicinal plants have been utilized to treat a variety of ailments, including cancer. However, there is little information available about the medicinal plants used to treat cancer in the Elgon subregion. As a result, the current study documented the plant species used in the management of cancer in the Elgon sub-region. Methods Data were gathered by observation, self-administered questionnaires, interview guides, and guided field trips. Analyzing descriptive statistics and creating graphs were done using SPSS (version 21.0) and GraphPad Prism® version 9.0.0, respectively. Well-established formulae were used to calculate quantitative indices. The narratives were interpreted using major theories and hypotheses in ethnobotany. Results A total of 50 plant species from 36 families were documented, and herbal knowledge was mainly acquired through inheritance. Fabaceae and Asteraceae comprised more plant species used in herbal preparation. Most plants were collected from forest reserves (63%); herbal therapies were made from herbs (45%); and leaves were primarily decocted (43%). The most frequently used plants were Tylosema fassoglensis, Hydnora abyssinica, Azidarachata indica, Prunus Africana, Kigelia africana, Syzygium cumini, Hydnora africana, Rhoicissus tridentata, Albizia coriaria, and Plectranthus cuanneus. All the most commonly used plants exhibited a high preference ranking (60–86%) and reliability level (74.1–93.9%). Generally, the ICF for all the cancers treated by medicinal plants was close to 1 (0.84–0.95). Conclusions The ten most commonly utilized plants were favored, dependable, and most important for treating all known cancers. As a result, more investigation is required to determine their phytochemistry, toxicity, and effectiveness in both in vivo and in vitro studies. This could be a cornerstone for the pharmaceutical sector to develop new anticancer medications.

Abstract Background There has been a lot of debate on genetically modified (GM) crops both globally and locally in Uganda. Whereas some of the debates have been informed by scientific research, many are not. The level of acceptance and attitudes of people towards GM crops is a function of their knowledge. However, there is a paucity of studies on the knowledge and attitudes of Ugandans on GM crops. This study aimed to assess the level of knowledge of GM crops in Uganda. Methods We carried out a mixed methods study in September 2021 in Bushenyi, Jinja and Wakiso districts. We conducted 18 focus group discussions (FGDs), 13 key informant interviews (KIIs) and 698 quantitative interviews. The quantitative interviews were conducted using structured questionnaires. The FGD and KI interviews were audio recorded and transcribed verbatim. The qualitative data were analyzed using framework analysis and the quantitative data were analyzed using modified Poisson regression to identify factors associated with the level of knowledge of GM crops in STATA 15. Results Out of the 698 respondents interviewed, only 273 (39.1%) had ever heard of GM crops. About 204 (74.7%) of the 273 respondents reported having a moderate–high understanding of GM crops and 62.3% (170) of the respondents further reported that GM crops are harmful to their health and environment, despite some of their intrinsic benefits, such as high productivity, improved income and resilience to pests and diseases. In addition, Out of the 698 respondents interviewed, only 37.7% were wary of the possibility of the emergence of super pests due to the development of resistance to some GM crops. Conclusions Most of the community members do not have adequate knowledge about GM crops, hence, there is need for sensitization and legislation on GM crops before their release to the public.

Background: Genetic polymorphisms within the gene loci of the promoter region of tumor necrosis factor (TNF) alpha have been associated with the pathogenesis of hepatitis B virus (HBV) infection. In Uganda, there is a wide variation in the HBV endemicity, ranging from low endemicity, through moderate endemicity, to hyper-endemicity. However, the underlying reasons for this disparity in HBV burden are not fully elucidated. Thus, we aimed to test the hypothesis that the TNF-α-863C/A and -1031T/C polymorphic sites may have an effect on the difference between the burden of HBV in our country. We screened 384 participants, from which a sample of 134 was drawn, to determine the HBV, TNF-α-863C/A, and TNF-α-863T/C genotypes. The nucleotide BLAST was used to match the unknown targeted sequence obtained from the Sanger sequence against the known deposited sequence. This process unveiled the base substitution mutation and the HBV genotypes. The odds ratio (OR) and Chi-square test of proportions were used for the analysis. All the analyses were performed using SPSS version 26.0 and MedCalc software version 20.010 at 95% CI. A p < 0.05 was considered statistically significant. Results: The prevalence of both the TNF-α-863C/A and the TNF-α-1031T/C genotypes and their alleles did not differ significantly by endemicity (p > 0.05). However, the prevalence of the nucleotide substitution mutations for TNF-α-863C>A and TNF-α-1031T>C was significantly low for all the study groups (p < 0.05). Conclusion: The TNF-α gene promoter at the TNF-α-863C/A and 1031T/C positions is conserved in our population and may not affect the endemicity of HBV infection. However, future research should focus on the use of nationwide samples in order to reach concreate determinations regarding the role of the TNF-α polymorphisms in the risk/resolution of HBV infections in an African or Black population.

