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BACKGROUND: Artisanal cheeses usually contain a highly diverse microbial community which can significantly impact their quality and safety. Here, we describe a detailed longitudinal study assessing the impact of ripening in three natural caves on the microbiome and resistome succession across three different producers of Cabrales blue-veined cheese. RESULTS: Both the producer and cave in which cheeses were ripened significantly influenced the cheese microbiome. Lactococcus and the former Lactobacillus genus, among other taxa, showed high abundance in cheeses at initial stages of ripening, either coming from the raw material, starter culture used, and/or the environment of processing plants. Along cheese ripening in caves, these taxa were displaced by other bacteria, such as Tetragenococcus, Corynebacterium, Brevibacterium, Yaniella, and Staphylococcus, predominantly originating from cave environments (mainly food contact surfaces), as demonstrated by source-tracking analysis, strain analysis at read level, and the characterization of 613 metagenome-assembled genomes. The high abundance of Tetragenococcus koreensis and Tetragenococcus halophilus detected in cheese has not been found previously in cheese metagenomes. Furthermore, Tetragenococcus showed a high level of horizontal gene transfer with other members of the cheese microbiome, mainly with Lactococcus and Staphylococcus, involving genes related to carbohydrate metabolism functions. The resistome analysis revealed that raw milk and the associated processing environments are a rich reservoir of antimicrobial resistance determinants, mainly associated with resistance to aminoglycosides, tetracyclines, and ß-lactam antibiotics and harbored by aerobic gram-negative bacteria of high relevance from a safety point of view, such as Escherichia coli, Salmonella enterica, Acinetobacter, and Klebsiella pneumoniae, and that the displacement of most raw milk-associated taxa by cave-associated taxa during ripening gave rise to a significant decrease in the load of ARGs and, therefore, to a safer end product. CONCLUSION: Overall, the cave environments represented an important source of non-starter microorganisms which may play a relevant role in the quality and safety of the end products. Among them, we have identified novel taxa and taxa not previously regarded as being dominant components of the cheese microbiome (Tetragenococcus spp.), providing very valuable information for the authentication of this protected designation of origin artisanal cheese. Video Abstract.
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Queijo , Microbiologia de Alimentos , Microbiota , Queijo/microbiologia , Queijo/normas , Microbiota/fisiologia , Transferência Genética Horizontal/genética , Metagenoma/genética , Resistência Microbiana a Medicamentos/genéticaRESUMO
This study investigated the effects of diet and stage of lactation (SOL) on sensory profiles, texture, volatile profiles, and colour of Cheddar cheese. Cheddar cheese was manufactured from early-, mid-, and late-lactation milk obtained from seasonally calved cows (n = 54). Cows were assigned a diet; group 1: perennial ryegrass (GRS), group 2: total mixed ration (TMR), and group 3: partial mixed ration (PMR). Instrumental analysis was performed at 270 days (mature Cheddar). Sensory evaluation took place after 548 days (extra mature Cheddar). Toluene was the only volatile compound that was significantly influenced by diet. The trained panel rated early-lactation cheese as stronger than mid- and late- for cowy/barny flavour and late-lactation cheese as sweeter than early- and mid-lactation cheese. Mid-lactation cheese was liked least overall. Early-lactation cheeses were rated higher for 'crumbly' texture than mid- and late. Diet affected consumer ratings, with GRS and PMR cheese rated as more intense than TMR for flavour, aftertaste, and saltiness. Consumers reported that TMR cheese was lighter in colour compared to GRS cheese, which was supported by instrumental analysis. Consumers perceived GRS as more springy and less crumbly than TMR and PMR, while Texture Profile Analysis indicated that TMR was harder than GRS. Consumer segmentation was observed with two clear preference groups, one preferring GRS and one preferring TMR. For both groups, 'taste' seemed to be the main driver of liking, highlighting that consumer preference is most impacted by individual taste preferences.
