Infant Formula Research Articles

Transcriptomics analysis of the role of SdiA in desiccation tolerance of Cronobacter sakazakii in powdered infant formula

The quorum-sensing receptor SdiA is vital for regulating the desiccation tolerance of C. sakazakii, yet the specific mechanism remains elusive. Herein, transcriptomics and phenotypic analysis were employed to explore the response of C. sakazakii wild type (WT) and sdiA knockout strain (ΔsdiA) under drying conditions. Following 20 days of drying in powdered infant formula (PIF), WT exhibited 4 log CFU/g higher survival rates compared to ΔsdiA. Transcriptome revealed similar expression patterns between csrA and sdiA, their interaction was confirmed both by protein-protein interaction analysis and yeast two-hybrid assays. Notably, genes associated with flagellar assembly and chemotaxis (flg, fli, che, mot regulon) showed significantly higher expression levels in WT than in ΔsdiA, indicating a reduced capacity for flagellar synthesis in ΔsdiA, which was consistent with cellular morphology observations. Similarly, genes involved in trehalose biosynthesis (ostAB, treYZS) and uptake (thuEFGK) exhibited similar expression patterns to sdiA, with higher levels of trehalose accumulation observed in WT under desiccation conditions compared to ΔsdiA. Furthermore, WT demonstrated enhanced protein and DNA synthesis capabilities under desiccation stress. Higher expression levels of genes related to oxidative phosphorylation were also noted in WT, ensuring efficient cellular ATP synthesis. This study offers valuable insights into how SdiA influences the desiccation tolerance of C. sakazakii, paving the way for targeted strategies to inhibit and control this bacterium.

Health risk assessment for dietary exposure to 3-monochloropropane-1,2-diol, 2-monochloropropane-1,2-diol, and glycidol for Italian consumers

3-Monochloropropane-1,2-diol (3-MCPD), 2-monochloropropane-1,2-diol (2-MCPD) and 2,3-epoxy-1-propanol (glycidol), in their free form or esterified to fatty acids, are food contaminants formed during the refinement of oils and fats. We conducted a survey to quantify the levels of these compounds in 130 food items, in order to assess the exposure to them in food and the consequent health risk for consumers. Food samples, including infant formula, were analysed by gas-chromatography mass spectrometry with the indirect method, and we used the latest open access food consumption database for the Italian population for a probabilistic assessment of exposure. We adopted an in silico approach to fill the gap for the toxicity of 2-MCPD. The occurrence values for the three contaminants in food were in most cases lower than or comparable to those reported in previous surveys. Exposure assessment for the most exposed individuals (95thpercentiles of consumers only) of different age groups, gave values below the tolerable daily intake recommended by the European Food Safety Authority for 3-MCPD and below the simulated or predicted toxicity thresholds for 2-MCPD, indicating a negligible risk due to dietary exposure to these contaminants. For glycidol, however, estimated exposure indicated a non-negligible increase in cancer risk, and a margin of exposure <25,000 for younger population groups, indicating a potential health concern.

Protein-bound and free glycation compounds in human milk: A comparative study with minimally processed infant formula and pasteurized bovine milk

The role of the Maillard reaction and the accumulation of non-enzymatic glycation compounds in human milk have been scarcely considered. In this study, we investigated the proteins most susceptible to glycation, the identity of the corresponding modified residues and the quantitative relationship between protein-bound and free glycation compounds in raw human milk and, for comparison, in minimally processed infant formula and pasteurized bovine milk. In human milk, total protein-bound lysine modifications were up to 10% of the counterparts in infant formula, while Nε-carboxymethyllysine reached up to 27% of the concentration in the other two products. We demonstrated that the concentration of free pyrraline and methylglyoxal-hydroimidazolone were of the same order of magnitude in the three milk types. Our results delineate how the occurrence of some glycation compounds in human milk can be an unavoidable part of the breastfeeding and not an exclusive attribute of infant formulas and pasteurized bovine milk.

Chloropropanols and Their Esters in Food: An Updated Review.

