Dynamic water profile in various types of cheese analyzed by means of nuclear magnetic resonance relaxometry.

The aim of the study was to enquire to which extend 1H spin-lattice nuclear magnetic resonance (NMR) relaxometry data collected over a broad range of resonance frequencies (from 10 kHz to 10 MHz) have the potential to be used for assessing quality and authenticity of different categories of cheese. The following cheeses were selected mozzarella, processed cheese, pizza cheese, pizza cheese with modified fat phase), low-fat cheese, and long ripened cheese. The cheeses from 3 different production plants and various cheese production batches were used in the study. The samples from each group were subjected to instrumental composition analysis (FoodScan analyzer type 78810, FOSS, Hillerod, Denmark), water activity assessment (Aqualab 4TEV analyzer, type S40001855) and nuclear magnetic resonance relaxation dispersion study (SMARtracer FFC relaxometer, Stelar S.r.l, Italy). The state and dynamics of water present in products as free and bound water largely determines the properties of food products, including cheeses. Nuclear magnetic resonance relaxometry studies of cheese enable to reveal relaxation features characteristic of specific categories of cheese. Consequently, the studies can be treated as a step toward exploiting NMR relaxometry for accessing quality and authenticity of cheese. It was shown that at low resonance frequencies, the lower the moisture, the larger the relaxation rate. The durability and quality of cheeses depend on the presence and condition of water, so it is necessary to find a relationship between the presence, condition and mobility of water in cheeses, to increase and improve the quality and extend the shelf life.

Influence of sustainable packaging material and packaging conditions on physicochemical, microbiological, and sensorial properties of cheeses.

The aim of this study was to evaluate the influence of different packaging materials [standard foil: BOPP (biaxially oriented polypropylene)/PET (polyester)/PE (polyethylene) for upper layer, and APET (polyethylene terephthalate)/PE for bottom layer; foil 1: PP (polypropylene)/PET/PE/EVOH (ethylene-vinyl alcohol copolymer)/PE upper layer, and PP/PE/EVOH/PE bottom layer; foil 2: PP/PET/PE/EVOH/PE upper layer, and PA (polyamide)/EVOH/PE bottom layer; foil 3: PP/PET/PE upper layer, and PA/EVOH/PE bottom layer; foil 4: PP/PET/PE upper layer, and PA/PE bottom layer; foil 5: PP upper layer, and PP/PP bottom layer] on the quality of 3 different ripening rennet cheeses packed under different modified atmosphere (MAP) conditions as reflected in particular physicochemical, microbiological, and sensorial changes. The changes were monitored during a period of 90 d of storage at 2°C or 8°C. For Gouda cheese, CO2 content of the headspace of the packages was in the range 35% to 45%, whereas for Maasdamer and Sielski Klasyczny cheeses it was 55% to 65%. Three-way ANOVA showed that the foil type influenced the moisture content of Gouda cheese stored for 90 d at 2°C and for Sielski Klasyczny cheese at 8°C, whereas the moisture content was not dependent on MAP conditions during storage. Moreover, the foil type had a significant effect on free fatty acid changes for Gouda and Sielski Klasyczny cheeses stored at 2°C for 90 d. Sensory attributes changed significantly over storage time at 2°C for all studied cheeses as affected by foil type, whereas there was no effect of MAP conditions. In general, the cheeses packed in standard foil and foil 4 were characterized by the highest values of mean sensory attributes. Time was the most significant factor influencing most changes in physicochemical and sensory attributes of cheeses stored at 2°C and 8°C. The storage temperature did not affect the moisture of the samples during storage. In general, we found an increase in the pH value during storage regardless of storage temperature. It was possible to decrease the thickness of the packaging material from initial 103 and 250 µm (standard foil; lid and bottom, respectively) to 98 and 100 µm (foil 4) without affecting sensory attributes of the product.

Application of tea polyphenols as additives in brown fermented milk: Potential analysis of mitigating Maillard reaction products.