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Queijo , Feminino , Animais , Bovinos , Lactação , Paladar , Percepção Gustatória , LeiteRESUMO
In order to safeguard authentic whiskey products from fraudulent or counterfeit practices, high throughput solutions that provide robust, rapid, and reliable solutions are required. The implementation of some analytical strategies is quite challenging or costly in routine analysis. Qualitative screening of whiskey products has been explored, but due to the nonspecificity of the chemical compounds, a more quantitative confirmatory technique is required to validate the result of the whiskey analysis. Hence, combining analytical and chemometric methods has been fundamental in whiskey sample differentiation and classification. A comprehensive update on the most relevant and current analytical techniques, including spectroscopic, chromatographic, and novel technologies employed within the last 5 years in whiskey analysis for authentication, discrimination, and quality control, are presented. Furthermore, the technical challenges in employing these analytical techniques, future trends, and perspectives are emphasized.
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Bebidas Alcoólicas , Bebidas Alcoólicas/análise , Análise Espectral , Controle de QualidadeRESUMO
In recent years, consumer demand for health benefitting, pleasant-tasting rapeseed oil has increased, and so has production. Ireland's climate and agricultural background can support the production of high-quality rapeseed oil. Volatile organic compounds (VOC) can give rise to highly distinctive flavours in rapeseed oils, produced during crop growth and generated during processing. This study performed VOC and sensory evaluation to determine if correlations exist. Samples of Irish rapeseed oils from 6 different producers were analysed. Compounds detected in the oil samples consisted of acids, alcohols, aldehydes, ketones, benzenes, esters, ether, terpenes, and sulphurs. While variations in whole volatile profiles were not considered significant, individual compounds and volatile classes were for hexanal, pentanal, ketones, acids, and sulphurs compounds. Correlations were observed between the VOCs detected and the sensory profile, which indicated the VOC content may influence an oil's sensory profile.
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This study evaluated the impact of three distinct diets; perennial ryegrass (GRS), perennial ryegrass/white clover (CLV) and total mixed ration (TMR), on the sensory properties and volatile profile of whole milk powder (WMP). The samples were evaluated using a hedonic sensory acceptance test (n = 99 consumers) and by optimised descriptive profiling (ODP) using trained assessors (n = 33). Volatile profiling was achieved by gas chromatography mass spectrometry using three different extraction techniques; headspace solid phase micro-extraction, thermal desorption and high capacity sorptive extraction. Significant differences were evident in both sensory perception and the volatile profiles of the WMP based on the diet, with WMP from GRS and CLV more similar than WMP from TMR. Consumers scored WMP from CLV diets highest for overall acceptability, flavour and quality, and WMP from TMR diets highest for cooked flavour and aftertaste. ODP analysis found that WMP from TMR diets had greater caramelised flavour, sweet aroma and sweet taste, and that WMP from GRS diets had greater cooked aroma and cooked flavour, with WMP derived from CLV diets having greater scores for liking of colour and creamy aroma. Sixty four VOCs were identified, twenty six were found to vary significantly based on diet and seventeen of these were derived from fatty acids; lactones, alcohols, aldehydes, ketones and esters. The abundance of δ-decalactone and δ-dodecalactone was very high in WMP derived from CLV and GRS diets as was γ-dodecalactone derived from a TMR diet. These lactones appeared to influence sweet, creamy, and caramelised attributes in the resultant WMP samples. The differences in these VOC derived from lipids due to diet are probably further exacerbated by the thermal treatments used in WMP manufacture.