Chloropropanols, their fatty acid esters, and glycidol and its fatty acid esters (GEs) are process contaminants in foods that pose potential health risks. These contaminants typically arise during the deodorization process of vegetable oils, particularly in high concentrations within oils like palm oil and products derived from them, such as margarine, baked goods, pastries, and infant formula. Chloropropanol esters and GE can hydrolyze under the influence of lipases, forming chloropropanols. Elevated temperatures during food production can lead to the release of free 3-chloro-1,2-propanediol (3-MCPD) or free 2-chloro-1,3-propanediol (2-MCPD) in products containing both fat and salt. The exposure to these contaminants, especially for infants and young children, raises concerns about potential health hazards. While extensive research has focused on 3-MCPD, 2-MCPD, and GE, knowledge regarding other chloropropanols such as 1,3-dichloro-2-propanol (1,3-DCP), 2,3-dichloro-1-propanol (2,3-DCP), and their fatty acid esters remains limited. This review aims to provide a comprehensive overview encompassing formation mechanisms, analysis methods, toxicological implications, occurrence patterns, exposure levels, mitigation strategies, and legislative considerations concerning these contaminants.

Human milk vs. Infant formula digestive fate: In vitro dynamic digestion and in vivo mini-piglet models lead to similar conclusions

Infant formula (IF), the only nutritionally adequate substitute for human milk (HM), still needs to be improved to be more biomimetic with HM, including in terms of digestive fate. The latter can be explored using different digestion models. The present study aimed to compare IF and HM digestion using in vivo (mini-piglet) and in vitro (dynamic system, DIDGI®) models. Fresh mature HM was collected and compared with a standard bovine IF. In vivo, 18 Yucatan mini-piglets (24-day-old) received HM or IF and were euthanized 30 min after the last meal. The entire digestive content was collected from the stomach to the colon. In vitro, the same meals were fed to an in vitro dynamic digestion model simulating the term infant at four weeks of age. Digesta were sampled regularly in the gastric and intestinal compartments. Structure (confocal microscopy and laser light scattering) and proteolysis (SDS-PAGE for residual intact proteins, OPA for hydrolysis degree, LC-MS/MS for peptides) were investigated along digestion. The digesta microstructure differed between HM and IF in a similar way between in vitro and in vivo digestion. In vitro gastric proteolysis of caseins and α-lactalbumin was significantly slower for HM than for IF, such as for the early intestinal proteolysis degree. In vitro bioaccessibility of free AAs explained only 30 % of the true ileal digestibility of AAs. Peptide mapping of caseins differed between HM and IF along their digestion. The relative peptide mapping data over six proteins from HM and IF were highly correlated between in vitro and in vivo digestion, particularly at 80 and 120 min of in vitro gastric digestion vs. in vivo stomach data and at 20 and 40 min of in vitro intestinal digestion vs. in vivo proximal jejunum data (r = 0.7–0.9, p < 0.0001, n = 1604). 40 to 50 % of the bioactive peptides identified in vivo were also found in vitro, with a good correlation of their abundances (r = 0.5, p < 0.0001, n = 61). Overall, in vitro and in vivo digestion were in good agreement, both indicating a different digestive fate for HM and IF.

Bifidobacterium longum subsp. infantis YLGB-1496-Toxicological evaluation.

Bifidobacterium infantis YLGB-1496, originally isolated from breast milk from a Taiwanese mother, is under study for use as a probiotic. As part of safety assessment, an Ames, in vivo mouse micronucleus, and in vivo mouse spermatocyte chromosome aberration assay were conducted along with a 13-week oral rat toxicity study. B.infantis YLGB-1496 had no activity in any of the genotoxicity assays. Administration of the bacteria to Sprague-Dawley rats at doses ranging from 0 to 1.5 g/kg bw/day had no treatment-related effects on any of the endpoints measured. There appear to be no concerns for translocation or pathogenicity of B.infantis YLGB-1496 based on extensive experience with the species in general. The results of the current investigations support potential use of B.infantis YLGB-1496 as a probiotic in infant formula.

Variations in the Sensory Attributes of Infant Formula among Batches and Their Impact on Maternal Consumer Preferences: A Study Combining Consumer Preferences, Pivot Profile, and Quantitative Descriptive Analysis.

The sensory quality of infant formula (IF) has a significant impact on the preferences and purchasing behavior of maternal consumers. Consumer-based rapid descriptive methods have become popular and are widely preferred over classical methods, but the application of Pivot Profile (PP) in IF is still little explored. In this study, both Pivot Profile (PP) and Quantitative Descriptive Analysis (QDA) were applied to characterize the sensory profile of 12 batches of one-stage and three-stage IF with different storage periods, respectively, along with consumer preference data to determine the flavors contributing to liking. The results of PP and QDA aligned moderately well, with the most perceptible differences identified as "fishy", "milky", and "T-sweet" attributes. IFs with shorter storage times were highly associated with "milky" aromas and "T-sweet" tastes, whereas IFs with longer storage times exhibited a strong correlation with "fishy" and "oxidation" aromas. External preference analysis highlighted that the occurrence of "fishy" and "oxidation" aromas during prolonged storage periods significantly reduced the consumer preference for IFs. Conversely, the perception of "milky" and "creamy" aromas and "T-sweet" tastes may be critical positive factors influencing consumer preference. This study provided valuable insights and guidance for enhancing the sensory quality and consumer preference of IF.

