Assessment of Enrofloxacin Usage and Residue Levels of Enrofloxacin-Ciprofloxacin in Breast and Liver Tissues of Commercial Broilers Sold in Kampala-Uganda.

Background: Human exposure to veterinary drugs like fluoroquinolones occurs due to the presence of their residues in foods from animal sources in varying concentrations. The existence of antibiotic residues in foodstuffs can pose great public health problems to consumers. This study aimed to assess enrofloxacin use patterns and assess residue levels of enrofloxacin/ciprofloxacin in breast muscle and liver tissues of broiler chickens sold for consumption in Kampala capital city. Methods: This was a cross-sectional study that involved both field survey and laboratory-based methods. The field study involved the use of qualitative and semi-quantitative data collection tools to interview 34 broiler farmers and 10 veterinary drugs vendors. For the determination of enrofloxacin/ciprofloxacin levels, 68 chicken breast and liver tissue samples were collected from main markets in Kampala over one month and analyzed using HPLC-UV. Results: Enrofloxacin was the most used antibiotic (100%) for the management of poultry diseases, majorly respiratory diseases (100%), salmonella infections (40%), and disease prevention (60%). Over 76% of the farmers knew the meat withdrawal time (WDT) for enrofloxacin, but none of them adhered to this. Over 70% of the farmers reported that the veterinary drugs vendors were not providing meat WDT information. Enrofloxacin/ciprofloxacin residues were identified in 35.3% (12/34) of the muscle and 38.2% (13/34) of the liver tissues analyzed. Of those muscle and liver tissue samples that tested positive, 25% (3/12) and 38.5% (5/13) respectively had drug concentrations higher than the recommended Maximum Residue Limits. The overall mean enrofloxacin/ciprofloxacin concentration in the chicken muscle and liver tissues was 83.6 (±34.5) µg/kg and 171.5 (±75.9) µg/kg. Conclusion: This observed presence of enrofloxacin/ciprofloxacin levels above safety requirements is attributable to inadequate medicines use information provided by veterinary drugs vendors to farmers and also to the non-compliance of some farmers to meat WDT due to the economic implications.

Application of the herbal chemical marker ranking system (Herb MaRS) to the standardization of herbal raw materials: a case study.

INTRODUCTION: Phytochemical standardization of herbal materials involves establishing consistent levels of one or more active ingredients or markers. It ensures the authenticity and quality of herbal materials, extracts, and their products. This research aimed to apply the herbal chemical marker ranking system (Herb MaRS) originally proposed for quality assurance of complex herbal products to establish markers for controlling the quality of herbal raw materials. METHODS: The assessment of compounds for suitability as markers was based on the Herb MaRS, with minor modifications as follows: for more objective scoring, evidence of biological activity of the potential marker compound(s) was determined at three levels based on the number of symptoms of the disease condition a compound can treat or alleviate: (i) one symptom (1 point), two symptoms (2 points), and 3 or more symptoms (3 points). The reported concentrations of the compounds were also scored as follows: concentration not determined (0 points), concentration ≥ 5 ppm (1 point), concentration ≥ 50 ppm (2 points) and availability of analytical standards (1 point). Finally, the compounds were scored for the availability of an analytical method (1 point). The compounds were scored from 0 to 8, where 8 indicated the most suitable chemical marker. RESULTS: The selected markers were as follows: aromadendrine, α-terpineol, globulol, and 1,8-cineol (in Eucalyptus globulus Labill. ); aloin, aloe emodin, acemannan (in Aloe barbadensis (L.) Burm.f. ), lupeol, lupenone, betulinic acid, betulin, and catechin (in Albizia coriaria Oliv.); mangiferin, catechin, quercetin, and gallic acid (in Mangifera indica L.); polygodial (in Warburgia ugandensis Sprague); azadirachtin, nimbin, nimbidin (in Azadirachta indica A. Juss. ); and 6,8,10-gingerols, and 6-shogaol (in Zingiber officinalis Roscoe). CONCLUSIONS: Herb MaRS can be efficiently applied to select marker compounds for quality control of herbal materials. However, for herbs whose phytochemicals have not been sufficiently researched, it is difficult to establish evidence of activity, and there are no analytical standards and/or methods; this is the case for plants exclusively used in Africa. The markers identified should be incorporated into chromatographic fingerprints, their quantitative methods developed, and evaluated for applicability at the various stages of the production chain of herbal medicines; then, they can be included in future local plant monographs. There is also a need to build local capacity to isolate marker compounds, particularly those that are not sold by current vendors.

