Acute and subacute toxicity profile of ethanolic stem bark extract of Albizia coriaria Welw. ex Oliv. in Wistar albino rats.

Albizia coriaria (Fabaceae) crude extracts are key ingredients of several licensed and unlicensed herbal products in East Africa. However, there is limited and often contradicting information regarding its toxicity. We therefore evaluated the acute and subacute toxicity of the ethanolic stem bark extract of A. coriaria in mature healthy Wistar albino rats following Lorke's method and OECD guidelines 407. The LD50 of the ethanolic stem bark extract of A. coriaria was 2000 mg/kg. The acute toxicity signs observed included piloerection, hyperventilation, lethargy, and loss of righting reflex. There was a significant increase in aspartate aminotransferase, alkaline phosphatase, red blood cells and haemoglobin in rats after 28 days at the dose of 500 mg/kg. Histological analyses revealed multifocal random parenchymal necrosis and scattered periportal mononuclear inflammatory cells infiltration in the liver, interstitial nephritis in the kidney and multifocal lymphoid accumulation in the peribronchiolar and perivascular lung tissue at 500 mg/kg. The ethanolic stem bark of A. coriaria was therefore moderately toxic to the rats when administered in a single high oral dose within 24 h. The extract caused a dose dependent toxicity with significant damage to the kidney, liver and lung tissues at a dose of 500 mg/kg after 28 days. Herbal medicines containing A. coriaria extracts should be consumed cautiously due to likelihood of toxicity particularly at higher doses greater than 500 mg/kg.

Immunological and biochemical biomarker alterations among SARS-COV-2 patients with varying disease phenotypes in Uganda.

Every novel infection requires an assessment of the host response coupled with identification of unique biomarkers for predicting disease pathogenesis, treatment targets and diagnostic utility. Studies have exposed dysregulated inflammatory response induced by the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) as significant predictor or cause of disease severity/prognosis and death. This study evaluated inflammatory biomarkers induced by SARS-CoV-2 in plasma of patients with varying disease phenotypes and healthy controls with prognostic or therapeutic potential. We stratified SARS-CoV-2 plasma samples based on disease status (asymptomatic, mild, severe, and healthy controls), as diagnosed by RT-PCR SARS-CoV-2. We used a solid phase sandwich and competitive Enzyme-Linked Immunosorbent Assay (ELISA) to measure levels of panels of immunological (IFN-γ, TNF-α, IL-6, and IL-10) and biochemical markers (Ferritin, Procalcitonin, C-Reactive Protein, Angiotensin II, Homocysteine, and D-dimer). Biomarker levels were compared across SARS-CoV-2 disease stratification. Plasma IFN-γ, TNF-α, IL-6, and IL-10 levels were significantly (P < 0.05) elevated in the severe SARS-CoV-2 patients as compared to mild, asymptomatic, and healthy controls. Ferritin, Homocysteine, and D-dimer plasma levels were significantly elevated in severe cases over asymptomatic and healthy controls. Plasma C-reactive protein and Angiotensin II levels were significantly (P < 0.05) higher in mild than severe cases and healthy controls. Plasma Procalcitonin levels were significantly higher in asymptomatic than in mild, severe cases and healthy controls. Our study demonstrates the role of host inflammatory biomarkers in modulating the pathogenesis of COVID-19. The study proposes a number of potential biomarkers that could be explored as SARS-CoV-2 treatment targets and possible prognostic predictors for a severe outcome. The comprehensive analysis of prognostic biomarkers may contribute to the evidence-based management of COVID-19 patients.

Comparison of the Nutritional Status of Swiss Albino Mice Fed on Either a Purified or Cereal-Based Diet for 15 weeks.

