The present study was aimed at understanding the high pressure (HP) processing-induced interactions of milk proteins in whey protein concentrate (WPC) solutions, in skim milk and in pure protein systems. The changes in milk proteins induced by heat treatments in the same systems under selected conditions were also evaluated. The main approach taken was to elucidate changes in the whey proteins in heat- and pressure-treated samples from common aliquots, under identical conditions, using various one-dimensional (1D) and two-dimensional (2D) polyacrylamide gel electrophoresis (PAGE) techniques in the absence or presence of a disulphide bond reducing agent. In some instances, the samples were also analysed using small deformation rheology, size exclusion chromatography (SEC) and transmission electron microscopy (TEM). The results of the present study indicated that, in general terms, heat treatment and HP treatment had common effects, i.e. denaturation and subsequent aggregation of whey proteins. Both heat treatment and HP treatment generated disulphide-bonded and hydrophobically bonded aggregates of whey proteins. However, the sensitivities of each of the whey proteins to heat treatment [immunoglobulin (Ig) > lactoferrin (LF) > bovine serum albumin (BSA) > β-laetoglobulin B (β-LG B) > β-LG A > α-lactalbumin (α-LA)] and pressure treatment (β-LG B > β-LG A > IgG > LF > BSA > α-LA) were considerably different. Also, HP treatment generated a comparatively greater proportion of smaller aggregates than did heat treatment. The effects of protein concentration, intensity of pressure treatment, holding time and pressurising temperature on whey protein aggregation in WPC solutions were investigated. The rate of aggregation of whey proteins increased with an increase in the concentration of protein in the WPC solution and the pressurising temperature. The combination of low protein concentration, mild pressure treatment (200 MPa) and low pressurising temperature (20°C) led to minimal loss of native-like and SDS-monomeric β-LG, whereas the combination of high protein concentration, severe pressure treatment (600 MPa) and higher pressuring temperature (40°C and higher) led to significant loss of both native-like and SDS-monomeric β-LG. The sensitivity of pressure-resistant whey proteins, such as α-LA and BSA, to the aggregation was significantly increased at pressurising temperatures of 40°C and higher. Self-supporting gels were formed when 8 or 12% (w/v) WPC solutions were pressure treated at 600-800 MPa. 20°C. Detailed analysis of the behaviour of the proteins during the formation of these gels revealed a novel pathway, suggesting that intermolecular disulphide bond formation occurred at high pressure but that hydrophobic association became important after the HP treatment. In the later part of the study, heat- and HP-induced interactions of caseins and whey proteins were studied in a more complex system, i.e. skim milk. With the application of modified PAGE techniques, it was possible to show that the high molecular weight disulphide-bonded aggregates that were formed by HP treatment of skim milk contained disulphide-linked complexes consisting of αS2-casein (αS2-CN) as well as κ-CN, β-LG and other whey proteins. The results showed that the effects of heat treatment and HP on the interactions of the caseins and whey proteins in milk were significantly different. The accessibility of αS2-CN and the formation of complexes involving αS2-CN, κ-CN and whey proteins in the HP-treated milk, as demonstrated using the modified 2D PAGE technique, and as explained by possible proposed reactions of the caseins and whey proteins in pressure-treated milk, was an important finding of the present study. Finally, a study on the effects of HP treatment in model systems using pure proteins in solution, both singly or in binary and ternary combinations, generated very useful information and clarified the role of each protein in pressure-induced aggregation and interactions of milk proteins in complex systems such as WPC and milk. It was found that the reactions of β-LG were not significantly affected by other proteins such as α-LA or BSA, but that the presence of β-LG in the system catalysed the reactions of other proteins such as α-LA or BSA.

