INTERNATIONAL DAIRY JOURNAL

Hydrolysis improves the inhibition efficacy of bovine lactoferrin against infection by SARS-CoV-2 pseudovirus
Patil D, Chen S, Fogliano V and Madadlou A
The entry of SARS-CoV-2 into host cells may involve the spike protein cleavage by cathepsin L (CTSL). Certain food proteins such as lactoferrin (Lf) inhibit CTSL. The current study investigated the impact of hydrolysis (0-180 min) by proteinase K on electrophoretic pattern, secondary structure, cathepsin inhibitory and SARS-CoV-2 pseudovirus infectivity inhibitory of bovine Lf. Gel electrophoresis indicated that hydrolysis cut Lf molecules to half lobes (∼40 kDa) and produced peptides ≤18 kDa. Approximation of the secondary structural features through analysis of the second-derivative amide I band collected by infra-red spectroscopy suggested a correlative-causative relationship between cathepsin inhibition and the content of helix-unordered structures in Lf hydrolysate. The half maximal inhibitory concentration (IC) of Lf hydrolysed for 90 min (H90) against CTSL was about 100 times smaller than that of the Lf hydrolysed for 0 min (H0). H90 had also double activity against SARS-CoV-2 pseudo-types infectivity compared with H0.
Antibodies against spike protein of SARS-CoV-2 variants in bovine whey IgG enriched fraction
Oshiro S, Mizutani N, Tada T, Sekiguchi JI, Takahashi M and Kirikae T
Bovine whey IgG enriched fraction contains IgG antibodies against bacterial and viral pathogens, including antibodies against the spike protein [amino acids (aa) 1-1274] of SARS-CoV-2 Wuhan strain (2019-nCoV WHU01). To date, 13 SARS-CoV-2 variants have been identified, including gamma, delta, kappa, and omicron, which contain 10, eight, seven, and over 30 mutations in the spike protein, respectively. We investigated whether bovine whey IgG enriched fraction contains antibodies against spike proteins of these variants, specifically recombinant partial length spike proteins (aa 177-512, aa 509-685, aa 177-324, aa 250-410 and aa 387-516) of these variants. Direct enzyme-linked immunosorbent assays revealed bovine whey IgG enriched fraction contained antibodies against all recombinant spike proteins of these variants with highest reactivity against aa 177-512 region of omicron spike protein. These results indicate bovine whey IgG enriched fraction contains antibodies against spike proteins of several SARS-CoV-2 variants, including omicron.
Higher yogurt intake is associated with lower blood pressure in hypertensive individuals: Cross-sectional findings from the Maine-Syracuse Longitudinal study
Wade AT, Guenther BA, Ahmed FS and Elias MF
Associations between fermented dairy products and blood pressure are unclear. The current study therefore examined the association between yogurt and blood pressure in hypertensive and non-hypertensive individuals. Cross-sectional analyses were undertaken on 915 community-dwelling adults from the Maine-Syracuse Longitudinal Study. Habitual yogurt consumption was measured using a food frequency questionnaire. The primary outcomes were systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial pressure (MAP), and pulse pressure. Secondary outcomes included BMI (kg m), total cholesterol (mg dL), glucose (mg dL), HDL (mg dL), LDL (mg dL), triglycerides (mg dL), and plasma homocysteine (μmol L). Multivariable regression analyses revealed significant inverse associations between yogurt and both SBP ( < 0.05) and MAP ( < 0.05) in hypertensive (n = 564) but not non-hypertensive participants (n = 351). Future observational and intervention studies should continue to focus on at-risk individuals to examine the potential benefits of yogurt.
Presence of antibodies against SARS-CoV-2 spike protein in bovine whey IgG enriched fraction
Oshiro S, Tada T, Mizutani N, Funatogawa K, Sekiguchi JI, Takahashi M and Kirikae T
Bovine whey IgG enriched fraction contains antibodies against various human bacterial pathogens. It contains antibodies against some viral antigens, including human respiratory syncytial virus and influenza virus. We investigated whether the IgG enriched fraction has cross-reactivity with IgG antibodies against SARS-CoV-2 spike (S) and nucleocapsid (N) proteins. The full-length and partial-length SARS-CoV-2 S, N, a recombinant protein of the receptor binding domain (RBD) and nine peptides covering the receptor binding motif (RBM) of S were prepared. Direct enzyme-linked immunosorbent assays were conducted using these recombinant proteins and peptides as coating antigens and revealed the IgG enriched fraction contained antibodies against partial-length S [amino acids (aa) 177-512, 288-512, 348-578, 387-516 and 408-664], full-length N (aa 1-419) and partial-length N (aa 1-120, 111-220, 1-220 and 210-419), two RBD peptides, covering aa 427-446 and 502-520 of S, and recombinant RBD of S. These results indicate IgG enriched fraction contains antibodies against SARS-CoV-2.
