• Salt content and minimum acceptable levels in whole-muscle cured meat products

      Delgado-Pando, Gonzalo; Fischer, Estelle; Allen, Paul; Kerry, Joesph; O'Sullivan, Maurice; Hamill, Ruth M; Department of Agriculture, Food and the Marine; 11F 026 (Elsevier, 2018-02-01)
      Reported salt levels in whole-muscle cured meat products differ substantially within and among European countries, providing substantial scope for salt reduction across this sector. The objective of this study was to identify the minimum acceptable salt levels in typical whole-muscle cured products in terms of physicochemical, microbial and sensorial properties. Salt levels in a small selection of commercial Irish meat products were determined to establish a baseline for reduction. Subsequently, eight different back bacon rasher and cooked ham products were produced with varying levels of salt: 2.9%, 2.5%, 2% and 1.5% for bacon, and 2%, 1.6%, 1.0% and 0.8% for ham. Salt reduction produced products with significantly harder texture and higher microbial counts, with no difference in the colour and affecting the sensory properties. Nonetheless, salt reduction proved to be feasible to levels of 34% and 19% in bacon and ham products, respectively, compared to baseline.
    • Saturation Mutagenesis of Lysine 12 Leads to the Identification of Derivatives of Nisin A with Enhanced Antimicrobial Activity

      Molloy, Evelyn M.; Field, Des; O'Connor, Paula M.; Cotter, Paul D.; Hill, Colin; Ross, R Paul; Irish Research Council for Science, Engineering and Technology; Enterprise Ireland; Science Foundation Ireland (PLOS, 11/03/2013)
      It is becoming increasingly apparent that innovations from the “golden age” of antibiotics are becoming ineffective, resulting in a pressing need for novel therapeutics. The bacteriocin family of antimicrobial peptides has attracted much attention in recent years as a source of potential alternatives. The most intensively studied bacteriocin is nisin, a broad spectrum lantibiotic that inhibits Gram-positive bacteria including important food pathogens and clinically relevant antibiotic resistant bacteria. Nisin is gene-encoded and, as such, is amenable to peptide bioengineering, facilitating the generation of novel derivatives that can be screened for desirable properties. It was to this end that we used a site-saturation mutagenesis approach to create a bank of producers of nisin A derivatives that differ with respect to the identity of residue 12 (normally lysine; K12). A number of these producers exhibited enhanced bioactivity and the nisin A K12A producer was deemed of greatest interest. Subsequent investigations with the purified antimicrobial highlighted the enhanced specific activity of this modified nisin against representative target strains from the genera Streptococcus, Bacillus, Lactococcus, Enterococcus and Staphylococcus.
    • Second generation GM foods: perspectives on likely future acceptance by Irish consumers

      O'Connor, Elaine; Cowan, Cathal; Williams, Gwilym; O'Connell, John J.; Boland, Maurice; Downey, Gerard; Teagasc Walsh Fellowship Programme; Safefood; Safefood (Teagasc, 2005-07)
      Two hypothetical second-generation genetically-modified (GM) products, a yogurt and a dairy spread, were evaluated by consumers. Second-generation GM food products offer specific consumer benefits - in the case of this study, health benefits.
    • Second Generation GM Foods: Perspectives on Likely Future Acceptance by Irish Consumers

      O'Connor, Elaine; Cowan, Cathal; Williams, Gwilym; O'Connell, John J.; Boland, Maurice; Downey, Gerard (Teagasc, 01/07/2005)
      Two hypothetical second-generation genetically-modified (GM) products, a yogurt and a dairy spread, were evaluated by consumers. Second-generation GM food products offer specific consumer benefits - in the case of this study, health benefits.
    • Selective enrichment of dairy phospholipids in a buttermilk substrate through investigation of enzymatic hydrolysis of milk proteins in conjunction with ultrafiltration