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Commercially available Irish edible brown (Himanthalia elongatasea spaghetti (SS), Alaria esculentaIrish wakame (IW)) and red (Palmaria palmatadulse (PP), Porphyra umbilicalisnori) seaweeds were incorporated into pork sausages at 1%, 2.5%, and 5%. Proximate composition, salt, water-holding (WHC), cook loss, instrumental colour analysis, texture profile analysis (TPA), and sensory analysis were examined. Protein (13.14−15.60%), moisture (52.81−55.71%), and fat (18.79−20.02%) contents of fresh pork sausages were not influenced (p > 0.05) by seaweed type or addition level. The ash content of pork sausages containing PP, SS, and IW at 2.5% and 5%, and nori at 5%, were higher (p < 0.05) than the control sample. In comparison to the control, sausages containing nori, SS, and IW at 5% displayed higher (p < 0.05) WHC. Cook loss was unaffected (p > 0.05) by the addition of seaweeds into sausage formulations, compared to the control and within each seaweed. The addition of seaweeds into sausages had an impact on the surface colour (L* a* b*) and texture profile analysis (TPA) at different inclusion levels. Overall, hedonic sensory acceptability decreased (p < 0.05) in cooked sausages containing PP at 2.5% and 5%, and SS and IW at 5%.
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Aroma-active compounds in raw bovine milk produced from cows fed perennial ryegrass (GRS) or total mixed ration (TMR) consisting of grass silage, maize silage, and concentrates were identified by direct immersion sorptive extraction (DI Hi-Sorb), coupled with gas-chromatography-mass spectrometry and olfactometry using odour intensity (OI) and aroma extraction dilution analysis (AEDA). Ninety-nine volatile organic compounds (VOC) were identified in these raw GRS and TMR milk samples; 33 of which were also present in the feed and rumen samples from these diets. Only the abundance of 13 VOC varied significantly based on diet. However, the odours of both raw milks were quite distinct as aroma perception is not influenced by abundance alone but also by the odour activity of each VOC. Approximately, 30% of the VOC influenced the aroma perception of these raw milks. This study clearly highlighted the significant impact of VOC transferring from the diet that influenced the aroma perception of both raw GRS and TMR milk. The aroma of the raw TMR milk was more complex than that of the raw GRS milk, and many of the key dietary-derived-odour-active VOC likely arose during the production of the TMR feed as most were either derived from Maillard reactions or impacted by heat. Seventeen of the 44 odour activities detected differed between both sample types. This study has clearly demonstrated the impact of diet on the aroma perception of raw bovine milk.
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The focus of the global cheese industry on accessing new markets for cheese is currently driving a greater need for innovation in cheese products. Research to date suggests that, for example, East Asian consumers prefer cheeses that have a soft texture, with mild and milky flavors. Strategies for achieving such cheese characteristics are reviewed in this article. For example, incorporation of polysaccharides into cheese results in cheese with higher moisture levels and softer textures; this also results in modification of other properties such as adhesiveness, meltability and flavor release. Hydrated polysaccharides may be considered as filler particles within cheese matrices, and therefore filled gel models with suitable filler particles can be used to establish the effect of filler volume, size and surface properties on the fractural and rheological properties of cheese matrices, thus guiding the use of polysaccharides. Addition of plant proteins such as soy and pea protein can also result in cheeses with softer texture. Furthermore, it has been suggested that heat-induced gelation of soy or pea protein with casein results in a gel structure consisting of two independent protein gels, thus facilitating the design of bespoke structures by adjusting the ratio of the two proteins. Finally, it is proposed that incorporation of ingredients with sensory properties familiar to East and Southeast Asian consumers and with the capacity to achieve bespoke textures offer potential for the development of cheese products for consumers in these markets.
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Queijo , Proteínas de Ervilha , Sudeste Asiático , Proteínas de Plantas , PolissacarídeosRESUMO
This study investigated the impact of the addition of exogenous enzymes (Accelerzyme CPG, Debitrase DBP20) or cellular preparations (FlavoGard), traditionally used in the cheese industry, to accelerate flavour development of dry fermented sausages with 6% of lactic derivatives content. Sausages were fermented to pH 5.0, dried for 32 days and vacuum packed stored under refrigeration for 60 days. Sausages were analysed for physicochemical parameters, technological microbiota and proteolysis after fermentation, drying/ripening and storage. Similar compositional results were obtained in all products (38-39% humidity in the final product; 38.2% fat and 40.7% protein as dry matter throughout the study). Debitrase application positively affected proteolysis by changing the free amino acid profile and increasing non-protein nitrogen and total free amino acids by 2.2 and 11.8-fold, respectively. Accelerzyme increased ripened cheese flavour and overall sensory quality from 5.1 to 5.8; Debitrase increased ripened cheese odour and flavour, bitterness, umami, adhesiveness, pastiness, and overall sensory quality from 5.0 to 5.9, and decreased acid and hardness. This study highlights the effects of adding some exogenous enzyme/bacterial preparations traditionally used in the cheese industry to enhance the flavour of dry fermented sausages with high content of lactic ingredients and increase its sensory quality.