Background: In the East African region, herbal plants are essential in the treatment and control of cancer. Given the diverse ecological and cultural makeup of the regional states, it is likely that different ethnic groups will use the same or different plants for the same or different diseases. However, since 2019, this has not been compiled into a single study. Purpose: The study aimed to compile and record the medicinal plants utilized in East Africa from April 2019 to June 2023 to treat various cancer types. Materials and methods: The study examined 13 original studies that included ethnobotanical research conducted in East Africa. They were retrieved from several internet databases, including Google Scholar, Scopus, PubMed/Medline, Science Direct, and Research for Life. The study retrieved databases on plant families and species, plant parts used, preparation methods and routes of administration, and the country where the ethnobotanical field surveys were conducted. Graphs were produced using the GraphPad Prism 8.125 program (GraphPad Software, Inc., San Diego, CA). Tables and figures were used to present the data, which had been condensed into percentages and frequencies. Results: A total of 105 different plant species from 45 different plant families were identified, including Asteraceae (14), Euphorbiaceae (12), Musaceae (8), and Apocynaceae (7). Uganda registered the highest proportion (46% of the medicinal plants used). The most commonly mentioned medicinal plant species in cancer management was Prunus africana . Herbs (32%), trees and shrubs (28%), and leaves (45%) constituted the majority of herbal remedies. Most herbal remedies were prepared by boiling (decoction) and taken orally (57%). Conclusion: East Africa is home to a wide variety of medicinal plant species that local populations and herbalists, or TMP, frequently use in the treatment of various types of cancer. The most frequently used families are Asteraceae and Euphorbiaceae, with the majority of species being found in Uganda. The most frequently utilized plant species is Prunus africana . Studies on the effectiveness of Prunus africana against other malignancies besides prostate cancer are required.

Mercury (Hg) has been used for millennia in artisanal and small-scale gold mining (ASGM) to extract gold from ore as an amalgam that is heated to recover gold. Since there is hardly any sustainable technology that can be used to recover it, the vast percentage of released Hg finds its way into the environment. Currently, ASGM is the largest source of mercury pollution on Earth. Mercury (Hg) is listed among the top 10 most harmful metals by the World Health Organization (WHO), and it is known to cause several neurological disorders in humans. Thus, Hg levels in environmental systems surrounding pollution hotspots such as ASGM mines need to be monitored to ensure their proper management and protect vulnerable ecosystems and human health. This work was aimed to study the level of Hg pollution in soils and plants thriving around ASGM mines in Eastern Uganda and to evaluate the Hg phytoremediation potential of the plants. The total Hg contents were determined for soils, food crops and wild plants growing around ASGM mines. The results for the pH, organic carbon (OC) and electric conductivity (EC) of the soils in the study area were comparable to those of the control soils. The soils in the studied ASGM areas exhibited high Hg concentrations ranging between 723 and 2067 folds more than those of the control soils. All soils were moderately to heavily contaminated according to geoaccumulation (Igeo) index values that ranged between 1.16 and 3.31. The results of this study also showed that the food crops and wild plants accumulated Hg levels that were above the 20 ng/g (0.02 ppm) permissible limit. This study revealed relatively higher levels of Hg in the aerial parts of the plants compared with the underground organs, which can be attributed to Hg deposition, entry through stomata and foliar adsorption. Mercury levels in 47% of the food crop samples were above the FAO/WHO permissible mercury limit of 0.5 µg/g. Similarly, medicinal plants accumulated Hg to levels that were several folds higher than the 0.2 ppm permissible limit of mercury in herbal materials of Canada. Interestingly, this study showed that some wild plant species, especially sedges, exhibited relatively higher levels of mercury accumulation than others thriving in the same environment, an indication that such plants could be utilized in the phytoremediation of Hg-contaminated sites.

