Search Results (350)

A2 milk has recently gained popularity, mainly due to its different beneficial effects on the human digestive system compared to popular A1 milk. Because of these structural differences, A2 milk exhibits characteristics that need to be investigated to design processes appropriately. The effects of β-casein genotype and heat treatment method (LTLT, HTST, and HTLT: a high-temperature preparatory treatment applied in fermented dairy production) on the fatty acid profile, volatile organic compounds (VOCs), composition, emulsion stability, acidification kinetics, and color parameters were investigated. Fatty acid composition was determined exclusively by milk type. A1 milk exhibited higher saturated fatty acids, higher total PUFA content, higher Σn-3 fatty acids, and a more favorable n-6/n-3 ratio, while A2 milk showed higher monounsaturated fatty acids and higher CLA (Conjugated Linoleic Acid) in a trait-dependent manner. VOC profiling identified 29 compounds. Significant genotype × treatment interactions were observed: A1 milk was characterized by high diacetyl in fresh samples and exclusive methanethiol formation upon pasteurization, while A2 milk showed enhanced Maillard-derived volatile formation and high ester accumulation after LTLT treatment. Emulsion stability was highest after LTLT treatment regardless of milk type. A1 milk reached the coagulation threshold of pH 4.6 within 45 min, whereas A2 exhibited delayed acidification. HTLT induced the greatest color changes in both types of milk. Full article

Mare milk is rich in nutrients and bioactive compounds, and its composition changes throughout lactation. This study investigated variations in the production, nutritional composition, and bioactive components of Yili mare milk across lactation stages. Twenty-six healthy grazing Yili mares were sampled on days 1, 10, 30, 60, 90, and 120 of lactation. Milk production, nutritional components, amino acids, fatty acids, minerals, vitamins, and immunologically active proteins were analyzed. Milk production peaked on day 30 and then declined. Colostrum contained significantly higher fat, protein, solids-not-fat, total solids, minerals, lactoferrin, lysozyme, and immunoglobulins than mature milk (p < 0.05), whereas lactose increased and stabilized after day 30. Essential amino acids peaked on day 30. As lactation progressed, saturated fatty acids decreased while polyunsaturated fatty acids increased. Vitamin profiles also varied across lactation, with ascorbic acid increasing during late lactation. β-casein content was higher during mid-lactation. In summary, colostrum is enriched in immunoactive proteins and minerals, whereas mature milk exhibits a more balanced amino acid and fatty acid profile. While these observed variations likely reflect the combined effect of lactation stage and seasonal pasture fluctuations under natural grazing, these findings provide practical insights into changes in milk composition in grazing Yili mares and may support the development of mare milk products under similar grazing systems. Full article

Genetic screening of monogenic traits is important for improving genetic health and increasing favorable milk protein in dairy cattle. This study developed and applied an amplification refractory mutation system PCR (ARMS-PCR) assay to simultaneously screen 21 causal variants underlying 18 monogenic traits in Holstein cattle, including 13 recessive genetic defects, 2 milk protein loci, and 3 morphological loci. The assay was designed as a unified multiplex PCR-capillary electrophoresis workflow, enabling clear detection of allele-specific products distinguished by fragment size and fluorescent color. Sanger sequencing validation of newly incorporated loci supported the accuracy of the assay. A total of 1656 cows from 12 commercial farms were genotyped using the multiplex ARMS-PCR panel, and amplicons were analyzed by capillary electrophoresis. Carriers were detected for all genetic defects except DUMPS, with carrier frequencies ranging from 0.12% to 6.64%. The highest frequencies were observed for HH5 (6.64%) and MWS (6.58%), whereas HH3, HH1, and HCD showed intermediate frequencies of 1.81% to 3.08%; all the remaining defects were below 1%. Overall, 22.10% of sampled cows carried at least 1 defect allele, including 20.47% carrying 1 defect, 1.51% carrying 2, and 0.12% carrying 3. For milk protein loci, the desirable β-casein A2A2 and κ-casein BB genotypes occurred at frequencies of 45.83% and 14.13%, respectively, and the favorable A2A2/BB combination was present in 6.22% of sampled cows. Dominant red, recessive red, and polled alleles were rare. These results indicate that multiplex fluorescent ARMS-PCR can serve as a practical targeted screening tool for the simultaneous management of known deleterious alleles and selection of favorable monogenic variants in dairy cattle breeding. Full article