Comparison of bovine milk oligosaccharides in native North European cattle breeds
Sunds AV, Bunyatratchata A, Robinson R, Glantz M, Paulsson M, Leskauskaite D, Pihlanto A, Inglingstad R, Devold TG, Vegarud GE, Birgisdottir BE, Gudjonsdottir M, Barile D, Larsen LB and Poulsen NA
Milk oligosaccharides are of high interest due to their bioactive properties. This study is the first to characterise milk oligosaccharides from native North European cattle breeds, as represented by 80 milk samples collected from eight native breeds originated from Norway (Norwegian Doela cattle and Norwegian Telemark cattle), Sweden (Swedish Mountain cattle), Denmark (Danish Red anno 1970), Iceland (Icelandic cattle), Lithuania (native Lithuanian Black and White) and Finland (Western Finncattle and Eastern Finncattle). Using high-performance liquid-chromatography chip/quadrupole time-of-flight mass-spectrometry, 18 unique monosaccharide compositions and a multitude of isomers were identified. No N-glycolylneuraminic acid was identified among these breeds. Western Finncattle milk was most abundant in neutral, acidic and fucosylated oligosaccharides. Further, Eastern Finncattle milk was significantly higher in acidic oligosaccharides and Icelandic cattle milk significantly higher in fucosylated oligosaccharides, compared to the mean. This study highlights specific native breeds of particular interest for future exploitation of milk oligosaccharides and breeding strategies.
Review of the prevalence of foodborne pathogens in milk and dairy products in Ethiopia
Keba A, Rolon ML, Tamene A, Dessie K, Vipham J, Kovac J and Zewdu A
Food safety is a significant barrier to social and economic development throughout the world, particularly in developing countries. Here, we reviewed the prevalence of major bacterial foodborne pathogens ( spp., , O157:H7 and spp.) in the rapidly growing Ethiopian dairy supply-chain. We identified 15, 9, 5 and 0 studies that had reported the prevalence of spp., O157:H7, and spp. in dairy foods, respectively. The studies reviewed reported a median prevalence of and O157:H7 of 6, 9 and 10%, respectively, in raw cow milk in Ethiopia, indicating a concerning occurrence of bacterial foodborne pathogens in raw milk. Implementation of good hygiene and production practices and assessment of interventions targeting the reduction of contamination in the dairy supply chain is needed to inform coordinated efforts focused on improvement of dairy food safety in Ethiopia.
Production of functional mimics of human milk oligosaccharides by enzymatic glycosylation of bovine milk oligosaccharides
Weinborn V, Li Y, Shah IM, Yu H, Dallas DC, German JB, Mills DA, Chen X and Barile D
Consumption of mothers' milk is associated with reduced incidence and severity of enteric infections, leading to reduced morbidity in breastfed infants. Fucosylated and sialylated human milk oligosaccharides (HMO) are important for both direct antimicrobial action - likely via a decoy effect - and indirect antimicrobial action through commensal growth enhancement. Bovine milk oligosaccharides (BMO) are a potential source of HMO-mimics as BMO resemble HMO; however, they have simpler and less fucosylated structures. BMO isolated at large scales from bovine whey permeate were modified by the addition of fucose and/or sialic acid to generate HMO-like glycans using high-yield and cost-effective one-pot multienzyme approaches. Quadrupole time-of-flight LC/MS analysis revealed that 22 oligosaccharides were synthesized and 9 had identical composition to known HMO. Preliminary anti-adherence activity assays indicated that fucosylated BMO decreased the uptake of enterohemorrhagic O157:H7 by human intestinal epithelial Caco-2 cells more effectively than native BMO.