      Barry, Kate M.; Dinan, Timothy G.; Kelly, Philip M.; Department of Agriculture, Food and the Marine, Ireland; Teagasc Walsh Fellowship Programme; 10RD/TMFRC/709 (Elsevier, 04/02/2017)
      Extensive enzymatic hydrolysis of milk proteins in reconstituted buttermilk powder was combined with ultrafiltration to generate a phospholipid (PL) enriched fraction with maximum permeation of hydrolysed peptides. Buttermilk, naturally high in PLs, is the ideal substrate for enrichment of these bio- and techno-functionally active compounds. A 7.8 fold increase in PL was achieved in the 50 kDa retentate; 6.16 ± 0.02% total PL compared with 0.79 ± 0.01% in the starting substrate, an increase considerably greater than previously reported. Total lipid content (% dry matter) increased 6.3 fold in the retentate, 43.43 ± 0.61%, from the starting substrate, 6.84 ± 0.17%. This combined strategic approach enabled maximum enrichment of PLs with no transmission of lipid material into the permeate, 0.09 ± 0.02% total lipid, and non-detectable levels of PLs recovered in the permeate, 0.00 ± 0.01% total PL.
    • Self-association of bovine β-casein as influenced by calcium chloride, buffer type and temperature

      Li, Meng; Auty, Mark; Crowley, Shane V.; Kelly, Alan L.; O'Mahoney, James A.; Brodkorb, Andre; Irish Dairy Levy Research Trust; Teagasc Walsh Fellowship Programme; MDDT 6261 (Elsevier, 2018-09-25)
      The aim of this study was to investigate the aggregation behaviour of a pure β-casein (β-CNpure) and a β-casein concentrate (β-CNconc) as a function of temperature, buffer type (pH 6.8) and the presence of CaCl2. The particle size distribution and turbidity of β-casein (β-CN) dispersions were measured by dynamic light-scattering (DLS) and UV/vis spectroscopy between 4 and 55 °C. Upon heating (4–55 °C), the particle size of both β-CN samples increased, indicating self-association via hydrophobic interactions. It was shown that the self-association of β-CN increased with increasing β-CN concentration and that β-CNpure self-associated at significantly lower concentration than β-CNconc. Both turbidity and particle size measurements showed that the β-CN samples had similar aggregation behaviour in water and imidazole buffer (pH 6.8) but differed in sodium phosphate buffer (pH 6.8), especially at higher ionic calcium concentrations. Fourier Transform Infrared (FTIR) spectroscopy revealed very little change in the secondary structure of β-CN during heating (4–55 °C). The microstructure of β-CN aggregates was monitored during heating from 10 to 55 °C, followed by cooling to 10 °C, using polarised light microscopy. Spherical and heterogeneous aggregates were observed when heated at temperatures above 37 °C, which were reversible upon cooling. This study confirmed that β-CN undergoes self-association on heating that reverses upon cooling, with the aggregation process being highly dependent on the purity of β-CN, the solvent type and the presence of ionic calcium.
    • Semi-supervised linear discriminant analysis

      Toher, Deirdre; Downey, Gerard; Murphy, Thomas Brendan; Science Foundation Ireland; Teagasc (Wiley, 02/07/2012)
      Fisher's linear discriminant analysis is one of the most commonly used and studied classification methods in chemometrics. The method finds a projection of multivariate data into a lower dimensional space so that the groups in the data are well separated. The resulting projected values are subsequently used to classify unlabeled observations into the groups. A semi-supervised version of Fisher's linear discriminant analysis is developed, so that the unlabeled observations are also used in the model fitting procedure. This approach is advantageous when few labeled and many unlabeled observations are available. The semi-supervised linear discriminant analysis method is demonstrated on a number of data sets where it is shown to yield better separation of the groups and improved classification over Fisher's linear discriminant analysis.
    • Sensory optimisation of salt-reduced corned beef for different consumer segments

      Conroy, Paula M.; O'Sullivan, Maurice; Hamill, Ruth M; Kerry, Joseph; Department of Agriculture, Food and the Marine; 11/F/045 (Elsevier, 2019-03-21)
      The study objectives were to determine assessors' (n = 256) preference for corned beef, produced with sequential reductions in NaCl concentrations and to determine if preference was affected by assessor age. The use of a salt replacer such as potassium lactate was also assessed. The youngest age cohort disliked samples containing the highest level of NaCl, whereas the oldest age cohort did not detect differences between samples. The most negatively perceived sample was the control, suggesting that NaCl levels added to commercial corned beef are currently too high for consumer acceptance. All age cohorts, with the exception of the 65–74 age cohort, accepted corned beef samples possessing NaCl levels closest to the FSAI target (1.63 g/100 g). No major sensory differences were noted between samples with and without potassium lactate by the ≥65 age cohort. Potassium lactate may be added to corned beef without affecting sensory attributes, whilst enhancing nutritional content. Assessors of varying age groups have differing preferences for certain NaCl levels and salt replacers.
    • Separation of Oligosaccharides from Lotus Seeds via Medium-pressure Liquid Chromatography Coupled with ELSD and DAD

