• Indicator organisms to determine the use of chilling as a critical point in beef slaughter HACCP

      Prendergast, Deirdre M.; Sheridan, James J.; Downey, Gerard (Teagasc, 01/11/2008)
      During chilling, temperatures of carcass surfaces at different sites change over time as do other parameters such as water activity (aw), the structure of the muscle and other tissues, as the carcass enters rigor mortis. Many of these factors are known to have a major effect on cell survival and growth and must be considered in determining the influence of chilling on bacterial survival on carcass surfaces. This study aimed to determine if chilling could be used as a critical control point (CCP) in beef slaughter in relation to pathogens such as E. coli O157:H7 and L. monocytogenes, using E. coli and Listeria innocua as pathogen indicators. The present study was designed to determine the influence of (a) chilling at 10oC for 72 h on the survival of E. coli and (b) chilling at 4oC for 72 h on the survival of L. innocua inoculated at different sites on beef carcasses. Three sites (neck, outside round and brisket) were inoculated (1) immediately after dressing while hot (E. coli and L. innocua) and (2) when cold after chilling (L. innocua). The influence of changes in surface aw was also considered and their relationship to the survival of E. coli and L. innocua over time was assessed. The data are discussed in relation to the use of chilling as a CCP in beef hazard analysis (HACCP) and the monitoring of neck temperature as the most suitable CCP.
    • Adding value to Beef Forequarter Muscles

      Kenny, Tony; Lennon, Ann; Ward, Patrick; Sullivan, Paul; McDonald, Karl; O'Neill, Eileen; Downey, Gerard (Teagasc, 01/11/2008)
      The forequarter constitutes 50% of the weight of a beef carcase but only about 25% of its value.To fulfill the objectives of this project, the work was organised into 4 parts as follows: 1.Characterisation of the available raw material, in terms of properties of individual muscles seamed out from carcasses of representative types of animals produced in Ireland. 2.Comparison of yields and operator time for seaming and conventional boning. 3.Utilisation of separated muscles in added-value products using appropriate tenderising, bonding and forming technology. 4.Transfer of the knowledge and technology to the industry.
    • New Product Development Opportunities for Irish Companies in the British Cheese Market

      Cowan, Cathal; Downey, Gerard (Teagasc, 01/11/2008)
      The primary objective of this research was to identify innovative cheese concepts appropriate for UK consumers and suitable for Irish industry to manufacture. It also aimed to identify personal, situational and market factors that influence consumers when purchasing cheese. This research study used existing market literature, in-depth interviews and consumer focus groups.
    • An examination of the molecular mechanisms controlling the tissue accumulation of conjugated linoleic acid (CLA) in cattle

      Waters, Sinead M.; Hynes, A.C.; Killeen, Aideen P.; Moloney, Aidan P; Kenny, David A. (Teagasc, 01/12/2008)
      Long chain n-3 polyunsaturated fatty acids (n-3 PUFA) and conjugated linoleic acid (CLA) have demonstrable and potential human health benefits in terms of preventing cancer, diabetes, chronic inflammation, obesity and coronary heart disease. Supplementation of cattle diets with a blend of oils rich in n-3 PUFA and linoleic acid have a synergistic effect on the accumulation of ruminal and tissue concentrations of trans vaccenic acid (TVA), the main substrate for ?-9 desaturase which is responsible for de novo tissue synthesis of the cis 9, trans 11 isomer of CLA. This dietary strategy translates into increases in milk concentrations of CLA in dairy cows; however, concentrations in the muscle of beef animals have not always been increased. There is an apparent paradox in that n-3 PUFA supplementation enhances ruminal synthesis of trans-vaccenic acid (TVA), but then inhibits its conversion to CLA possibly through altering the activity of ?-9 desaturase. Recently, the promoter regions of the bovine ?- 9 desaturase gene has been isolated and analysed and has been shown to contain a conserved PUFA response region.
    • Formation of non-native β-lactoglobulin during heat-induced denaturation