Xanthomonas campestris pv. musacearum (Xcm) bacteria cause banana Xanthomonas wilt (BXW), the most destructive disease of bananas in East and Central Africa. During early stages of infection in susceptible banana cultivars, incomplete systemic movement of Xcm limits bacterial colonization in the upper organs. The mechanistic basis of this delayed movement is unknown. We hypothesized that Xcm infection triggers basal pattern-triggered immune (PTI) responses whose spatial and temporal variability along the banana's anatomical structure accounts for initially limiting Xcm in upper organs. Hence, we examined PTI responses such as callose deposition and hydrogen peroxide (H2O2) production in different organs in response to Xcm infection in BXW-susceptible Kayinja and Mbwazirume banana cultivars and wild resistant progenitor Musa balbisiana. Xcm-induced callose increased and peaked at 14 days postinoculation (dpi) and 28 dpi as assessed by fluorescence microscopy and enzyme-linked immunosorbent assays, respectively. The levels of Xcm-induced H2O2 and callose were highest in the pseudostems and corms, respectively, and were independent of host susceptibility or resistance to BXW. H2O2 production showed a biphasic transient pattern with an initial increase at 1-hour post Xcm inoculation (hpi), followed by a decline 3 to 6 hpi and then a second increase by 12 hpi. Our findings point to organ-specific responses to Xcm infection in bananas. The corm, which doubles as a subterranean perennating organ and interface between mother plants and lateral shoots, was the most responsive organ in callose production, whereas the pseudostem was the most responsive organ in H2O2 production, suggesting the significance of these organs in banana response to BXW. [Figure: see text] Copyright © 2022 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Background. Hepatitis B virus (HBV) is the leading cause of liver-related diseases. In Uganda, there is a regional disparity in the HBV burden. Our study was aimed at establishing the circulating genotypes in a low and a high endemic region to give plausible explanations for the differences in regional burden and guide the future management of the disease. Methods. A total of 200 HBsAg-seropositive subjects were recruited into the study by convenience sampling. The HBsAg Rapid Test Strip (Healgen Scientific Limited Liability Company, Houston, TX77047- USA) was used to screen for HBsAg while the Roche machine (Roche, Basel Switzerland/Abbot Technologies (USA)) was used to determine the viral load. The Chemistry Analyzer B120 (Mindray, China) was used for chemistry analysis. For HBV genotyping, total DNA was extracted from whole blood using the QIAamp® DNA extraction kit. Nested PCR amplification was performed using Platinum Taq DNA Polymerase (Invitrogen Corporation, USA) to amplify the 400 bp HBV polymerase gene. Purification of nested PCR products was performed using Purelink PCR product purification kit (Life Technologies, USA). Automated DNA sequencing was performed using BigDye Terminator v3.1 Cycle Sequencing Kit on 3130 Genetic Analyzer (Applied Biosystems, USA). The NCBI HBV genotyping tool (https://www.ncbi.nlm.nih.gov/projects/genotyping/formpage.cgi) was used for determination of genotype for each HBV sequence. Pearson’s chi-square, multinomial logistic regression, and Mann–Whitney U tests were used for the analysis. All the analyses were done using SPSS version 26.0 and MedCalc software version 19.1.3 at 95% CI. A p

The existing methods of callose quantification include epifluorescence microscopy and fluorescence spectrophotometry of aniline blue-stained callose particles, immuno-fluorescence microscopy and indirect assessment of both callose synthase and β-(1,3)-glucanase enzyme activities. Some of these methods are laborious, time consuming, not callose-specific, biased and require high technical skills. Here, we describe a method of callose quantification based on Sandwich Enzyme-Linked Immunosorbent Assay (S-ELISA). Tissue culture-derived banana plantlets were inoculated with Xanthomonas campestris pv. musacearum (Xcm) bacteria as a biotic stress factor inducing callose production. Banana leaf, pseudostem and corm tissue samples were collected at 14 days post-inoculation (dpi) for callose quantification. Callose levels were significantly different in banana tissues of Xcm-inoculated and control groups except in the pseudostems of both banana genotypes. The method described here could be applied for the quantification of callose in different plant species with satisfactory level of specificity to callose, and reproducibility. Additionally, the use of 96-well plate makes this method suitable for high throughput callose quantification studies with minimal sampling and analysis biases. We provide step-by-step detailed descriptions of the method.

Abstract Background Azolla is an important aquatic fern whose agronomic potential has not been fully exploited in Uganda. This study aimed at determining the genetic variation in the Azolla species existing in Uganda and the factors influencing their distribution in the different agro-ecological zones of Uganda. Molecular characterization was preferred in this study because of its efficiency in detecting variations among closely related species. Results Four species of Azolla were identified in Uganda with 100, 93.36, 99.22 and 99.39% sequence identities to the reference database sequences of; Azolla mexicana, Azolla microphylla, Azolla filiculoides and Azolla cristata, respectively. These different species were distributed in four out of the ten agro-ecological zones of Uganda which are situated in close vicinity to large water masses. The principal component analysis (PCA) results revealed that maximum rainfall and altitude significantly accounted for the variations in the distribution of Azolla with factor loadings of 0.921 and 0.922, respectively. Conclusion Massive destruction coupled with prolonged disturbance of Azolla’s habitat negatively affected its growth, survival and distribution in the country. Therefore, there is a need to develop standard methods that can preserve the various species of Azolla, so as to salvage them for future use, research and reference.

Abstract Background Oil spills are ranked among the greatest global challenges to humanity. In Uganda, owing to the forthcoming full-scale production of multi-billion barrels of oil, the country’s oil pollution burden is anticipated to escalate, necessitating remediation. Due to the unsuitability of several oil clean-up technologies, the search for cost-effective and environmentally friendly remediation technologies is paramount. We thus carried out this study to examine the occurrence of metabolically active indigenous bacterial species and chemical characteristics of soils with a long history of oil pollution in Uganda that can be used in the development of a bacterial-based product for remediation of oil-polluted sites. Results Total hydrocarbon analysis of the soil samples revealed that the three most abundant hydrocarbons were pyrene, anthracene and phenanthrene that were significantly higher in oil-polluted sites than in the control sites. Using the BIOLOG EcoPlate™, the study revealed that bacterial species richness, bacterial diversity and bacterial activity (ANOVA, p