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Queijo , Produtos da Carne , Aminoácidos , Queijo/análise , Dessecação , Fermentação , Produtos da Carne/análise , PaladarRESUMO
Irish edible brown (Himanthalia elongata-sea spaghetti, Alaria esculenta-Irish wakame) and red seaweeds (Palmaria palmata-dulse, Porphyra umbilicalis-nori) were assessed for nutritional (proximate composition; salt; pH; amino acid; mineral and dietary fibre contents); bioactive (total phenolic content (TPC) and in vitro antioxidant activity (DPPH and FRAP)); thermal (thermogravimetric analysis (TGA)); and technological (water holding capacity (WHC), oil holding capacity (OHC) and swelling capacity (SC)) properties. Red seaweeds had higher (p < 0.05) protein levels, whereas brown seaweeds possessed higher (p < 0.05) moisture, ash, insoluble and total dietary fibre contents. Nori had the lowest (p < 0.05) salt level. Seaweed fat levels ranged from 1 to 2% DW. Aspartic and glutamic acids were the most abundant amino acids. The total amino acid (TAA) content ranged from 4.44 to 31.80%. Seaweeds contained numerous macro (e.g., Na) and trace minerals. The TPC, DPPH and FRAP activities followed the order: sea spaghetti ≥ nori > Irish wakame > dulse (p < 0.05). TGA indicated maximum weight loss at 250 °C. Dulse had the lowest (p < 0.05) WHC and SC properties. Dulse and nori had higher (p < 0.05) OHC than the brown seaweeds. Results demonstrate the potential of seaweeds as functional food product ingredients.
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Lipid oxidation (LO) is a primary cause of quality deterioration in fat-containing dairy powders and is often used as an estimation of a products shelf-life and consumer acceptability. The LO process produces numerous volatile organic compounds (VOC) including aldehydes, ketones and alcohols, which are known to contribute to the development of off-flavours in dairy powders. The main factors influencing the oxidative state of dairy powders and the various analytical techniques used to detect VOC as indicators of LO in dairy powders are outlined. As the ability to identify and quantify specific VOC associated with LO improves this review highlights how these techniques can be used in conjunction with olfactory and sensory analysis to better understand product specific LO processes with the aim of maximizing shelf-life without compromising quality.
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This study explores the experiences of a cohort of young, educated, internationally mobile Chinese consumers with cheese and other dairy products, and how these experiences shape their behavior toward cheese products. In total, 41 Chinese students studying at an Irish university participated in 5 focus groups (n = 41, n = 7-10). Thematic analysis identified important factors that influence consumer behaviors regarding cheese products. Individuals' expectations toward cheese were embedded in their knowledge structures, which were constructed from previous experience. Participants had general positive expectations toward cheese due to associations with western-style foods and nostalgia; however, direct eating experience determined long-term behavior. When making a purchase decision, choice motives were weighed and negotiated to establish a fundamental driving factor for purchase. Perceived probability of choice motive fulfillment was important in determining purchase decisions, with many participants having low perceived ability to select cheese and limited motivation to engage with cheese due to limited perceived relevance of cheese to their daily food life. Individuals' innovativeness was an important factor that influences their openness to cheese products when moving beyond familiar foods. Opportunities exist such as using nostalgic cues as marketing tools to increase consumers' interest in cheese or combining cheese with Chinese food to increase perceived relevance of cheese to their daily food life. Providing information at point of purchase could reduce the disconnect between expectation and actual experience, and innovative cheese products may be developed to better fulfill important choice motives.