Infant formula donations and code violations during earthquake relief efforts in Türkiye in 2023: an observational study

BackgroundOn 6 February 2023, an Mw 7.8 earthquake struck southern and central Türkiye and north-western Syria, affecting the lives of 4.6 and 2.5 million children, respectively. In such crises, infants who are dependent on commercial milk formula (CMF) face increased vulnerability to diseases and malnutrition as safe preparation of CMF becomes difficult and sometimes impossible. The Operational Guidance on Infant and Young Child Feeding in Emergencies (OG-IFE) provides guidance on protecting and supporting recommended infant and young child feeding and minimizing the risks that come with CMF feeding. In addition, the International Code of Marketing of Breastmilk Substitutes (the Code) ensures adequate nutrition for infants by protecting and promoting breastfeeding and ensuring the proper usage of CMF. This study aims to document violations of the Code and the OG-IFE during the earthquake relief efforts to help strengthen infant and young child feeding emergency responses and inform future disaster relief policies.MethodsData was collected from 6 February to 10 March 2023 through Internet sources. Social media, news websites, and the Emergency Nutrition Network forum were used for data collection. Turkish content was translated into English for analysis, with a focus on donation-related information and Code violations related to baby food and infant feeding.ResultsA total of 40 reports on CMF, complementary food, and feeding equipment donations were collected. Three main types of violations of the OG-IFE and the Code were identified, with the majority of them being incidences of individuals, humanitarian organizations, and government agencies seeking or accepting donations. Infant formula companies continued to violate the Code by donating large quantities of CMF and feeding equipment to the Turkish Red Crescent, government agencies, and disaster relief infant and young child feeding (IYCF) coordination authorities.ConclusionsThese incidents reflected a systematic violation of the Code and non-compliance with the OG-IFE. Globally accepted IYCF standards and recommendations were not consistently followed due to fragmented early responses. There is a critical need to step up efforts to ensure appropriate and safe IYCF practice protecting and supporting breastfed and non-breastfed infants in emergencies.

Assessment of the reliability of at-home caregiver-collected anthropometric measurements.

Anthropometric measurements provide valuable information about infant growth patterns and can help identify nutrition, growth, and developmental concerns. With the increasing use of telehealth and decentralized clinical trial approaches, there is potential for caregivers to collect anthropometric measurements at home via teleconference with healthcare providers (HCPs) to monitor infant growth, which indirectly reflects health status. This study aimed to evaluate whether telehealth-guided caregivers can utilize standardized methods and home-use measurement equipment to collect reliable anthropometric measurements compared to HCPs and study nurses. The study compared the weight, length, and head circumference measurements collected by caregivers (n = 8 pairs), pediatric HCPs (n = 7), and study nurses (n = 4), who served as the gold standard comparator group. Four silicone dolls with varied anthropometrics were used as surrogates for human infants. Caregiver inter- and intra-observer technical errors of measurement (TEMs) were all equal to or below the maximum allowed error (MAE). For HCPs, only intra-observer TEM for length and inter-observer TEM for HC and length were within the MAE. There was no evidence of bias for either caregiver or HCP measurements compared to the gold standard. Coefficients of reliability (R) were greater than 0.96 for all measurements. Preliminary results from this study demonstrate that telehealth-guided caregivers can capture accurate and reliable anthropometric measurements compared to HCPs. The results suggest that remote measurement collection allows for more frequent monitoring while reducing the burden on patients and caregivers in primary care and clinical trials such as infant formula growth monitoring studies.

