Infant milk formula, produced by membrane filtration, promotes mucus production in the upper small intestine of young pigs.

Human breast milk promotes maturation of the infant gastrointestinal barrier, including the promotion of mucus production. In the quest to produce next generation infant milk formula (IMF), we have produced IMF by membrane filtration (MEM-IMF). With a higher quantity of native whey protein, MEM-IMF more closely mimics human breast milk than IMF produced using conventional heat treatment (HT-IMF). After a 4-week dietary intervention in young pigs, animals fed a MEM-IMF diet had a higher number of goblet cells, acidic mucus and mucin-2 in the jejunum compared to pigs fed HT-IMF (P < 0.05). In the duodenum, MEM-IMF fed pigs had increased trypsin activity in the gut lumen, increased mRNA transcript levels of claudin 1 in the mucosal scrapings and increased lactase activity in brush border membrane vesicles than those pigs fed HT-IMF (P < 0.05). In conclusion, MEM-IMF is superior to HT-IMF in the promotion of mucus production in the young gut.

Comparative Structural and Compositional Analyses of Cow, Buffalo, Goat and Sheep Cream.

Factors affecting milk and milk fraction composition, such as cream, are poorly understood, with most research and human health application associated with cow cream. In this study, proteomic and lipidomic analyses were performed on cow, goat, sheep and Bubalus bubalis (from now on referred to as buffalo), bulk milk cream samples. Confocal laser scanning microscopy was used to determine the composition, including protein, lipid and their glycoconjugates, and the structure of the milk fat globules. BLAST2GO was used to annotate functional indicators of cream protein. Functional annotation of protein highlighted a broad level of similarity between species. However, investigation of specific biological process terms revealed distinct differences in antigen processing and presentation, activation, and production of molecular mediators of the immune response. Lipid analyses revealed that saturated fatty acids were lowest in sheep cream and similar in the cream of the other species. Palmitic acid was highest in cow and lowest in sheep cream. Cow and sheep milk fat globules were associated with thick patches of protein on the surface, while buffalo and goat milk fat globules were associated with larger areas of aggregated protein and significant surface adsorbed protein, respectively. This study highlights the differences between cow, goat, sheep, and buffalo milk cream, which can be used to support their potential application in functional foods such as infant milk formula.

Immunoglobulin G from bovine milk primes intestinal epithelial cells for increased colonization of bifidobacteria.

A bovine colostrum fraction (BCF) was recently shown to enhance the adherence of several commensal organisms to intestinal epithelial cells through modulating the epithelial cell surface. In this study, the main components of the BCF were examined to investigate the active component/s responsible for driving the changes in the intestinal cells. The adherence of various bifidobacteria to HT-29 cells was increased when the intestinal cells were pre-incubated with immunoglobulin G (IgG). Modulation of the intestinal cells by IgG was concentration dependent with 16 mg/mL IgG resulting in a 43-fold increase in the adhesion of Bifidobacterium longum NCIMB 8809 to HT-29 cells. Periodate treatment of colostral IgG prior to performing the colonization studies resulted in a reduction in the adhesion of the strain to the intestinal cells demonstrating that the glycans of IgG may be important in modulating the intestinal cells for enhanced commensal adhesion. IgG isolated from mature milk also resulted in significant increases in adhesion of the Bifidobacterium strains tested albeit at reduced levels (3.9-fold). The impact of IgG on the HT-29 cells was also visualised via scanning electron microscopy. This study builds a strong case for the inclusion of IgG ingredients sourced from cow's milk in functional foods aimed at increasing numbers of health promoting bacteria in the human gut.

Microstructure and fracture properties of semi-hard cheese: Differentiating the effects of primary proteolysis and calcium solubilization.

