This study evaluated the preventive potential of perinatal supplementation with human breast milk-derived Lactiplantibacillus plantarum WLPL04-mCherry to mitigate Staphylococcus aureus infection via prophylactic intervention using a murine mastitis model. Through comprehensive analyses including bacterial enumeration, histopathology, immunohistochemistry, immunofluorescence, and TUNEL staining, we demonstrated that perinatal administration of L. plantarum WLPL04-mCherry significantly reduced S. aureus loads in mammary glands, mesenteric lymph nodes, and liver. The probiotic alleviated S. aureus-induced mammary tissue damage, suppressed neutrophil infiltration and myeloperoxidase activity, downregulated pro-inflammatory cytokines (IL-6, IL-1β, and TNF-α), and preserved tight junction proteins (Claudin-1, Occludin, and ZO-1), thereby reinforcing the blood-milk barrier and reducing mammary epithelial apoptosis. In vitro co-culture experiments confirmed its direct antagonistic effect against S. aureus through pH-dependent growth inhibition. These findings demonstrate that L. plantarum WLPL04-mCherry prevents S. aureus-induced mastitis through multimodal actions including pathogen inhibition, inflammation modulation, barrier protection, and apoptosis reduction. This study highlights L. plantarum WLPL04 as a promising non-antibiotic preventive agent for perinatal mastitis prophylaxis, offering a sustainable alternative for addressing antibiotic resistance and supporting lactation health.

ABSTARCTStarch-based delivery systems (SBDSs) have emerged as promising carriers for bioactive compounds (BCs) due to their biodegradability, low cost, and biocompatibility. However, the limited emulsifying capacity, poor physicochemical stability, and rapid digestibility of native starches restrict their applications. Yet critical gaps persist in understanding how starch modifications impact digestibility, release kinetics, and functional performance. This review critically evaluates how the modifications induce changes in molecular structure and supramolecular structure, which influence the encapsulation efficiency (EE), gastrointestinal (GI) release, and stability of BCs. A unified structure-property-function framework is provided by integrating parameters such as crystallinity, cross-link density, amphiphilicity, and digestibility fractions, including rapidly digestible starch (RDS), slowly digestible starch (SDS), and resistant starch (RS), with digestion behavior, release mechanisms, and bioaccessibility. The review further highlights emerging concepts such as digestion-tailored carrier engineering and synergistic dual-modification approaches. Additionally, it addresses key translation challenges, including clean-label requirements, regulatory constraints, scalability, process complexity, and inconsistencies between in vitro and in vivo performance. Overall, this review provides mechanistic and translation insights for designing advanced starch-based carriers with improved controlled-release performance and practical applicability.

Deoxynivalenol (DON) is a common mycotoxin in crops that could induce intestinal inflammation, affecting the susceptibility of intestinal epithelial cells (IECs) to pathogen infection. This study aimed to investigate DON's effects on mucin and cytokine production as part of the local immune system and how it affected intestinal susceptibility to pathogen infection. Caco-2 cells were exposed to DON followed by acute enteroinvasive Escherichia coli (EIEC) infection. An increase in EIEC attachment to DON-exposed cells was observed, probably in part, mediated by secretory MUC5AC mucins and membrane-bound MUC4 and MUC17 mucins. Additionally, DON with EIEC posttreatment led to significant changes in the gene expression of several proinflammatory cytokines (IL1 alpha, IL1 beta, IL6, IL8, TNF alpha, and MCP-1), which may be in part, mediated by NK-kappa B and/or MAPK signaling pathways. These data suggested DON may exert immunomodulatory effects on IECs, altering the IEC susceptibility to bacterial infection.Practical ApplicationThe results suggested that DON might modulate immune responses by affecting mucus and cytokine production, which may affect the susceptibility of intestinal epithelial cells to pathogen infection.