      Kehoe, Joseph James; Wang, Lizhe; Morris, Edwin R; Brodkorb, Andre; Teagasc Walsh Fellowship Programme; Department of Agriculture, Food and the Marine (Springer, 01/12/2011)
      A mechanism describing the denaturation and aggregation behavior during heat-treatment of pure β-lactoglobulin and β-lactoglobulin in whey protein isolate (WPI) under selected conditions (20 to 90 gL−1 in water at pH 7.0, 78 °C) is presented. A combination of reversed-phase and gel permeation chromatography was used to study the disappearance of native β-lactoglobulin and the formation of non-native intermediates in the aggregation process. The mean reaction order for pure β-lactoglobulin and β-lactoglobulin in WPI were the same, 1.4. While the rate of β-lactoglobulin denaturation was greater in WPI there was less aggregation compared to that of pure β-lactoglobulin. More of the β-lactoglobulin in WPI remained in a non-native monomer intermediate state after 30 min of heating. After an initial lag period, during which non-native monomers appeared, aggregates formed and rapidly reached a plateau in terms of their size. These aggregates were visualized using atomic force microscopy. There was no significant effect of protein concentration on either aggregate size or the number of exposed sulfhydryls in the heated solutions.
    • 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.
    • Alternative uses for co-products: Harnessing the potential of valuable compounds from meat processing chains

      Mullen, Anne Maria; Álvarez García, Carlos; Zeugolis, Dimitrios; Henchion, Maeve; O'Neill, Eileen; Drummond, Liana; Department of Agriculture, Food and the Marine, Ireland; 11/F/043 (Elsevier, 03/05/2017)
      Opportunities for exploiting the inherent value of protein-rich meat processing co-products, in the context of increased global demand for protein and for sustainable processing systems, are discussed. While direct consumption maybe the most profitable route for some, this approach is influenced greatly by local and cultural traditions. A more profitable and sustainable approach may be found in recognizing this readily available and under-utilised resource can provide high value components, such as proteins, with targeted high value functionality of relevance to a variety of sectors. Applications in food & beverages, petfood biomedical and nutrition arenas are discussed. Utilization of the raw material in its entirety is a necessary underlying principle in this approach to help maintain minimum waste generation. Understanding consumer attitudes to these products, in particular when used in food or beverage systems, is critical in optimizing commercialization strategies.
    • 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; 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.
    • Tryptophan-Mediated Denaturation of β-Lactoglobulin A by UV Irradiation

      Kehoe, Joseph James; Remondetto, Gabriel E; Subirade, Muriel; Morris, Edwin R; Brodkorb, Andre (American Chemical Society, 04/06/2008)
      β-Lactoglobulin A, a genetic variant of one of the main whey proteins, was irradiated at 295 nm for 24 h. After irradiation, 18% of the protein was denatured (determined by reverse-phase chromatography). The fluorescence spectrum of the irradiated protein was red-shifted compared to that of the native protein, indicating a change in protein folding. Sulfhydryl groups, which are buried in native β-lactoglobulin, were exposed following irradiation and became available for quantification using the Ellman assay. The quantity of exposed sulfhydryls increased, but the number of total sulfhydryl groups decreased. Gel permeation chromatography showed that some protein aggregation occurred during irradiation. Fourier transform infrared (FTIR) spectroscopy of irradiated β-lactoglobulin revealed changes in the secondary structure, comparable to that of early events during heat-induced denaturation. There was evidence for some photo-oxidation of tryptophan.
    • Transcriptome analysis of porcine M. semimembranosus divergent in intramuscular fat as a consequence of dietary protein restriction