The antimicrobial activity of bovine milk xanthine oxidase
Ozturk G, Shah IM, Mills DA, German JB and de Moura Bell JMLN
Mammalian milk is a source of antimicrobial compounds such as xanthine oxidase (XO). The interplay of infant saliva, which contains the substrates for XO activity, and human milk containing XO has been recently shown to inhibit the growth of pathogenic bacteria. Based on the complex and protective mechanism observed in human milk, we hypothesized that bovine milk XO operates similarly, thus representing an opportunity to investigate its functionality in broader health implications. We demonstrated that bovine milk-hypoxanthine mixture (0 to 400 μM) inhibited several Gram-negative and -positive bacterial pathogens in a dose-dependent manner. Kinetic experiments revealed that XO catalyzed hypoxanthine reduction (K, 58.0 μM; V, 5.1 μmol min mg) resulted in the production of antimicrobial hydrogen peroxide. These results demonstrate that the antimicrobial properties of bovine milk XO are similar to those of human milk XO with significant implications for the development of novel products targeting infant health.
Dairy food intake, diet patterns, and health: Findings from the Maine-Syracuse Longitudinal Study
Crichton GE, Bogucki OE and Elias MF
This study compared the intake of yogurt and other dairy foods in relation to other dietary habits, and associations with cardiovascular health indicators. Comparative analyses of dietary intakes and cardiometabolic health indicators were conducted in participants from the Maine-Syracuse Longitudinal Study, conducted in 2001-2006 ( = 699). Participants (10.2% of sample) who consumed yogurt more frequently (≥ 5 serves per week) also consumed more fruit, vegetables, nuts, and fish, but fewer sweets, sugar-sweetened soft drinks, and alcohol. The yogurt-healthy food score was significantly inversely associated with fasting plasma glucose ( = 0.044), waist circumference ( = 0.004), and smoking ( = 0.004), and significantly positively associated with physical activity ( < 0.001). In addition, the yogurt-healthy food score was associated with protection against MetS abdominal obesity (OR: 0.80, 95% CI: 0.66-0.07, = 0.02). The yogurt-healthy dietary pattern was significantly associated with a better cardiometabolic profile.
An improved method for the purification of milk oligosaccharides by graphitised carbon-solid phase extraction
Robinson RC, Colet E, Tian T, Poulsen NA and Barile D
Milk oligosaccharides (OS) are bioactive molecules that impart a variety of health benefits to the consumer. Techniques commonly used to analyse and quantify OS require optimised extraction methods to separate the OS from more abundant milk components. Solid phase extraction (SPE) is frequently used to isolate milk OS from lactose; however, the literature contains no formal studies on its efficacy in this application. In this study, established SPE conditions were modified to improve the technique's effectiveness in purifying OS from lactose. Low concentrations of acetonitrile (ACN) and trifluoroacetic acid (TFA) were tested for solid phase washing. Lactose removal and retention of many OS were significantly improved when using 4% ACN/0.1% TFA compared with the more common water washing technique. Different behaviours between acidic and neutral OS were evident. The new SPE technique improves extraction efficiency for bovine milk OS in applications that do not require prior lactose hydrolysis.
Role of pH in the recovery of bovine milk oligosaccharides from colostrum whey permeate by nanofiltration
Cohen JL, Barile D, Liu Y and de Moura Bell JMLN
Milk oligosaccharides are associated with improved health outcomes in infants. Nanofiltration (NF) is used for isolation of bovine milk oligosaccharides (BMO). The study aim was to improve the recovery of BMO from lactose-hydrolyzed colostrum whey permeate. The retention factors of carbohydrates at various pH and transmembrane pressures were determined for a nanofiltration membrane, which was used at pilot scale to purify BMO. Carbohydrates were quantified by liquid chromatography and characterized using nano-LC-Chip-QToF mass spectrometry. BMO purity was improved from an initial 4% in colostrum whey permeate to 98%, with 99.8% permeation of monosaccharides and 96% recovery of oligosaccharides, represented by 23 unique BMO compounds identified in the final retentate. The pH during NF was a determining factor in the selectivity of carbohydrate separation. This NF method can be applied to conventional cheese-whey permeate and other milk types for extraction of bioactive oligosaccharides providing new options for the dairy industry.
Peptidomic analysis of healthy and subclinically mastitic bovine milk
Guerrero A, Dallas DC, Contreras S, Bhandari A, Cánovas A, Islas-Trejo A, Medrano JF, Parker EA, Wang M, Hettinga K, Chee S, German JB, Barile D and Lebrilla CB
A variety of proteases release hundreds of endogenous peptide fragments from intact bovine milk proteins. Mass spectrometry-based peptidomics allows for high throughput sequence assignment of a large number of these peptides. Mastitis is known to result in increased protease activity in the mammary gland. Therefore, we hypothesized that subclinically mastitic milks would contain higher concentrations of released peptides. In this work, milks were sampled from three cows and, for each, one healthy and one subclinically mastitic teat were sampled for milk. Peptides were analyzed by nano-liquid chromatography quadrupole time of flight tandem mass spectrometry and identified with database searching. In total, 682 peptides were identified. The total number of released peptides increased 146% from healthy to subclinically mastitic milks ( < 0.05), and the total abundance of released peptides also increased significantly ( < 0.05). Bioinformatic analysis of enzyme cleavage revealed increases in activity of cathepsin D and elastase ( < 0.05) with subclinical mastitis.