Chemometric Classification of Mangifera indica L. Leaf Cultivars, Based on Selected Phytochemical Parameters; Implications for Standardization of the Pharmaceutical Raw Materials.

Introduction: Mangifera indica leaves are among the most common materials employed in manufacturing herbal medicinal products. Despite the phytochemical variation of M. indica cultivars, there are no monographs to guide the cultivation, processing, and authentication of the materials. Methods: This study characterized 15 Ugandan M. indica leaf varieties, with reference to extraction index (EI), total phenolic content (TPC), antioxidant activity (AOA), and mangiferin concentration (MC). In addition, HPLC fingerprints were established to evaluate the overall phytoequivalence of the materials. Then, using hierarchical clustering (HC) and principal component analysis (PCA), the materials were assigned quality grades. Results: The mean EI was 9.39 ± 1.64% and varied among the varieties (P=0.001); the TPC varied significantly (P < 0.0001), from 183.29 ± 2.36 mg/g (Takataka) to 79.47 ± 0.58 mg/g (Apple mango). AOA ranged from 16.81 ± 2.85 µg/mL (Doodo red) to 87.85 µg/mL (Asante). MC varied significantly (P < 0.0001), from 105.75 ± 0.60 mg/g (Kate) to 39.53 ± 0.30 mg/g (Asante). HC gave four major grades: A to D (A, varieties with the highest TPC, MC, and AOA). These parameters reduced to below average from group B to group D. The chromatographic fingerprints were visually similar, but the number of peaks varied, from 19 (Kawanda green) to 29 (Kawanda wide), with 23.5 ± 2.9 average peaks. Whole fingerprints were less similar (r < 0.8) than common peak fingerprints (r > 0.9, P < 0.001). PCA grouped the fingerprints into five clusters; loading plots for PC 1 and 2 revealed two important compounds, one at Rt = 15.828 minutes (mangiferin) and the other at 6.021 minutes. Using the standardized common fingerprints, unknown field samples clustered closely with Koona, Kate, and Kawanda green varieties. Conclusions: The EI, TPC, MC, and AOA values can be utilized to monitor consistency in the quality of materials and the production process. The grades generated can be used to select materials for cultivation and manufacturing. Where minimum concentrations are set, materials of different concentrations are used to dilute or concentrate each other. The HPLC fingerprints can be utilized to authenticate the materials. More samples from different agroecological regions of the country should be tested to cater to climatic variations in order to develop GMP-compliant botanical identification methods.

Ethnobotanical Survey of Medicinal Plants Used in Breast Cancer Treatment by Traditional Health Practitioners in Central Uganda.

Purpose: The study aimed to document the existing knowledge and practices related to breast cancer recognition and treatment using medicinal plants by traditional health practitioners in Central Uganda. Methods: This cross-sectional exploratory survey, conducted between February and August 2020, applied a mixed methods research approach. A semi-structured questionnaire was administered to 119 traditional health practitioners (THPs) in Kampala, Wakiso and Mukono. Content analysis of qualitative data was done. Quantitative ethnobotanical survey indices, namely user reports (Nur), percentage of respondents with knowledge (PRK), informant consensus factor (Fic), fidelity level (FL), preference ranks (PR) and direct matrix ranking (DMR) were determined. Results: Most THPs recognized breast cancer by breast swelling (n=74, 62.2%) and breast pain (n=29, 24.4%). They cited 30 plants from 30 genera in 23 families (Fic 0.75 on breast cancer). Asteraceae, Apocynaceae, Euphorbiaceae, Fabaceae, Lamiaceae and Rutaceae were the predominant families. The ten most cited plants were Annona muricata L. (Nur=24), Rhoicissus tridentata (L.f.) Wild & R.B.Drumm (Nur =19), Erythrococca bongensis Pax (Nur=11), Ficus sp. (Nur=10), Cannabis sativa L. (Nur=8), Ipomoea wightii (Wall.) Choisy (Nur=7), Erythrina abyssinica DC. (Nur=5), Leucas martinicensis (Jacq.) R.Br. (Nur=4), Abelmoschus esculentus (L.) Moench (Nur=4) and Zanthoxylum chalybeum Engl. (Nur=3). Annona muricata L. was highly preferred by THPs (PR 1), Ficus sp. had highest fidelity level (FL=77%) and Zanthoxylum chalybeum Engl. ranked as the highest multipurpose plant (DMR 1). Herbs (n=14, 47%) were the most commonly used life forms besides trees (n=11, 37%) and shrubs (n=5, 16%). THPs mostly used leaves (46%), prepared decoctions (82%) and applied residues on the breast. Conclusion: THPs in Central Uganda recognized breast cancer by symptoms. Medicinal plants applied in its folk treatment have been documented and the claims of cure by THPs merit further investigation.