The aim of this study was to explore the characteristics of a non-gelling exopolysaccharide (EPS) obtained from Lactobacillus delbrueckii subsp. bulgaricus NCFB 2483 and a gelling EPS obtained from Sphingomonas elodea ATCC 31461 (31461). The EPSs were isolated from the two bacterial strains grown in milk permeate-based media. They were purified and then characterised using light scattering and viscometric techniques. A greater emphasis of this research was placed on 2483 EPS since its physical characteristics have not been reported to date. In the case of 31461 EPS. a model for gelation of the sodium gellan was proposed based on rheological and light scattering measurements. The rheological properties of the two EPSs were also compared with several commercial polysaccharides. Microscopy examination of 2483 EPS was carried out using confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM). In CLSM, the lectin SBA (from Glycine max) Alexa Fluor 488 conjugate was used to stain the EPS since it has affinity for galactopyranosyl residues present in 2483 EPS. The CLSM micrographs showed a random distribution of EPS aggregates in the culture medium. At high magnification, the SEM micrographs showed web-like EPS structures. These structures were formed during the critical point drying process, when the EPS, which filled the interstices and channels of the protein aggregates, dehydrated. The 2483 EPS aggregates were found to be stable at neutral or low pH (~3.9) but were disrupted at high pH (pH 8-10). Procedures commonly used to quantify EPS from culture medium were found to be unreliable. In the development of an improved EPS assay, each of the processing steps was examined. Key improvements included the use of Flavourzyme for protein hydrolysis; optimising ethanol concentration to prevent lactose crystallisation yet allowing complete EPS precipitation; and a suitable centrifugation regime to minimise EPS loss. The improved EPS assay gave reproducible results (5% coefficient of variation). The isolation of 2483 EPS from milk media proved to be a difficult task because of interference from non-EPS components. An effective and simple approach allowing maximum EPS recovery involved the use of a hydrolysed milk medium which was ultrafiltered (UF) to remove molecular species larger than 2.5 x 105 Da. The UF permeate was suitable for the growth of 2483 with an EPS yield of ~400mg/L. Two EPS fractions (namely a soluble and an insoluble fraction) were isolated by ethanol precipitation and the soluble 'ropy' fraction was further purified to achieve ~98% purity. The elemental analysis of the purified fraction revealed the presence of nitrogen (~2.7% w/w). This could be due to the interaction of some peptides (from the growth medium) with the EPS. The polysaccharide composition of the soluble EPS fraction comprised of galactose, glucose, rhamnose and mannose residues (5:1:0.6:0.5). Traces of glucosamine were also found in the fraction. The purified fraction of 2483 EPS was characterised. Using a capillary viscometer, an intrinsic viscosity of ~2013mL/g was determined. The flow curves of the 2483 EPS solutions obtained using a rotational viscometer showed shear-thinning behaviour and an exponent value of ~0.76 (based on the Cross-type model) is typical of random coil polymers. The concentration dependence of the viscosity plot produced gradients of ~1.1 in the dilute domain and ~3.3 in the semi-dilute to concentrated domain. The coil overlap parameters at three concentration domains (c*[ŋ],ccr[ŋ] and c**[ŋ]) were 0.55, 2.86 and 5.67 respectively. The molecular parameters of the 2483 EPS were found via static light scattering measurements to have a weight-average molar mass (Mw.) of ~2 x 106 Da, a z-average root-mean-square radius ((r2g)z1/2) of ~165nm and a low polydispersity index (Mw/Mn ~1.15). The plot of Mw versus (r2g)z1/2 gave a gradient of approximately 0.5, which also suggested that the EPS polymer adopted a random coil conformation. The second part of the research involved gellan gum. Two gellan samples were studied. The first gellan sample was obtained from the fermentation of Sphingomonas elodea ATCC 31461 using milk permeate-based medium (31461). The second sample was a commercial high acyl gellan (LT100). Both gellan samples were converted to their sodium forms (Na-31461 and Na-LT100 gellan) using cation exchange resin and purified. The Na-gellan samples were highly sensitive to changes in Na+ concentrations. From oscillatory measurements, it was found that the complex moduli of the two Na-gellan samples superimposed closely at a specific Na+ concentration. The model for the conformational changes of Na-gellan molecules from a solution to a gel was proposed based on rheological and light scattering data. At very low Na+ concentrations (<19mM, in the case of Na-LT100). Na-gellan molecules were single-stranded (Mw ~2.5 x 10 5 Da) and adopted random coil conformation (exponent value based on the Cross-type model of ~0.76). At a slightly higher Na+ concentration (~19-24mM), Na-gellan molecules formed double-helices which led to a two-fold increase in molecular weight (M w ~5.2 x 105 Da). The double-stranded molecule appeared to be stiffer (exponent value of the Cross-type model ~0.82) and the mechanical spectra (G',G”) demonstrated 'weak ge' characteristics. A further increase in the Na+ concentration (>24mM) resulted in the formation of a gel network. The study also found that at low Na+concentration, both single-stranded and double-stranded Na-gellan molecules had a tendency to form aggregates under zero-shear conditions. The interactions involved in these aggregates were considered weak and transient, according to the Cox-Merz plot and light scattering data.