Mastitis limits yak dairy production and is associated with lipopolysaccharide (LPS)-mediated inflammation in yak mammary epithelial cells (YMECs). This study aimed to investigate the protective effect of caffeic acid (CA) against LPS-induced cellular injury and to elucidate the underlying mechanisms, with a particular focus on autophagy regulation via the NF-κB signaling pathway. LPS exposure strikingly reduced cellular viability and increased intracellular reactive oxygen species (ROS) levels, accompanied by activation of the NF-κB pathway. Furthermore, it increased the expression of pro-inflammatory cytokines (TNF-α, IL-8, and IL-1β). In addition, LPS enhanced endoplasmic reticulum (ER) stress and Ca²⁺ dysregulation, increased LC3-II/LC3-I ratio, and reduced synthesis of α-casein and β-casein. Pretreatment with CA resulted in the effective alleviation of these alterations by restoring cellular viability, suppressing inflammatory responses, and normalizing autophagy-related markers. Additionally, inhibition of Nrf2 reversed the partial reversal of the protective effects of CA, resulting in increased ROS accumulation and autophagy activation, but did not impact NF-κB suppression. These findings indicate that CA attenuates LPS-induced inflammatory injury in YMECs involved in both Nrf2-dependent and independent pathways. These findings provide a mechanistic analysis of yak mastitis pathogenesis and CA potential as a natural therapeutic for improving mammary health and milk quality in yak dairy systems. Full article

β-Casein A1 and A2 (β-CN-A1 and β-CN-A2) are the two predominant β-CN proteoforms in bovine milk. β-CN-A1 has been associated with a greater propensity to release opioid peptides, such as β-casomorphin-7 (β-CM-7) and β-casomorphin-5 (β-CM-5), during gastrointestinal (GI) digestion, which may have adverse biological effects. This has stimulated growing interest in milk from cows carrying the β-CN A2A2 genotype (A2 milk), which requires reliable characterization methods. In this work, we developed a rapid, selective, and sensitive capillary electrophoresis–mass spectrometry (CE-MS) method for the accurate identification and quantification of β-CM-7 and β-CM-5 in milk hydrolysates from in vitro GI digestion of bovine milk. The method showed good linearity (R² > 0.99, over 0.5–100 mg/L for β-CM-7 and 0.25–100 mg/L for β-CM-5), limits of detection (0.25 and 0.10 mg/L), and repeatability (<0.2% for times and <1.4% for areas), and tandem mass spectrometry (MS/MS) allowed confirmation. The method was applied to A1A1 and A2A2 milk digested using the standardized INFOGEST protocol, followed by solid-phase extraction. β-CM-7 was detected and quantified only in A1A1 digests (0.98 mg/L), whereas β-CM-5 was not detected (<0.10 mg/L). These results indicate a differential release of β-CMs from A1 and A2 milk and support the method’s suitability for β-CM profiling, which may help assess A2 milk quality control and β-CM health impact. Full article

Buffalo milk is a rich source of various nutritional components and bioactive peptides, offering significant health benefits. Endogenous peptides, which occur naturally in milk, represent a valuable source of bioactive peptides with potential nutraceutical applications. However, research on endogenous peptides in buffalo milk remains limited. This study employed a quantitative peptidomic approach to characterize endogenous peptides across different lactation stages. A total of 2099, 2946, and 4418 peptides were identified in colostrum, transitional milk, and mature milk, respectively. The majority of these peptides were derived from β-casein, followed by α_S1-casein, κ-casein, and other proteins. Notably, variations in precursor proteins contributing to peptide production were observed throughout lactation. Phosphorylation levels of endogenous peptides were highest in mature milk, with serine residues predominating. Enzymatic cleavage analysis identified cathepsin D as the key enzyme involved in endogenous peptide production, while proline endopeptidase and plasmin exhibited stage-specific activities. Bioinformatics analysis revealed differentially expressed precursor proteins linked to complement cascades and NF-κB signaling, emphasizing the immune protective role of colostrum. Furthermore, 54 potentially bioactive peptides with favorable water solubility were identified in colostrum, of which 17 were predicted to possess anti-inflammatory properties. These findings contribute to a deeper understanding of the molecular basis of buffalo milk’s functional properties, highlighting its potential as a source of bioactive peptides for both nutritional and pharmaceutical applications. Full article