ADMET profiling and molecular docking of potential antimicrobial peptides previously isolated from African catfish, Clarias gariepinus.

Amidst rising cases of antimicrobial resistance, antimicrobial peptides (AMPs) are regarded as a promising alternative to traditional antibiotics. Even so, poor pharmacokinetic profiles of certain AMPs impede their utility necessitating, a careful assessment of potential AMPs' absorption, distribution, metabolism, excretion, and toxicity (ADMET) properties during novel lead exploration. Accordingly, the present study utilized ADMET scores to profile seven previously isolated African catfish antimicrobial peptides (ACAPs). After profiling, the peptides were docked against approved bacterial protein targets to gain insight into their possible mode of action. Promising ACAPs were then chemically synthesized, and their antibacterial activity was validated in vitro utilizing the broth dilution method. All seven examined antimicrobial peptides passed the ADMET screening, with two (ACAP-IV and ACAP-V) exhibiting the best ADMET profile scores. The ACAP-V had a higher average binding energy (-8.47 kcal/mol) and average global energy (-70.78 kcal/mol) compared to ACAP-IV (-7.60 kcal/mol and -57.53 kcal/mol), with the potential to penetrate and disrupt bacterial cell membrane (PDB Id: 2w6d). Conversely, ACAP-IV peptide had higher antibacterial activity against E. coli and S. aureus (Minimum Inhibitory Concentration, 520.7 ± 104.3 µg/ml and 1666.7 ± 416.7 µg/ml, respectively) compared to ACAP-V. Collectively, the two antimicrobial peptides (ACAP-IV and ACAP-V) are potential novel leads for the food, cosmetic and pharmaceutical industries. Future research is recommended to optimize the expression of such peptides in biological systems for extended evaluation.

Safety and Efficacy of Medicinal Plants Used to Manufacture Herbal Products with Regulatory Approval in Uganda: A Cross-Sectional Study.

Introduction: The Uganda National Drug Authority requires phytochemical screening, freedom from microbial contamination, and evidence of safety and efficacy of the constituent plants to register herbal products. Since Uganda has no pharmacopeia, safety, efficacy, and plant processing information are not readily available. We documented the plant materials used to manufacture products in Uganda and established evidence of their safety and efficacy and availability of monographs. Methods: The NDA register of herbal products was reviewed, and a product list was extracted. The herbal products were purchased from local pharmacies, and their labels were studied to identify plant ingredients and drug use. Literature was reviewed to document evidence of the safety and efficacy of the plant materials concerning manufacturer's claims. Also, the WHO and available African Pharmacopeia were searched to establish the availability of the plant monographs. Results: Of the 84 NDA-registered local products, only 18 were obtained from the market; 82% were indicated for respiratory tract disorders. Thirty-three plant materials were listed with Eucalyptus globulus Labill, being the commonest. Several in vitro and in vivo studies demonstrate efficacy, thus supporting the use of the selected plant species for empirical treatment as stated on the product label. While most plants were safe, some species such as Albizia coriaria Oliv. had dose-dependent toxicities that cannot be predicted in combinations. The WHO, African Pharmacopoeia, and West African Herbal Pharmacopoeia had only 16 plant monographs of the 33 plants of interest. Nevertheless, Aloe vera (L.) Burm.f., Azadirachta indica A.Juss., Zingiber officinale Roscoe, and Allium sativum L. monographs were published by all three pharmacopoeias. Conclusions: Preclinical evidence of safety and efficacy exists in the literature for most of the plants used to manufacture registered herbal products in Uganda. More specific bioassays and clinical trials are required for the products to provide conclusive evidence of safety and toxicity. Monographs are urgently needed for the Ugandan plants.