Background: Laboratory animals are commonly fed on cereal-based diets (CBDs) whose nutrient composition is unknown and may confound the metabolic response to study interventions. Purified diets such as AIN-93M are therefore recommended, as their nutrient composition is known. However, few studies have evaluated their use as adequate control diets. The aim of this study was to compare the nutrition status of Swiss albino mice fed on either CBD or AIN-93M for 15 weeks. Methods: Twenty Swiss albino mice aged 6-8 weeks and weighing 21.7 g ± 0.6 were fed on either CBD or AIN-93M diet for 15 weeks. Their nutritional status was evaluated using anthropometric and hematological indices, serum glucose, total protein, albumin, and total cholesterol to select an appropriate normal control diet. Results: The CBD had low-calorie content (2.57 kcal/g) and protein (11 ± 3.8 g/100 g) compared to AIN-93M (3.8 kcal/g and 14 g/100 g, respectively). The BMI of male mice fed on CBD and AIN-93M diets was significantly higher (P=0.0139 and P=0.0325, respectively) compared to that of females fed on similar diets. Animals in the CBD group had lower hemoglobin (15.1-16.9 g/dl) compared to those in the AIN-93M group (18.1-20.8 g/dl). Serum albumin levels were higher in both male (P=0.001) and female (P=3 × 10-6) mice fed on AIN-93M compared to those fed on CBD. Females in the AIN-93M group had higher cholesterol (P=0.026) than those in the CBD group. Conclusion: The AIN-93 diet of caloric value 3.85 kcal/g (total protein 14 g, total fat 4 g of soy bean oil, fibre 5 g, and total carbohydrate 42 g per 100 g) can be safely used as a normal control diet in long-term research studies using Swiss albino mice.

Mapping hepatitis B virus genotypes on the African continent from 1997 to 2021: a systematic review with meta-analysis.

Hepatitis B virus (HBV) has ten genotypes (A-J) and over 40 sub-genotypes based on the divergence of ≥ 8% and 4 to < 8% in the complete genome respectively. These genotypes and sub-genotypes influence the disease prognosis, response to therapy and route of viral transmission. Besides, infection with mixed genotypes and recombinant genotypes has also been reported. This study aimed at mapping the de novo genotypes and correlate them with the immigration trends in order to inform future research on the underlying reasons for the relative distribution of HBV genotypes from a large sample size pooled from many primary studies. Data was extracted from 59 full research articles obtained from Scopus, PubMed, EMBASE, Willy library, African Journal Online (AJOL) and Google Scholar. Studies that investigated the genotypes, sub-genotypes, mixed genotypes and recombinant were included. The Z-test and regression were used for the analysis. The study protocol is registered with PROSPERO under the registration number CRD42022300220. Overall, genotype E had the highest pooled prevalence significantly higher than all the other genotypes (P < 0.001). By region, genotype A posted the highest pooled prevalence in eastern and southern Africa, E in west Africa and D in north Africa (P < 0.0001). Regarding the emerging genotypes B and C on the African continent, genotype B was significantly higher in south Africa than C (P < 0.001). In contrast, genotype C was significantly higher in east Africa than west Africa (P < 0.0001). The A1 and D/E were the most diverse sub-genotypes and genotype mixtures respectively. Finally, we observed a general progressive decrease in the prevalence of predominant genotypes but a progressive increase in the less dominant by region. Historical and recent continental and intercontinental migrations can provide a plausible explanation for the HBV genotype distribution pattern on the African continent.

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.

Polycyclic aromatic hydrocarbons in breast milk of nursing mothers: Correlates with household fuel and cooking methods used in Uganda, East Africa.

Maternal breast milk, which is a complete food for the infant's growth, development, and health, contains fats and lipids making it susceptible to accumulation of lipophilic compounds like polycyclic aromatic hydrocarbons (PAHs). This study aimed at analyzing correlates of measured levels of PAHs in breast milk of nursing mothers to frequently used household fuels and cooking methods in Uganda, and estimate the potential health risks of PAHs to infants through breastfeeding. Sixty breast milk samples were collected from healthy and non-smoking mothers who had lived in Kampala capital city (urban area) and Nakaseke district (rural area) for at least five years. Sample extracts were analyzed for PAHs using a gas chromatograph coupled with a triple quadrupole mass spectrometer. ∑13PAHs in samples from Kampala ranged from 3.44 to 696 ng/g lw while those from Nakaseke ranged from 0.84 to 87.9 ng/g lw. PAHs with 2-3 rings were more abundant in the samples than PAHs with 4-6 rings. At least 33 % of the variance in the levels of ∑13PAHs in the breast milk samples was attributable to the fuel type and cooking methods used. Nursing mothers who used charcoal for cooking accumulated higher levels of ∑13PAHs in their breast milk samples compared to those who used firewood. Levels of ∑13PAHs in breast milk of mothers increased depending on the cooking methods used in the order; boiling< grilling< deep-frying. In all samples, hazard quotients for PAHs were <1 and estimated incremental cancer risks were all between 10-6 and 10-4, indicating that the health risks to infants due to the ingestion of PAHs in breast milk was tolerable. Further studies with large datasets on PAHs and their derivatives and, larger samples sizes are needed to confirm these findings.