This narrative review summarizes evidence on the bovine β-casein (CSN2) A1/A2 polymorphism as a case study of how intensive dairy selection and global gene flow can reshape allele frequencies in ways that matter for consumers, processing and agroecological resilience. We draw together evidence from (i) population-genetic surveys of CSN2 in contrasting cattle populations, including a descriptive summary of published genotype-frequency studies; (ii) controlled human studies that separate A1-containing from A2-only dairy exposure; and (iii) dairy technology and the authenticity literature relevant to identity-preserved A2 value chains. Across intensively selected Holstein-Friesian populations, A1 was consistently present at substantial frequency (approximately one-third), whereas indigenous, beef and zebu-adjacent populations were typically A2-enriched, highlighting the role of historical breed formation and modern introgression in shaping apparent geographic and climatic patterns. Human intervention studies most consistently support improved short-term gastrointestinal tolerance with A2-only milk in susceptible individuals, while evidence for longer-horizon systemic outcomes remains mixed and insufficient for causal disease claims. Processing and analytical studies suggest that β-casein genotype can modestly affect coagulation and product behavior in a context-dependent manner and that validated proteoform quantification coupled with traceability is essential for credible A2 labeling at scale. We discuss implications for breeding programs, including staged A2 selection that avoids performance trade-offs, and emphasize governance of artificial insemination and supply-chain segregation as levers to limit inadvertent allele diffusion while supporting climate-relevant genetic resources in locally adapted breeds. Collectively, the reviewed evidence suggests that A1/A2 β-casein can be usefully interpreted within a One Health framework spanning animal genetics, dairy systems and human tolerance research. Full article

Background: The first 1000 days of a child’s life, from a woman’s pregnancy to her child’s second birthday, represent a critical window during which nutritional and environmental exposures shape long-term health. Gut microbiota play an important role in metabolic and overall health. Although pet exposure during pregnancy affects neonatal microbiota, immunity, and development, its effects on maternal health remain unclear. This study investigated the associations of pet exposure with gestational health, maternal and infant microbiota, and breast milk composition in overweight/obese pregnant women. Methods: Fecal samples and breast milk samples were collected from pet-exposed participants (n = 22) and non-exposed controls (n = 32) for 16S rRNA sequencing. Breast milk lipids and proteins were also quantified. Results: Pet exposure before conception, during pregnancy, and postpartum was not associated with gestational diabetes mellitus or gestational weight gain. In the maternal gut, the relative abundances of Proteobacteria, Verrucomicrobia, Sutterellaceae, Enterobacteriaceae, Akkermansia muciniphila, and Parabacteroides were higher, whereas that of Ruminococcus was lower, in the pet-exposed group. In breast milk, the relative abundance of Escherichia-Shigella and the concentrations of phosphatidylinositol 36:2, phosphatidylethanolamine 38:3, lysine, and β-casein were higher, whereas the abundance of Rothia was lower, in the pet-exposed group. The relative abundance of Escherichia-Shigella was also lower in the infant gut of the pet-exposed group. Conclusions: In overweight/obese pregnant women, pet exposure was associated with differences in maternal gut and breast milk microbiota, higher concentrations of selected breast milk phospholipids and β-casein, and lower Escherichia-Shigella abundance in the infant gut. Full article

Background/Objectives: Gastroesophageal reflux disease (GERD) is prevalent in infants and frequently managed with acid-suppressive medications that elevate gastric pH. This pilot study aimed to evaluate how varying gastric pH levels (2.5, 4.0 and 6.0) influence the hydrolysis of milk proteins in human milk (HM), cow’s milk-based infant formula (CMF), and goat milk-based infant formula (GMF). Methods: Samples were subjected to a 30 min in vitro gastric digestion using pediatric human gastric juice obtained from clinical donors. Protein degradation was analyzed via SDS-PAGE densitometry, comparing digested aliquots to undigested controls. Results: At pH 2.5, caseins were highly digested in all samples, especially in HM and GMF. At pH 4.0, GMF displayed an apparent 51% greater casein degradation relative to CMF and HM in this pilot analysis. α-lactalbumin degradation was markedly higher in GMF at all pH levels; notably, at pH 4.0 and 6.0, only GMF exhibited digestion of this protein. Albumin showed almost complete degradation in HM and GMF at pH 2.5, and GMF maintained greater degradation at higher pH levels. β-lactoglobulin (absent in HM) was better digested in GMF at pH 2.5, whereas CMF showed higher hydrolysis observed at pH 4.0 and 6.0. Lactoferrin digestion was most efficient in HM and GMF at pH 2.5, with no differences observed at higher pH levels. Conclusions: These preliminary findings suggest that GMF may offer digestive advantages for infants with GERD under pharmacological acid suppression, particularly regarding casein and α-lactalbumin breakdown at higher pH. The distinct digestion kinetics of CMF and GMF at different pH levels provide a physiological basis for targeted infant feeding strategies. Further large-scale studies are required to validate these exploratory observations. Full article

