Background: Artisanal Colonial Cheese (ACC), a traditional Brazilian dairy product, represents a complex interplay between microbial ecology, seasonal climate and artisanal practices. ACC has few studies related to its physicochemical and microbiological composition, which would assist in the standardisation and legislation of this product.Aim: This study investigated how seasonal variations (summer vs. winter) influence ACC's microbial dynamics, biochemical profile and nutritional quality during 21 days (1, 7, 14, 21) of ripening.Methods: Using high‐throughput sequencing, gas chromatography and capillary electrophoresis analyses, the microbial communities, fatty acids profiles, aliphatic organic acids (AOA) and volatile compounds of eight different cheeses were characterised.Major Findings: Summer ACCs exhibited Lactococcus‐driven dominance (64.95%–69.73%), correlating with elevated unsaturated fatty acids (UFAs: 18.035–32.900 g/100 g oleic acid), favourable hypocholesterolemic/hypercholesterolemic (H/H) ratios (0.861 vs. winter 0.330) and ester‐rich volatile profiles (e.g. ethyl butanoate). Winter samples showed dynamic microbial succession: early Raoultella (42.28%–39.74%) and Rahnella (11.96%–12.78%) transitioned to Leuconostoc dominance (9.33%–16.37%), yielding higher short‐chain fatty acids (SCFAs: caproic, caprylic acids) and branched alcohols (e.g. 3‐methylbutanol, 60% of W21 volatiles). Fungal communities further differentiated seasons, with Debaryomyces genus suppressing spoilage fungi in winter and Diutina genus enhancing lipolysis in summer. Nutritional indices linked summer ACCs to improved anti‐atherogenic properties, while winter's microbial diversity showed more complex AOA pathways (propionic and succinic acids).Scientific Implications: These findings underscore ACC's microbial terroir, where seasonal microbiota govern its biochemical identity, providing a scientific basis for geographical indication protocols. Furthermore, information on biochemical and microbiological transformations promoted a better understanding of the maturation period of this product.

Artisanal colonial cheese (ACC) produced from raw milk is a rich reservoir of autochthonous lactic acid bacteria (LAB), but strain-level evidence supporting safe downstream application and technological stability remains limited. In this study, 10 LAB isolates from ACC were screened for phenotypic safety, antimicrobial susceptibility, and probiotic-related traits, and their viability was further assessed after inulin-based spray-drying microencapsulation under different storage temperatures. All isolates showed no hemolytic or mucinolytic activity and did not produce gelatinase, supporting an initial safety profile, and all strains were sensitive to at least two antimicrobial classes. Strain prioritization identified Lacticaseibacillus casei LAB06, LAB09, and LAB10 and Lactiplantibacillus plantarum LAB03 as the most robust candidates for downstream development because they maintained stable cell counts throughout simulated gastrointestinal digestion. Inulin spray-drying yielded structurally stable microcapsules and supported refrigerated storage, with substantially lower viability losses at 4 °C than at 25 °C; notably, L. plantarum LAB01 and LAB02 showed the best refrigerated shelf-life, remaining above 6.0 log CFU/g after 45 days. Together, these results position ACC as a source of promising LAB candidates and highlight cold-chain-compatible microencapsulation as a strategy to support safe functional food development with potential public health benefits.

Artisanal colonial cheese (ACC), produced from raw milk, retains its native microbiota throughout maturation, with lactic acid bacteria (LAB) being predominant, many of which exhibit probiotic potential. In this study, ten LAB strains isolated from ACC matured for 14 and 21 days were characterized and identified as Lacticaseibacillus casei, Lactiplantibacillus plantarum, Levilactobacillus brevis, and Pediococcus acidilactici. The strains were Gram-positive, catalase-negative, lacked phenotypic virulence factors (hemolysin, mucinase, and gelatinase), and showed sensitivity to at least two classes of antibiotics. They also exhibited high tolerance to simulated gastrointestinal conditions, along with strong auto-aggregation and hydrophobicity. The selected strains were microencapsulated by spray drying using inulin as a carrier agent. The resulting microparticles displayed predominantly spherical morphology and diameters ranging from 1.19 to 5.71 µm. Post-processing viability ranged from 76% to 99%, with cell counts remaining above 6 log CFU/g for up to 45 days under refrigeration. Storage at 25 °C significantly reduced cell survival. FTIR, XRD, and TGA/DTGA analyses confirmed the physicochemical stability of the microcapsules. Overall, the findings highlight ACC as a relevant source of safe and potentially probiotic LAB. Microencapsulation with inulin proved to be an effective strategy for preserving strain viability and stability, supporting their potential application in functional foods.