The technique of micro-oxygenation involves the deliberate addition of continuous, metered amounts of oxygen into a vessel of bulk wine during the maturation period (between the end of fermentation and bottling). The aim of the process is to improve the sensory properties of red wine, particularly the mouthfeel characteristics associated with the various polyphenol constituents. The success of the process appears to depend strongly on the ability to control the rate of oxygen dosage. The effect of dosage rate on the chemical and corresponding sensory changes of a red wine is the central theme of this thesis. A method of dosing oxygen (at typical micro-oxygenation rates) into small volumes of wine (<100 litres) was developed using a dense polymer membrane diffuser. It was clearly demonstrated that wine could be reliably oxygenated at very low rates using a coiled length of FEP as the diffuser material. Oxygen dosage was regulated by adjusting the oxygen pressure inside the tube. The advantage with a dense polymer diffuser is that no bubbles are generated and the oxygenation efficiency is 100%. The diffuser was fully modeled and characterised for use in the laboratory scale trials detailed in Chapters Four and Six. The small scale oxygenation equipment was used to conduct a fully replicated experiment to investigate the evolution of a Cabernet Sauvignon wine under four oxygenation treatments at dosage rates of 0, 10, 23 and 36 mg/L/mth. The total period of the trial was 105 days. HPLC analysis indicated that the rate change of low molecular weight polyphenols is directly related to the oxygen dosage rate. The concentration of the majority of the identifiable monomers, most notably the anthocyanins decreased throughout the course of the trial. The rate of decrease was directly related to oxygen dosage rate. Thiolysis results showed an increase in mDP for all treatments over the course of the trial until day 77 when they were observed to decrease for all treatments. The decrease in mDP coincided with an addition of SO2 which was investigated in a subsequent trial. Spectrophotometric results indicated that the rate of formation of non-bleachable pigments was directly related to the rate of oxygen dosage with significant differences between the high rates (23 and 36 mg/L/mth) and the low rates (0 and 10 mg/L/mth). The trend for all treatments was for increased levels of stable pigments. The sensory results show that the measured organoleptic temporal development exhibits a similar oscillatory behaviour compared to the anecdotally derived curve presented in figure 1-2. The distinction between the respective phases described in section 1.1.1 was, however less clear. The most significant factor in the model weighting was mouthfeel and astringency which correlates with the observed changes occurring in the wine polypenols during maturation. Overall the laboratory scale trial showed that the chemical polyphenol development was directly related to the oxygen dosage rate. The sensory evolution also appeared to be accelerated with higher oxygen dosage rates, although the oscillatory nature of the sensory response given a single linear input indicates a complex underlying mechanism driving the changes. The effect of SO2 on the development of wine polyphenols with and without oxygen was also investigated. The presence of SO2 was found to have a significant effect on both mDP and the concentration of non-bleachable pigments. mDP was observed to decrease over the six week trial period irrespective of whether oxygen had been added or not. The mDP for the treatments without SO2 increased steadily over the course of the trial. Similarly the formation of non-bleachable pigments was suppressed and even retarded with SO2 present whereas for the treatments without SO2 a steady increase was observed. The implication of these results is that SO2 may have a much larger effect on tannin development than oxygen. The use of electrochemical micro-oxidation (or ELMOX) was examined ostensibly to determine proof of concept and also compare the performance of glassy carbon and titanium as electrode materials against traditional micro-oxygenation. Notable transformations occurred with titanium showing higher levels of ethanal than the other treatments both chemically and by sensory measure. A greater rate of stable pigment formation was also observed for the titanium compared to the other treatments. The respective dosage rates for the glassy carbon ELMOX and traditional micro-oxygenation treatments were too low to be able to discriminate any significant differences compared to the control wine. / AGMARDT Doctoral Scholarship

A shelf life trial using a fully balanced factorial experimental design was used to analyse the effects of acidity and light on broccoli juice made on a semi commercial scale over an eight week period in simulated retail refrigerated storage conditions. The research focused on making broccoli juice on a pilot scale, and what happens to the colour, composition and flavour during storage. A pilot scale production of pasteurised broccoli juice was conducted and the juice satisfied microbiological safety limits for the eight week shelf life trial in retail storage conditions. The stability of the green colour of fresh broccoli through processing and storage was assessed. Neutral broccoli juice remained green for four weeks before the colour became more yellow. The acidified juice became yellow on acidification and did not change significantly during storage. Dietary fibre and pectin levels did not change during storage. Chlorophyll and carotenoids levels decreased during storage and were directly influencing the colour changes in the juices. Ascorbic acid