This study aimed to determine lactoferrin (LF) in breast milk-based powders and formulas. Lactoferrin is an important whey protein in all mammalian milks and is responsible in large part for the known antimicrobial effects of human milk in particular. As breast feeding is not always possible, formulas based on cows milk have been developed in order to meet the nutritional needs of the newborn, while more recently human breast milk-based powders have been introduced to offer the biological functionality of human milk to pre-term and critically ill babies. In the present work, the amount of LF in commercial breast milk-based powders was tested by a validated RF-HPLC method for the determination of LF in breast milk in order to examine both the applicability of the method but at a second level the amount of LF in these commercial products. The detection of LF was possible but the complexity of the matrix lead us to the use the standard addition methodology in order to achieve quantification. The results indicated that breast milk-based powders had higher amount of LF than cows milk-based formulas, both non-fortified and fortified.

We evaluated the possibility of increasing the storage temperature of raw milk for Provolone Valpadana cheesemaking, to identify the most suitable conditions of time and temperature for a pre-maturation process. We used Principal Component Analysis (PCA) to analyze the overall effects of different storage conditions on chemical, nutritional and technological characteristics of the raw milk. Four different thermal storage cycles, two at fixed temperature/time (6 and 12°C for 60 h) and two with two-phase thermal cycle (10 and 12°C for 15 h, followed by refrigeration at 4°C for 45 h) were studied. Although a moderate heterogeneity among raw milks from the 11 producers of Provolone Valpadana cheese was observed, PCA revealed the critical aspects of the extreme storage conditions (60 h of refrigeration). Some samples resulted in anomalous behaviors, probably related to unexpected fermentation phenomena occurring with increasing storage temperature. The acidification and the increase in the contents of lactic acid, soluble calcium, and degree of retinol isomerization observed in the anomalous samples can compromise the technological functionality of milk. Conversely, the storage with a two-phase thermal cycle did not lead to variations in any measured characteristic, suggesting that mild refrigeration conditions (10 or 12°C for 15 h followed by 4°C for 45 h) could be a good compromise in favoring milk pre-maturation without altering its quality characteristics.

Reuterin is a broad-spectrum antimicrobial substance produced by lactic acid bacteria, and most previous studies have reported that reuterin is only produced under anaerobic conditions. If there are lactic acid bacteria that also produce it under aerobic conditions, it could be applied to fermented foods. In this study, it was found that Lactobacillus coryniformis WBB05 showed optimal reuterin production (123 mM reuterin from 200 mM glycerol) when incubated aerobically at 20°C. Furthermore, the minimum inhibitory concentration (MIC) of reuterin was determined for starter lactic acid bacteria strains and cheese moulds. MIC toward Penicillium camemberti was 0.125 mM and the white mould starter was much more sensitive than other moulds.

Functions of mucin, as the major macromolecular component in saliva or gastric fluids, are drawing increasing attention in the context of understanding the oral processing or digestion of dairy foods at the molecular level. This study was designed to investigate the interactions between β-lactoglobulin (BLG)-lactose, mucin-lactose and BLG-lactose-mucin at the molecular level under different temperature and pH conditions using fluorescence spectroscopy in combination with scanning electron microscopes (sem). It is the first study of its kind. There was no lactose-dependent quenching on BLG fluorophore in the range of 0–10 mM lactose concentration. On the contrary, there was a continuous increase in the fluorescence intensity of the BLG protein when the lactose concentration increased, especially at 25°C. BLG-lactose complex became thermally unstable at 37 and 45°C. Moreover, BLG exhibited a pH dependent conformational change and had higher fluorescence intensity at pH 3 than pH 6.8. The fluorescence result was in correspondence with sem images where we observed lactose crystals gathering around and on the BLG molecule, but lactose molecules could not be seen in the presence of mucin. It was anticipated that mucin molecules interacted with BLG-lactose complex via electrostatic attraction and formed an extra protective layer around the BLG molecules to avoid solvent exposure.

This Research Communication aims to compare the effect of A1A2 and A2A2 cow milk diets on the biochemical and histological parameters of rats. The rats were divided into four groups and fed with a normal diet, A2 milk powder, A1A2 or A2A2 cow milk diets for 90 d. Blood glucose, kidney function, liver function and lipid profile were examined during the experimental period. The study showed an increase in the body weight of the A1A2 group whereas a slight decrease in the A2A2 group, and blood glucose levels increased from d 0 to day 90 in all experimental groups. However, none of these changes were found to be statistically insignificant (P > 0.05). Moreover, no significant changes were recorded in other parameters (serum glutamic pyruvic transferase and serum glutamic-oxaloacetic transaminase for liver function, bilirubin direct, cholesterol, triglycerides, creatinine and uric acid). The histology of the liver, kidney and pancreas also showed no changes in all groups. Overall, this study revealed no significant difference in the nutritional values of A1A2 and A2A2 milk types and hence equally beneficial for health. Although the present study showed no significant difference in the effect of both milk types in 90 d, further studies might be conducted to evaluate their longer term effects.