Evaluation of the Clinical Use of Ceftriaxone among In-Patients in Selected Health Facilities in Uganda.

Ceftriaxone has a high propensity for misuse because of its high rate of utilization. In this study, we aimed at assessing the appropriateness of the clinical utilization of ceftriaxone in nine health facilities in Uganda. Using the World Health Organization (WHO) Drug Use Evaluation indicators, we reviewed a systematic sample of 885 patients' treatment records selected over a three (3)-month period. Our results showed that prescriptions were written mostly by medical officers at 53.3% (470/882). Ceftriaxone was prescribed mainly for surgical prophylaxis at 25.3% (154/609), respiratory tract infections at 17% (104/609), and sepsis at 11% (67/609), as well as for non-recommended indications such as malaria at 7% (43/609) and anemia at 8% (49/609). Ceftriaxone was mostly prescribed once daily (92.3%; 817/885), as a 2 g dose (50.1%; 443/885), and for 5 days (41%; 363/885). The average score of inappropriate use of ceftriaxone in the eight indicators was 32.1%. Only 58.3% (516/885) of the ceftriaxone doses prescribed were administered to completion. Complete blood count and culture and sensitivity testing rates were 38.8% (343/885) and 1.13% (10/885), respectively. Over 85.4% (756/885) of the patients improved and were discharged. Factors associated with appropriate ceftriaxone use were gender, pregnancy status, days of hospitalization, health facility level of care, health facility type, and type of prescriber.

Identification of Antimicrobial Peptides Isolated From the Skin Mucus of African Catfish, Clarias gariepinus (Burchell, 1822).

Antimicrobial peptides (AMPs) constitute a broad range of bioactive compounds in diverse organisms, including fish. They are effector molecules for the innate immune response, against pathogens, tissue damage and infections. Still, AMPs from African Catfish, Clarias gariepinus, skin mucus are largely unexplored despite their possible therapeutic role in combating antimicrobial resistance. In this study, African Catfish Antimicrobial peptides (ACAPs) were identified from the skin mucus of African Catfish, C. gariepinus. Native peptides were extracted from fish mucus scrapings in 10% acetic acid (v/v) and ultra-filtered using 5 kDa molecular weight cut-off membrane. The extract was purified using C18 Solid-Phase Extraction. The antibacterial activity was determined using the Agar Well Diffusion method and broth-dilution method utilizing Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922). Thereafter, Sephadex G-25 gel filtration was further utilized in bio-guided isolation of the most active fractions prior to peptide identification using Orbitrap Fusion Lumos Tribrid Mass Spectrometry. The skin mucus extracted from African Catfish from all the three major lakes of Uganda exhibited antimicrobial activity on E. coli and S. aureus. Lake Albert's C. gariepinus demonstrated the best activity with the lowest MIC of 2.84 and 0.71 µg/ml on S. aureus and E. coli, respectively. Sephadex G-25 peak I mass spectrometry analysis (Data are available via ProteomeXchange with identifier PXD029193) alongside in silico analysis revealed seven short peptides (11-16 amino acid residues) of high antimicrobial scores (0.561-0.905 units). In addition, these peptides had a low molecular weight (1005.57-1622.05 Da) and had percentage hydrophobicity above 54%. Up to four of these AMPs demonstrated α-helix structure conformation, rendering them amphipathic. The findings of this study indicate that novel AMPs can be sourced from the skin mucus of C. gariepinus. Such AMPs are potential alternatives to the traditional antibiotics and can be of great application to food and pharmaceutical industries; however, further studies are still needed to establish their drug-likeness and safety profiles.