levels decreased significantly during processing resulting in low ascorbic acid levels (12 - 15 mg /100ml of juice) at the start of the shelf life trial and dropped further to 2-6 mg /100ml of juice after eight weeks. Acidification and storage in the dark had a protective effect on the degradation of ascorbic acid with only a 58% reduction in ascorbic acid levels compared to an 84% reduction in neutral light stored broccoli juice. The effect of processing and storage on the flavour of the beverage was assessed using a trained sensory panel providing descriptive analysis. The sensory profiles for neutral and acidified juices were extremely different with the unbalanced acidity suppressing the perception of the basic tastes, sweet, salty and bitter. The neutral juice sensory profile only changed slightly in aroma attributes during storage for seven weeks. The astringent aftertaste of the acidified juice increased while the broccoli smell decreased during storage. The results from this research indicate that the production of a broccoli juice with a yellow green colour and some retained nutritional components is achievable with a refrigerated (4 °C) shelf life of 30 days in light excluding glass packaging. The neutral juice is recommended as it was greener and had a broccoli flavour.A shelf life trial using a fully balanced factorial experimental design was used to analyse the effects of acidity and light on broccoli juice made on a semi commercial scale over an eight week period in simulated retail refrigerated storage conditions. The research focused on making broccoli juice on a pilot scale, and what happens to the colour, composition and flavour during storage. A pilot scale production of pasteurised broccoli juice was conducted and the juice satisfied microbiological safety limits for the eight week shelf life trial in retail storage conditions. The stability of the green colour of fresh broccoli through processing and storage was assessed. Neutral broccoli juice remained green for four weeks before the colour became more yellow. The acidified juice became yellow on acidification and did not change significantly during storage. Dietary fibre and pectin levels did not change during storage. Chlorophyll and carotenoids levels decreased during storage and were directly influencing the colour changes in the juices. Ascorbic acid levels decreased significantly during processing resulting in low ascorbic acid levels (12 - 15 mg /100ml of juice) at the start of the shelf life trial and dropped further to 2-6 mg /100ml of juice after eight weeks. Acidification and storage in the dark had a protective effect on the degradation of ascorbic acid with only a 58% reduction in ascorbic acid levels compared to an 84% reduction in neutral light stored broccoli juice. The effect of processing and storage on the flavour of the beverage was assessed using a trained sensory panel providing descriptive analysis. The sensory profiles for neutral and acidified juices were extremely different with the unbalanced acidity suppressing the perception of the basic tastes, sweet, salty and bitter. The neutral juice sensory profile only changed slightly in aroma attributes during storage for seven weeks. The astringent aftertaste of the acidified juice increased while the broccoli smell decreased during storage. The results from this research indicate that the production of a broccoli juice with a yellow green colour and some retained nutritional components is achievable with a refrigerated (4 °C) shelf life of 30 days in light excluding glass packaging. The neutral juice is recommended as it was greener and had a broccoli flavour.

Emulsions containing three different types of milk fat fractions (MF13, MF27 and MF42) and anhydrous milk fat (AMF) were prepared at oil to water (O/W) ratios of 1:9, 3:7, 5:5 and 7:3 using rhamnolipids as emulsifiers. The prepared emulsions were analyzed for their storage stability and properties (colour, particle size, zeta potential and rheology). The effects of various factors (freezing/thawing, heating, pH, salts and ionic strength) on the stability of emulsions were also investigated. All emulsions prepared with an O/W ratio of 7:3, regardless of the type of milk fat, rendered a highly condensed, semi solid and cream-like substance whereas other emulsions containing less oil were in a liquid form. Among the four different O/W ratios tested, the highest emulsion stability during the storage of 12 weeks was observed from the emulsions containing 1:9 O/W ratios, due to a combine effect of smaller emulsion particle size and lower collision frequency between droplets. Interestingly, the emulsions with 7:3 O/W ratios were found to be more stable than the ones with 5:5 O/W ratios. This might be due to the limited movements of closely-packed emulsion droplets induced by the high oil concentration of 7:3 O/W ratios. The emulsion stability was significantly affected by low pH, especially at lower than pH 4, due to the loss of electrostatic repulsions between droplets leading to droplet coalescence and also possibly due to hydrolysis of rhamnolipid molecules. The presence of salts (NaCl, KCl and CaCl2) also rendered the emulsion unstable. The degree of instability was gradually increased with increasing salt concentrations. CaCl2 had the most significant effect even at a very low concentration. The viscosity of emulsions increased with increasing oil concentration but was not affected by the types of milk fats. Emulsions with 3:7, 5:5 and 7:3 O/W ratios exhibited non-Newtonian and shear thinning flow behaviour. At 7:3 O/W ratios, MF13 exhibited gel-like properties whereas both MF42 and AMF emulsions became more solid-like at higher frequency.