Brown fermented milk (BFM) is favored by consumers in the dairy market for its unique burnt flavor and brown color. However, Maillard reaction products (MRP) from high-temperature baking are also noteworthy. In this study, tea polyphenols (TP) were initially developed as potential inhibitors of MRP formation in BFM. The results showed that the flavor profile of BFM did not change after adding 0.08% (wt/wt) of TP, and its inhibition rates on 5-hydroxymethyl-2-furaldehyde (5-HMF), glyoxal (GO), methylglyoxal (MGO), Nε-carboxymethyl lysine (CML), and Nε-carboxyethyl lysine (CEL) were 60.8%, 27.12%, 23.44%, 57.7%, and 31.28%, respectively. After 21 d of storage, the levels of 5-HMF, GO, MGO, CML, and CEL in BFM with TP were 46.3%, 9.7%, 20.6%, 5.2%, and 24.7% lower than the control group, respectively. Moreover, a smaller change occurred in their color and the browning index was lower than that of the control group. The significance of this study was to develop TP as additives to inhibit the production of MRP in brown fermented yogurt without changing color and flavors, thereby making dairy products safer for consumers.

Synergy between oligosaccharides and probiotics: From metabolic properties to beneficial effects.

Synbiotic is defined as the dietary mixture that comprises both probiotic microorganisms and prebiotic substrates. The concept has been steadily gaining attention owing to the rising recognition of probiotic, prebiotics, and gut health. Among prebiotic substances, oligosaccharides demonstrated considerable health beneficial effects in varieties of food products and their combination with probiotics have been subjected to full range of evaluations. This review delineated the landscape of studies using microbial cultures, cell lines, animal model, and human subjects to explore the functional properties and host impacts of these combinations. Overall, the results suggested that these combinations possess respective metabolic properties that could facilitate beneficial activities therefore could be employed as dietary interventions for human health improvement and therapeutic purposes. However, uncertainties, such as applicational practicalities, underutilized analytical tools, contradictory results in studies, unclear mechanisms, and legislation hurdles, still challenges the broad utilization of these combinations. Future studies to address these issues may not only advance current knowledge on probiotic-prebiotic-host interrelationship but also promote respective applications in food and nutrition.

Organelle 16S rRNA amplicon sequencing enables profiling of active gut microbiota in murine model.

High-throughput sequencing of ribosomal RNA (rRNA) amplicons has served as a cornerstone in microbiome studies. Despite crucial implication of organelle 16S rRNA measurements to host gut microbial activities, genomic DNA (gDNA) was overwhelmingly targeted for amplicon sequencings. Although gDNA could be a reliable resource for gene existing validation, little information is revealed in regard to the activity of microorganisms owing to the limited changes gDNA undertaken in inactive, dormant, and dead bacteria. We applied both rRNA- and gDNA-derived sequencings on mouse cecal contents. Respective experimental designs were verified to be suitable for nucleic acid (NA) purification. Via benchmarking, mainstream 16S rRNA hypervariable region targets and reference databases were proven adequate for respective amplicon sequencing study. In phylogenetic studies, significant microbial composition differences were observed between two methods. Desulfovibrio spp. (an important group of anaerobic gut microorganisms that has caused analytical difficulties), Pediococcus spp., and Proteobacteria were drastically lower as represented by gDNA-derived compositions, while microbes like Firmicutes were higher as represented by gDNA-derived microbiome compositions. Also, using PICRUSt2 as an example, we illustrated that rRNA-derived sequencing might be more suitable for microbiome function predictions since pathways like sugar metabolism were lower as represented by rRNA-derived results. The findings of this study demonstrated that rRNA-derived amplicon sequencing could improve identification capability of specific gut microorganisms and might be more suitable for in silico microbiome function predictions. Therefore, rRNA-derived amplicon sequencings, preferably coupled with gDNA-derived ones, could be used as a capable tool to unveil active microbial components in host gut. KEY POINTS: • Conventional pipelines were adequate for the respective amplicon sequencing study • Groups, such as Desulfovibrio spp., were differently represented by two methods • Comparative amplicon sequencings could be useful in host active microbiota studies.

Calcium (Ca2+)-regulated exopolysaccharide biosynthesis in probiotic Lactobacillus plantarum K25 as analyzed by an omics approach.