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Queijo , Comportamento do Consumidor , Animais , China , Comportamento de Escolha , Preferências Alimentares , Motivação , PaladarRESUMO
Volatile profiling of whole milk powder is valuable for obtaining information on product quality, adulteration, legislation, shelf life, and aroma. For routine analysis, automated solventless volatile extraction techniques are favored due their simplicity and versatility, however no single extraction technique can provide a complete volatile profile due to inherent chemical bias. This study was undertaken to compare and contrast the performance of headspace solid phase microextraction, thermal desorption, and HiSorb (a sorptive extraction technique in both headspace and direct immersion modes) for the volatile analysis of whole milk powder by gas chromatography mass spectrometry. Overall, 85 unique volatiles were recovered and identified, with 80 extracted and identified using a non-polar gas chromatography column, compared to 54 extracted, and identified using a polar gas chromatography column. The impact of salting out was minimal in comparison to gas chromatography column polarity and the differences between the extraction techniques. HiSorb extracted the most and greatest abundance of volatiles, but was heavily influenced by the number and volume of lactones extracted in comparison to the other techniques. HiSorb extracted significantly more volatiles by direct immersion than by headspace. The differences in volatile selectivity was evident between the techniques and highlights the importance of using multiple extraction techniques in order to obtain a more complete volatile profile. This study provides valuable information on the volatile composition of whole milk powder and on differences between extraction techniques under different conditions, which can be extrapolated to other food and beverages.
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The in vitro antioxidant effects of the most potent antioxidants of rosemary, namely carnosol, carnosic acid and rosmarinic acid (c: ca: ra) were assessed in fat-filled milk powders (FFMPs) under accelerated conditions (40 °C and relative humidity (RH) 23%) over 90 days. Lipid oxidation was assessed in FFMPs by measuring peroxide values (PVs), thiobarbituric acid reactive substances (TBARS) and aroma volatiles using headspace (HS) solid-phase microextraction (SPME) coupled to gas-chromatography-mass spectrometry (GC-MS). The antioxidant potency of c: ca: ra exhibited a concentration-related effect (308 ppm > 200 ppm > 77 ppm), with the highest concentration being the most effective at controlling the formation of TBARS and PVs. At a concentration of 308 ppm c: ca: ra were particularly effective (p < 0.05) in inhibiting all the evaluated oxidation indices (primary and secondary) compared to the control samples, but in some cases less effectively (p < 0.05) than butylated hydroxyanisole: butylated hydroxytoluene (BHA: BHT) (200 ppm).
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The impact of barley variety and its geographical growth location (environment) on the flavour of new make spirit was investigated to determine if "terroir" can be applied in the production of single malt whisk(e)y. New make spirits were produced at laboratory scale under controlled conditions from two different barley varieties (Olympus and Laureate) grown at two distinct environments (Athy, Co Kildare and Bunclody, Co Wexford) in Ireland over two consecutive seasons (2017 and 2018). The spirit samples were analysed by gas chromatography mass spectrometry olfactometry and descriptive sensory analysis. Forty-two volatiles were detected with eight deemed as very influential and fifteen deemed as influential to the aroma of new make spirit. Sensory attributes were influenced by barley variety, environment, and the interactions thereof over both seasons, with environment and the interaction of variety x environment having a greater impact than variety alone. Chemometric analysis of the olfactometry and sensory data found that both environment and season had a greater impact on the aromatic sensory perception of the new make spirits than variety alone. Therefore, this study clearly demonstrates a "terroir" impact on the flavour of new make spirit and highlights its potential importance especially in relation to single malt whisk(e)y.