Hepatitis B virus (HBV) serological patterns among the HBsAg negative hospital attendees screened for immunization.

The Hepatitis B virus (HBV) is a highly infectious virus and is endemic in Uganda. It is one of the major etiological agents for liver diseases including liver cancer. In this work, we evaluated the prevalence of the HBV serological markers and the associated socio-demographic factors among hepatitis B surface antigen (HBsAg) seronegative persons screened during routine immunization against the virus in eastern Uganda. Data on the socio-demographic characteristics were collected using a structured questionnaire, while that on the serological markers were obtained from serum samples and evaluated by using the 5-panel HBV One Step Hepatitis B Virus Combo Test Device (FastepR, HBV-P43M). The following markers were evaluated by the panel: HBsAg, HBsAb, HBcAb, and HBeAb. Data were analyzed using SPSS (version 26), and multinomial logistic regression was used to elicit the adjusted odds ratio. All the analysis were performed at a 95% confidence limit, and a P value ≤ 0.05 was considered significant. The 424 participants included in this study were mainly female (62.3%), married (55.4%) and aged 30 years and above (54.2%). The seropositivity of the HBsAb, HBeAb, HBcAb marker prevalence rates was 48(11.3%), 73(17.2%) and 45(10.6%) respectively. The majority of the participants (327, 77.1%) did not present with any marker. Married paricipants were significantly associated with reduced HBsAb seropositvity rate, whereas young people aged 18-29 years were associated the with increased odds of HBsAb seropositivity (p < 0.05). Male participants were significantly associated with the HBeAb and HBcAb seropositivity (p < 0.05). Similarly, contact with an HBV infected person was significantly associated with HBeAb and HBcAb seropositivity (p < 0.05). Further still, blood transfusion was significantly associated with the increased risk of HBcAb seropositivity (P < 0.05). This study has revealed a prevalence of HBV serological markers among the HBsAg seronegative persons in this community and an increased risk of transmission of the virus in the community. Our findings have key consequences pertaining the interventions that are pertinent in the control and prevention of the spread of the virus among apparently health persons.

Disproportionate Distribution of HBV Genotypes A and D and the Recombinant Genotype D/E in the High and Low HBV Endemic Regions of Uganda: A Wake-Up Call for Regional Specific HBV Management.

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 < 0.05 was considered statistically significant. RESULTS: Majority of our study subjects were female (64.5%), youth (51.0%), and married (62.0%). Overall, genotype A was the most prevalent (46%). Genotype D and the recombinant genotype D/E were proportionately more distributed in the high endemic (38.2%) and low endemic (36.5%) regions, respectively. Genotype D was significantly more prevalent in the high endemic region and among the elderly (p < 0.05). Genotype D was significantly associated with elevated viral load and direct bilirubin (p < 0.05). The recombinant genotype D/E was significantly associated with elevated viral load (p < 0.05). Similarly, genotype A was significantly associated with elevated AST and GGT, lowered viral load, and normal direct bilirubin levels (p < 0.05). CONCLUSION: There is disproportionate distribution of genotypes A and D and the recombinant genotype D/E in the low and high endemic regions of Uganda. This probably could explain the differences in endemicity of HBV in our country signifying the need for regional specific HBV management and control strategies.

Knowledge, perceptions and effects of Ebola virus disease outbreak on the pig value chain in the agro-pastoralist district of Luwero, Central Uganda.