Isolation and purification of bovine N‐glycans from whey protein concentrate (WPC 70)

N‐glycans present in milk glycoproteins (e.g. lactoferrin and immunoglobulin G) can provide infants with nutritional and protective effects similar to breast milk. In this study, we enriched N‐glycoproteins from WPC 70 using ultrafiltration and anion exchange chromatography on DEAE Sepharose Fast Flow column, with N‐glycan content and N‐glycoprotein purity as evaluation indicators. The results showed that changing feed flow rate and molecular weight cut‐off was more effective to improve ultrafiltration separation than changing feed concentration. When using a 100 kDa ultrafiltration membrane with a feed flow rate of 10 rpm and a feed concentration of 10 mg/mL, N‐glycan content could be increased by 200%. However, anion exchange chromatography was more efficient for purification, increasing N‐glycan content by 700%. This study provided a reliable method for N‐glycan enrichment and a scientific basis for further research and potential application of N‐glycans in infant formula and whey products.

Exploring the relationship between child temperament, maternal psychiatric symptoms, family environment and infant feeding.

Infant temperament is assumed to be primarily innate. However, newer research suggests that maternal affection impacts ratings of temperament and environmental factors, including feeding method, can also influence infant temperament. This study investigates child temperament and its relationships with maternal psychiatric symptoms, environmental variables and feeding method longitudinally in a cohort of children followed from 6 to 72 months. Differences in temperament by feeding group are also investigated. We hypothesized that maternal psychiatric symptoms, environmental stressors, and impaired family dynamics would have negative impact on child temperament, whereas breastfeeding would have a positive impact on child temperament. Mothers' ratings of child's temperament, own psychiatric symptomatology, environmental stresses and family cohesion were obtained in 504 mother-infant dyads via rating scales completed by mothers. Infants were breastfeed (BF), fed soy-based infant formula (SF) or dairy-based infant formula (MF). Linear mixed effect models investigated the relationship of variables on child's temperament while controlling for significant covariates and repeated measurements. Mothers in this study did not endorse clinical-level psychiatric symptomatology; however, when adjusted for significant covariates, higher psychiatric symptomatology significantly correlated with environmental stressors, impaired family dynamics and elevations in temperament ratings of infants' adaptability and mood. There were no lasting differences for temperament between feeding groups. However, some significant transient increases in rhythmicity and adaptability were found between SF and BF children. Positive relationships between family environment stressors and maternal psychiatric ratings were found. Transient differences were found in child temperament based upon feeding method.

Multi-Omics Profiles of Small Intestine Organoids in Reaction to Breast Milk and Different Infant Formula Preparations.

Ensuring optimal infant nutrition is crucial for the health and development of children. Many infants aged 0-6 months are fed with infant formula rather than breast milk. Research on cancer cell lines and animal models is limited to examining the nutrition effects of formula and breast milk, as it does not comprehensively consider absorption, metabolism, and the health and social determinants of the infant and its physiology. Our study utilized small intestine organoids induced from human embryo stem cell (ESC) to compare the nutritional effects of breast milk from five donors during their postpartum lactation period of 1-6 months and three types of Stage 1 infant formulae from regular retail stores. Using transcriptomics and untargeted metabolomics approaches, we focused on the differences such as cell growth and development, cell junctions, and extracellular matrix. We also analyzed the roles of pathways including AMPK, Hippo, and Wnt, and identified key genes such as ALPI, SMAD3, TJP1, and WWTR1 for small intestine development. Through observational and in-vitro analysis, our study demonstrates ESC-derived organoids might be a promising model for exploring nutritional effects and underlying mechanisms.

Changes in whey protein produced by different sterilization processes and lactose content: Effects on glycosylation degree and whey protein structure

In this study, the interaction patterns between whey protein and lactose were investigated after different ratios of whey protein and lactose (1:2.5; 1:3.0; 1:3.5; 1:4.0; 1:4.5) were treated with two typical sterilization process parameters in powdered products (68 °C, 20 min; 72 °C, 15 min) and two typical sterilization process parameters in liquid products (121 °C, 10 min and 121 °C, 15 min). It was found that the degree of the Maillard reaction of whey protein-lactose solution increased with increasing lactose content, increasing sterilization temperature, or extending sterilization time, which was accompanied by changes in the protein structure of whey protein. In addition, glycosylation accelerates the decrease in free sulfhydryl content, decreasing surface hydrophobicity and decreasing fluorescence intensity. These findings highlight the advantages of designing protein-carbohydrate formulations with a high ratio of whey protein-lactose (1:2.5) to meet the requirements of the product standards for preterm/low-birth-weight infant formulas. When comparing only the sterilization processes of powder and liquid products, the pasteurization process of powder products significantly reduced the degree of the Maillard reaction caused by heat treatment and reduced protein denaturation.

