Unlocking the nutritional and functional potential of legume waste to produce protein ingredients.

Worldwide, many production supply chains generate a considerable amount of legume by-products (e.g., leaves, husks, broken seeds, defatted cakes). These wastes can be revalorized to develop sustainable protein ingredients, with positive economic and environmental effects. To separate protein from legume by-products, a broad spectrum of conventional (e.g., alkaline solubilization, isoelectric precipitation, membrane filtration) and novel methodologies (e.g., ultrasound, high-pressure homogenization, enzymatic approaches) have been studied. In this review, these techniques and their efficiency are discussed in detail. The present paper also provides an overview of the nutritional and functional characteristics of proteins extracted from legume by-products. Moreover, existing challenges and limitations associated with the valorization of by-product proteins are highlighted, and future perspectives are proposed.

The effect of whey source on heat-induced aggregation of casein and whey protein mixtures of relevance to infant nutritional product formulation.

Sweet and, to a lesser extent, acid whey protein ingredients can be used for the formulation of infant nutritional products. Unlike acid whey, sweet whey contains caseinomacropeptide (CMP), a heat-stable peptide liberated from κ-casein during cheese and rennet casein manufacture. Four protein systems-sweet whey (SW) and acid whey (AW), with or without standardization for CMP protein content-were added to skim milk (50/50, wt/wt) and unheated or heated to 85 or 110°C. These 12 samples were assessed for physicochemical stability in the presence of added calcium at pH 6.8. The effect of CMP content on the physicochemical properties of the protein systems was also assessed. Without preheat treatment, mixtures of AW and skim milk (SM) were more heat stable than SW and SM, demonstrating the effect of whey protein type on heat stability. Preheat treatment of the SW in the presence of SM significantly improved the heat stability of the resultant protein systems on subsequent heating. All of the protein systems had significantly lower heat stability with the addition of Ca, although the reduction was significantly smaller for the heated protein systems than the unheated controls. The findings can help identify heating parameters and ingredients for optimizing processing stability and physicochemical characteristics of nutritional beverages such as infant formulations.

Physicochemical properties of whole milk powder derived from cows fed pasture or total mixed ration diets.

This study examined the effect of dietary factors on compositional and functional properties of whole milk powder (WMP) produced from bovine milk. Raw milk samples were obtained from 3 groups of 18 Holstein Friesian spring-calving cows randomly assigned to diets based on perennial ryegrass (GRS), perennial ryegrass/white clover sward (CLV), and total mixed ration (TMR). Raw milks obtained in late lactation were subsequently standardized for fat, heat-treated (90°C for 30 s), evaporated, and homogenized before spray drying. The WMP produced from each diet were analyzed to determine differences in color, particle size distribution, heat coagulation time, yogurt gelation, texture profile, and protein profile due to each diet. Significant differences in heat coagulation time were observed between the CLV and TMR samples, whereas color values were significantly different between GRS and TMR samples. No significant differences in gross composition, protein profile, or whey protein nitrogen index were found between the 3 WMP samples. Average D90 values (the particle size at which 90% of the particles were smaller than the specified size) for fat globules were significantly lower in the TMR sample compared with the GRS and CLV samples. Yogurts produced from GRS- and CLV-derived WMP had significantly higher elastic moduli (G') than those produced from TMR-derived WMP. Similarly, texture profile analysis revealed significantly higher firmness values in yogurt samples derived from CLV compared with TMR samples. Our data characterize the effect of these diets on the composition and functional properties of fat-standardized WMP, suggesting better yogurt functionality and thermal stability in WMP derived from pasture-based bovine diets.

Dairy cow feeding system alters the characteristics of low-heat skim milk powder and processability of reconstituted skim milk.