BACKGROUND: Ebola Virus Disease (EVD) outbreaks have a significant impact on the health and wellbeing, and livelihoods of communities. EVD response interventions particularly affect the food value chain, and income security of pig farmers in agro-pastoral communities. Despite the enormous effort of EVD response interventions, there is paucity of information towards EVD among those involved in the pig value chain, as well as the effect of EVD outbreaks on the pig value chain. This study therefore, assessed the knowledge, perceptions on the occurrence of Ebola and its effects on the pig value chain in the agro-pastoral district of Luweero, Central Uganda. METHODS: A cross sectional study was conducted in two parishes of Ssambwe and Ngalonkulu, Luwero district. A total of 229 respondents were included in the study. Structured questionnaires, key informant interviews and focus group discussions were conducted to collect data. Quantitative data was analysed using SPSS version 22 while qualitative data was analysed using thematic content analysis. RESULTS: Of the 229 respondents, 95.6% could recall the occurrence of the last EVD outbreak in their locality. About 24.5% associated EVD with touching pigs or eating pork. Regarding knowledge, 194 (84.7%) correctly associated EVD with handling Ebola infected persons, 191 (83.4%) with migration of people from endemic areas, 148 (64.9%) eating monkey meat, 127 (55.5%) with eating bats, and 198 (64.9%) with conducting public meetings where there is an Ebola infected person. Out of 142 farmers, 55 (38.7%) believed that Ebola outbreaks affected demand and sale of pigs. The EVD outbreak significantly led to a reduction in the average number of pigs sold (P = 0.001), the average number of pigs bought by traders (P = 0.04), and the number of pigs sold/ slaughtered by butcher men at pork eating places (P = 0.03). CONCLUSION: This study showed that EVD outbreak negatively affected the pig value chain i.e., the demand and supply of pigs and pork. Therefore, there is need to sensitize the stakeholders in the pig value chain on EVD in order to minimize the negative economic impacts associated with EVD outbreaks.

A Descriptive-Multivariate Analysis of Community Knowledge, Confidence, and Trust in COVID-19 Clinical Trials among Healthcare Workers in Uganda.

Background-misinformation and mistrust often undermines community vaccine uptake, yet information in rural communities, especially of developing countries, is scarce. This study aimed to identify major challenges associated with coronavirus disease 2019 (COVID-19) vaccine clinical trials among healthcare workers and staff in Uganda. Methods-a rapid exploratory survey was conducted over 5 weeks among 260 respondents (66% male) from healthcare centers across the country using an online questionnaire. Twenty-seven questions assessed knowledge, confidence, and trust scores on COVID-19 vaccine clinical trials from participants in 46 districts in Uganda. Results-we found low levels of knowledge (i.e., confusing COVID-19 with Ebola) with males being more informed than females (OR = 1.5, 95% CI: 0.7-3.0), and mistrust associated with policy decisions to promote herbal treatments in Uganda and the rushed international clinical trials, highlighting challenges for the upcoming Oxford-AstraZeneca vaccinations. Knowledge, confidence and trust scores were higher among the least educated (certificate vs. bachelor degree holders). We also found a high level of skepticism and possible community resistance to DNA recombinant vaccines, such as the Oxford-AstraZeneca vaccine. Preference for herbal treatments (38/260; 14.6%, 95% CI: 10.7-19.3) currently being promoted by the Ugandan government raises major policy concerns. High fear and mistrust for COVID-19 vaccine clinical trials was more common among wealthier participants and more affluent regions of the country. Conclusion-our study found that knowledge, confidence, and trust in COVID-19 vaccines was low among healthcare workers in Uganda, especially those with higher wealth and educational status. There is a need to increase transparency and inclusive participation to address these issues before new trials of COVID-19 vaccines are initiated.

Multiple persistent organic pollutants in mothers' breastmilk: Implications for infant dietary exposure and maternal thyroid hormone homeostasis in Uganda, East Africa.

Persistent organic pollutants (POPs) are ubiquitous contaminants with adverse health effects in the ecosystem. One of such effects is endocrine disruption in humans and wildlife even at background exposure concentrations. This study assessed maternal breastmilk concentrations of POPs; brominated flame retardants (BFRs), polychlorinated biphenyls (PCBs) and polychlorinated dibenzo-p-dioxins/furans (PCDD/Fs), and the potential health risks posed to the nursing infants. We also evaluated the association of these POPs with total 3,3',5-triiodo-L-thyronine (T3), L-thyroxine (T4), and 3,3',5'-triiodo-L-thyronine (rT3) levels measured in human breast milk. Thirty breastmilk samples were collected from Kampala, Uganda between August and December 2018. Hexabromobenzene was not detected while the maximum level of 2,2',4,4',5,5'-hexabrombiphenyl was 64.7 pg/g lw. The median levels of total indicator PCBs, PBDEs, dioxin-like PCBs, and PCDD/Fs in the samples were 159 pg/g lw, 511 pg/g lw, 1.16 pg TEQ/g lw, and 0.4 pg TEQ/g lw, respectively. These levels were lower than those reported in other countries. Owing to their bio accumulative nature, PCBs -81, -169, and ∑PCDD/Fs increased with increase in maternal age. Estimated dietary intakes for dioxin-like PCBs and PCDD/Fs were lower than those reported elsewhere but were higher than the WHO tolerable daily intakes suggesting potential health risks to nursing infants. In adjusted single pollutant models, PCB-126, PCB-169, and ∑PCBTEQ were negatively associated with T3, while 1,2,3,4,5,7,8-HpCDF was positively associated with rT3. Although these associations did not persist in multipollutant models, our findings suggest potential thyroid hormone disruption by POPs in mothers. This may reduce the levels of thyroid hormones transferred from the mother to the neonates and, hence, adversely influence infant growth. A temporal study with a bigger sample size is required to corroborate these findings.