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A rich sugar diet has negative health implications so it is necessary to reduce sugar where possible. The objective of this study was to determine if different sugar size fractions could increase the sweetness intensity of Shortbread biscuits and therefore, permit sugar reduction. For this, the unground commercial sugar (Control, 102 to 378 µm) and two of its sieved separates, (Coarse (C), 228 to 377 µm and Fine (F), 124 to 179 µm) were investigated in biscuit formulation with the following content: Control 100% or 50%; (C), 100 or 50% and (F), 100 or 50% or its initial content. Biscuits were tested using sensory (hedonic and intensity), physical (dimensions, fracture properties, color), and compositional analyses. Trends showed that samples containing C-sugar with its 50% content were more preferable than samples containing the Control and F- sugar fractions at the same level without impact upon acceptability of the final product in all three sugar fraction sizes. As sweetness intensity scores correlated directly with flavor liking scores, these findings promote the use of this sugar fraction in the formulation of low-sugar baked biscuits. Sugar particle size manipulation could be used as a viable, cheap, technological approach to reduce sugar in baked goods and therefore promote consumer acceptable and commercially available baked biscuits products. PRACTICAL APPLICATION: In order to reduce sucrose content in products one of the approaches might be via the utilization of small sugar particles which has been shown to increase the intensity of sweetness in chocolate brownies, which has been previously shown by Richardson et al. in 2018. This study investigates the use of a clean label approach in sugar reduction of short bread biscuits. This approach involves decreasing the sugar particles size and demonstrates how reducing the content to half of its initial content formulation will affect the sensory perception and physical properties.
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Culinária , Sacarose/química , Humanos , Tamanho da Partícula , Sensação , PaladarRESUMO
Determining minimum levels of fat and sucrose needed for the sensory acceptance of sponge cake while increasing the nutritional quality was the main objective of this study. Sponge cakes with 0, 25, 50 and 75% sucrose replacement (SR) using a combination of inulin and Rebaudioside A (Reb A) were prepared. Sensory acceptance testing (SAT) was carried out on samples. Following experimental results, four more samples were prepared where fat was replaced sequentially (0, 25, 50 and 75%) in sucrose-replaced sponge cakes using pureed butter beans (Pbb) as a replacer. Fat-replaced samples were investigated using sensory (hedonic and intensity) and physicochemical analysis. Texture liking and overall acceptability (OA) were the only hedonic sensory parameters significantly affected after a 50% SR in sponge cake (p < 0.05). A 25% SR had no significant impact on any hedonic sensory properties and samples were just as accepted as the control sucrose sample. A 30% SR was chosen for further experiments. After a 50% fat replacement (FR), no significant differences were found between 30% sucrose-replaced sponge cake samples in relation to all sensory (hedonic and intensity) parameters investigated. Flavour and aroma intensity attributes such as buttery and sweet and, subsequently, liking and OA of samples were negatively affected after a 75% FR (p < 0.05). Instrumental texture properties (hardness and chewiness (N)) did not discriminate between samples with increasing levels of FR using Pbb. Moisture content increased significantly with FR (p < 0.05). A simultaneous reduction in fat (42%) and sucrose was achieved (28%) in sponge cake samples without negatively affecting OA. Optimised samples contained significantly more dietary fibre (p < 0.05).
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BACKGROUND: Information pertaining to the aromatic profile of seaweeds and seaweed extracts can provide evidence regarding their potential suitability as ingredients in processed foods. To date only limited material has been available on the volatile profiles of some seaweed species. Others in this study have not previously been described. The volatile profiles of dried brown (Himanthalia elongata, Undaria pinnatifida, Alaria esculenta) and red (Porphyra umbilicalis, Palmaria palmata) seaweeds, and a brown seaweed extract (fucoxanthin) from Laminaria japonica were investigated using a chemometric approach to collate data from volatile gas chromatography - mass spectrometry (GC-MS), direct sensory aroma evaluation, and gas-chromatography - olfactometry (GC-O) to obtain a better understanding of their volatile profile and sensory perception. RESULTS: More than 100 volatile compounds were identified by static headspace solid phase micro-extraction (HS-SPME) and thermal desorption gas chromatography - mass spectrometry (TD GC-MS). Brown seaweeds were characterized by 'grassy/herbal/floral', 'fruity', and 'fatty' aromas, red seaweeds by 'green/vegetable', 'mushroom/earthy' and 'sweet/buttery' aromas, and the fucoxanthin extract by 'rancid' and 'nutty' aromas with an overall lower intensity. Heptanal appeared to be a major odor-active compound in all samples. Other volatiles were more characteristic of each individual seaweed: hexanal, (E,Z)-2,6-nonadienal and 2-pentylfuran for H. elongata; ethyl butanoate and 2,3-butanedione for U. pinnatifida; 6-dimethylpyrazine, (E,Z)-2,6-nonadienal and sulactone for P. palmata; 1-octen-3-ol for P. umbilicalis, heptanone for A. esculenta, and 2-furanmethanol for fucoxanthin. CONCLUSION: Brown and red seaweeds had distinct sensory properties with individual seaweeds having differing volatiles and odorants. This study provides additional information that can contribute to the development of products incorporating dried seaweeds / extracts that are more acceptable to the consumer. © 2020 Society of Chemical Industry.