The individual roles of hydrolysis of αS1- and ß-caseins, and calcium solubilization on the fracture properties of semi-hard cheeses, such as Maasdam and other eye-type cheeses, remain unclear. In this study, the hydrolysis patterns of casein were selectively altered by adding a chymosin inhibitor to the curd/whey mixture during cheese manufacture, by substituting fermentation-produced bovine chymosin (FPBC) with fermentation-produced camel chymosin (FPCC), or by modulating ripening temperature. Moreover, the level of insoluble calcium during ripening was quantified in all cheeses. Addition of a chymosin inhibitor, substitution of FPBC with FPCC, or ripening of cheeses at a consistent low temperature (8 °C) decreased the hydrolysis of αS1-casein by ~95%, ~45%, or ~30%, respectively, after 90 d of ripening, whereas ~35% of ß-casein was hydrolysed in that time for all cheeses, except for those ripened at a lower temperature (~17%). The proportion of insoluble calcium as a percentage of total calcium decreased significantly from ~75% to ~60% between 1 and 90 d. The rigidity or strength of the cheese matrix was found to be higher (as indicated by higher fracture stress) in cheeses with lower levels of proteolysis or higher levels of intact caseins, primarily αS1-casein. However, contrary to the expectation that shortness of cheese texture is associated with αS1-casein hydrolysis, fracture strain was significantly positively correlated with the level of intact ß-casein and insoluble calcium content, indicating that the cheeses with low levels of intact ß-casein or insoluble calcium content were more likely to be shorter in texture (i.e., lower fracture strain). Overall, this study suggests that the fracture properties of cheese can be modified by selective hydrolysis of caseins, altering the level of insoluble calcium or both. Such approaches could be applied to design cheese with specific properties.

Heat and Chemical Treatments Affect the Viability, Morphology, and Physiology of Staphylococcus aureus and Its Subsequent Antibody Labeling for Flow Cytometric Analysis.

The effects of heat and chemical treatments on Staphylococcus aureus viability and physiology and their subsequent effects on antibody binding ability and cell morphology were measured. Treatments included lethal and sublethal heat; exposure to organic acids, salt, and sodium hydroxide; and freeze-thawing. Strain-related differences in viability were noted depending on treatment and were reflected in changes in physiology as monitored by flow cytometry (FCM) using three different staining protocols: SYTO 9/propidium iodide (PI), DiOC2(3), or calcein acetoxymethyl ester (calcein-AM)/PI. Treatments that resulted in significant losses in viability as measured by plate counting were reflected better by the first two staining combinations, as intracellular calcein-AM uptake may have been impaired by certain treatments. FCM analysis using labeling by commercial anti-S. aureus antibodies indicated that differences in cell physiology as a result of treatments influenced immunofluorescence detection. The ratio of the mean fluorescence intensities of stained cells to those of unstained cells [MFI/MFI(us)] varied with treatment, five of these treatments, including freeze-thaw, citric acid, oxalic acid, NaCl, and NaOH treatments, resulted in significantly lower fluorescence values compared to controls.IMPORTANCE FCM data indicated that cells conventionally considered to be dead and which would not give rise to CFU in a plate count assay, e.g., cells heated to 80°C, were labeled by antibody staining. This finding suggests that without the inclusion of a live/dead discriminating dye, these cells would be erroneously detected as viable within an FCM assay. Reductions in antibody staining due to physicochemical treatment were strain related, reflecting the complexity of the phenomenon under study and illustrating that substantial validation of any new antibody detection-based method, including physiological staining and cell sorting, should be undertaken. Researchers should be aware of physicochemical treatments causing false-negative results: in this study, freeze-thawing severely reduced antibody binding without affecting the viability of a substantial percentage of cells. Scanning electron microscopy carried out on treated cells revealed a range of morphological changes resulting from physicochemical treatments which may have hindered antibody binding.

Cereal type significantly affects the composition and reconstitution characteristics of dried fermented milk-cereal composites.

