Phage banks as potential tools to rapidly and cost-effectively manage antimicrobial resistance in the developing world.

Lower and middle-income countries seldom develop vaccines and therapeutics for their own populations and are dependent on supplies from industrialized countries, which are often hampered by financial or supply chain limitations. This has resulted in major delays in delivery with significant loss of life, as seen with the coronavirus pandemic. Since the vast majority of deaths from the antimicrobial resistance crisis are expected to occur in developing countries, there is an urgent need for in-country production of antibacterial therapies such as phages. Nationally controlled phage banks might provide such a solution since locally developed phage therapies tailored to endemic bacterial strains could offer cost-effective antibiotic alternatives.

Herbal medicine used for the treatment of diarrhea and cough in Kampala city, Uganda.

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.

Medically important bacteria isolated from commercial herbal medicines in Kampala city indicate the need to enhance safety frameworks.

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.

Antibiotic-resistance in medically important bacteria isolated from commercial herbal medicines in Africa from 2000 to 2021: a systematic review and meta-analysis.

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 (I2 = 99.17%; p < 0.0001), with no evidence of publication bias. The most prevalent multidrug-resistant species was Escherichia coli (24.0%). The most highly resisted drug was Ceftazidime with a pooled prevalence of 95.10% (95% CI = 78.51-99.87%), while the drug-class was 3rd generation cephalosporins; 91.64% (95% CI = 78.64-96.73%). None of the eligible studies tested isolates for Carbapenem resistance. Extended Spectrum ß-lactamase genes were detected in 89 (37.2%) isolates, mostly Salmonella spp., Proteus vulgaris, and K. pneumonia. Resistance plasmids were found in 6 (5.8%) isolates; the heaviest plasmid weighed 23,130 Kilobases, and Proteus vulgaris harbored the majority (n = 5; 83.3%). CONCLUSIONS: Herbal medicines in Africa harbor bacterial contaminants which are highly resistant to conventional medicines. This points to a potential treatment failure when these contaminants are involved in diseases causation. More research on this subject is recommended, to fill the evidence gaps and support the formation of collaborative quality control mechanisms for the herbal medicine industry in Africa.

Prevalence and dynamics of clinically significant bacterial contaminants in herbal medicines sold in East Africa from 2000 to 2020: a systematic review and meta-analysis.

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 I2 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 (I2 = 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.

Prevalence of pathogenic Klebsiella pneumoniae based on PCR capsular typing harbouring carbapenemases encoding genes in Uganda tertiary hospitals.

BACKGROUND: Klebsiella pneumoniae is an opportunistic pathogen that has been implicated as one of commonest cause of hospital and community acquired infections. The K. pneumoniae infections have considerably contributed to morbidity and mortality in patients with protracted ailments. The capacity of K. pneumoniae to cause diseases depends on the presence of an array virulence factors. Coexistence and expression of virulence factors and genetic determinants of antibiotic resistance complicates treatment outcomes. Thus, emergence of pathogenic MDR K. pneumoniae poses a great threat to the healthcare system. However, the carriage of antibiotic resistance among pathogenic K. pneumoniae is yet to be investigated in Uganda. We sought to investigate the carbapenem resistance profiles and pathogenic potential based on capsular serotypes of K. pneumoniae clinical isolates. METHODS: This was a cross sectional study involving use of archived Klebsiella pneumoniae isolates collected between January and December, 2019 at four tertiary hospitals in Uganda. All isolates were subject to antimicrobial susceptibility assays to determine phenotypic antibiotic resistance, pentaplex PCR to detect carbapenemases encoding genes and heptaplex PCR to identify capsular serotypes K1, K2, K3, K5, K20, K54 and K57. RESULTS: The study found an overall phenotypic carbapenem resistance of 23.3% (53/227) and significantly higher genotypic resistance prevalence of 43.1% (98/227). Over all, the most prevalent gene was blaOXA-48-like (36.4%), followed by blaIMP-type (19.4%), blaVIM-type (17.1%), blaKPC-type (14.0%) and blaNDM-type (13.2%). blaVIM-type and blaOXA-48-like conferred phenotypic resistance in all isolates and 38.3% of isolates that harbored them respectively. Capsular multiplex PCR revealed that 46.7% (106/227) isolates were pathogenic and the predominantly prevalent pathotype was K5 (18.5%) followed by K20 (15.1%), K3 (7.1%), K2 (3.1%) and K1 (2.2%). Of the 106 capsular serotypes, 37 expressed phenotypic resistance; thus, 37 of the 53 carbapenem resistant K. pneumoniae were pathogenic. CONCLUSION: The high prevalence of virulent and antibiotic resistant K. pneumoniae among clinical isolates obtained from the four tertiary hospital as revealed by this study pose a great threat to healthcare. Our findings underline the epidemiological and public health risks and implications of this pathogen.