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Odorantes/análise , Phaeophyceae/química , Extratos Vegetais/química , Rodófitas/química , Alga Marinha/química , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Extratos Vegetais/isolamento & purificação , Paladar , Verduras/químicaRESUMO
Perception and liking among Irish, German and USA consumers of salted butter produced from different feed systems-outdoor grass (FS-GRSS), grass/clover (FS-CLVR), and indoor concentrate (FS-TMR)-was investigated. A consumer study was conducted in all three countries. Irish and German assessors participated in ranking descriptive analysis (RDA), whereas descriptive analysis (DA) was carried out by a trained panel in the USA. Volatile analysis was conducted to identify differences in aroma compounds related to cow diet. Overall, there was no significant difference in overall liking of the butters, among USA, German and Irish consumers, although cross-cultural preferences were evident. Sensory attribute differences based on cow diet were evident across the three countries, as identified by German and Irish assessors and trained USA panelists, which are likely influenced by familiarity. The abundance of specific volatile aromatic compounds, especially some aldehydes and ketones, were significantly impacted by the feed system and may also contribute to some of the perceived sensory attribute differences in these butters.
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Macrococcus caseolyticus subsp. caseolyticus is a Gram-positive, commensal organism documented to be present as a component of the secondary microflora in fermented foods such as Ragusano and Fontina cheeses and Cantonese sausage. In these products, the organism appears to play a role in ripening and the development of the final organoleptic qualities. However, the role of this organism in flavor generation is not well understood. Therefore, the objective of this study was to investigate the role of M. caseolyticus subsp. caseolyticus in flavor compound formation through an examination of enzymatic, metabolomic and genomic data. A bank of M. caseolyticus subsp. caseolyticus strains derived from a variety of niches were examined. Enzyme activities analyzed comprised those of the proteolytic and lipolytic cascades including cell-envelope proteinase (CEP), peptidases, esterases, lipases, aminotransferases and glutamate dehydrogenase (GDH). Strain to strain variation was observed, often associated with niche. All strains, except those isolated from non-dairy sources, demonstrated high CEP activity. Such high CEP activity associated with dairy strains implies the importance of this characteristic in the adaptation of these strains to a dairy-specific niche. However, limited downstream peptidolytic activity, in addition to a limited ability to generate free amino acids (FAA) was observed across all strains, indicating weak ability of this organism to generate amino-acid derived flavor compounds. Interestingly, the strains with high CEP activity also demonstrated high esterase activity and gas chromatography-mass spectrometry (GC-MS) analysis of the volatile compounds produced when these strains were grown in lactose-free milk demonstrated differences in the range and types of volatiles produced. In contrast to this metabolic versatility, comparative genome analysis revealed the distribution of components of the proteolytic and lipolytic system in these strains to be conserved. Overall, this study demonstrates the potential of M. caseolyticus subsp. caseolyticus to generate diverse volatile flavor compounds. Additionally, the identification of the highly active strain-specific cell wall bound caseolytic proteases deriving extensive casein hydrolysis, serves as a promising avenue which can be potentially harnessed in the future to produce greater and more diverse flavor compounds.