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.

New Putative Antimicrobial Candidates: In silico Design of Fish-Derived Antibacterial Peptide-Motifs.

Antimicrobial resistance remains a great threat to global health. In response to the World Health Organizations' global call for action, nature has been explored for novel and safe antimicrobial candidates. To date, fish have gained recognition as potential source of safe, broad spectrum and effective antimicrobial therapeutics. The use of computational methods to design antimicrobial candidates of industrial application has however, been lagging behind. To fill the gap and contribute to the current fish-derived antimicrobial peptide repertoire, this study used Support Vector Machines algorithm to fish out fish-antimicrobial peptide-motif candidates encrypted in 127 peptides submitted at the Antimicrobial Peptide Database (APD3), steered by their physico-chemical characteristics (i.e., positive net charge, hydrophobicity, stability, molecular weight and sequence length). The best two novel antimicrobial peptide-motifs (A15_B, A15_E) with the lowest instability index (-28.25, -22.49, respectively) and highest isoelectric point (pI) index (10.48 for each) were selected for further analysis. Their 3D structures were predicted using I-TASSER and PEP-FOLD servers while ProSA, PROCHECK, and ANOLEA were used to validate them. The models predicted by I-TASSER were found to be better than those predicted by PEP-FOLD upon validation. Two I-TASSER models with the lowest c-score of -0.10 and -0.30 for A15_B and A15_E peptide-motifs, respectively, were selected for docking against known bacterial-antimicrobial target-proteins retrieved from protein databank (PDB). Carbapenam-3-carboxylate synthase (PDB ID; 4oj8) yielded the lowest docking energy (-8.80 and -7.80 Kcal/mol) against motif A15_B and A15_E, respectively, using AutoDock VINA. Further, in addition to Carbapenam-3-carboxylate synthase, these peptides (A15_B and A15_E) were found to as well bind to membrane protein (PDB ID: 1by3) and Carbapenem synthetase (PDB: 1q15) when ClusPro and HPEPDOCK tools were used. The membrane protein yielded docking energy scores (DES): -290.094, -270.751; coefficient weight (CW): -763.6, 763.3 for A15_B and A15_E) whereas, Carbapenem synthetase (PDB: 1q15) had a DES of -236.802, -262.75 and a CW of -819.7, -829.7 for peptides A15_B and A15_E, respectively. Motif A15_B of amino acid positions 2-19 in Pleurocidin exhibited the strongest in silico antimicrobial potentials. This segment could be a good biological candidate of great application in pharmaceutical industries as an antimicrobial drug candidate.

Fluoroquinolone resistant bacterial isolates from the urinary tract among patients attending hospitals in Bushenyi District, Uganda.

INTRODUCTION: bacterial resistance to fluoroquinolones is on the rise globally, bacteria causing urinary tract infections (UTIs) are no exception to this fact. Judicious use of the current antibiotics by clinicians is therefore deemed necessary to combat development of resistance. This study determined fluoroquinolone resistant profiles, multiple antibiotic resistance indices (MARI), factors associated with fluoroquinolone resistance and their strength among patients attending hospitals in Bushenyi District, Uganda. METHODS: this was a cross-sectional study in which a total of 86 bacterial uropathogens isolated previously by standard microbiological methods were subjected to antibiotic susceptibility testing using Kirby Bauer disk diffusion method. Data for factors suspected to be associated with fluoroquinolone resistant UTI were obtained by use of questionnaires. RESULTS: the most resisted fluoroquinolone was ofloxacin with 29/83 (34.9%), followed by moxifloxacin 27/83 (32.5%), levofloxacin 24/86 (27.9%) and ciprofloxacin 23/86 (26.7%). The bacterial uropathogens that exhibited the highest frequency of fluoroquinolone resistant strains were P. mirabilis with 2/3 (66.7%) and E. faecalis with 2/3 (66.7%), followed by E. coli 19/36 (52.8%), S. aureus 13/27 (48.1%), K. oxytoca 2/6 (33.3%), K. pneumoniae 2/10 (20.0%) and P. vulgaris 0/1 (0.0%). All the bacterial uropathogens tested showed MARI of ≥ 0.2. Hospitalization, history of fluoroquinolones use in the last 12 months and wrong prescription of antibiotics were found to bear statistically significant relationships (p < 0.05) with fluoroquinolone resistant UTI. CONCLUSION: antibiotic susceptibility testing of the first generation quinolones such as nalidixic acid in hospitalized patients, patients with history of fluoroquinolones' use in the last 12 months and wrong prescription of antibiotics should be adopted to avoid fluoroquinolone abuse. For empiric treatment of UTIs in Bushenyi District, ciprofloxacin still remains the first line of choice among the fluoroquinolone class of antibiotics.