Patients with type 2 diabetes mellitus (T2DM) often exhibit reduced Lactobacillus abundance, dysregulated immune responses, disrupted intestinal barrier integrity, and increased insulin resistance. Consumption of non-digestible oligosaccharides has been shown to support the persistence of Lactobacillus in the gut and improve gut homeostasis. Lactiplantibacillus plantarum ZDY2013, a probiotic capable of metabolizing various oligosaccharides, serves as a potent regulator of intestinal mucosal immunity. In this study, we investigated the potential ameliorative effects of xylooligosaccharides combined with L. plantarum ZDY2013 (synbiotic) on T2DM-induced intestinal injury and explored the underlying mechanisms. Our results showed that synbiotic improved glucose metabolism, reduced lipid accumulation, and alleviated insulin resistance in T2DM rats. Moreover, synbiotic outperformed L. plantarum ZDY2013 alone in restoring intestinal barrier integrity by suppressing oxidative stress and intestinal inflammation, while significantly enhancing the colonization of L. plantarum ZDY2013 and altering the abundance of key bacterial genera. Interestingly, synbiotic treatment also increased the production of short-chain fatty acids (SCFAs), which were strongly associated with specific bacterial taxa. Furthermore, gut microbiota-derived SCFAs were confirmed to ameliorate insulin resistance by promoting glucose uptake and glycogen synthesis in IR-HepG2 cells. Collectively, these findings suggest the potential use of synbiotics as a clinical intervention to ameliorate T2DM. This study provides a rationale for exploring dietary approaches as a mitigating strategy for managing long-standing diabetes.

A waxy and a non-waxy proso millet flour were each fermented by Lactobacillus amylovorus, Lactobacillus fermentum, and Lactobacillus plantarum. The samples were fermented for one to five days, and starch was isolated from the fermented flours. The pH of fermented proso millet flour ranged from 3.27 to 3.6. The starch morphology of fermented samples differed from that of raw starches, with surface indentations and small pores leading to granule channels observed on the granule. The gelatinization temperatures were significantly decreased, whereas the enthalpies were not affected by fermentation. Peak and final viscosities were decreased after fermentation. The hardness of Lb. fermentum and Lb. plantarum fermented waxy starch gels was decreased, but the non-waxy samples fermented by Lb. amylovorus had significantly increased hardness. The adhesiveness of the starch gels from fermented samples was significantly increased. Lactic acid fermentation had significant effects on the morphology and physicochemical properties of proso millet starch.

Deoxynivalenol (DON) is a mycotoxin that commonly occurs in crops. It was hypothesized that DON could trigger intestinal inflammation and increase the susceptibility of intestinal epithelial cells (IECs) to pathogen infection. Accordingly, the aim of this study was to investigate the effects of DON on intestinal susceptibility to pathogen infection. Semiconfluent Caco-2 cells were exposed to DON followed by acute entero-invasive Escherichia coli (EIEC) infection. The effects of DON and EIEC contamination on mucin, cytokines and related signal transduction pathways were examined as part of the local immune system. Caco-2 cells were able to generate a rapid immune response against DON with or without EIEC post-challenge. An increase in EIEC attachment to DON-exposed cells was observed, probably in part, mediated by modulation of secretory MUC5AC mucins and membrane bound MUC4 and MUC17 mucins. Cells were also able to express and produce important mediators of inflammation, such as cytokines as a result of activation of toll-like receptors signalling cascades, modulation of nuclear factor κ-light chain-enhancer of activated B cells (NK-κB) and/or mitogen-activated protein kinase (MAPK) pathways. These data indicate that DON may exert immunomodulatory effects on intestinal epithelial cells, which might thereby modify the susceptibility to bacterial infection. ### Competing Interest Statement The authors have declared no competing interest.

Fermented cassava is obtained through fermentation process where the main role is played by various lactic acid bacteria (LAB), while the by-products (effluents) are discarded. The objective of this study was to isolate, identify and analyze potential probiotics in fermented cassava by-products/effluents using molecular techniques. We quantified the LAB's metabolites and tested their antimicrobial properties. We isolated 13 LAB strains which are all Gram-positive, and catalase, oxidase, indole and coagulase negative. Only 6 out of 13 selected isolates survived the low pH of 1.5 and 0.3% bile salt. These 6 isolates showed different levels of antimicrobial activity against the tested pathogens with zones of inhibition ranging from 5.10 mm to 22.80 mm. The isolated LAB were identified as Lactiplantibacillus plantarum and Limosilactobacillus fermentum. The results indicated that the selected LAB strains have potential as probiotics due to their ability to survive in acidic and bile-rich environments as well as their antimicrobial properties.