Exopolysaccharide (EPS)-producing lactic acid bacteria have been widely used in dairy products, but how calcium, the main metal ion component in milk, regulates the EPS biosynthesis in lactic acid bacteria is not clear. In this study, the effect of Ca2+ on the biosynthesis of EPS in the probiotic Lactobacillus plantarum K25 was studied. The results showed that addition of CaCl2 at 20 mg/L in a semi-defined medium did not affect the growth of strain K25, but it increased the EPS yield and changed the microstructure of the polymer. The presence of Ca2+ also changed the monosaccharide composition of the EPS with decreased high molecular weight components and more content of rhamnose, though the functional groups of the polymer were not altered as revealed by Fourier transform infrared spectral analysis. These were further confirmed by analysis of the mRNA expression of cps genes, 9 of which were upregulated by Ca2+, including cps4F and rfbD associated with EPS biosynthesis with rhamnose. Proteomics analysis showed that Ca2+ upregulated most of the proteins related to carbon transport and metabolism, fatty acid synthesis, amino acid synthesis, ion transport, UMP synthesis. Specially, the increased expression of MelB, PtlIIBC, EIIABC, PtlIIC, PtlIID, Bgl, GH1, MalFGK, DhaK, and FBPase provided substrates for the EPS synthesis. Meanwhile, metabolomics analysis revealed significant change of the small molecular metabolites in tricarboxylic acid cycle, glucose metabolism and propionic acid metabolism. Among them the content of active small molecules such as polygalitol, lyxose, and 5-phosphate ribose increased, facilitating the EPS biosynthesis. Furthermore, Ca2+ activated HipB signaling pathway to inhibit the expression of manipulator repressor such as ArsR, LytR/AlgR, IscR, and RafR, and activated the expression of GntR to regulate the EPS synthesis genes. This study provides a basis for understanding the overall change of metabolic pathways related to the EPS biosynthesis in L. plantarum K25 in response to Ca2+, facilitating exploitation of its EPS-producing potential for application in probiotic dairy products.

Gouda Cheese with Modified Content of ß-Casein as a Source of Peptides with ACE- and DPP-IV-Inhibiting Bioactivity: A Study Based on In Silico and In Vitro Protocol.

In silico and in vitro methods were used to analyze ACE- and DPP-IV-inhibiting potential of Gouda cheese with a modified content of ß-casein. Firstly, the BIOPEP-UWM database was used to predict the presence of ACE and DPP-IV inhibitors in casein sequences. Then, the following Gouda cheeses were produced: with decreased, increased, and normative content of ß-casein after 1 and 60 days of ripening each (six variants in total). Finally, determination of the ACE/DPP-IV-inhibitory activity and the identification of peptides in respective Gouda-derived water-soluble extracts were carried out. The identification analyses were supported with in silico calculations, i.e., heatmaps and quantitative parameters. All Gouda variants exhibited comparable ACE inhibition, whereas DPP-IV inhibition was more diversified among the samples. The samples derived from Gouda with the increased content of ß-casein (both stages of ripening) had the highest DPP-IV-inhibiting potency compared to the same samples measured for ACE inhibition. Regardless of the results concerning ACE and DPP-IV inhibition among the cheese samples, the heatmap showed that the latter bioactivity was predominant in all Gouda variants, presumably because it was based on the qualitative approach (i.e., peptide presence in the sample). Our heatmap did not include the bioactivity of a single peptide as well as its quantity in the sample. In turn, the quantitative parameters showed that the best sources of ACE/DPP-IV inhibitors were all Gouda-derived extracts obtained after 60 days of the ripening. Although our protocol was efficient in showing some regularities among Gouda cheese variants, in vivo studies are recommended for more extensive investigations of this subject.

Flux and transmission of ß-casein during cold microfiltration of skim milk subjected to different heat treatments.