Structured Triacylglycerol with Optimal Arachidonic Acid and Docosahexaenoic Acid Content for Infant Formula Development: A Bio-Accessibility Study.

Polyunsaturated fatty acids (PUFAs), especially arachidonic acid (ARA) and docosahexaenoic acid (DHA), are extremely important fatty acids for brain development in the fetus and early childhood. Premature infants face challenges obtaining these two fatty acids from their mothers. It has been reported that supplementation with triacylglycerols (TAGs) with an ARA:DHA (w/w) ratio of 2:1 may be optimal for preterm infants, as presented in commercial formulas such as Formulaid™. This study explored methods to produce TAGs with a 2:1 ratio (ARA:DHA), particularly at the more bioavailable sn-2 position of the glycerol backbone. Blending and enzymatic acidolysis of microalgae oil (rich in DHA) and ARA-rich oil yielded products with the desired ARA:DHA ratio, enhancing sn-2 composition compared to Formulaid™ (1.6 for blending and 2.3 for acidolysis versus 0.9 in Formulaid™). Optimal acidolysis conditions were 45 °C, a 1:3 substrate molar ratio, 10% Candida antarctica lipase, and 4 h. The process was reproducible, and scalable, and the lipase could be reused. In vitro digestion showed that 75.5% of the final product mixture was bio-accessible, comprising 19.1% monoacylglycerols, ~50% free fatty acids, 14.6% TAGs, and 10.1% diacylglycerols, indicating better bio-accessibility than precursor oils.

P01-14 In vitro bioaccessibility of mycotoxins co-occurrence in infant formulas and complementary foods

P01-14 In vitro bioaccessibility of mycotoxins co-occurrence in infant formulas and complementary foods

Scientific and technical assistance report on the evaluation of human‐identical milk oligosaccharides (HiMOs) as novel foods

Abstract EFSA was asked by the European Commission to provide scientific and technical assistance on the evaluation of human‐identical milk oligosaccharides (HiMOs) as novel foods (NFs). In recent years, the number of authorisations of HiMOs as NFs has markedly increased, which may lead to situations of multiple concurrent uses. Since the safety assessment of HiMOs is based on the comparison with the ‘natural’ intake of human milk oligosaccharides (HMOs), i.e. the HMO intake from human milk in exclusively breastfed infants, a scoping literature review was outsourced to update the database of mean concentrations of single and total HMOs in human milk, and relevant HMO intakes were thus updated. It is noted that in infants up to 16 weeks of age exclusively fed with infant formula, when adding all the assessed HiMOs at their highest maximum use levels across production methods, the resulting highest daily intakes of single HiMOs and the sum of HiMOs are within the natural HMO intake range. These intakes were also estimated (DietEx) for the most exposed population groups (i.e., infants and young children) considering the highest maximum use levels in those food categories with the highest contribution to the overall HiMO intake (i.e., infant and follow‐on formulae, ready‐to‐eat meals, yoghurt and cow milk). The resulting highest P95 daily HiMO intakes were unlikely to be higher than the highest mean daily HMO intakes. However, any possible higher intake would not necessarily imply a safety concern since such natural intake estimates represent an upper end of average natural intakes. It can be concluded that currently there are no safety concerns resulting from the authorised or assessed HiMO intakes from their concurrent combined uses. Nonetheless, in consideration of the increased interest in the use of HiMOs as NFs, a simplified but realistic approach for the intake assessment is proposed in the case of new HiMOs or extensions of use of already authorised HiMOs.

Is it possible to obtain substitutes for human milk oligosaccharides from bovine milk, goat milk, or other mammal milks?

Considering the current level of chemical and biological synthesis technology, it was a sensible selection to obtain milk oligosaccharides (MOs) from other mammals as the potential substitute for human MOs (HMOs) that possessed various structural features in the infant formula. Through a comprehensive analysis of the content, structure, and function of MOs in six distinct varieties of mammal milk, it has been shown that goat milk was the most suitable material for the preparation as a human milk substitute. Goat MOs (GMOs) had a relatively high content and diverse structural features compared to those found in other mammalian milks. The concentration of GMOs in colostrum ranged from 60 to 350mg/L, whereas in mature milk, it ranged from 200 to 24,00mg/L. The acidic oligosaccharides in goat milk have attracted considerable attention due to their closeness in acidic content and structural diversity with HMOs. Simultaneously, it was discovered that some structures, like N-glycolylneuraminic acid, were found to have a certain content in GMOs and served essential functional properties. Moreover, studies focused on the extraction of MOs from goat milk indicated that the production of GMOs on an industrial scale was viable. Furthermore, it is imperative to do further study on GMOs to enhance the preparation process, discover of new MOs structures and bioactivity evaluation, which will contribute to the development of both the commercial production of MOs and the goat milk industry.