      Lu, Xu; Zheng, Zhichang; Miao, Song; Li, Huang; Guo, Zebin; Zhang, Yi; Zheng, Yafeng; Zheng, Baodong; Xiao, Jianbo; National Natural Science Foundation of China; et al. (Springer Science, 2017-03-09)
      Lotus seeds were identified by the Ministry of Public Health of China as both food and medicine. One general function of lotus seeds is to improve intestinal health. However, to date, studies evaluating the relationship between bioactive compounds in lotus seeds and the physiological activity of the intestine are limited. In the present study, by using medium pressure liquid chromatography coupled with evaporative light-scattering detector and diode-array detector, five oligosaccharides were isolated and their structures were further characterized by electrospray ionization-mass spectrometry and gas chromatography-mass spectrometry. In vitro testing determined that LOS3-1 and LOS4 elicited relatively good proliferative effects on Lactobacillus delbrueckii subsp. bulgaricus. These results indicated a structure-function relationship between the physiological activity of oligosaccharides in lotus seeds and the number of probiotics applied, thus providing room for improvement of this particular feature. Intestinal probiotics may potentially become a new effective drug target for the regulation of immunity.
    • Sequencing of the Cheese Microbiome and Its Relevance to Industry

      Yeluri Jonnala, Bhagya R.; McSweeney, Paul L. H.; Sheehan, Jeremiah J.; Cotter, Paul D.; Teagasc Walsh Fellowship Programme (Frontiers, 2018-05-23)
      The microbiota of cheese plays a key role in determining its organoleptic and other physico-chemical properties. It is essential to understand the various contributions, positive or negative, of these microbial components in order to promote the growth of desirable taxa and, thus, characteristics. The recent application of high throughput DNA sequencing (HTS) facilitates an even more accurate identification of these microbes, and their functional properties, and has the potential to reveal those microbes, and associated pathways, responsible for favorable or unfavorable characteristics. This technology also facilitates a detailed analysis of the composition and functional potential of the microbiota of milk, curd, whey, mixed starters, processing environments, and how these contribute to the final cheese microbiota, and associated characteristics. Ultimately, this information can be harnessed by producers to optimize the quality, safety, and commercial value of their products. In this review we highlight a number of key studies in which HTS was employed to study the cheese microbiota, and pay particular attention to those of greatest relevance to industry.
    • Sequencing of the Cheese Microbiome and Its Relevance to Industry

      Yeluri Jonnala, Bhagya R.; McSweeney, Paul L. H.; Sheehan, Jeremiah J.; Cotter, Paul D.; Teagasc Walsh Fellowship Programme (Frontiers, 2018-05-23)
      The microbiota of cheese plays a key role in determining its organoleptic and other physico-chemical properties. It is essential to understand the various contributions, positive or negative, of these microbial components in order to promote the growth of desirable taxa and, thus, characteristics. The recent application of high throughput DNA sequencing (HTS) facilitates an even more accurate identification of these microbes, and their functional properties, and has the potential to reveal those microbes, and associated pathways, responsible for favorable or unfavorable characteristics. This technology also facilitates a detailed analysis of the composition and functional potential of the microbiota of milk, curd, whey, mixed starters, processing environments, and how these contribute to the final cheese microbiota, and associated characteristics. Ultimately, this information can be harnessed by producers to optimize the quality, safety, and commercial value of their products. In this review we highlight a number of key studies in which HTS was employed to study the cheese microbiota, and pay particular attention to those of greatest relevance to industry.
    • Sequencing-Based Analysis of the Bacterial and Fungal Composition of Kefir Grains and Milks from Multiple Sources