The purpose of the research paper was firstly to identify bacteriocin-producing lactic acid bacteria characterizing strains with anti-listeria activity and, secondly, to characterize bacteriocin evaluating its in vitro efficiency as a natural preservative and, thirdly, to evaluate the anti-listeria effect of the bacteriocinogenic strain of Lactiplantibacillus plantarum in cheeses and produce an edible film with anti-listerial effect. Of 355 lactic acid bacteria strains tested, two were able to produce bacteriocin against Listeria monocytogenes and were identified as Lactiplantibacillus plantarum and Lactiplantibacillus pentosus. A bactericidal effect of strain QS494 (Lactiplantibacillus plantarum) was observed in the first 8 h, with a reduction of 1.7 log, using cell-free supernatant with Listeria monocytogenes, where viable cells were counted on listeria selective agar. Both strains showed good technological characteristics and were without production of virulence factors. Changes in the pH of the cell-free supernatant obtained from Lactiplantibacillus plantarum did not affect its antimicrobial activity, which remained stable after heat treatments for up to 15 min at 121°C. Inhibitory activity was also observed after 12 weeks of storage at −20°C. In the evaluation of the anti-listeria effect in cheeses, a 3 log reduction in the Listeria monocytogenes count was observed in 120 h in cheeses produced with bacteriocinogenic lactic acid bacteria, while in cheeses produced with non-bacteriocinogenic culture, we observed a 2 log increase in the count. Edible films produced with the addition of precipitate from the cell free supernatant showed an antimicrobial effect against Listeria monocytogenes. Thus, the two strains studied have technological and biosafety potential.

We investigated four different production methods of Künefe cheese, emphasizing their industrial importance. The four methods used fresh Künefe cheese (FKC), salted Künefe cheese (SKC), Boru type Künefe cheese (BKC) and culture-added processed Künefe cheese (CPKC), all used in the production of Künefe, a syrupy dessert unique to the Middle East. FKC was produced as a result of curd formation from raw milk with rennet and then the fermentation of curd. SKC was manufactured using the salting method in addition to FKC production. BKC was obtained using the dry cooking process with emulsifying salts applied to cheese curd. CPKC was produced by applying heat treatment to raw milk and using starter culture before adding rennet that was different to Boru-type Künefe cheese. The composition, color, meltability, texture and sensory properties of the Künefe cheeses were examined. As a result of statistical analysis, it was determined that the effect of different production methods on the composition, meltability, texture (except springiness and cohesiveness) and sensory properties was significant in all cheeses (P < 0.05). CKPC was the most appropriate cheese in various properties.

This study was conducted to determine the effect of acidophilus yoghurt (yoghurt fortified with Lactobacillus acidophilus) in comparison to traditional plain yoghurt (St. thermophilus and L. bulgaricus starter cultures) on the survival of three pathogenic Escherichia coli strains; Shiga toxigenic O157 (STx O157), non-toxigenic O157 (Non-STx O157) and Shiga toxigenic non-O157 (STx O145). After six days of refrigerated storage of laboratory-manufactured yoghurt inoculated with the three strains of E. coli separately, all were eliminated in acidophilus yoghurt, while their survival extended in the traditional yoghurt along the storage period (17 d). Reduction percentages of the tested strains in acidophilus yoghurt were 99.93, 99.93 and 99.86%, with log reduction of 3.176, 3.176, and 2.865 cfu/g for Stx O157, Non-Stx O157, and Stx O145 E. coli, respectively, in comparison to 91.67, 93.33 and 93.33%, with log reduction of 1.079, 1.176 and 1.176 cfu/g in traditional yoghurt. Statistical analysis showed a significant effect of acidophilus yoghurt in reducing the count of Stx E. coli O157 (P = 0.001), Non-Stx E. coli O157 (P < 0.01) and Stx E. coli O145 (P < 0.01) compared to the traditional yoghurt. These findings emphasize the potential use of acidophilus yoghurt as a biocontrol alternative method for eliminating pathogenic E. coli, as well as other similar applications in the dairy industry.