In vitro regeneration of Ugandan passion fruit cultivars from leaf discs.

OBJECTIVE: Passion fruit improvement efforts by conventional breeding have had limited success calling for research into alternative approaches such as tissue culture and genetic engineering. An efficient and reproducible regeneration system is a prerequisite for successful genetic engineering. Currently, there is no reliable regeneration system for Uganda's passion fruit varieties owing to the high heterogeneity of the Passiflora genus. Therefore, this study aimed at establishing an efficient and reproducible regeneration system for Uganda's Passiflora edulis f. flavicarpa (yellow passion fruit) and Passiflora edulis f. edulis (purple passion fruit) for routine utilization with an ultimate goal of improving its agronomic value. RESULTS: The study successfully induced shoots by both direct and indirect organogenesis for the yellow passion fruit variety. Highest shoot induction frequency (14.85%) was achieved on 8.9 µM BAP while 7.9 µM BAP did not initiate any shoots. Optimal shoot elongation and rooting was achieved on 0.44 µM BAP and 5.37 µM α-naphthaleneacetic (NAA) respectively. Rooted yellow passion fruit plantlets were successfully weaned with over 65% survival rates. It took approximately 6 months to produce a weaned healthy passion fruit plant. The purple passion fruit variety proved to be recalcitrant to tissue culture with no successful shoot or callus induction.

Influence of physicochemical parameters on storage stability: Microbiological quality of fresh unpasteurized fruit juices.

Fresh juices rich in health and nutritional benefits are valued for their fresh flavor, taste, and aroma. These juices' quality however is affected by factors like temperature, light, and microbiological contamination significantly changing physicochemical parameters and storage stability. Physicochemical and microbiological analyses of passion fruit, pineapple, and mango juices in dark and light bottles at 24°C and 4°C were conducted in Kampala, Uganda for 12 days. Physicochemical parameters significantly reduced (p < .05) storage stability of fresh juices, while no significant changes (p > .05) were observed for the microbiological analyses. pH values ranged from 3.0 to 4.2 (dark) bottles and 2.9 to 4.0 (light) bottles for juices at 24°C and 4°C. °Brix values were from 1.0 to 5.5 for dark and clear bottles at 24°C and 4°C. TTA (%) values ranged from 1.1 to 7.2 (dark) bottles and 1.1 to 7.4 for (light) bottles at 24°C and 4°C. Ascorbic acid content ranged from 3.5 to 61.0 mg/ 100 ml and 5.5 to 56.7 mg/100 ml for juices in dark and clear bottles, respectively. total plate counts ranged from 1.3 × 10___ to 3.3 × 107 CFU/ml (dark bottles at 24°C) to 3.5 × 10³ to 3.3 × 108 CFU/ml (dark bottles at 4°C). For juices in light bottles, total plate counts ranged from 1.8 × 10___ to 8.0 × 107 CFU/ml (24°C) and 2.7 × 10___ to 1.5 × 108 CFU/ml (4°C). High microbial loads suggest the use of poor processing techniques and lack of good hygiene which lower quality and reduce storage stability of juices. Storage temperature greatly reduces physicochemical parameters both at ambient and refrigeration temperatures. This implies that temperature control for unpasteurized juices is critical in order to inhibit microorganism metabolic activities which accelerate biodeterioration leading to spoilage and short shelf life. Fresh unpasteurized juices stored at 24°C and 4°C may safely be consumed within 1 and 2 days, respectively.