Optimisation of template preparation and laboratory evaluation of the Loopamp™ Trypanosoma brucei kit for detection of parasite DNA in blood.

The Loopamp™ Trypanosoma brucei Detection Kit is the latest addition of molecular techniques for amplification of parasite DNA in biological materials. We have evaluated the kit on a number of preparations of crude templates from the blood of experimentally infected rodents, to provide the best option that can be extrapolated to resource-poor healthcare settings where human African trypanosomiasis (HAT) is endemic. We used rodent blood spiked with T. b. brucei at various serial dilutions to test whole blood, that was concentrated by differential lysis of red blood cells (RBCs) followed by centrifugation, or buffy coat samples recovered from whole blood after centrifugation. We also tested crude templates produced after lysis of blood with sodium dodecyl sulphate (SDS) or Triton X, and storage for up to 28 days at room temperature after spotting on filter paper or glass slides. Concentration by RBC lysis provided the highest analytical sensitivity (0.04 trypanosomes/ml), closely followed by the much cheaper SDS at 0.1 trypanosomes/ml sensitivity. We also monitored the persistence of DNA in lysed blood dried onto filter papers by testing them weekly with the LAMP kit and by PCR for the 177bp repeats characteristic of the T. brucei subspecies. At a concentration of 100 trypanosomes/ml, signals indicating presence of parasite DNA could be detected up to week 10, while at 10 trypanosomes/ml detection of signals was limited to week 4. Thus, an ordinary filter paper provides a convenient medium for preservation of trypanosome DNA at ambient conditions for use with the LAMP kit in the short run. Lysis of samples with SDS enhanced sensitivity by facilitating parasite DNA availability. This opens the avenue to incorporate LAMP in routine algorithms for HAT diagnosis and surveillance, as well as for monitoring elimination programs.

Comparison of HIV drug resistance profiles across HIV-1 subtypes A and D for patients receiving a tenofovir-based and zidovudine-based first line regimens in Uganda.

BACKGROUND: Resistance to antiretroviral drugs is a major challenge among Human Immunodeficiency Virus (HIV) positive patients receiving antiretroviral therapy (ART). Mutations that arise as a result of this are diverse across the various drugs, drug classes, drug regimens and subtypes. In Uganda, there is a paucity of information on how these mutations differ among the different drug regimens and the predominant HIV-1 subtypes. The purpose of this study was to determine mutation profile differences between first-line drug regimens: TDF/3TC/EFV and AZT/3TC/EFV and HIV-1 subtypes: A and D in Uganda. The study also investigated the potential usage of rilpivirine, doravirine and etravirine in patients who failed treatment on efavirenz. METHODS: A retrospective study was conducted on 182 archived plasma samples obtained from patients who were experiencing virological failure between 2006 and 2017 at five Joint Clinical Research Center (JCRC) sites in Uganda. Sanger sequencing of the Reverse Transcriptase (RT) gene from codons 1-300 was done. Mutation scores were generated using the Stanford University HIV Drug Resistance Database. A Chi-square test was used to determine the association between drug resistance mutations (DRMs) and drug regimens or HIV-1 subtypes. RESULTS: The prevalence of DRMs was 84.6% among patients failing a first-line efavirenz (EFV)-based regimen. The most prevalent Nucleoside Reverse Transcriptase Inhibitor (NRTI) mutations were M184V/I (67.3%), K219/Q/E (22.6%) and K65R (21.1%). While K103N (50.8%) and G190A/S/E/G (29.1%) were the most prevalent Non-Nucleoside Reverse Transcriptase Inhibitor (NNTRI) mutations. As expected, discriminatory DRMs such as K65R, L74I, and Y115F were noted in Tenofovir (TDF) containing regimens while the Thymidine Analogue Mutations (TAMs) L210W and T215 mutations were in Zidovudine (AZT)-based regimens. No significant difference (p = 0.336) was found for overall DRMs between HIV-1 subtypes A and D. Among the patients who had resistance to EFV, 37 (23.6%) were susceptible to newer NNRTIs such as Rilpivirine and Etravirine. CONCLUSION: Accumulation of DRMs between AZT/3TC/EFV and TDF/3TC/EFV is comparable but individual mutations that confer resistance to particular drugs should be considered at virological failure. Having either HIV-1 subtype A or D is not associated with the acquisition of DRMs, therefore HIV diversity should not determine the choice of treatment. Rilpivirine, etravirine and doravirine had minimal benefits for patients who failed on efavirenz.