Compliance of private pharmacies in Uganda with controlled prescription drugs regulations: a mixed-methods study.

BACKGROUND: Controlled prescription drug use disorders are a growing global health challenge in Sub-Saharan Africa. Effective supply chain regulations on dispensing and stock control are important for controlling this epidemic. Since compliance with these regulations in resource-limited countries is poor, there is need to understand its predictors in order to reduce the risk of prescription drug use disorders. METHODS: A mixed-methods study utilizing a structured questionnaire and a simulated client guide was undertaken in Kampala and Mbale towns in Uganda. The questionnaire recorded self-reported dispensing and verified stock control practices and their covariates from 101 private pharmacies. The guide recorded actual dispensing practices from 27 pharmacies. Snowball sampling was done to enrich the sample with pharmacies that stock opioids. The mean compliance with good dispensing and stock control practices was calculated. Multivariate logistic regression analyses were applied to identify predictors of compliance. RESULTS: The mean compliance with dispensing and stock control requirements was 82.9% and 23%, respectively. Twenty percent and 40% of the pharmacies dispensed pethidine without a prescription and with invalid prescriptions, respectively. Having a pharmacist on duty (OR = 5.17; p = 0.02), prior in-service training on narcotics regulations (OR = 3.51; p = 0.04), and previous narcotics audits by the regulator (OR = 5.11; p = 0.01) were independent predictors of compliance with stock control requirements. Pharmacies with a previous history of poor compliance with dispensing requirements were less likely to demonstrate good compliance (OR = 0.21; p = 0.01). CONCLUSIONS: There is suboptimal compliance to controlled prescription drug regulations among Uganda's pharmacies. A previous history of poor compliance to dispensing requirements predicted low compliance in subsequent assessments. Training and regulatory audits increased compliance in stock control but not dispensing. Expansion of training and audits to more pharmacies and/or incentives for compliance are necessary.

Why regulatory indifference towards pharmaceutical pollution of the environment could be a missed opportunity in public health protection. a holistic view.

The last generation has witnessed bludgeoning of the world's population, a spike in disease burden, and unprecedented levels of pharmaceutical consumption and production. Unfortunately, pharmaceuticals have left their industrial and household confines and leaked into the environment. Pharmaceuticals are now major environmental pollutants, and are ubiquitous in waters and soils. Unlike other environmental contaminants, pharmaceutical pollutants are not yet regulated globally, simply because acute risk assessments show insignificant human health hazard. But the pitfalls of pharmaceutical pollutants extend beyond acute effects to delayed effects from bioaccumulation, amplified effects from drug-drug interactions, exacerbation of drug resistance, and reduction in aquatic and terrestrial food production. Therefore, ignoring pharmaceutical pollutants deprives society of holistic public health protection.

Threats posed by stockpiles of expired pharmaceuticals in low- and middle-income countries: a Ugandan perspective.

In some low- and middle-income countries, the national stores and public-sector health facilities contain large stocks of pharmaceuticals that are past their expiry dates. In low-income countries like Uganda, many such stockpiles are the result of donations. If not adequately monitored or regulated, expired pharmaceuticals may be repackaged and sold as counterfeits or be dumped without any thought of the potential environmental damage. The rates of pharmaceutical expiry in the supply chain need to be reduced and the disposal of expired pharmaceuticals needs to be made both timely and safe. Many low- and middle-income countries need to: (i) strengthen public systems for medicines' management, to improve inventory control and the reliability of procurement forecasts; (ii) reduce stress on central medical stores, through liberalization and reimbursement schemes; (iii) strengthen the regulation of drug donations; (iv) explore the salvage of officially expired pharmaceuticals, through re-analysis and possible shelf-life extension; (v) strengthen the enforcement of regulations on safe drug disposal; (vi) invest in an infrastructure for such disposal, perhaps based on ultra-high-temperature incinerators; and (vii) include user accountability for expired pharmaceuticals within the routine accountability regimes followed by the public health sector.