      Marsh, Alan J.; O'Sullivan, Orla; Hill, Colin; Ross, R Paul; Cotter, Paul D.; Science Foundation Ireland; 07/CE/B1368 (PLOS, 19/07/2013)
      Kefir is a fermented milk-based beverage to which a number of health-promoting properties have been attributed. The microbes responsible for the fermentation of milk to produce kefir consist of a complex association of bacteria and yeasts, bound within a polysaccharide matrix, known as the kefir grain. The consistency of this microbial population, and that present in the resultant beverage, has been the subject of a number of previous, almost exclusively culture-based, studies which have indicated differences depending on geographical location and culture conditions. However, culture-based identification studies are limited by virtue of only detecting species with the ability to grow on the specific medium used and thus culture-independent, molecular-based techniques offer the potential for a more comprehensive analysis of such communities. Here we describe a detailed investigation of the microbial population, both bacterial and fungal, of kefir, using high-throughput sequencing to analyse 25 kefir milks and associated grains sourced from 8 geographically distinct regions. This is the first occasion that this technology has been employed to investigate the fungal component of these populations or to reveal the microbial composition of such an extensive number of kefir grains or milks. As a result several genera and species not previously identified in kefir were revealed. Our analysis shows that the bacterial populations in kefir are dominated by 2 phyla, the Firmicutes and the Proteobacteria. It was also established that the fungal populations of kefir were dominated by the genera Kazachstania, Kluyveromyces and Naumovozyma, but that a variable sub-dominant population also exists.
    • Shelf-life extension ingredient and processing technologies applied to Atlantic salmon (Salmo salar)

      Fogarty, Colin; Department of Agriculture Food and the Marine; 13F458 (2020)
      These current studies explore the possibility of improving analytical methods to assess freshness in seafood and to investigate the antimicrobial potential of a range of natural ingredients both alone and in combination with packaging and chilled storage temperatures to extend the shelf life of Atlantic salmon. The aims of this research study were; • To investigate bacterial growth on Atlantic salmon stored under chilled aerobic conditions thus providing data which may be used to assess which bacterial groups and concentrations are most appropriate for shelf-life determination. • To characterize the microbiota present in the GI tract of Atlantic salmon, using Miseq Illumina high throughput sequencing • To develop and validate rapid sensory (QIM and QDA) and ATP derivative based methods for assessing the freshness of Atlantic salmon. • To investigate the effects of a natural antimicrobial immersion treatment on microbial growth for Atlantic salmon fillets during chilled storage. • To examine the effects of either a natural antimicrobial immersion or spray treatment on mean bacterial counts in combination with packaging technologies for Atlantic salmon fillets during chilled storage. • To assess the effects of skin packaging with retail and sub-zero temperatures on the mean bacterial counts for Atlantic salmon fillets.
    • Shelf-life extension of herring (Clupea harengus) using in-package atmospheric plasma technology

      Albertos, Irene; Martin-Diana, A. B.; Cullen, Patrick J.; Tiwari, Brijesh; Ojha, K. Shikha; Bourke, Paula; Rico, D.; Regional Government of Castille and Leon, Spain; NUGAFU-PEP 2011/769 (Elsevier, 2017-09-17)
      Atmospheric cold plasma is a green and emerging technology, highly interesting to the food industry for its application. Dielectric Barrier Discharges (DBD) can generate atmospheric cold plasma inside sealed packages filled with air through the use of high voltages. This study investigated the use of a large gap DBD design to generate a plasma discharge within the headspace of packaged herring fillets, and its effects on microbiological and quality markers of the fish stored for 11 days at 4 °C. DBD plasma treatment conditions were 70 kV or 80 kV for 5 min treatment time. Results showed that the microbial load (total aerobic mesophilic, total aerobic psychrotrophics, Pseudomonas, lactic acid bacteria and Enterobacteriaceae) were significantly (p < 0.05) lower in the treated samples compared to the control samples. However, samples exposed to the lowest applied voltage better retained key quality factors (lower oxidation and less colour modification). DBD-treatment caused a reduction in trapped water in the myofibrillar network (T21), identified using low-field nuclear magnetic resonance of protons (1H LF NMR). The results indicate that in-package plasma treatment could be employed as an effective treatment for reducing spoilage bacteria in fish.
    • Short communication: Multi-component interactions causing solidification during industrial-scale manufacture of pre-crystallized acid whey powders