Application of one health approach in training at Makerere University: experiences from the one health workforce project in Uganda.

The interconnections of humans, domestic animals, wildlife and the environment have increasingly become complex, requiring innovative and collaborative approaches (One Health approach) for addressing global health challenges. One Health is a multidisciplinary and multi-sectoral collaborative approach to human, animal, plant and environmental health. The role of academia in training professionals oriented in One Health is critical in building a global workforce capable of enhancing synergies of various sectors in improving health. Makerere University, Uganda has implemented pre-service capacity building initiatives aimed to foster One Health competencies among students who are future practitioners. In addition to incorporating the One Health concept in didactic curricula, Student One Health Innovation Clubs, undergraduate field placements in 11 demonstration sites, graduate fellowships, small grants to support research and innovations, and cross-college collaborative training approaches have greatly aided the assimilation of One Health into the fabric of university offerings. Partnerships with government ministries, private sector and international agencies were initiated to benefit the students, as well as chart a path for experiential learning and in-service offerings in the future. One major challenge, however, has been the tendency to focus on infectious diseases, especially zoonoses, with less consideration of other health issues. The opportunity for improvement, nonetheless, lies in the increasing emerging and re-emerging health concerns including epidemics, environmental pollution and related challenges which justify the need for countries and institutions to focus on building and strengthening multidisciplinary health systems.

Plasma cytokine profiles associated with rhodesiense sleeping sickness and falciparum malaria co-infection in North Eastern Uganda.

BACKGROUND: Immunological Human African Trypanosomiasis (HAT) studies often exclude malaria, although both infections overlap in specific endemic areas. During this co-infection, it is not known whether this parasitic interaction induces synergistic or antagonistic cytokine response among humans. This study determined prevalence of Plasmodium falciparum malaria among Trypanosoma brucei rhodesiense HAT and plasma cytokine profile levels associated with HAT and/or malaria infections. METHODS: Participants were recruited at Lwala hospital in north eastern Uganda: healthy controls (30), malaria (28), HAT (17), HAT and malaria (15) diagnosed by microscopy and PCR was carried out for parasite species identification. Plasma cytokine levels of Interferon-gamma (IFN-γ), Tumour Necrosis Factor-alpha (TNF-α), Interleukin (IL)-6, IL-10 and Transforming Growth Factor-beta (TGF-ß) were measured by sandwich Enzyme-Linked Immuno Sorbent Assay and data statistically analysed using Graphpad Prism 6.0. RESULTS: The prevalence of P. falciparum malaria among T. rhodesiense HAT cases was high (46.8%). Malaria and/or HAT cases presented significant higher plasma cytokine levels of IFN-γ, TNF-α, IL-6, IL-10 and TGF-ß than healthy controls (P < 0.05). Levels of IFN-γ, IL-6 and IL-10 were significantly elevated in HAT over malaria (P < 0.05) but no significant difference in TNF-α and TGF-ß between HAT and malaria (P > 0.05). Co-infection expressed significantly higher plasma IFN-γ, IL-6, and IL-10 levels than malaria (P < 0.05) but no significant difference with HAT mono-infection (P > 0.05). The TNF-α level was significantly elevated in co-infection over HAT or malaria mono-infections (P < 0.05) unlike TGF-ß level. Significant positive correlations were identified between IFN-γ verses TNF-α and IL-6 verses IL-10 in co-infection (Spearman's P < 0.05). CONCLUSIONS: The T. b. rhodesiense significantly induced the cytokine response more than P. falciparum infections. Co-infection led to synergistic stimulation of pro-inflammatory (IFN-γ, TNF-α), and anti-inflammatory (IL-6, and IL-10) cytokine responses relative to malaria mono-infection. Level of TNF-α partially indicates the effect induced by T. b. rhodesiense and P. falciparum mono-infections or a synergistic interaction of co-infections which may have adverse effects on pathogenesis, prognosis and resolution of the infections.Trial registration VCD-IRC/021, 26/08/2011; HS 1089, 16/01/2012.