Low-heat skim milk powder (LHSMP) was manufactured on 3 separate occasions in mid lactation (ML, July 4-20) and late lactation (LL, September 27 to October 7) from bulk milk of 3 spring-calving dairy herds on different feeding systems: grazing on perennial ryegrass (Lolium perenne L.) pasture (GRO), grazing on perennial ryegrass and white clover (Trifolium repens L.) pasture (GRC), and housed indoors and offered total mixed ration (TMR). The resultant powders (GRO-SMP, GRC-SMP, and TMR-SMP) were evaluated for composition and color and for the compositional, physicochemical, and processing characteristics of the reconstituted skim milk (RSM) prepared by dispersing the powders to 10% (wt/wt) in water. Feeding system significantly affected the contents of protein and lactose, the elemental composition, and the color of the LHSMP, as well as the rennet gelation properties of the RSM. The GRO and GRC powders had a higher protein content; lower levels of lactose, iodine, and selenium; and a more yellow-green color (lower a* and higher b* color coordinates) than TMR powder. On reconstitution, the GRO-RSM had higher concentrations of protein, casein, and ionic calcium, and lower concentrations of lactose and nonprotein nitrogen (% of total N). It also produced rennet gels with a higher storage modulus (G') than the corresponding TMR-RSM. These effects were observed over the combined ML and LL period but varied somewhat during the separate ML and LL periods. Otherwise, feeding system had little or no effect on proportions of individual caseins, concentration of serum casein, casein micelle size, casein hydration, heat coagulation time, or ethanol stability of the RSM at pH 6.2 to 7.2, or on the water-holding capacity, viscosity, and flow behavior of stirred yogurt prepared by starter-induced acidification of RSM. The differences in the functionality of the LHSMP may be of greater or lesser importance depending on the application and the conditions applied during the processing of the RSM.

Mesophilic Sporeformers Identified in Whey Powder by Using Shotgun Metagenomic Sequencing.

Spoilage and pathogenic spore-forming bacteria are a major cause of concern for producers of dairy products. Traditional agar-based detection methods employed by the dairy industry have limitations with respect to their sensitivity and specificity. The aim of this study was to identify low-abundance sporeformers in samples of a powdered dairy product, whey powder, produced monthly over 1 year, using novel culture-independent shotgun metagenomics-based approaches. Although mesophilic sporeformers were the main target of this study, in one instance thermophilic sporeformers were also targeted using this culture-independent approach. For comparative purposes, mesophilic and thermophilic sporeformers were also tested for within the same sample using culture-based approaches. Ultimately, the approaches taken highlighted differences in the taxa identified due to treatment and isolation methods. Despite this, low levels of transient, mesophilic, and in some cases potentially pathogenic sporeformers were consistently detected in powder samples. Although the specific sporeformers changed from one month to the next, it was apparent that 3 groups of mesophilic sporeformers, namely, Bacillus cereus, Bacillus licheniformis/Bacillusparalicheniformis, and a third, more heterogeneous group containing Brevibacillus brevis, dominated across the 12 samples. Total thermophilic sporeformer taxonomy was considerably different from mesophilic taxonomy, as well as from the culturable thermophilic taxonomy, in the one sample analyzed by all four approaches. Ultimately, through the application of shotgun metagenomic sequencing to dairy powders, the potential for this technology to facilitate the detection of undesirable bacteria present in these food ingredients is highlighted.IMPORTANCE The ability of sporeformers to remain dormant in a desiccated state is of concern from a safety and spoilage perspective in dairy powder. Traditional culturing techniques are slow and provide little information without further investigation. We describe the identification of mesophilic sporeformers present in powders produced over 1 year, using novel shotgun metagenomic sequencing. This method allows detection and identification of possible pathogens and spoilage bacteria in parallel. Strain-level analysis and functional gene analysis, such as identification of toxin genes, were also performed. This approach has the potential to be of great value with respect to the detection of spore-forming bacteria and could allow a processor to make an informed decision surrounding process changes to reduce the risk of spore contamination.

Pilot-scale formation of whey protein aggregates determine the stability of heat-treated whey protein solutions-Effect of pH and protein concentration.