Milk is a complex biochemical mixture in which proteins significantly influence the behaviour of xenobiotics and bioactive compounds. Interactions between milk proteins and substances such as veterinary drugs or natural bioactives can modify molecular stability, binding dynamics, and exposure pathways, affecting food safety and the One Health concept. This study presents a comparative, matrix-focused investigation on how three chemically distinct ligand classes, sulfanilamide antibiotics, naturally occurring phenolic compounds and zinc–polyphenol complexes, interact with major milk proteins, β-lactoglobulin and casein. Protein–ligand interactions were examined using steady-state fluorescence spectroscopy to assess quenching behaviour and comparative interaction trends. Molecular docking was employed as a qualitative tool to provide structural context. Distinct interaction patterns were observed across ligand classes, reflecting differences in molecular structure, hydrophobicity, and coordination chemistry. Importantly, zinc coordination modified interaction profiles relative to the corresponding free ligands, indicating that metal coordination can affect ligand–protein interactions within the milk matrix. These findings support the concept that milk proteins may function as matrix-dependent modulators of ligand behaviour. The study is positioned as a hypothesis-generating framework highlighting the importance of food matrices as active biochemical environments. Herein, we provide a foundation for hypothesising how the milk matrix affects residue behaviour and bioactive interactions, with relevance to veterinary pharmacology and food safety risk assessment. Full article

This study presents a structured narrative review integrating methodological and decision-oriented perspectives. Milk proteins, particularly β-casein, have attracted increasing scientific and commercial attention due to their genetic variability and role in dairy production and product differentiation. Among β-casein variants, the A1 and A2 alleles of the CSN2 gene are of particular relevance, as their single-nucleotide difference has influenced breeding strategies and the expansion of A2-oriented dairy markets. Although multiple validated molecular genotyping approaches are available for CSN2 A1/A2 discrimination, guidance on their context-appropriate deployment in agricultural systems remains largely technique-centric. The present framework integrates analytical performance, sample complexity, and operational constraints to support the selection of fit-for-purpose methods across breeding, diagnostic, and dairy authentication contexts. Classical and advanced approaches, including polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP), allele-specific polymerase chain reaction (AS-PCR) and amplification refractory mutation system PCR (ARMS-PCR), high-resolution melting (HRM) analysis, sequencing-based methods, single nucleotide polymorphism (SNP) arrays, and digital polymerase chain reaction (dPCR), are comparatively evaluated not only in terms of sensitivity and throughput but also with respect to scalability, reproducibility, and decision risk. This framework provides a practical decision-support tool for aligning genotyping strategies with application-specific risk profiles, thereby improving reliability, transparency, and regulatory compliance in modern dairy systems. Full article

MDM4 (Murine Double Minute 4), also known as MDMX, is a crucial negative regulator of the tumor suppressor p53. MDM4 heterodimerizes with MDM2 to enhance MDM2-mediated ubiquitination and degradation of p53, thereby promoting tumorigenesis. Beyond its canonical role in inhibiting p53 activity, recent studies have revealed diverse p53-independent functions. MDM4 interacts with various proteins, including p73, E2F1, casein kinase 1α, PPARα, and TRIM21 to regulate cell cycle progression, β-catenin-mediated pre-leukemic progression, and ferroptosis independent of p53. In addition, MDM4 functions independently of both p53 and MDM2 by interacting with proteins, such as SMAD family members 3/4, retinoblastoma protein (pRB), p21, Nbs1 (also known as Nibrin), mTOR complex 1 (mTORC1), and the Polycomb Repressive Complexes (PRCs) complex, to control cell proliferation and survival, as well as protein degradation, double-strand break (DSB) repair, and replication fork progression. Intriguingly, multiple studies suggest that MDM4 exhibits oncogenic activity independent of p53; however, other reports highlight a potential tumor-suppressive role for MDM4 in the absence of p53. Thus, MDM4’s functions extend well beyond the canonical p53–MDM2 axis. A deeper understanding of MDM4 biology may facilitate the development of novel targeted therapies for various cancers. Full article