Hu sheep is an important sheep breed in China whose intestines are rich in probiotics. In this study, a total of fifty-nine lactic acid bacteria strains were isolated from Hu sheep feces and twelve strains were identified as Lactiplantibacillus plantarum. Among them, six L. plantarum strains were selected and further evaluated for their probiotic properties. Our results suggested that L. plantarum ELPL14 possess potential probiotic properties. The application of ELPL14 against L. monocytogenes in the pasteurized milk was investigated and the results showed that the growth of L. monocytogenes was significantly decreased when simultaneously incubated with ELPL14 in pasteurized milk at 37 degrees C. Meanwhile, in the ELPL14 fermented pasteurized milk, the growth of L. monocytogenes was also significantly decreased at 28 degrees C and 4 degrees C. We further tested the plantaricin genes by PCR, and found that eleven plantaricin genes (plnEF, plnR, plnJ, plnK, plnL, plnN, plnB, plnC, plnD, plnI, and plnH) were detected in the genome of ELPL14, suggesting that the inhibition of ELPL14 against L. monocytogenes might be due to the production of bacteriocin. Our results demonstrated that L. plantarum ELPL14 from Hu sheep has the potential as a natural candidate for dairy additives to inhibit food-borne pathogens, especially L. monocytogenes.

The emergence of antimicrobial resistance (AMR) in lactic acid bacteria (LAB) raises questions on qualified presumptive safety status and poses challenge of AMR transmission in food milieu. This study focuses on isolation, identification and characterization of AMR in LAB prevalent in traditional fermented Indian food products. The analysis of 16SrRNA based phylogenetic tree showed placements of isolates among four different genera Lactobacillus, Enterococcus, Weissella and Leuconostoc. In E-strip gradient test of susceptibility to 14 different antibiotics, over 50% of isolates showed resistance to ampicillin, chloramphenicol, ciprofloxacin, erythromycin, kanamycin, linezolid, streptomycin, trimethoprim and vancomycin. A multivariate principal component analysis, an antibiogram and multiple antibiotic resistance index-values (> 0.2) indicated presence of multidrug-resistance among the isolates. This study reports prevalence of an alarmingly high rate of AMR LAB strains in traditional fermented foods and is important to regulators and public health authorities for developing strategies to control transmission in food systems.

Barley is one of the most widely consumed grains in sundry societies owing to its impressive health benefits. In this context, yogurt drink supplemented with different levels (0%, 0.5%, 1%, 1.5%, 2%, 2.5%, and 3%) of roasted barley powder was prepared and evaluated. The effects of minimal addition on the physicochemical properties, rheological parameters, color, and sensory evaluation were investigated. An example case of the production cost and profits of yogurt drink supplemented with roasted barley powder was also provided. The use of barley powder increased yogurt acidity, total phenolic compounds content, antioxidant activity, and water-binding capacity. Yogurt drinks containing barley powder showed lower fat content, pH, and flow behavior index than the control sample. Treatments prepared by using 1.5% and 2% of roasted barley powder had higher averages of texture and flavor, whereas the treatment containing 3% exhibited higher appearance and color scores.

Fatty acids and amino acids are recognized to have a significant impact on the bioactivity, aroma, and flavor of ripened cheese. In this study, fatty acid profiles, lipid quality parameters, and amino acid composition of ripened and unripened cheeses produced from different milk sources were investigated. Fatty acids and amino acids were identified by using the gas chromatography-flame ionization detector and high-performance liquid chromatography, respectively. Lipid quality indicators, including atherogenicity index, thrombogenicity index, desired hypocholesterolemic fatty acids, undesired hypercholesterolemic fatty acids, hypocholesterolemic and hypercholesterolemic fatty acids ratio, and the nutritive value index of cheese samples were calculated. The results revealed that the major fatty acids detected were myristic, palmitic, stearic, and oleic. Cheese made from goat and sheep milk, such as Kashkaval, Chevrette Frico, and goat cheese with honey exhibited higher contents of capric acid representing 8.47 %, 9.56 %, and 10.07 %, respectively. In addition, fresh Roumy cheese showed higher contents of oleic (37.15 %) and linoleic (9.18 %) acids than other cheese types. The atherogenicity index of cheese fat varied between 1.13 and 3.97 for the different samples. Furthermore, cheese samples showed similar nutritive value indices. Each cheese type had its distinctive amino acid profile, which was attributed to the enzymatic activity. Milk source and ripening were the main reason for the variations in fatty acid and amino acid contents. The findings of this study could provide interesting knowledge to the dairy industry sector and could be used as a database in the field of food composition and analysis.