Denaturation and consequent aggregation in whey protein solutions is critical to product functionality during processing. Solutions of whey protein isolate (WPI) prepared at 1, 4, 8, and 12% (wt/wt) and pH 6.2, 6.7, or 7.2 were subjected to heat treatment (85°C × 30 s) using a pilot-scale heat exchanger. The effects of heat treatment on whey protein denaturation and aggregation were determined by chromatography, particle size, turbidity, and rheological analyses. The influence of pH and WPI concentration during heat treatment on the thermal stability of the resulting dispersions was also investigated. Whey protein isolate solutions heated at pH 6.2 were more extensively denatured, had a greater proportion of insoluble aggregates, higher particle size and turbidity, and were significantly less heat-stable than equivalent samples prepared at pH 6.7 and 7.2. The effects of WPI concentration on denaturation/aggregation behavior were more apparent at higher pH where the stabilizing effects of charge repulsion became increasingly influential. Solutions containing 12% (wt/wt) WPI had significantly higher apparent viscosities, at each pH, compared with lower protein concentrations, with solutions prepared at pH 6.2 forming a gel. Smaller average particle size and a higher proportion of soluble aggregates in WPI solutions, pre-heated at pH 6.7 and 7.2, resulted in improved thermal stability on subsequent heating. Higher pH during secondary heating also increased thermal stability. This study offers insight into the interactive effects of pH and whey protein concentration during pilot-scale processing and demonstrates how protein functionality can be controlled through manipulation of these factors.

Outdoor grazing of dairy cows on pasture versus indoor feeding on total mixed ration: Effects on gross composition and mineral content of milk during lactation.

The influence of feeding system and lactation period on the gross composition, macroelements (Ca, P, Mg, and Na), and trace elements (Zn, Fe, Cu, Mo, Mn, Se, and Co) of bovine milk was investigated. The feeding systems included outdoor grazing on perennial ryegrass pasture (GRO), outdoor grazing on perennial ryegrass and white clover pasture (GRC), and indoors offered total mixed ration (TMR). Sixty spring-calving Holstein Friesian dairy cows were assigned to 3 herds, each consisting of 20 cows, and balanced with respect to parity, calving date, and pre-experimental milk yield and milk solids yield. The herds were allocated to 1 of the 3 feeding systems from February to November. Milk samples were collected on 10 occasions over the period June 17 to November 26, at 2 or 3 weekly intervals, when cows were on average 119 to 281 d in lactation (DIL). The total lactation period was arbitrarily sub-divided into 2 lactation periods based on DIL, namely mid lactation, June 17 to September 9 when cows were 119 to 203 DIL; and late lactation, September 22 to November 26 when cows were 216 to 281 DIL. With the exception of Mg, Na, Fe, Mo, and Co, all other variables were affected by feeding system. The GRO milk had the highest mean concentrations of total solids, total protein, casein, Ca, and P. The TMR milk had the highest concentrations of lactose, Cu, and Se, and lowest level of total protein. The GRC milk had levels of lactose, Zn, and Cu similar to those of GRO milk, and concentrations of TS, Ca, and P similar to those of TMR milk. Lactation period affected all variables, apart from the concentrations of Fe, Cu, Mn, and Se. On average, the proportion (%) of total Ca, P, Zn, Mn, or Se that sedimented with the casein on high-speed ultracentrifugation at 100,000 × g was ≥60%, whereas that of Na, Mg, or Mo was ≤45% total. The results demonstrate how the gross composition and elemental composition of milk can be affected by different feeding systems.

Grazing of dairy cows on pasture versus indoor feeding on total mixed ration: Effects on low-moisture part-skim Mozzarella cheese yield and quality characteristics in mid and late lactation.

This study investigated the effects of 3 dairy cow feeding systems on the composition, yield, and biochemical and physical properties of low-moisture part-skim Mozzarella cheese in mid (ML; May-June) and late (LL; October-November) lactation. Sixty spring-calving cows were assigned to 3 herds, each consisting of 20 cows, and balanced on parity, calving date, and pre-experimental milk yield and milk solids yield. Each herd was allocated to 1 of the following feeding systems: grazing on perennial ryegrass (Lolium perenne L.) pasture (GRO), grazing on perennial ryegrass and white clover (Trifolium repens L.) pasture (GRC), or housed indoors and offered total mixed ration (TMR). Mozzarella cheese was manufactured on 3 separate occasions in ML and 4 in LL in 2016. Feeding system had significant effects on milk composition, cheese yield, the elemental composition of cheese, cheese color (green to red and blue to yellow color coordinates), the extent of flow on heating, and the fluidity of the melted cheese. Compared with TMR milk, GRO and GRC milks had higher concentrations of protein and casein and lower concentrations of I, Cu, and Se, higher cheese-yielding capacity, and produced cheese with lower concentrations of the trace elements I, Cu, and Se and higher yellowness value. Cheese from GRO milk had higher heat-induced flow and fluidity than cheese from TMR milk. These effects were observed over the entire lactation period (ML + LL), but varied somewhat in ML and LL. Feeding system had little, or no, effect on gross composition of the cheese, the proportions of milk protein or fat lost to cheese whey, the texture of the unheated cheese, or the energy required to extend the molten cheese. The differences in color and melt characteristics of cheeses obtained from milks with the different feeding systems may provide a basis for creating points of differentiation suited to different markets.