Milk proteins are known to bind volatile flavour compounds to varying extents, depending on the nature of the protein and flavour compound. Processing conditions, such as temperature and pH, are also known to have an influence on the interactions between milk proteins and flavour compounds. These interactions cause a great challenge for flavour scientists because they influence the perceived aroma profile of food products significantly, in particular in low-fat food products. The objectives of this research were to develop a headspace solid-phase microextraction (SPME) method followed by gas chromatography with flame ionisation detection (GC-FID) for the investigation of protein-flavour interactions, and to determine binding parameters of the hydrophobic flavour compound, 2-nonanone, to individual milk proteins - namely, β-lactoglobulin (β-lg), α-lactalbumin (α-la), bovine serum albumin (BSA), αs1-casein, and β-casein -, whey protein isolate (WPI), and sodium caseinate. Secondly, it was the aim to compare the binding of the structurally similar flavour compounds - 2-nonanone, 1-nonanal, and trans-2-nonenal – to WPI in aqueous solution, and to investigate the effect of heat and high pressure treatment, and pH on the extent of protein-flavour binding. The final objective was to investigate the in vivo release of the reversibly bound flavour compound, 2-nonanone, from WPI and sodium caseinate using proton transfer-reaction mass spectrometry (PTR-MS), and to understand the effect of viscosity on flavour release in vivo. The binding of the model flavour compound 2-nonanone to individual milk proteins, WPI, and sodium caseinate in aqueous solutions was investigated, using headspace SPME followed by GC-FID. The 2-nonanone binding capacities decreased in the order: BSA > β-lg > α-la > αs1-casein > β-casein, and the binding to WPI was stronger than the binding to sodium caseinate. All proteins appeared to have one binding site for 2-nonanone, except for BSA which possessed two classes of binding sites. The influence of heat treatment, high pressure processing and pH of the protein solutions on the binding of 2-nonanone, 1-nonanal, and trans-2-nonenal to WPI was determined. The binding of these compounds to WPI decreased in the order: trans-2-nonenal > 1-nonanal > 2-nonanone. The binding of 2-nonanone appears to involve hydrophobic interactions only, whereas the aldehydes, in particular trans-2-nonenal, also react through covalent binding. Upon both heat and high pressure denaturation, the binding of 2-nonanone to WPI decreased, the binding of 1-nonanal remained unchanged, while the binding of trans-2-nonenal increased. The binding affinity of the flavour compounds and WPI increased with increasing pH, which is likely to result from pH dependent conformational changes of whey proteins. The in vivo flavour (2-nonanone) release from solutions of WPI and sodium caseinate was investigated using proton-transfer-reaction mass spectrometry. During consumption, 2-nonanone was partly released from WPI, whereas there was no significant release from sodium caseinate. Even after swallowing of the samples, a substantial amount of flavour was detected in the breath, suggesting that the milk proteins interact with the mucosa in the mouth and throat, resulting in a further release of flavour from mucosa-bound proteins. An increase in viscosity of the protein solutions by the addition of carboxymethylcellulose enhanced the release of 2-nonanone from WPI, and resulted in 2-nonanone release from sodium caseinate. This may be due to a thicker coating of the mucosa with the sample solution after swallowing due to the higher viscosity, resulting in additional release of protein-bound flavour. These findings contribute to the knowledge of the interactions that occur between flavour compounds and proteins, which is required to improve food flavouring and to make protein based foods, e.g., low-fat dairy products, sensorily more acceptable to the consumer. The results also emphasize a careful choice of food processing conditions, such as temperature, high pressure or pH to obtain a desirable flavour profile.