Gamma-aminobutyric acid (GABA) is a non-protein amino acid with various physiological functions. Levilactobacillus brevis NPS-QW 145 strains active in GABA catabolism and anabolism can be used as a microbial platform for GABA production. Soybean sprouts can be treated as a fermentation substrate for making functional products. This study demonstrated the benefits of using soybean sprouts as a medium to produce GABA by Levilactobacillus brevis NPS-QW 145 when monosodium glutamate (MSG) is the substrate. Based on this method, a GABA yield of up to 2.302 g L−1 was obtained with a soybean germination time of one day and fermentation of 48 h with bacteria using 10 g L−1 glucose according to the response surface methodology. Research revealed a powerful technique for producing GABA by fermentation with Levilactobacillus brevis NPS-QW 145 in foods and is expected to be widely used as a nutritional supplement for consumers.

Lactobacillus is a widely used bacteria and consumed through various fermented foods and beverages. Strains have been shown to carry resistance genes and mobile genetic elements with their ability to transfer the resistance to sensitive pathogenic strains. To study this, 4 cultures of Lactobacillus were isolated from traditional fermented milk. The isolates were able to grow up to 4% (w/v) NaCl concentration and 45 °C temperature, and showed > 97% 16S rRNA gene similarities with Lactobacillus fermentum. All the isolates were phenotypically screened for the presence of antibiotic resistance. Minimum inhibitory concentration (MIC) as microbiological breakpoints were observed against a varied class of antibiotics. Isolates AKO 94.6, DVM 95.7, and NIFTEM 95.8 were explicitly resistant to ampicillin, ciprofloxacin and vancomycin with MIC well beyond the maximum range of 256 µg/ml in the E-strip test. While isolate SKL1 was sensitive to ampicillin and showed MIC at 0.25 µg/ml but resistant to streptomycin and trimethoprim (MIC > 256 µg/ml). Molecular characterization showed the presence of tet(M) gene in three isolates SKL1, DVM 95.7, and NIFTEM 95.8 which was chromosomally associated resistance determinants while erm(B) resistance gene was detected in isolates DVM 95.7 and NIFTEM 95.8 only which was a plasmid associated gene and could be transferrable conjugally. Gene for Tn916 family (xis) was also observed in isolates DVM 95.7 and NIFTEM 95.8. Transferability of antibiotic resistance to pathogenic recipient strains was examined in isolates DVM 95.7 and NIFTEM 95.8 in different food matrices. The highest conjugation frequency with ~ 10–1 was obtained in alfalfa seed sprouts. This study reports the presence of acquired gene resistance in Lactobacillus species and dissemination to susceptible strains of bacteria in different food matrices. 16S rRNA gene sequences of isolates were uploaded to the NCBI GenBank database to retrieve the accession number.

Probiotics are microorganisms (including bacteria, yeasts and moulds) that confer various health benefits to the host, when consumed in sufficient amounts. Food products containing probiotics, called functional foods, have several health-promoting and therapeutic benefits. The significant role of yeasts in producing functional foods with promoted health benefits is well documented. Hence, there is considerable interest in isolating new yeasts as potential probiotics. Survival in the gastrointestinal tract (GIT), salt tolerance and adherence to epithelial cells are preconditions to classify such microorganisms as probiotics. Clear understanding of how yeasts can overcome GIT and salt stresses and the conditions that support yeasts to grow under such conditions is paramount for identifying, characterising and selecting probiotic yeast strains. This study elaborated the adaptations and mechanisms underlying the survival of probiotic yeasts under GIT and salt stresses. This study also discussed the capability of yeasts to adhere to epithelial cells (hydrophobicity and autoaggregation) and shed light on in vitro methods used to assess the probiotic characteristics of newly isolated yeasts.