BACKGROUND: Dairy and cereal products are frequently combined to create composites with enhanced nutritional benefits. Commercially available dried dairy-cereal composites are typically reconstituted and cooked to produce porridge or soup. RESULTS: Dried fermented milk-cereal composites (FMCC) with ∼193 g kg-1 protein were prepared by blending fermented milk with parboiled oats (FMCCo), wheat (FMCCw), or barley (FMCCb), incubating the blend, drying, and milling. Cereal type significantly affected the composition of the FMCC and the properties of the reconstituted, cooked FMCC (R-FMCC). The FMCCo had a higher starch and fat content and lower levels of lactose, lactic acid, and amylose than FMCCb. The R-FMCCo had higher viscosity during cooking at 95 °C and cooling to 35 °C, and higher values of yield stress (σ0 ), consistency index (K) and viscosity on shearing from 20 to 120 s-1 at 60 °C than R-FMCCb. The FMCCw had lower levels of fat and ß-glucan than FMCCo or FMCCb, but was otherwise closer to FMMCb with respect to composition, cooking properties and flow behavior. CONCLUSION: Differences in composition and consistency associated with cereal type are likely to affect the nutritional value of the FMCC. © 2018 Society of Chemical Industry.

Application of Flow Cytometry to the Detection of Pathogenic Bacteria.

Outbreaks of infections have emphasized the necessity for rapid and economic detection methods for pathogens in samples ranging from those of clinical origin to food products during production and retail storage, and increasingly, in environmental samples. Flow cytometry (FCM) allows the rapid acquisition of multi-parametric data regarding cell populations within fluidised samples. However, the application of FCM to pathogen detection depends on the availability of specific fluorescent probes such as antibodies and RNA probes capable of detecting and isolating pathogens from these diverse samples. A particular issue for FCM methodology is the ability to recover and discriminate bacteria from the sample matrix which may pose a major technical hurdle towards accurate and sensitive analysis. This review article focuses on detection of pathogens using FCM in samples originating from food, water, environmental and clinical sources and outlines the current state of the art and potential future applications.

Emulsification properties of pea protein isolate using homogenization, microfluidization and ultrasonication.

Pea protein isolate (PPI) is used in many food formulations, due to its low cost, commercial availability and excellent amino acid profile. The objective of this study was to determine the emulsification properties of PPI. Particle size of PPI powders showed neither temperature (25-65°C) nor time (up to 24h) increased solubilisation of powder particles during mixing. Heating PPI dispersions (10%, w/w, protein) from 45 to 90°C led to an increase in storage modulus (G'; Pa) at 71°C, indicating the onset of protein aggregation. Gel formation occurred at 79°C (G'>1Pa). Pea protein-stabilised emulsions made using homogenization (15MPa; 1 pass) or microfluidization (50MPa; 1 pass) resulted in the formation of cold-set gels, with gel strength increasing with increasing oil concentration and fluidic pressure. Droplet size and viscosity of pea protein-stabilised emulsions decreased and increased, respectively, with increasing ultrasonication time. Overall, ultrasonication (<50°C) can create a uniform droplet size emulsion, while, homogenization and microfluidization can produce cold-set gels for use in a wide-range of food applications.

Responses of Escherichia coli, Listeria monocytogenes, and Staphylococcus aureus to simulated food processing treatments, determined using fluorescence-activated cell sorting and plate counting.

Three common food pathogenic microorganisms were exposed to treatments simulating those used in food processing. Treated cell suspensions were then analyzed for reduction in growth by plate counting. Flow cytometry (FCM) and fluorescence-activated cell sorting (FACS) were carried out on treated cells stained for membrane integrity (Syto 9/propidium iodide) or the presence of membrane potential [DiOC2(3)]. For each microbial species, representative cells from various subpopulations detected by FCM were sorted onto selective and nonselective agar and evaluated for growth and recovery rates. In general, treatments giving rise to the highest reductions in counts also had the greatest effects on cell membrane integrity and membrane potential. Overall, treatments that impacted cell membrane permeability did not necessarily have a comparable effect on membrane potential. In addition, some bacterial species with extensively damaged membranes, as detected by FCM, appeared to be able to replicate and grow after sorting. Growth of sorted cells from various subpopulations was not always reflected in plate counts, and in some cases the staining protocol may have rendered cells unculturable. Optimized FCM protocols generated a greater insight into the extent of the heterogeneous bacterial population responses to food control measures than did plate counts. This study underlined the requirement to use FACS to relate various cytometric profiles generated by various staining protocols with the ability of cells to grow on microbial agar plates. Such information is a prerequisite for more-widespread adoption of FCM as a routine microbiological analytical technique.