Chlorate and Other Oxychlorine Contaminants Within the Dairy Supply Chain.

The presence of chlorate in milk and dairy products can arise from the use of chlorinated water and chlorinated detergents for cleaning and sanitation of process equipment at both farm and food processor level. Chlorate and other oxychlorine species have been associated with inhibition of iodine uptake in humans and the formation of methemoglobin, with infants and young children being a high-risk demographic. This comprehensive review of chlorate and chlorine derivatives in dairy, highlights areas of concern relative to the origin and/or introduction of chlorate within the dairy supply chain. This review also discusses the associated health concerns, regulations, and chemical behavior of chlorate and chlorine-derived by-products, and provides a summary of mechanisms for their detection and removal.

Determining the influence of fava bean pre-processing on extractability and functional quality of protein isolates.

In this study, fava bean protein (FPI) was isolated from flours prepared from dehulled seeds and compared to FPI extracted from whole flours; in the latter case, flours were prepared either by dry- or wet-milling. Significant differences in composition and functionality were observed between the three FPIs produced. Dehulling maximized protein purity, oil-absorption capacity, solubility, foamablity and minimized both starchy and non-starchy carbohydrate contents. Protein isolated from wet-milled whole beans provided higher mass and extraction yields, better water-absorption capacity, and exhibited higher surface charge (zeta potential) compared to other samples. The protein extracted from dry-milled whole seed exhibited a higher least gelation concentration, emulsifying activity and zeta value compared to its dehulled counterpart. Dehulling was also found to be a useful process to increase the lightness/whiteness of protein powder. Overall, the present findings provide useful technological information relating to the production of FPI with and without a dehulling step.

Seasonal and geographical impact on the Irish raw milk microbiota correlates with chemical composition and climatic variables.

Season and location have previously been shown to be associated with differences in the microbiota of raw milk, especially in milk from pasture-based systems. Here, we further advance research in this area by examining differences in the raw milk microbiota from several locations across Ireland over 12 months, and by investigating microbiota associations with climatic variables and chemical composition. Shotgun metagenomic sequencing was used to investigate the microbiota of raw milk collected from nine locations (n = 241). Concurrent chemical analysis of the protein, fat, lactose, total solids, nonprotein nitrogen contents, and titratable acidity (TA) of the same raw milk were performed. Although the raw milk microbiota was highly diverse, a core microbiota was found, with Pseudomonas_E, Lactococcus, Acinetobacter, and Leuconostoc present in all samples. Microbiota diversity significantly differed by season and location, with differences in seasonality and geography corresponding to 11.8% and 10.5% of the variation in the microbiota. Functional and antibiotic resistance profiles also varied across season and location. The analysis of other metadata revealed additional interactions, such as an association between mean daily air and grass temperatures with the abundance of spoilage taxa like Pseudomonas species. Correlations were identified between pathogenic, mastitis-related species, fat content, and the number of sun hours, suggesting a seasonal effect. Ultimately, this study expands our understanding of the interconnected nature of the microbiota, environment/climate variables, and chemical composition of raw milk and provides evidence of a season- and location-specific microbiota. IMPORTANCE: The microbiota of raw milk is influenced by many factors that encourage or prevent the introduction and growth of both beneficial and undesirable microorganisms. The seasonal and geographical impacts on the microbial communities of raw milk have been previously seen, but the relationships with environmental factors and the chemical composition has yet to be investigated. In this year-long study, we found that while raw milk is highly diverse, a core microbiota was detected for Irish raw milk, with strong evidence of seasonal and geographical influence. We also found associations between groups of microorganisms, environmental factors, and milk composition, which expand current knowledge on the relationships between microbial and chemical composition and the climate. These results provide evidence for the development of a tool to allow for the prediction of raw milk quality and safety.