Seroprevalence of brucellosis and risk factors associated with its seropositivity in cattle, goats and humans in Iganga District, Uganda.

INTRODUCTION: The burden of brucellosis among smallholder farmers is poorly-documented in Uganda. The disease burden is likely to be high, given the high levels of endemicity, lots of exposures and due to lack of control measures. In order to designate appropriate control measures, the magnitude and risk factors for brucellosis need to be known. We established the burden of and risk factors for Brucella seropositivity in cattle, goats, and humans in Iganga district, eastern Uganda. METHODS: A cross-sectional study was conducted in in Kigulamo Parish, Iganga District. We enrolled 226 households and administered a structured questionnaire to heads of households to capture data on socio-demographic characteristics, human brucellosis-related risk factors, and livestock farming practices. Human, cattle, and goat blood samples were collected and tested serologically using commercial indirect-ELISA kits manufactured by USDA, USA. RESULTS: Of 451 human blood samples, 20 (4.4%) were positive. Among 345 cattle blood samples, 4 (1.2%) were positive and among 351 goat blood samples, one (0.3%) was positive. Persons who reported consuming locally-made dairy products had 4 times higher odds of Brucella seropositivity (OR = 4.0, CI = 1.14-14.03, p = 0.031) than those who did not. None of the risk factors we asked about were significantly associated with seropositivity in cattle and goats. CONCLUSION: The seroprevalence of brucellosis in humans in smallholder households in Kigulamo was relatively low and associated with consumption of locally made dairy products. No risk factors were significantly associated with seropositivity in livestock, likely due to the small number of seropositive animals. We recommend a One Health approach to control brucellosis simultaneously in animals and humans needed to sustainably reduce the burden of brucellosis in Uganda and beyond.

Prevalence of Dermatophytosis and Antifungal Activity of Ethanolic Crude Leaf Extract of Tetradenia riparia against Dermatophytes Isolated from Patients Attending Kampala International University Teaching Hospital, Uganda.

Dermatophyte infections are a global health problem but neglected in Uganda. This work aimed at determining prevalence of dermatophytosis and antifungal activity of ethanolic crude leaf extract of Tetradenia riparia against dermatophytes isolated from patients attending Kampala International University Teaching Hospital (KIU-TH), Uganda. A total of 100 samples of skin and nail scrapings were collected and processed using standard microscopy (KOH) and cultural methods. T. riparia leaves were collected and processed with 95% ethanol using standard extraction method. The crude leaves ethanolic extract was tested against three dermatophytes: Trichophyton tonsurans, T. mentagrophyte, and Microsporum audouinii using modified agar well diffusion method. Minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of the ethanolic leaves crude extract were also determined using broth tube dilution and culture, respectively. Out of 100 samples collected, 49 (49%, 95%CI: 0.3930-0.5876) were found positive for microscopy. The prevalence of dermatophytosis was significantly (p=0.001) associated with age groups of participants with higher infection among those aged 11-20 and 21-30 years with 75.0% each. Out of the 49 that were positive by microscopy, 28 (57.15%, 95% CI: 0.1987-0.3739) were positive by culture. Thirty-one (31) fungal isolates were obtained which included both dermatophyte and non-dermatophyte fungi. T. verrucosum had highest distribution 6 (19.35%) among dermatophytes species while Aspergillus spp. were found to have highest distribution 7 (22.58%) among non-dermatophyte species. The result of the antidermatophytic test showed that T. riparia ethanolic crude leaves extract had activity against tested dermatophytes at 1 g/ml. MIC and MFC of the crude extract of T. riparia against tested dermatophytes ranged from 62.5 to 250 mg/ml and 125 to 500 mg/ml, respectively. The findings of this study reported the presence of dermatophytes causing dermatophytosis among patients attending KIU-TH. The results of the current study showed that T. riparia leaves ethanolic crude extract has antidermatophytic activity against tested dermatophytes.