%0 Journal Article %T Presence of rpoS Gene and Environmental Stress-Dependent Biofilm Formation in Gram-Negative Bacteria in Mexican Artisanal Dairy Establishments %A Ericka Vá %A zquez-Ojeda %A Emmanuel Castillo-Quiñ %A ones %A Bertha Landeros-Sá %A nchez %A Lilia Hurtado-Ayala %J Advances in Microbiology %P 297-312 %@ 2165-3410 %D 2025 %I Scientific Research Publishing %R 10.4236/aim.2025.156021 %X Artisanal Mexican cheeses are frequently produced without pasteurization, favoring microbial contamination and potential biofilm formation. Aim: To characterize Gram‑negative isolates from artisanal cheese outlets in Tijuana, Mexico, evaluate their biofilm‑forming ability under different environmental conditions and detect the stress‑response gene rpoS. Methods: Seventeen Gram‑negative and 13 Gram‑positive bacteria were isolated from cheese, contact surfaces and air. Curves of growth (BHI, 37 ˚C), antimicrobial susceptibility (VITEK 2‑AST), biofilm formation in microtiter plates (four media/temperature combinations) and PCR detection of rpoS were performed. Results: Escherichia coli displayed the highest growth rate (µ  =  0.28  h1, g  =  149  min), while Citrobacterfreundii formed the only “strong” biofilm in all conditions. Two Pantoeaagglomerans isolates were resistant to cefoxitin and one E. coli to ampicillin/sulbactam; tetracycline resistance appeared in a single airborne Staphylococcus epidermidis. Biofilm biomass (OD630) differed significantly by genus and condition (two‑way ANOVA, p  <  0.05). All Gram‑negative isolates were positive for rpoS, but gene presence alone did not predict biofilm intensity. Conclusions: Gram‑negative biofilm‑forming bacteria carryingrpoS are common in Tijuana artisanal cheeses, yet antimicrobial resistance remains sporadic. Targeted sanitation against mixed‑species biofilms and continued AMR surveillance are warranted to enhance food safety. %K Artisanal Cheese %K Biofilm %K rpoS Gene %K Antimicrobial Resistance %U http://www.scirp.org/journal/PaperInformation.aspx?PaperID=143451