Effect of gelatinisation of starch with casein proteins on incretin hormones and glucose transporters in vitro.

Foods that have a low glycaemic index or foods that contain slowly digestible starch are beneficial in controlling fluctuations in blood glucose and insulin levels. The study hypothesis is that gelatinisation of starch in structured casein networks provides a method for decreasing the digestion rate of the starch and, hence, minimising postprandial glucose fluctuations. This study examined the effect of starch gelatinisation with or without casein on (1) gene expression and peptide secretion levels of the incretin hormones glucagon-like peptide 1 and glucose-independent insulinotropic polypeptide and (2) gene expression of the sodium-glucose cotransporter and GLUT-2 in intestinal cell culture systems. The intestinal epithelial cell line, STC-1, and the enteroendocrine colonic cell line, Caco-2, were exposed to in vitro digested foods (starch gelatinised with α-casein, starch gelatinised with ß-casein and gelatinised starch alone). The encapsulation of starch with casein before in vitro digestion lowers levels of incretin hormone secretion. Digestion of starch gelatinised with casein also releases less glucose than starch alone as indicated by significantly (P < 0·05) lower levels of glucose transporter mRNA transcripts. Some subtle cellular response differences were observed following exposure to starch gelatinised with α- compared to ß-casein. Fractionation of α-casein and ß-casein by reverse-phase HPLC identified that fractions that differed in hydrophobicity differed significantly (P < 0·05) in their ability to promote secretion of the incretin hormones. Evidence suggests that gelatinisation of starch with casein may be a functional food ingredient that minimises blood glucose fluctuations.

Spectroscopic technologies and data fusion: Applications for the dairy industry.

Increasing consumer awareness, scale of manufacture, and demand to ensure safety, quality and sustainability have accelerated the need for rapid, reliable, and accurate analytical techniques for food products. Spectroscopy, coupled with Artificial Intelligence-enabled sensors and chemometric techniques, has led to the fusion of data sources for dairy analytical applications. This article provides an overview of the current spectroscopic technologies used in the dairy industry, with an introduction to data fusion and the associated methodologies used in spectroscopy-based data fusion. The relevance of data fusion in the dairy industry is considered, focusing on its potential to improve predictions for processing traits by chemometric techniques, such as principal component analysis (PCA), partial least squares regression (PLS), and other machine learning algorithms.

Powder and Reconstituted Properties of Commercial Infant and Follow-On Formulas.

The physical properties of 15 commercially available infant formulas (IF) and follow-on (FO) formulas were analysed. Powders made with intact milk proteins were classified into two groups; Type I-homogenous mixtures of milk powder particles (n = 6); and Type II-heterogeneous mixtures of milk powder particles and tomahawk-shaped α-lactose monohydrate crystals (n = 6). Powders made using hydrolysed proteins were classified as Type III powders (n = 3). Type II powders exhibited similar flow characteristics to Type I powders despite having significantly (p < 0.05) smaller particle size, lower circularity, and greater elongation. Type III powders exhibited lowest particles size, highest surface free fat, and poorest flow properties (p < 0.05 for all). Upon reconstitution of powders (12.5% w/w), no significant difference (p < 0.05) in apparent viscosity was observed between Type I and II powders. Reconstituted Type III powders had relatively poor stability to separation compared to Type I and II powders, caused by large starch granules and/or poor emulsification by hydrolysed proteins. Overall, this study illustrated the range of physical behaviour and structures present in commercial IF powders. In particular, the effect of dry addition of lactose and the hydrolysis of protein were found to have major effects on physical properties.

Tracking the Dairy Microbiota from Farm Bulk Tank to Skimmed Milk Powder.

