Aerobic culturable bacterial burden, biofilm formation, antimicrobial susceptibility, and fungal recovery from household and kindergarten toys in Misurata, Libya: a cross-sectional study

Abstract Background Children’s toys are frequently handled surfaces that may retain culturable microorganisms and facilitate repeated environmental exposure. Evidence from Libya integrating quantitative aerobic culturable bacterial burden, biofilm-forming capacity, and antimicrobial susceptibility of microorganisms recovered from household and kindergarten toys remains limited. This study compared these characteristics among a quota-convenience sample of children’s toys from participating sites in Misurata, Libya; the design was not intended to estimate citywide prevalence. Results A total of 800 toys from 40 households and 20 kindergartens were sampled across four regions of Misurata. Kindergarten toys had a higher mean aerobic culturable bacterial burden than household toys (3.140 versus 2.807 log₁₀ CFU/25 cm²; mean difference, 0.333; 95% CI, 0.286–0.380; P < 0.001). In a multivariable linear mixed-effects model accounting for clustering within establishments, kindergarten setting remained associated with higher burden (β = 0.341; 95% CI, 0.298–0.383; P < 0.001). Shared use, fabric material, and poorer observed cleanliness were also associated with higher burden. One selected representative culturable morphotype was characterized per toy; 664 records were bacterial and 136 fungal. These proportions reflect the operational one-morphotype-per-toy selection procedure and do not represent community prevalence, total within-sample diversity, or absolute bacterial-versus-fungal abundance. Strong biofilm formation occurred in 312/664 (47.0%) selected bacterial isolates. Among interpretable bacterial organism–antimicrobial combinations, cefoxitin resistance occurred in 28/140 (20.0%) Staphylococcus aureus isolates and meropenem resistance in 16/52 (30.8%) Pseudomonas aeruginosa isolates. Among Candida albicans isolates, 41/44 (93.2%) were fluconazole susceptible, one was susceptible-dose dependent, and two were resistant according to the applicable CLSI disk-diffusion criteria. Conclusions Kindergarten and shared-use toys had greater aerobic culturable bacterial burdens, while several selected representative bacterial isolates showed substantial in vitro biofilm-forming capacity and antimicrobial-resistant phenotypes. Taxonomic frequencies describe selected representative culturable morphotypes rather than organism-specific surface prevalence. Findings apply to the sampled establishments and study period and support risk-based toy hygiene and cautious consideration of childcare surfaces within community environmental antimicrobial-resistance surveillance.