Hoslundia opposita vahl; a potential source of bioactive compounds with antioxidant and antibiofilm activity for wound healing.

BACKGROUND: Biofilms and oxidative stress retard wound healing. The resistance of biofilms to antibiotics has led to a search for alternative approaches in biofilm elimination. Antioxidants work synergistically with antibacterial agents against biofilms. Hence recent research has suggested plants as candidates in the development of new alternatives in biofilm treatments and as antioxidants due to the presence of phytocompounds which are responsible for their bioactivities. Hoslundia opposita Vahl is one of the plants used by traditional healers to treat wounds and other infections, this makes it a potential candidate for drug discovery hence, in this study, we investigate the antibiofilm and antioxidant activity of methanolic extract of hoslundia opposita Vahl from Uganda. We also identify phytochemicals responsible for its bioactivity. METHOD: the plant was extracted by maceration using methanol, and the extract was investigated for antioxidant activity using 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) assay. The antibiofilm activity using microtiter plate assay (MTP) assay where the Minimum biofilm inhibitory concentration required to inhibit 50% or 90% of the biofilm (MBIC50 and MBIC90) and Minimum biofilm eradication concentration required to remove 50% or 90% of the biofilm (MBEC50 and MBEC90) were measured. It was further analysed for its phytochemical composition using quantitative screening, as well as Gas chromatography-mass spectrometry (GC-MS) and Liquid chromatography mass-spectrometry (LC-MS). RESULTS: H. Opposita Vahl extract showed good antioxidant activity with of 249.6 mg/mL. It inhibited the growth of P. aeruginosa and S. aureus biofilms with MBIC50 of 28.37 mg/mL and 10 mg/mL, respectively. It showed the ability to eradicate P. aeruginosa and S. aureus biofilms with MBEC50 of 23.85 and 39.01 mg/mL respectively. Phytochemical analysis revealed the presence of alkaloids, tannins, flavonoids, and phenols. GC-MS analysis revealed 122 compounds in the extract of which, 23 have evidence of antioxidant or antibiofilm activity in literature. The most abundant compounds were; 1,4- Citric acid, Tetracontane-1,40-diol (43.43.3%, 1, Olean-12-en-28-oic acid, 3-hydroxy-, methyl ester, (3.beta) (15.36%) 9-Octadecenamide (12.50%), Squalene (11.85%) Palmitic Acid 4TMS (11.28%), and alpha Amyrin (11.27%). The LC-MS identified 115 and 57 compounds in multiple reaction mode (MRM) and scan modes respectively. CONCLUSION: H. opposita Vahl showed antibiofilm and antioxidant activity due to bioactive compounds identified, hence the study justifies its use for wound healing. It can be utilised in further development of new drugs as antibiofilm and antioxidants.

In Vitro Evaluation of the Virulence Attributes of Oropharyngeal Candida Species Isolated from People Living with Human Immunodeficiency Virus with Oropharyngeal Candidiasis on Antiretroviral Therapy.

Background: Oropharyngeal Candida species are part commensal microflora in the the oral cavity of health individuals. Commensal Candida species can become opportunist and transition to pathogenic causes of oropharyngeal candidiasis (OPC) in individuals with impaired immunity through ecological cues and expression of virulence factors. Limited studies have evaluated virulence attributes of oropharyngeal Candida species among people living with human immunodeficiency virus (PLHIV) with OPC on antiretroviral therapy (ART) in Uganda. Objective: Evaluation of the Virulence Attributes of Oropharyngeal Candida Species Isolated from People Living with Human Immunodeficiency Virus with Oropharyngeal Candidiasis on Antiretroviral Therapy. Methods: Thirty-five (35) Candida isolates from PLHIV with OPC on ART were retrieved from sample repository and evaluated for phospholipase activity using the egg yolk agar method, proteinase activity using the bovine serum albumin agar method, hemolysin activity using the blood agar plate method, esterase activity using the Tween 80 opacity test medium method, coagulase activity using the classical tube method and biofilm formation using the microtiter plate assay method in vitro. Results: Phospholipase and proteinase activities were detected in 33/35 (94.3%) and 31/35 (88.6%) of the strains, respectively. Up to 25/35 (71.4%) of the strains exhibited biofilm formation while esterase activity was demonstrated in 23/35 (65.7%) of the strains. Fewer isolates 21/35 (60%) of the strains produced hemolysin and coagulase production was the least virulence activity detected in 18/35 (51.4%). Conclusion: Phospholipase and proteinase activities were the strongest virulence attributes of oropharyngeal Candida species.

Phenotypic Characterization and Antibiograms of Extended-Spectrum Beta-Lactamase-Producing Escherichia coli Isolated at the Human-Animal-Environment Interface Using a One Health Approach Among Households in Wakiso District, Uganda.

Background: The occurrence of extended spectrum beta-lactamase (ESBL) producing bacteria such as Escherichia coli has increasingly become recognized beyond hospital settings. Resistance to other types of antibiotics limits treatment options while the existence of such bacteria among humans, animals, and the environment is suggestive of potential zoonotic and reverse-zoonotic transmission. This study aimed to establish the antibiotic susceptibility profiles of the ESBL-producing Escherichia coli (ESBL-EC) from human, animal, and environmental isolates obtained among farming households within Wakiso district using a One Health approach. Methods: A total of 100 ESBL-EC isolates from humans 35/100 (35%), animals 56/100 (56%), and the environment 9/100 (9%) were tested for susceptibility to 11 antibiotics. This was done using the Kirby-Bauer disk diffusion method according to Clinical and Laboratory Standards Institute (CLSI) guidelines. Data were analyzed in STATA ver. 16 and graphs were drawn in Microsoft excel ver. 10. Results: Most of the ESBL-EC isolates (98%) were resistant to more than two antibiotics. ESBL-EC isolates were most susceptible to meropenem (MEM) (88.0%), and imipenem (82.0%) followed by gentamicin (72%). ESBL-EC isolates from humans were most susceptible to meropenem (MEM) followed by imipenem (IPM)> gentamicin (CN)> ciprofloxacin (CIP). Animal samples were more susceptible to MEM, IPM, and CN but were highly resistant to cefotaxime (CTX)> cefepime (FEP)>other antibiotics. Multidrug resistance (MDR) was mostly reported among households keeping goats under intensive husbandry practices. Seven percent of the isolates exhibited carbapenem resistance while 22% showed aminoglycoside resistance. Similar resistance patterns among humans, animals, and environmental samples were also reported. Conclusion: Our study provides baseline information on non-hospital-based MDR caused by ESBL-EC using a One Health approach. ESBL-EC isolates were prevalent among apparently healthy community members, animals, and their environment. It is important to conduct more One Health approach studies to generate evidence on the drivers, resistance patterns, and transmission of ESBL-producing organisms at the human-animal-environmental interface.

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.