Butter keeping quality and pallet physical stability during transport and storage are dependent on the temperature distribution through the product. Understanding these temperature changes are of vital importance for the dairy industry with regard to butter manufacture, storage and shipping. Three dimensional mathematical models of heat transfer were developed to predict thawing and freezing in butter products. These models require accurate thermophysical data as an input. Specific heat capacity and enthalpy of butter with different composition was measured using Differential Scanning Calorimetry. The specific heat capacity of butter differs for cooling and heating operations due to significant supercooling and delayed crystallization of the fat fraction of butter at temperatures well below the equilibrium phase change temperature during cooling. This reduces the heat capacity for cooling relative to that for heating. Thawing of individual blocks of butter was accurately predicted by the conduction only model (no mass transfer limitations) with equilibrium thermal properties giving accurate predictions when the butter was completely frozen before thawing. For partially frozen butter the conduction model with the measured temperature dependent specific heat capacity data for unfrozen butter including melting of some of the fat fraction gave accurate predictions. For freezing it was observed that water in the butter supercools many degrees below its initial freezing point before freezing due to its water in oil structure. Experiments suggested that during freezing release of latent heat observed as a temperature rebound is controlled as much by the rate of crystallisation of water in each of the water droplets as by the rate of heat transfer. A conduction only model including water crystallization kinetics based on the Avrami Model predicted freezing in butter successfully. Simple models with equilibrium thermal properties and nucleation only kinetics (based on homogenous nucleation theory) or the sensible heat only model (no release of latent heat) gave poor predictions. The models for individual blocks were extended to predict heat transfer in butter pallets. A butter pallet contains product, packaging material and the air entrapped between the packaging and butter cartons. Measurements were made for freezing and thawing of full and half pallets at a commercial storage facility and in the University laboratory. Thawing and freezing in wrapped tightly stacked pallets was predicted accurately by the conduction only model with effective thermal properties (incorporating butter, packaging and air) estimated by the parallel model. For unwrapped tightly stacked or loosely stacked pallets there is potential for air flow between the adjacent cartons of butter. An alternative approach was developed which consisted of modeling the pallet on block by block basis using effective heat transfer coefficients for each surface. Different heat transfer coefficients were used on different faces of the blocks depending on the location of the block in the pallet. This approach gave good predictions for both unwrapped tightly stacked and loosely stacked pallets using the estimated effective heat transfer coefficients from the measured data. Further experimental and/or modelling work is required in order to develop guidelines for estimating effective heat transfer coefficient values for internal block face for industrial scenarios.

Lactose represents approximately one third of the total solids in bovine milk. In the dairy industry, lactose is recovered from whey and whey permeates using a crystallisation process that involves both evaporation and a cooling stage. A good understanding of the lactose crystallisation kinetics enables both these processes to be operated at conditions that maximise the yield and minimise capital and processing costs. This study has looked at the nucleation and growth kinetics of the lactose crystallisation process. A model has been produced that can accurately predict the changing concentration profile as lactose is removed, via growth, from an industrial solution. This model incorporated the available literature information and expanded on it where required. The primary nucleation of alpha lactose monohydrate was investigated on the laboratory scale. The work identified the changing relationship, which occurs with increasing supersaturation, as lactose nucleation moves from being dominated by the heterogeneous mechanism to the homogenous mechanism. The absolute supersaturation at which the mechanism changes was found not to be affected by the solution temperature and agitation rate; however the presence of impurities lowered the supersaturation required for homogeneous nucleation. The effect of mixing on the primary nucleation rate was studied in a Rushton turbine agitated vessel and through a Venturi. Increasing the agitation rate increased the frequency of activated molecular collisions but the critical nucleus size remained constant. A strong correlation was found, for both mixing systems, between the nucleation rate and the frequency of vortex shedding.