Microorganisms from the environment can enter the dairy supply chain at multiple stages, including production, milk collection, and processing, with potential implications for quality and safety. The ability to track these microorganisms can be greatly enhanced by the use of high-throughput DNA sequencing (HTS). Here HTS, both 16S rRNA gene amplicon and shotgun metagenomic sequencing were applied to investigate the microbiomes of fresh mid- and late-lactation milk collected from farm bulk tanks, collection tankers, milk silos, skimmed milk silos, a cream silo, and powder samples to investigate the microbial changes throughout a skim milk powder manufacturing process. 16S rRNA gene analysis established that the microbiota of raw milks from farm bulk tanks and in collection tankers were very diverse but that psychrotrophic genera associated with spoilage, Pseudomonas and Acinetobacter, were present in all samples. Upon storage within the whole-milk silo at the processing facility, the species Pseudomonas fluorescens and Acinetobacter baumannii became dominant. The skimmed milk powder generated during the mid-lactation period had a microbial composition that was very different from that of raw milk; specifically, two thermophilic genera, Thermus and Geobacillus, were enriched. In contrast, the microbiota of skimmed milk powder generated from late-lactation milk more closely resembled that of the raw milk and was dominated by spoilage-associated psychrotrophic bacteria. This study demonstrates that the dairy microbiota can differ significantly across different sampling days. More specifically, HTS can be used to trace microbial species from raw milks through processing to final powdered products.IMPORTANCE Microorganisms can enter and persist in dairy at several stages of the processing chain. Detection of microorganisms within dairy food processing is currently a time-consuming and often inaccurate process. This study provides evidence that high-throughput sequencing can be used as an effective tool to accurately identify microorganisms along the processing chain. In addition, it demonstrates that the populations of microbes change from raw milk to the end product. Routine implementation of high-throughput sequencing would elucidate the factors that influence population dynamics. This will enable a manufacturer to adopt control measures specific to each stage of processing and respond in an effective manner, which would ultimately lead to increased food safety and quality.

Effect of Diet on the Vitamin B Profile of Bovine Milk-Based Protein Ingredients.

The influence of diet on the water-soluble vitamin composition of skim milk powder and whey protein ingredients produced from the milk of cows fed pasture or concentrate-based diets was examined. Fifty-one Holstein-Friesian cows were randomly assigned into three diets (n=17) consisting of outdoor grazing of perennial ryegrass (GRS), perennial ryegrass/white clover (CLV), or indoor feeding of total mixed ration (TMR) for an entire lactation. Raw mid-lactation milk from each group was processed into skim milk powder and further processed to yield micellar casein whey and acid whey. Sweet whey was also produced by renneting of pasteurised whole milk from each system. The water-soluble vitamin profile of each sample was analysed using a combination of direct injection mass spectrometry and reverse-phase liquid chromatography-mass spectrometry. Vitamin B3 and B3-amide concentrations were significantly higher (p < 0.05) in TMR-derived samples than in those from CLV and GRS, respectively. Vitamin B1, B2, and B7 concentrations were significantly higher in GRS and CLV-derived samples than those from TMR. Significant differences in vitamins B1, B2, and B3-amide were also observed between protein ingredient types. This study indicates that bovine feeding systems have a significant effect on B vitamin composition across a range of protein ingredient types.

Dephosphorylation of caseins in milk protein concentrate alters their interactions with sodium hexametaphosphate.

This study investigated the effects of dephosphorylation and sodium hexametaphosphate (SHMP) salt addition on the viscosity of milk protein concentrate (MPC) solutions. Dephosphorylation (DP) of casein was performed using bovine alkaline phosphatase. Nuclear magnetic resonance (NMR) spectra showed that dephosphorylation depleted the casein-bound phosphate region (CNP). SHMP addition (5 mM) had no impact on the 31P NMR spectra of DP-MPC; addition of 5 mM SHMP to control MPC (C-MPC) resulted in a shift in peaks associated with the CNP region, possibly caused by SHMP sequestering calcium, leading to swelling of micelles. DP-MPC exhibited a lower viscosity compared to C-MPC, with SHMP addition at 12.5 and 25 mM causing gelation of C-MPC and DP-MPC solutions. This work confirmed the role that phosphate residues have in maintaining micelle structural stability and provides new insights into controlling viscosity of MPC solutions.

Influence of herd diet on the metabolome of Maasdam cheeses.