Dans certains pays à revenu faible et intermédiaire, les pharmacies nationales et les centres publics de santé détiennent d'énormes stocks de produits pharmaceutiques périmés. Dans les pays à revenu faible comme l'Ouganda, nombre de ces stocks proviennent de dons. Or, s'ils ne sont pas correctement contrôlés et réglementés, les produits pharmaceutiques périmés peuvent être reconditionnés et revendus en tant que contrefaçons ou bien jetés sans considération du danger pour l'environnement. Dans la chaîne d'approvisionnement, le pourcentage des produits pharmaceutiques périmés doit diminuer, et les produits périmés doivent être éliminés au bon moment et de façon sûre. De nombreux pays à revenu faible et intermédiaire doivent: (i) renforcer les systèmes publics pour la gestion des médicaments, afin d'améliorer le contrôle des stocks et la fiabilité des prévisions d'approvisionnement; (ii) réduire la pression à laquelle les pharmacies centrales sont soumises, grâce à des programmes de libéralisation et de remboursement; (iii) renforcer la régulation des dons de médicaments; (iv) étudier les options envisageables pour réemployer les produits pharmaceutiques officiellement périmés mais dont la durée de conservation pourrait éventuellement être prolongée après la réalisation de nouveaux tests; (v) renforcer l'application des réglementations pour une élimination sans risques des médicaments; (vi) investir dans des infrastructures d'élimination sans risques des médicaments, éventuellement au moyen d'incinérateurs à ultra-haute température; et (vii) responsabiliser les utilisateurs, en intégrant dans les programmes d'encadrement du secteur de la santé publique une obligation de rendre compte pour les produits périmés.

En algunos países con ingresos bajos y medios, los almacenes nacionales y las instalaciones del sector sanitario público contienen grandes cantidades de existencias de medicamentos que han superado su fecha de caducidad. En países con ingresos bajos como Uganda, muchas de estas existencias son el resultado de donaciones. Si no se controlan o regulan adecuadamente, puede ser que los medicamentos caducados sean empaquetados de nuevo y vendidos como falsificaciones o que se depositen sin tener conciencia del potencial daño medioambiental. Deben reducirse las tasas de caducidad de los medicamentos en la cadena de suministro y los medicamentos caducados deben depositarse a tiempo y de forma segura. Muchos países con ingresos bajos y medios necesitan: (i) fortalecer los sistemas públicos para la gestión de medicamentos para mejorar el control del inventario y la fiabilidad de la adquisición de provisiones; (ii) reducir el estrés en los almacenes médicos centrales a través de sistemas de reembolso y liberación; (iii) aumentar la regulación de las donaciones de medicamentos; (iv) investigar la recuperación de medicamentos oficialmente caducados a través de análisis y una posible extensión de la vida útil; (v) fortalecer la aplicación de regulaciones sobre la eliminación segura de medicamentos; (vi) invertir en una infraestructura para dicha eliminación, tal vez basada en incineradores de alta temperatura; e (vii) incluir la responsabilidad por parte de los usuarios en relación con los medicamentos caducados en los regímenes de responsabilidad rutinaria seguidos por el sector sanitario público.

Threats posed by stockpiles of expired pharmaceuticals in low- and middle-income countries: a Ugandan perspective

In some low- and middle-income countries, the national stores and public-sector health facilities contain large stocks of pharmaceuticals that are past their expiry dates. In low-income countries like Uganda, many such stockpiles are the result of donations. If not adequately monitored or regulated, expired pharmaceuticals may be repackaged and sold as counterfeits or be dumped without any thought of the potential environmental damage. The rates of pharmaceutical expiry in the supply chain need to be reduced and the disposal of expired pharmaceuticals needs to be made both timely and safe. Many low- and middle-income countries need to: (i)strengthen public systems for medicines' management, to improve inventory control and the reliability of procurement forecasts; (ii) reduce stress on central medical stores, through liberalization and reimbursement schemes; (iii) strengthen the regulation of drug donations; (iv) explore the salvage of officially expired pharmaceuticals, through re-analysis and possible shelf-life extension; (v) strengthen the enforcement of regulations on safe drug disposal; (vi) invest in an infrastructure for such disposal, perhaps based on ultra-high-temperature incinerators; and (vii) include user accountability for expired pharmaceuticals within the routine accountability regimes followed by the public health sector