This study investigates the immunomodulatory effects and underlying mechanisms of KEMPFPK, a peptide derived from bovine β-casein, using integrated in vitro, in silico, and in vivo approaches. In RAW264.7 macrophages, KEMPFPK enhanced proliferation, phagocytosis, and migration and selectively upregulated the chemokine MCP-1. Under LPS-induced inflammation, KEMPFPK suppressed pro-inflammatory cytokines (IL-1β, TNF-α) and NO production while promoting the anti-inflammatory cytokine IL-10. These effects were mediated through the inhibition of NF-κB and MAPK signaling pathways. Molecular docking predicted high-affinity binding of KEMPFPK to Toll-like receptors (TLR2 and TLR4), suggesting a potential mechanism for its immunomodulatory activity. In cyclophosphamide (CTX)-induced immunosuppressed mice, KEMPFPK administration restored immune organ indices, rebalanced serum cytokine levels, and modulated humoral immunity. Importantly, KEMPFPK was associated with a significantly reshaped gut microbiota profile, characterized by the promotion of beneficial genera (e.g., Ligilactobacillus, Adlercreutzia) and the suppression of opportunistic pathogens (e.g., Escherichia–Shigella). These findings establish KEMPFPK as a dual-phase immunomodulator and suggest that its effects may involve direct immune cell regulation coupled with indirect microbiota remodeling. This study provides a scientific foundation for the application of KEMPFPK in immunomodulatory functional foods. Full article

This investigation focused on developing nutrient-dense Medjool date-based bars (MDBs) formulated with Medjool date paste, milk protein concentrate, whey proteins, and other functional ingredients. Comprehensive proximate analysis, mineral profiling, amino acid determination, and instrumental assessments, including color measurement, scanning electron microscopy, differential scanning calorimetry, and texture analysis, were performed, followed by organoleptic evaluation. Medjool date paste served as the primary carbohydrate source (76.44%), while whey protein isolate and milk protein concentrate contributed substantially to the protein fraction (89.26% and 81.62%, respectively). The resulting bars contained 19.32–26.78% crude protein, 10.96% fat, and 12.35–12.71% moisture, delivering 414.72–416.04 Kcal 100 g⁻¹. Sugar profiles remained consistent across formulations (total sugars: 36.77–36.98%), with appreciable mineral content including potassium (884–923 mg 100 g⁻¹), calcium (418–585 mg 100 g⁻¹), and phosphorus (402–459 mg 100 g⁻¹). The essential amino acid composition equaled or surpassed that of hen’s egg, establishing the product as a superior protein source. Antioxidant analysis demonstrated total phenolic content of 452.22–554.12 mg GAE 100 g⁻¹ and total flavonoids of 358.06–374.24 mg QE 100 g⁻¹, with consistent radical scavenging capacity, reduced browning via protein–polyphenol binding (ΔG −58 to −72 kJ mol⁻¹), a balanced texture (hardness 157–189 N), and consistent sensory scores (87.63–93.28% acceptability), without significant differences among formulations. Molecular docking confirmed β-lactoglobulin’s tight antioxidant shielding and caseinate’s flexible bioavailability boost, yielding shelf-stable functional snacks that advance date palm valorization. The results demonstrate the successful development of functional MDBs with an excellent nutritional profile and strong panelist acceptance. Full article

The CSN2 gene encoding β-casein has gained increasing attention in dairy cattle breeding due to the global adoption of A2-oriented selection strategies. However, robust large-scale evidence assessing potential unintended effects on functional traits, particularly fertility, under intensive commercial conditions remains limited. This study evaluated whether selection for the CSN2 A2 β-casein variant is associated with biologically relevant differences in fertility traits in Holstein cows. Reproductive and genomic data from 7826 lactation records of 2773 Holstein cows collected between 2022 and 2025 in a large commercial dairy herd were analyzed. Fertility indicators included days open, number of services per conception, calving interval, first-service conception rate, and pregnancy by 100 days in milk. Mixed-effects models accounting for repeated lactations and cow- and sire-level clustering were applied, and predefined equivalence margins were used to distinguish statistical non-significance from biological irrelevance. Across all evaluated fertility traits, differences among CSN2 genotypes (A1A1, A1A2, and A2A2) were consistently small, biologically negligible, and well within predefined equivalence margins. Differences in days open were within ±2 days, and effect sizes for count and binary traits were close to unity. Parity and calving year significantly influenced reproductive performance, whereas no CSN2 genotype × parity interactions were detected. These findings indicate that selection for the CSN2 A2 β-casein variant does not compromise reproductive performance under intensive commercial management conditions. From a breeding and industry perspective, the results support the implementation of A2-oriented selection strategies without biologically meaningful adverse effects on fertility. Full article