Eggnog, a dairy-based beverage, comprises both milk and egg proteins. We aimed at optimizing the eggnog formulation using Box–Behnken design of response surface methodology. The combined effects of milk (60–75), cream (25–35) and eggnog base (6–8, all three as g 100/ml) were investigated on heat coagulation time, viscosity and thermal gelation temperature. ANOVA indicated that experimental data were well explained by a quadratic model with high check values (R2 > 0.94) and non-significant lack of fit tests. Based on the responses, an optimized formulation of eggnog with 60.0 milk, 25.0 cream and 6.50 eggnog base (as g 100/ml), could be considered best for manufacturing eggnog with desired attributes. This optimized formulation was characterized for physico-chemical, microbial and sensory attributes and the results indicated significantly higher fat and protein content than control formulation, but lesser lactose and total sugar content. Significantly higher viscosity, heat stability and lower thermal gelation temperature were also observed for the optimized formulation. Coliform, yeast and mold, E. coli and Salmonella counts were not detected in any sample but a significantly lower total plate count was observed for the optimized formulation.

Health-conscious consumers are increasingly paying attention to healthy diets and focusing on natural bioactive compounds in foods and their effects on mental health. This opens new opportunities for the study of artisanal cheeses as biofunctional foods. In the work described in this Research Communication, the gamma-aminobutyric acid (GABA) content of seven different Portuguese cheeses produced from unpasteurized cow, sheep, and goat milk and granted with protected designation of origin (PDO) status was analysed. The PDO cheeses made from cow milk analysed in this study were São Jorge (3, 4, 7, 12 and 24 months of maturation) and Pico cheeses. PDO cheeses made from sheep milk were Serra da Estrela, Serpa, Nisa and Azeitão. Cheeses made from sheep and goat milk included Beira Baixa yellow cheese. The GABA content in the Azorean PDO cheeses (made from cow milk) ranged from 1.23 to 2.64 g/kg of cheese. Higher variations in GABA content were observed in cheeses made from sheep and goat milk (0.73–2.31 g/kg). This study provides information on the GABA content in different Portuguese PDO cheeses and shows that hard or semi-hard ripened cheeses are a suitable matrix for GABA production by lactic acid bacteria.

The aim of the research described here was to investigate the in vitro immunomodulatory effects of 3RS, 7R, 11R-phytanic acid (3RS-PHY) from the perspective of efficacy against autoimmune diseases. 3RS-PHY is a milk component with strong agonist activity at the peroxisome proliferator activated receptor (PPAR). As PPAR is a therapeutic target for several human diseases, 3RS-PHY intake may have possible health benefits. Recently, we chemically synthesized a preparation of 3RS-PHY and demonstrated that 3RS-PHY inhibited T-cell production of interferon (IFN)-γ. However, the overall immunomodulatory effects were not evaluated. In this study, mouse splenocytes, purified T-cells and B-cells were stimulated by mitogens and incubated with 3RS-PHY, followed by evaluation of cytokine and antibody production. A macrophage-like cell line J774.1 was also incubated with 3RS-PHY to evaluate nitric oxide production. 3RS-PHY decreased mRNA levels not only of IFN-γ but also of interleukin (IL)-2, IL-10 and IL-17A in splenocytes and similar effects were confirmed at the protein level. In addition, 3RS-PHY had a direct action on T-cells with preferential inhibitory effects on Th1 and Th17 cytokines such as IFN-γ and IL-17A. Furthermore, 3RS-PHY suppressed antibody secretion by B-cells and nitric oxide production by J774.1 almost completely, indicating that 3RS-PHY is a bioactive fatty acid with anti-inflammatory properties. These findings encourage further investigations, including in vivo experiments, to evaluate whether 3RS-PHY actually shows the potential to prevent autoimmune diseases, and provide basic information to produce milk and dairy products with an increased 3RS-PHY concentration.

This research was conducted to evaluate the knowledge and purchasing behaviour of consumers regarding lactose-free foods. Study data were collected from 1882 individuals with the help of a questionnaire form. Attitudes towards animal welfare, healthy nutrition and gastrointestinal symptoms were also evaluated. The majority (86.6%) of the participants reported that they had heard of the concept of lactose-free foods before, but more than half (67.0%) did not report buying them because of their higher costs compared to regular products and lack of sufficient information. Not causing gas/bloating (32.5%) and being healthier (28.3%) were shown to be the most significant motivational factors in purchasing lactose-free foods (P < 0.05). The factors affecting purchase of lactose-free products were education, employment status, history of having a food intolerance test and animal welfare perception together with higher scores from the gastrointestinal symptom rating scale, and attitude scale for healthy nutrition (P < 0.05). Body mass index was not found to be an effective factor determining purchase intention. The results provide a general overview of consumers, however, further studies on other demographic groups are needed.