Antibacterial Activities Of Apis And Meliponin Honey From Marigat Against Isolated Wound Bacterial Strains Among Patients Attending Nakuru CTRH

Abstract

Wound or burn infections refer to the colonization and invasion of bacteria in human skin or the underlying soft tissues in the body that ignite an immune response and lead to inflammation and injury of tissues. This study sought to establish the efficacy of natural honey produced from Baringo County Apis and Meliponines bees against some isolated and characterized microorganisms from 34 samples obtained from wound exudate from adult outpatients and surgical wards of The Nakuru County Referral Hospital. Honey samples were purchased from various known beekeepers in Marigat, Baringo County, and their initial qualitative, quantitative and proximate contents were determined using standard procedures. The targeted patients in this experimental study design attended The Nakuru County Referral Hospital with a mean ± SD age of 38.38 ± 16.88, utilizing a self-administered questionnaire to collect data and consent. According to Smith (2013), the value of ‘n’ sample size was 34 in the sampling technique. The sample size in honey (n=26) was determined according to Okur et al. (2020). The data from this study was analyzed using the IBM SPSS (Statistical Package for Social Sciences) statistics 23 and the variables were analyzed which were found to have a very highly significant difference (P< 0.005) through two-way ANOVA analysis using the Graph pad prism (version 7. The means were then compared using a parametric difference test–the T-test. These honey samples collected showed bacteriostatic and bactericidal effects compared to conventional antibiotics against the identified wound isolates by disc diffusion method. In the present study, Staphylococcus aureus was the most frequently isolated bacteria, with 15 (34.8%), followed by Pseudomonas aeruginosa – 12 (27.9%), Klebsiella pneumoniae – 10 (23.3%), and Escherichia coli – 6 (14.0%). Additional identification tests of the isolates involved the amplification of virulence factors genes encoded by specific primers that were Staphylococcus aureus (16Rrna and hla genes), Pseudomonas aeruginosa (gyrB and lasl genes), Klebsiella pneumoniae (magA and rmpA genes) and Escherichia coli (cnf1 and hlyA genes). The stingless bee honey provided mean values of 89.85 ± 5. 07g/100g, 3.86 ± 0.11 and 81.75 ± 10.35mg/g for sugar, pH and moisture, respectively, compared to the honeybee honey, which indicated 90.13 ± 5.76g/100g, 4.07 ± 0.08 and 114.28 ± 26.66 mg/g respectively. The phenolic compounds that acted as antioxidants were in the mean value of total phenolic compounds (92.18 ± 51.20mgGAE/100g), total flavonoids (23.70 ± 5.87 mg RE/100g) and total carotenoids (6.57 ± 0.21 mgβ–carotene/kg) for stingless bee honey compared to honeybee 81 ± 36.25mgGAE/100g, 21.83 ± 6. 16mgRE/100g and 4.41 ± 2.07mgβ –carotene/kg respectively. The isolates showed a differing level of resistance to honey-incorporated discs (10 x 104, 20 x 104, 50 x 104 and 75 x 104 μg/ml) with a mean zone of inhibition of 18.23 ± 0.4mm (Staphylococcus aureus), 17.49 ± 0.3mm (Pseudomonas aeruginosa), 16.05 ± 0.6mm (Klebsiella pneumoniae) and 10.19 ± 0.5mm (Escherichia coli) with a mean range of 14.54 ± 2.0mm to 17.58 ± 3mm and 10.81 ± 2.5mm to 19.4 ± 4.3mm for both honeybee and stingless bee honey respectively. The mean inhibition diameters of the used antibiotics (Gentamycin, Levofloxacin, Ampicillin, Tazobactum, Meropenem and Chloramphenicol) were 17.6 ± 0.5mm (Staphylococcus aureus), 8.9 ± 0.2mm (Pseudomonas aeruginosa), ix 9.3 ± 0.2mm (Klebsiella pneumoniae) and 11.5 ± 0.3mm (Escherichia coli). Hence, the high prevalence of reduced antibiotic susceptibility amongst most bacterial wound isolates implies that better strategies should be deployed to improve wound treatment and healing.’