This paper investigated the ability of lactic acid bacteria (LAB) and Bifidobacterium to remove acrylamide (AA). The mechanism(s) of the AA removal was examined by various microscopic techniques. After screening 40 isolates, Bifidobacterium breve and Lactiplantibacillus plantarum exhibited higher acrylamide (AA) reduction by 35% and 36%, respectively. pH (4.5-6.5), incubation temperature (32-42 degrees C), incubation time (14-22 h), and NaCl (0-3 g/100 g) were employed using Box-Behnken design (BBD) to investigate the AA removal. As a result, the AA removal ranged from 47.7 to 65.0% for B. breve and 5.8%-8.0% for Lb. plantarum. Under the in vitro gastrointestinal conditions, B. breve removed up to 41.2% of acrylamide while Lb. plantarum reduced around 53.5% of acrylamide. In addition, both strains tolerated low pH levels. TEM images showed that cells of both bacteria increased after culturing with acrylamide. Elements analysis of Lb. plantarum and the most dominant elements are C, N, and O with atomic % of 41.95, 17.63, and 31.66, respectively. FTIR exhibited that C--O, C-O, and N-H were main functional groups associated with AA adsorption in Lb. plantarum and B. breve. The zeta potential values of B. breve and Lb. plantarum are -15.47 and -25.87 mV, respectively.

The populations of Lactobacillus acidophilus, Bifidobacterium spp. and Lactobacillus casei were examined in 26 commercial fermented milk products represented by 14 companies. Six products contained L. acidophilus only, 12 products contained L. acidophilus and bifidobacteria, 6 products claimed L. acidophilus, bifidobacteria, and L. casei, and 2 products contained L. casei only as the probiotic organism. The initial bacterial count was enumerated immediately after purchase and the final count at the expiry date. L. acidophilus was enumerated on MRS-salicin agar and MRSsorbitol agar, bifidobacteria on MRS-NNLP (nalidixic acid, neomycin sulfate, lithium chloride, paromomycin sulfate) agar, and L. casei on LC agar. The counts of L. acidophilus decreased below 106 cfu/g in all the 6 products that contained L. acidophilus only. The counts of L. acidophilus decreased below 106 cfu/g in 75 % of the products that contained L. acidophilus and Bifidobacterium spp. at the expiry date. The population of bifidobacteria dropped below 106 cfu/g in 94 % of the products that claimed L. acidophilus, Bifidobacterium spp. or L. acidophilus, Bifidobacterium spp. and L. casei. Similarly, 50% of the products that contained L. acidophilus, Bifidobacterium spp. and L. casei showed L. casei count of < 106 cfu/g. The counts of L. casei in one of the two products that contained L. casei only dropped below 102 at the expiry date. The pH in all the products decreased during storage.

The human gastrointestinal (GI) tract contains a diverse mixture of commensal and pathogenic microbes, forming the gut microbiome. These gut microbes and their potential to improve human health are a topic of great interest to the scientific community. Many intestinal and age-related complications are linked to dysbiosis of the gut microbiome, often associated with a weakened immune system. A decrease in beneficial microbes, generally, along with decreased microbial diversity in the gut, can, in many cases, result in disease, particularly in older individuals. Probiotics, which are ingestible beneficial microorganisms, have the potential to positively modulate the indigenous gut microbiota. There are two predominant and conventional classes of lactic acid bacterial probiotics, lactobacilli and bifidobacteria, which have been confirmed for their health benefits and role in preventing certain gut-related disorders. The proper use of probiotics and/or supplements, along with a consistently healthy lifestyle, is a promising holistic approach to maintaining or improving gut health and minimizing other age-linked disorders. There are many properties that bacterial probiotics possess, which may allow for these beneficial effects in the gut. For instance, probiotics have adhesion capacities (capability to stay in GI tract) that are effective in excluding pathogens, while other probiotics have the potential to stimulate or modulate the intestinal immune system by regulating genes that reside within and outside of the gut environment. This review discussed the possible underlying mechanics of probiotics, evidence of probiotic-based mitigation of age-related disease, and the role of probiotics in modulating gut health and, in turn, maintaining brain health.