Raw skim milk was subjected to different heat treatments: thermization (65°C, 20 s), pasteurization (72°C, 15 s), and no heat treatment (milk was microfiltered using 1.4-µm membranes at 50°C for bacteria removal; 1.4 MF). The milk (thermized, pasteurized, and 1.4 MF) was cooled and stored at 2°C until processing (at least 24 h) with cold (∼6°C) microfiltration using a benchtop crossflow pilot unit (Pall Membralox XLAB 5, Pall Corp., Port Washington, NY) equipped with 0.1-µm nominal pore diameter ceramic Membralox membrane (ET1-070, α-alumina, Pall Corp.). The flux was monitored during the process, and ß-casein transmission and removal were calculated. The study aimed to indicate the conditions that should be applied to maximize ß-casein passage through the membrane during cold microfiltration (5.6 ± 0.4°C) of skim milk. The proper selection of heat treatment parameters (temperature, time) of the feed before the cold microfiltration process will increase ß-casein removal. It is not clear whether the difference in ß-casein transmission between 1.4 MF, thermized, and pasteurized milk results from the effect of heat treatment conditions on ß-casein dissociation from the casein micelles or on passage of ß-casein through the membrane. The values of the major parameters (permeation flux and tangential flow velocity, through the wall shear stress) responsible for a proper membrane separation process were considerably lower than the critical values. It seems that the viscosity of the retentate has a great effect on the performance of the microfiltration membranes for protein separation at refrigerated temperatures.

Short communication: Calcium partitioning during microfiltration of milk and its influence on rennet coagulation time.

Pasteurized skim milk was subjected to (1) microfiltration (MF) at 50°C and (2) MF at 6°C after storage at 2°C. The products of these treatments were retentate (RMF50) and permeate (PMF50), and retentate (RMF6) and permeate (PMF6), respectively. Additionally, RMF50 was subjected to (3) cold MF after water dilution to produce retentate (RMF6R) and permeate (PMF6R). Calcium migration was monitored by analyzing ionic, soluble, and total calcium content in feed, retentates, and permeates. The influence of calcium partitioning and calcium addition to feed, retentates, and retentates diluted with water was determined. Without CaCl2 addition, only skim milk, RMF50, and RMF6 coagulated after rennet addition. Higher true protein and casein content of RMF50 and RMF6 resulted in shorter time of renneting. The retentates diluted with water showed no signs of coagulation within 40 min. The addition of PMF6R to RMF50 did not affect rennet coagulation time within the observed 40 min in comparison to RMF50 + water. In general, higher CaCl2 addition resulted in shorter rennet coagulation time. Special attention should be paid to calcium partitioning during membrane processing of cheesemilk. The level of calcium addition should be adopted to calcium content in such cheesemilk, which is affected by conditions of the filtration process (i.e., concentration factor and temperature).

Assessment of changes in crystallization properties of pressurized milk fat.

The aim of the study was to demonstrate the use of fractal image analysis as a possible tool to monitor the effect of pressurization on the crystallization pattern of anhydrous milk fat. This approach can be useful when developing new products based on milk fat. The samples were subjected to different hydrostatic pressure (100, 200, 300, and 400 MPa) and temperature (10 and 40 °C) treatments. The crystallization microphotographs were taken with a scanning electron microscope. The image analysis of scanning electron microscope photographs was done to determine a fractal dimension. Milk-fat pressurization under the applied parameters resulted in slight, but statistically significant, changes in the course of crystallization curves, related to the triacylglycerol fraction crystallizing in the lowest temperature (I exothermic effect). These changes were dependent on the value of pressure but not dependent on the temperatures applied during the process of pressurization (at either 10 or 40 °C). In turn, significant differences were observed in crystallization images of milk-fat samples subjected to this process compared with the control sample. The results of additional fractal analysis additionally demonstrated the highest degree of irregularity of the surface of the crystalline form for the nonpressurized sample and the samples pressurized at 200 and 300 MPa at 10 °C. The lowest value of fractal dimension-indicative of the least irregularity-was achieved for the fat samples pressurized at 400 MPa, 10 °C and at 100 MPa, 40 °C. The possibilities of wider application of the fractal analysis for the evaluation of effects of parameters of various technological processes on crystallization properties of milk fat require further extensive investigations.