Relation between dairy food intake and cognitive function: The Maine-Syracuse Longitudinal Study
Crichton GE, Elias MF, Dore GA and Robbins MA
Diet modification to alter the course of age-related cognitive decline is becoming increasingly important. Few observational findings suggest that dairy food intake may be positively related to cognitive function, but research in this novel area is limited. The aim of this study was to investigate whether dairy food intake is associated with cognitive function, before and after adjustment for cardiovascular, lifestyle and dietary factors. To do this, a cross-sectional analyses of a subset of the community-based Maine-Syracuse Longitudinal Study (MSLS) sample ( = 972) was undertaken. It was determined that participants who consumed dairy products at least once per day had significantly higher scores on multiple domains of cognitive function compared with those who never or rarely consumed dairy foods, adjusting for cardiovascular risk factors, lifestyle and dietary factors. Frequent dairy food intake is associated with better cognitive performance but underlying causal mechanisms are still to be determined.
Molecular mechanisms underlying nutrient detection by incretin-secreting cells
Reimann F
The hormones glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-1 (GLP-1) are secreted postprandially from intestinal K- and L-cells, respectively. As incretins, these hormones stimulate insulin secretion from the pancreatic beta-cell, and have independently been implicated in the control of food intake and lipid metabolism. Whilst the enteroendocrine cells producing GIP and GLP-1 are therefore attractive targets for the treatment of diabetes and obesity, our understanding of their physiology is fairly limited. The mechanisms employed to sense the arrival of carbohydrate, fat and protein in the gut lumen have been investigated using organ perfusion techniques, primary epithelial cultures and cell line models. The recent development of mice with fluorescently labeled GIP or GLP-1-expressing cells is now enabling the use of single cell techniques to investigate stimulus-secretion coupling mechanisms. This review will focus on the current knowledge of the molecular machinery underlying nutrient sensing within K- and L-cells.
Permeate from cheese whey ultrafiltration is a source of milk oligosaccharides
Barile D, Tao N, Lebrilla CB, Coisson JD, Arlorio M and German JB
Previously undescribed oligosaccharides in bovine cheese whey permeate were characterized by a combination of nanoelectrospray Fourier Transform Ion Cyclotron Resonance (nESI-FTICR) mass spectrometry and matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance (MALDI-FTICR) mass spectrometry. Oligosaccharide composition was elucidated by collision-induced dissociation within the ICR cell. In addition to sialyllactose (the most abundant oligosaccharide in bovine colostrum), we identified 14 other oligosaccharides, half of which have the same composition of human milk oligosaccharides. These oligosaccharides could potentially be used as additives in infant formula and products for the pharmaceutical industry. Because whey permeate is a by-product from the production of whey protein concentrate (WPC) and is readily available, it is an attractive source of oligosaccharides for potential application in human nutrition.
Milk immunoglobulins for health promotion
Mehra R, Marnila P and Korhonen H
The biological function of bovine colostral immunoglobulins is to provide the newborn calf with adequate passive immune protection against microbial infections. Immunoglobulin preparations designed for farm animals are commercially available, and some colostrum-based products are marketed also for humans as dietary supplements. The concentration of specific antibodies against a certain pathogenic microorganism can be raised in colostrum and milk by immunizing cows with this pathogen or its antigen. Advances in bioseparation and chromatographic techniques have made it possible to fractionate and enrich these antibodies and formulate so-called hyperimmune colostral or milk preparations. Their efficacy in prevention and treatment of various microbial infections has been evaluated in numerous studies. Immune milk preparations have proven effective in prophylaxis against infections caused by a variety of gastrointestinal pathogens. Their therapeutic efficacy, however, seems more limited. A few commercial immune milk products are already on market and more applications can be expected in the coming years. This article reviews the recent progress made in isolation techniques of bovine immunoglobulins and the application of colostral and immune milk preparations in fighting various microbial infectious diseases in humans.