Combining 2'-fucosyllactose and galactooligosaccharides exerts anti-inflammatory effects and promotes gut health

This study investigated the potential of 2'-Fucosyllactose (2'-FL) and galactooligosaccharides (GOS) combinations as a novel and cost-effective substitute for human milk oligosaccharides (HMOs) in promoting gut health and reducing inflammation. In vitro studies using Caco-2 cells showed that 2'-FL and GOS combinations (H1: GOS:2'-FL ratio of 1.8:1; H2: ratio of 3.6:1) reduced lipopolysaccharide-induced inflammation by decreasing pro-inflammatory markers, while individual treatments had no significant effects. In a mouse model of dextran sulfate sodium (DSS)-induced colitis, combined 2'-FL and GOS supplementation alleviated symptoms, improved gut permeability, and enhanced intestinal structure, with the GH1 group (H1 combo with DSS) being the most effective. 2'-FL and GOS combinations also enhanced short-chain fatty acid production in infant fecal batch fermentation and mouse fecal analysis, with GH1 showing the most promising results. GH1 supplementation altered gut microbiota in mice with DSS-induced colitis, promoting microbial diversity and a more balanced Firmicutes to Bacteroidota ratio. Infant formula products (IFPs) containing 2'-FL and GOS combinations (IFP2: 174 mg GOS and 95 mg 2'-FL per 14 g serving, 1.8:1 ratio; IFP3: 174 mg GOS and 48 mg 2'-FL per 14 g serving, 3.6:1 ratio) demonstrated gastrointestinal protective and anti-inflammatory properties in a coculture model of Caco-2 and THP-1 cells. These findings suggest that 2'-FL and GOS combinations have potential applications in advanced infant formulas and supplements to promote gut health and reduce inflammation.

Longitudinal investigation of mineral composition in human milk and its correlation with infant anthropometric outcomes among Tibetan mother-infant dyads during the first 6 months postpartum

Nutrients in human milk, including minerals, relate growth and development of breast-fed infants. Tibetan mother-infant dyads possess unique characteristics on early nutrition due to their featured long-lasting life-style. This study longitudinally investigated the relationship between the mineral composition in human milk and the Z-scores of infants among Tibetan mother-infant dyads during their first 6 months postpartum through a prospective cohort study. The results show that the minerals of Na, Mg, K, Ca, Cu, Zn, and Se were of higher levels in colostrum than other lactation stages. Several minerals were below the recommended values for infants according to Chinese dietary guidelines. Besides, a large proportion of infant Z-scores were below −2 as lactation period continued. Multivariate statistical analysis revealed that classifications and correlations in varying degrees were observed between minerals in human milk and infant Z-scores. These findings will be advantageous for research upon Chinese early nutrition and progress of tailor-made infant formula.

Transcriptomic Analyses to Unravel Cronobacter sakazakii Resistance Pathways.

The proliferation of antibiotic usage has precipitated the emergence of drug-resistant variants of bacteria, thereby augmenting their capacity to withstand pharmaceutical interventions. Among these variants, Cronobacter sakazakii (C. sakazakii), prevalent in powdered infant formula (PIF), poses a grave threat to the well-being of infants. Presently, global contamination by C. sakazakii is being observed. Consequently, research endeavors have been initiated to explore the strain's drug resistance capabilities, alterations in virulence levels, and resistance mechanisms. The primary objective of this study is to investigate the resistance mechanisms and virulence levels of C. sakazakii induced by five distinct antibiotics, while concurrently conducting transcriptomic analyses. Compared to the susceptible strains prior to induction, the drug-resistant strains exhibited differential gene expression, resulting in modifications in the activity of relevant enzymes and biofilm secretion. Transcriptomic studies have shown that the expression of glutathione S-transferase and other genes were significantly upregulated after induction, leading to a notable enhancement in biofilm formation ability, alongside the existence of antibiotic resistance mechanisms associated with efflux pumps, cationic antimicrobial peptides, and biofilm formation pathways. These alterations significantly influence the strain's resistance profile.