Abstract
Background:
Honey is a natural antimicrobial agent with activity attributed to acidity, osmotic effects, hydrogen peroxide, and bioactive compounds. However, evidence regarding the antibacterial activity of Aceh linot honey against Gram-negative foodborne pathogens remains limited.

Aim:
This study aimed to evaluate the in vitro antibacterial activity of Aceh linot honey against Escherichia coli and Salmonella typhimurium.

Methods:
A laboratory-based experimental study was conducted using the Kirby–Bauer disc diffusion assay as a preliminary screening method. Linot honey was tested at concentrations of 20%, 40%, 60%, and 80% against reference strains of E. coli ATCC 25922 and S. typhimurium ATCC 14028. Ciprofloxacin (5 µg/disc) and chloramphenicol (30 µg/disc) were used as positive controls, and sterile distilled water was used as a negative control. Antibacterial activity was assessed by measuring the mean inhibition zone diameter and analyzed using one-way analysis of variance.

Results:
Linot honey exhibited concentration-dependent antibacterial activity against E. coli, with inhibition zones observed at ≥40%. The highest activity was recorded at 80% (8.8 ± 0.95 mm), indicating weak inhibition. No inhibitory effect was observed against S. typhimurium at any concentration. Statistical analysis showed significant differences among treatments for E. coli (P < 0.05).

Conclusion:
Aceh linot honey demonstrates selective antibacterial activity against E. coli but not against S. typhimurium under in vitro conditions. These findings indicate that linot honey may serve as a complementary natural antimicrobial agent. However, further studies using quantitative methods, such as minimum inhibitory concentration (MIC) and chemical characterization are required to confirm its antibacterial potency.

Key words: Antibacterial-activity; Escherichia coli; Linot honey; Salmonella typhimurium; Trigona itama.