Levilactobacillus brevis NPS-QW-145 isolated from kimchi is deficient in glutamate dehydrogenase-encoding gene ( gdhA ) to form glutamate, hence it required exogenous supplementation of glutamate/monosodium glutamate (MSG) for decarboxylation reaction to produce γ-aminobutyric acid (GABA). However, GABA conversion rate from MSG was relatively low. The individual effect of 20 amino acids on regulating GABA biosynthesis was investigated. Cysteine was selected to significantly improve GABA production from MSG. It was found that Lb. brevis was capable of producing H 2 O 2 , cysteine protected Lb. brevis against H 2 O 2 -induced oxidative damage to increase cell viability for the enhancement of GABA production. Moreover, cysteine promoted glucose consumption to produce acetyl-CoA for synthesizing long-chain fatty acids to significantly up-regulate GABA biosynthesis. These findings deciphered antioxidative capability of cysteine in Lb. brevis 145 and provided a theoretical basis for fatty acids synthesis-mediated GABA synthesis in Lb. brevis 145, and possibly in other lactic acid bacteria.

Previous research has showed that nonproteolytic Levilactobacillus brevis 145 (L) in coculture with Streptococcus thermophilus 1275 (S), not Lactobacillus delbrueckii ssp. bulgaricus (Lbu), was able to produce γ-aminobutyric acid (GABA) during milk fermentation in the presence of monosodium glutamate (MSG). It was assumed that differences of casein hydrolysis patterns between Strep. thermophilus 1275 and L. bulgaricus caused the phenomenon. Moreover, the GABA content was low and residual MSG was high in SL-fermented milk. In our research, comparison of peptide profiles determined by liquid chromatography/tandem mass spectrometry showed that αS2-casein, β-casein, and κ-casein degradation by L. bulgaricus and Strep. thermophilus varied. Importantly, the peptide number in the L and Lbu coculture group increased compared with the Lbu monoculture group, whereas the peptide number in the SL coculture group decreased in comparison with S monoculture group, suggesting that L. bulgaricus was not able to provide peptides for the growth of Lb. brevis 145. Furthermore, we found that after supplementation with cysteine (50 mg/L) during milk fermentation by SL, 10 g/L MSG was converted into 4.8 g/L GABA with a minimum level of residual MSG, viable cell counts of Lb. brevis and lactic acid production were increased, and the casein hydrolysis pattern was not influenced. Moreover, sulfhydryl group-containing chemicals including cystine, reduced glutathione, and oxidized glutathione showed effects similar to that of cysteine in improving GABA production. Finally, when L. bulgaricus YIB2 was combined with SL, supplementation of cysteine was also able to significantly improve GABA production.

The selection of potential probiotic strains that pos-sess the physiological capacity of performing success-fully in the gastrointestinal tract (GIT) is a critical challenge. Probiotic microorganisms must tolerate the deleterious effects of various stresses to survive pas-sage and function in the human GIT. Adhesion to the intestinal mucosa is also an important aspect. Recently, numerous studies have been performed concerning the selection and evaluation of novel probiotic microorgan-isms, mainly probiotic bacteria isolated from dairy and nondairy products. Therefore, it would be crucial to critically review the assessment methods employed to select the potential probiotics. This article aims to re-view and discuss the recent approaches, methods used for the selection, and outcomes of the evaluation of novel probiotic strains with the main purpose of supporting future probiotic microbial assessment studies. The find-ings and approaches used for assessing acid tolerance, bile metabolism and tolerance, and adhesion capability are the focus of this review. In addition, probiotic bile deconjugation and bile salt hydrolysis are explored. The selection of a new probiotic strain has mainly been based on the in vitro tolerance of physiologically related stresses including low pH and bile, to ensure that the potential probiotic microorganism can survive the harsh conditions of the GIT. However, the varied experimental conditions used in these studies (different types of media, bile, pH, and incubation time) hamper the comparison of the results of these investigations. Therefore, standardization of experimental conditions for characterizing and selecting probiotics is warranted.