      Drapala, Kamil P.; Murphy, Kevin M.; Ho, Quang Tri; Crowley, Shane V.; Mulcahy, Shane; McCarthy, Noel; O'Mahony, James A.; Technology Centres Programme; TC/2014/0016 (Elsevier, 2018-10-03)
      Acid whey (AW) is the liquid co-product arising from acid-induced precipitation of casein from skim milk. Further processing of AW is often challenging due to its high mineral content, which can promote aggregation of whey proteins, which contributes to high viscosity of the liquid concentrate during subsequent lactose crystallization and drying steps. This study focuses on mineral precipitation, protein aggregation, and lactose crystallization in liquid AW concentrates (∼55% total solids), and on the microstructure of the final powders from 2 independent industrial-scale trials. These AW concentrates were observed to solidify either during processing or during storage (24 h) of pre-crystallized concentrate. The more rapid solidification in the former was associated with a greater extent of lactose crystallization and a higher ash-to-protein ratio in that concentrate. Confocal laser scanning microscopy analysis indicated the presence of a loose network of protein aggregates (≤10 µm) and lactose crystals (100–300 µm) distributed throughout the solidified AW concentrate. Mineral-based precipitate was also evident, using scanning electron microscopy, at the surface of AW powder particles, indicating the formation of insoluble calcium phosphate during processing. These results provide new information on the composition- and process-dependent physicochemical changes that are useful in designing and optimizing processes for AW.
    • Short Communication: The effect of dry period duration and dietary energy density on the rennet gelation properties of milk in early lactation

      Butler, Stephen T.; de Feu, M.A.; O'Brien, Bernadette; Guinee, Timothy P.; Murphy, J.J.; National Development Plan (Dublin, Ireland) (American Dairy Science Association and Elsevier Inc., 2010-02)
      This study was carried out to examine the effects of decreasing dry period duration (DP) and altering the energy density of the diet during early lactation on the rheological characteristics of milk. Forty mature Holstein-Friesian cows were used in a completely randomized design with a 2 × 2 factorial arrangement of treatments. Cows were randomly assigned to one of two dry period treatments and one of two nutritional treatments. The dry period treatments were continuous milking (CM) or an 8-week standard dry period (SDP), and the nutritional treatments were a standard energy diet (SE) or a high energy diet (HE). Actual dry period lengths were 6.3 ± 1.7 days and 62.1 ± 1.9 days for cows for the CM and SDP treatments, respectively. Milk samples were collected at 2, 6 and 10 weeks postpartum. The concentration of fat, protein and lactose was determined in each sample. The rennet gelation properties were measured at 31 ° C using dynamic low-amplitude strain oscillatory rheometry. The following parameters were obtained from the resultant elastic shear modulus (G′): gelation time (GT), maximum curd firming rate (CFRmax) and gel strength (GS). Reducing dry period duration from 62 to 6 days resulted in increases in milk protein concentration (31.8 vs. 34.7 g/kg; P < 0.001), CFRmax (2.58 vs. 3.60 Pa/min; P < 0.001) and GS (69.4 vs. 90.5 Pa; P = 0.003). Raising the dietary energy density decreased percentage milk fat (43.1 vs. 37.7 g/kg; P < 0.001) but otherwise had no effect. GS was correlated with CFRmax (r = 0.98; P < 0.001), and both variables were correlated with milk protein concentration (r = 0.71; P < 0.001, and r = 0.73; P < 0.001, respectively). The results indicate that decreasing the duration of DP increased milk protein concentration and improved the rennet gelation properties of milk, but that dietary energy density had little effect.
    • A Simple Method for the Purification of Nisin

      Gough, Ronan; Gomez-Sala, Beatriz; O'Connor, Paula M.; Rea, Mary C.; Miao, Song; Hill, Colin; Brodkorb, Andre (Springer, 29/05/2017)
      Nisin, an antimicrobial peptide showing activity against a broad range of Gram-positive bacteria, is widely used as a food preservative and has potential as a therapeutic for a range of infectious diseases. Here, we present a simple purification method, based on a salting-out approach, which can produce a powder containing ∼33% nisin, from a nisin-producing culture in a whey permeate-based medium. This process removes over 99% of the lactic acid, NaCl, lactose and non-nisin proteins from the cell-free culture supernatant. The approach can also enrich a commonly used commercial nisin preparation over 30-fold to a purity of ∼58%. These are higher purities than comparable published methods. The simplicity of this approach facilitates its use in research and also its scale-up.
    • Simulated gastrointestinal digestion of nisin and interaction between nisin and bile