<p>Entomophagy, which is the habit of eating insects, has become relevant in the past few years as it could potentially help reduce current and future food insecurity, due to the highly nutritious and sustainable characteristics of edible insects. In addition to the nutritional content of insects, research on the potential bioactive components of insect extracts has also gained popularity. In this study, extracts from house cricket (<i>Acheta domesticus</i>) from two farms and their corresponding feeds were obtained using a microwave-assisted extraction. Further phenolic characterization led to the identification and quantification of 4-hydroxybenzoic acid, <i>p</i>-coumaric acid, ferulic acid and syringic acid as major phenolic compounds in both <i>A. domesticus </i>extracts as well as both feed extracts. Additionally, <i>in vitro</i> antioxidant activity was evaluated using 2,2-diphenyl-1-picrylhydrazyl radical cation (DPPH) and 2,2’-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical assays. In both <i>in vitro</i> assays, <i>A. domesticus</i> extracts showed higher antioxidant activity compared to the feeds. Antibacterial activity against <i>E. coli</i> and <i>L. innocua</i> was also evaluated using the microwell method. The <i>A. domesticus </i>extracts showed a selective inhibition (p<0.05) towards the gram-positive bacteria <i>L. innocua </i>between a period of 4 to 8 h. This inhibition is thought to have occurred as a result of the presence of phenolic acids and antimicrobial peptides, while the feed extracts did not exhibit any inhibitory activity towards any of the bacteria. The finding of the same phenolic acids in <i>A. domesticus</i> and their corresponding feed could imply the capacity of <i>A. domesticus</i> to absorb and sequester dietary phenolics that may provide additional health benefits when the insect is consumed, unveiling new benefits of entomophagy. </p>

<p>Within the last decade, the popularity and interest in dry-aging have constantly increased among both consumers and producers. Dry-aging is a natural value-adding process where meat is exposed to a controlled refrigerated environment without any protective barrier during the aging process. This process leads to the development of unique flavors in the final meat product. Although the prevalence of this process is increasing, there are inconsistent reports regarding the impacts of dry-aging on meat sensory attributes, especially on the flavor aspect. Given that flavor generation is dependent on the composition and availability of flavor precursors, the presence or absence of these precursors may contribute to the inconsistency observed. Thus the main objective of the research described here was to characterize the flavor precursors in dry-aged meat and elucidate potential factors or mechanisms favoring to their production.</p> <p> To achieve this objective, metabolomics analysis was conducted in conjunction with various chemical analyses (free amino acids, fatty acids, sugar content and volatile analysis), microbiome profiling and meat quality analysis (tenderness, water holding capacity, color stability, oxidative stability, microbial attributes and sensory analysis) to identify the essential flavor precursors and their production process. In addition, similar analyses were conducted using multiple meat sources (grass-fed beef loins, cull cow beef loins and pork loins) aged by wet-aging (WA), conventional dry-aging (DA), dry-aging in bag (DWA) and UV-light dry-aging (UDA) to elucidate the impact of the different aging treatments on meat quality, sensory attributes and flavor precursor availability.</p> <p>Regardless of the meat source, the results demonstrated that dry-aging altered the meat flavor precursor compositions, primarily by increasing the presence of protein-derived precursors (e.g., free amino acids and dipeptides), especially glutamine and glutamate compounds. Additionally, nucleotide and carbohydrate-derived compounds such as adenosine and reducing sugars were greatly increased after the dry-aging process. While the fatty acid profile was minimally affected, metabolomics analysis revealed a decrease in sterol and terpenoid lipids following dry-aging, which could potentially reduce off-flavors development in the meat. Other compounds such as vitamin B and vitamin C were also detected in the dry-aged product, which potentially could contribute to the flavor development.</p> <p>Analysis of the liberation mechanisms demonstrated that dehydration played a role in increasing the concentration of the flavor precursors in the dry-aged product, potentially promoting greater (e.g., Maillard reaction) during cooking. Furthermore, microorganisms might be responsible for further increasing the availability of flavor precursors in dry-aged meat, especially free amino acids, along with the dehydration process. Microbiome profiling found that <i>Pseudomonas</i> spp. are the most prominent bacterial species in microbial communities found on dry-aged meat which could affect the precursor release in dry-aged meat. Metabolomics analysis also indicated increased glutathione metabolism during dry-aging, which could lead to the liberation of glutamine-related compounds. The analysis also identified other compounds such as porphyrin rings (iron-related) and shikimic acid (bacterial metabolism), providing further examples of how metabolomics can identify dry-aged flavor precursors and reveal other potential mechanisms related to flavor development mechanisms.</p> <p>These outcomes demonstrate that dry-aging alters meat flavor precursor composition, mainly by increasing the availability of protein-, nucleotide- and carbohydrate-derived compounds. Such results indicate that the Maillard reaction is likely be the main mechanism in flavor generation in dry-aged meat. The current results provided more insights into the dry-aging flavor development, especially highlighting important flavor precursor such as glutamate and glutamine containing products, likely to contribute to the dry-aged flavor. Future study to identify the impact of different microorganism (especially mold and yeast) on dry-aging flavor development would be of interest. Additionally, impact of different cooking process should also be studies to maximize the dry-aged flavor potential from the product.</p>