The untargeted metabolic profiles of ripened Maasdam cheese samples prepared from milk derived from three herd groups, fed: (1) indoors on total mixed ration (TMR), or outdoors on (2) grass only pasture (GRA) or (3) grass and white clover pasture (CLO) were studied using high resolution nuclear magnetic resonance (1H NMR), high resolution magic angle spinning nuclear magnetic resonance (1H HRMAS NMR) and headspace (HS) gas chromatography mass spectrometry (GC-MS). A total of 31 compounds were identified using 1H NMR and 32 volatile compounds including 7 acids, 5 esters, 4 alcohols, 4 ketones, 4 sulfur compounds, 2 aldehydes, 3 hydrocarbons, 2 terpenes and a lactone were identified using GC-MS in Maasdam cheeses ripened for 97-d. On comparing the 1H NMR metabolic profiles, TMR-derived cheese had higher levels of citrate compared to GRA-derived cheese. The toluene content of cheese was significantly higher in GRA or CLO compared to TMR cheeses and dimethyl sulfide was identified only in CLO-derived cheese samples as detected using HS GC-MS. These compounds are proposed as indicator compounds for Maasdam cheese derived from pasture-fed milk. Clear differences between outdoor or indoor feeding systems in terms of cheese metabolites were detected in the lipid phase, as indicated by principal component analysis (PCA) from 1H HRMAS NMR spectra, although differences based on PCA of all 1H NMR spectra and HS-GC-MS were less clear. Overall, this study presented the metabolite profile and identified specific compounds which may be useful for discriminating between ripened Maasdam cheese and related cheese varieties manufactured from indoor or outdoor herd-feeding systems.

The "Grass-Fed" Milk Story: Understanding the Impact of Pasture Feeding on the Composition and Quality of Bovine Milk.

Milk is a highly nutritious food that contains an array of macro and micro components, scientifically proven to be beneficial to human health. While the composition of milk is influenced by a variety of factors, such as genetics, health, lactation stage etc., the animal's diet remains a key mechanism by which its nutrition and processing characteristics can be altered. Pasture feeding has been demonstrated to have a positive impact on the nutrient profile of milk, increasing the content of some beneficial nutrients such as Omega-3 polyunsaturated fatty acids, vaccenic acid, and conjugated linoleic acid (CLA), while reducing the levels of Omega-6 fatty acids and palmitic acid. These resultant alterations to the nutritional profile of "Grass-Fed" milk resonate with consumers that desire healthy, "natural", and sustainable dairy products. This review provides a comprehensive comparison of the impact that pasture and non-pasture feeding systems have on bovine milk composition from a nutritional and functional (processability) perspective, highlighting factors that will be of interest to dairy farmers, processors, and consumers.

Impact of Bovine Diet on Metabolomic Profile of Skim Milk and Whey Protein Ingredients.

The influence of bovine diet on the metabolome of reconstituted skim milk powder (SMP) and protein ingredients produced from the milk of cows fed on pasture or concentrate-based diets was investigated. Cows were randomly assigned to diets consisting of perennial ryegrass only (GRS), perennial ryegrass/white clover sward (CLV), or indoor total mixed ration (TMR) for an entire lactation. Raw milk obtained from each group was processed at pilot scale, to produce SMP and sweet whey, and SMP was further processed at laboratory scale, to yield ideal whey and acid whey. The total amino acid composition and metabolome of each sample were analyzed, using high-performance cation exchange and a targeted combination of direct-injection mass spectrometry and reverse-phase liquid chromatography-tandem mass spectrometry (LC-MS/MS), respectively. The nitrogen composition of the products from each of the diets was similar, with one exception being the significantly higher nonprotein nitrogen content in TMR-derived skim milk powder than that from the GRS system. Total amino acid analysis showed significantly higher concentrations of glycine in GRS- and CLV-derived sweet whey and acid whey than in those from TMR. The cysteine contents of CLV-derived ideal whey and acid whey were significantly higher than for TMR, while the valine content of GRS-derived acid whey was significantly higher than TMR. The phenylalanine content of GRS-derived ideal whey was significantly higher than that from CLV. Metabolomic analysis showed significantly higher concentrations of the metabolites glutamine, valine, and phosphocreatine in each ingredient type derived from TMR than those from GRS or CLV, while the serine content of each GRS-derived ingredient type was significantly higher than that in TMR-derived ingredients. These results demonstrate that the type of bovine feeding system used can have a significant effect on the amino acid composition and metabolome of skim milk and whey powders and may aid in the selection of raw materials for product manufacture, while the clear separation between the samples gives further evidence for distinguishing milk products produced from different feeding systems based on LC-MS/MS.