      Gough, Ronan; O'Connor, Paula M.; Rea, Mary C.; Gomez-Sala, Beatriz; Miao, Song; Hill, Colin; Brodkorb, Andre; Department of Agriculture, Food and the Marine; Teagasc Walsh Fellowship Programme; 10/RD/TMFRC/701; et al. (Elsevier, 14/08/2017)
      Nisin, an antimicrobial peptide showing activity against many Gram positive bacteria, is widely used as a food preservative. The simulated gastrointestinal digestion of nisin (variant A) was studied using the in vitro INFOGEST digestion method. Following oral, gastric and small intestinal digestion, there was no intact nisin in the system and the nisin was primarily digested by pancreatin. After digestion, six nisin fragments (1–11, 1–12, 1–20, 1–21, 1–29 and 1–32) were identified by reversed phase high performance liquid chromatography and mass spectroscopy and four of these nisin fragments (1–20, 1–21, 1–29 and 1–32) demonstrated low antibacterial activity against Lactococcus lactis HP in agar diffusion activity assays. Additionally, it was observed that bile salts form a complex with nisin. This was examined by atomic force microscopy, turbidity and dynamic light scattering, which showed that this interaction resulted in significantly larger bile salt micelles. The presence of bile salts at physiological levels significantly altered the relative amounts of the nisin fragments 1–12, 1–20 and 1–29 produced during an in vitro digestion. This study highlights the importance of including bile in simulated digestions of antimicrobial peptides in order to obtain a more accurate simulation of the in vivo digestion products and their activity.
    • Simulated gastrointestinal digestion of nisin and interaction between nisin and bile

      Gough, Ronan; O'Connor, Paula M.; Rea, Mary C.; Gomez-Sala, Beatriz; Miao, Song; Hill, Colin; Brodkorb, Andre; Department of Agriculture, Food and the Marine; Teagasc Walsh Fellowship Programme; 10/RD/TMFRC/701 (Elsevier, 2017-08-14)
      Nisin, an antimicrobial peptide showing activity against many Gram positive bacteria, is widely used as a food preservative. The simulated gastrointestinal digestion of nisin (variant A) was studied using the in vitro INFOGEST digestion method. Following oral, gastric and small intestinal digestion, there was no intact nisin in the system and the nisin was primarily digested by pancreatin. After digestion, six nisin fragments (1–11, 1–12, 1–20, 1–21, 1–29 and 1–32) were identified by reversed phase high performance liquid chromatography and mass spectroscopy and four of these nisin fragments (1–20, 1–21, 1–29 and 1–32) demonstrated low antibacterial activity against Lactococcus lactis HP in agar diffusion activity assays. Additionally, it was observed that bile salts form a complex with nisin. This was examined by atomic force microscopy, turbidity and dynamic light scattering, which showed that this interaction resulted in significantly larger bile salt micelles. The presence of bile salts at physiological levels significantly altered the relative amounts of the nisin fragments 1–12, 1–20 and 1–29 produced during an in vitro digestion. This study highlights the importance of including bile in simulated digestions of antimicrobial peptides in order to obtain a more accurate simulation of the in vivo digestion products and their activity.
    • Sinapinic and protocatechuic acids found in rapeseed: isolation, characterisation and potential benefits for human health as functional food ingredients

      Quinn, Leah; Gray, Stephen G.; Meaney, Steven; Finn, Stephen; Kenny, Owen; Hayes, Maria (Teagasc (Agriculture and Food Development Authority), Ireland, 13/12/2017)
      Rapeseed is one of the world’s major oilseeds, and rapeseed oil is produced by pressing of the seeds. This process results in the production of a low-economic-value by-product, rapeseed meal, which is commonly used as animal feed. Rapeseed meal is rich in bioactive phenolic compounds, including sinapinic acid (SA) and protocatechuic acid (PCA). Isolation of these bioactive compounds from a by-product of rapeseed oil production is largely in agreement with the current concept of the circular economy and total utilisation of crop harvest using a biorefinery approach. In this review, current information concerning traditional and novel methods to isolate phenolic compounds – including SA and PCA – from rapeseed meal, along with in vitro and in vivo studies concerning the bioactivity of SA and PCA and their associated health effects, is collated. These health effects include anti-inflammatory, anti-cancer, anti-diabetes activities, along with histone deacetylase inhibition and protective cardiovascular, neurological and hepatic effects. The traditional extraction methods include use of solvents and/or enzymes. However, a need for simpler, more efficient methodologies has led to the development of novel extraction processes, including microwave-assisted, ultrasound-assisted, pulsed electric field and high-voltage electrical discharge extraction processes.