      Hamill, Ruth M; Aslan, Ozlem; Mullen, Anne Maria; O'Doherty, John V.; McBryan, Jean; Morris, Dermot G.; Sweeney, Torres; Department of Agriculture, Food and the Marine, Ireland (Biomed Central, 06/07/2013)
      Background: Intramuscular fat (IMF) content is positively correlated with aspects of pork palatability, including flavour, juiciness and overall acceptability. The ratio of energy to protein in the finishing diet of growing pigs can impact on IMF content with consequences for pork quality. The objective of this study was to compare gene expression profiles of Musculus semimembranosus (SM) of animals divergent for IMF as a consequence of protein dietary restriction in an isocaloric diet. The animal model was derived through the imposition of low or high protein diets during the finisher stage in Duroc gilts. RNA was extracted from post mortem SM tissue, processed and hybridised to Affymetrix porcine GeneChip® arrays. Results: IMF content of SM muscle was increased on the low protein diet (3.60 ± 0.38% versus 1.92 ± 0.35%). Backfat depth was also greater in animals on the low protein diet, and average daily gain and feed conversion ratio were lower, but muscle depth, protein content and moisture content were not affected. A total of 542 annotated genes were differentially expressed (DE) between animals on low and high protein diets, with 351 down-regulated and 191 up-regulated on the low protein diet. Transcript differences were validated for a subset of DE genes by qPCR. Alterations in functions related to cell cycle, muscle growth, extracellular matrix organisation, collagen development, lipogenesis and lipolysis, were observed. Expression of adipokines including LEP, TNFα and HIF1α were increased and the hypoxic stress response was induced. Many of the identified transcriptomic responses have also been observed in genetic and fetal programming models of differential IMF accumulation, indicating they may be robust biological indicators of IMF content. Conclusion: An extensive perturbation of overall energy metabolism in muscle occurs in response to protein restriction. A low protein diet can modulate IMF content of the SM by altering gene pathways involved in lipid biosynthesis and degradation; however this nutritional challenge negatively impacts protein synthesis pathways, with potential consequences for growth.
    • Comparative Proteomic Profiling of Divergent Phenotypes for Water Holding Capacity across the Post Mortem Ageing Period in Porcine Muscle Exudate

      Di Luca, Alessio; Hamill, Ruth; Mullen, Anne Maria; Slavov, Nikolai; Giuliano, Elia; Department of Agriculture, Food and the Marine, Ireland; 06RDNUIG470 (PLOS, 07/03/2016)
      Two dimensional Difference Gel Electrophoresis (2-D DIGE) and mass spectrometry were applied to investigate the changes in metabolic proteins that occur over a seven day (day 1, 3 and 7) post mortem ageing period in porcine centrifugal exudate from divergent meat quality phenotypes. The objectives of the research were to enhance our understanding of the phenotype (water holding capacity) and search for biomarkers of this economically significant pork quality attribute. Major changes in protein abundance across nine phenotype-by-time conditions were observed. Proteomic patterns were dominated by post mortem ageing timepoint. Using a machine learning algorithm (l1-regularized logistic regression), a model was derived with the ability to discriminate between high drip and low drip phenotypes using a subset of 25 proteins with an accuracy of 63%. Models discriminating between divergent phenotypes with accuracy of 72% and 73% were also derived comparing respectively, high drip plus intermediate phenotype (considered as one phenotype) versus low drip and comparing low drip plus intermediate phenotype (considered as one phenotype) versus high drip. In all comparisons, the general classes of discriminatory proteins identified include metabolic enzymes, stress response, transport and structural proteins. In this research we have enhanced our understanding of the protein related processes underpinning this phenotype and provided strong data to work toward development of protein biomarkers for water holding capacity.
    • Milk Quality Handbook. Practical Steps to Improve Milk Quality

      Moorepark Dairy Production Research Centre; O'Brien, Bernadette (Teagasc, 07/09/2008)
      The overall objective of this booklet is to provide guidelines and recommendations for the on-farm milk production process such that the milk produced may achieve increasingly stringent milk hygiene and processing quality standards. It is intended that the genuine problem areas in relation to milk quality will be targeted and an increased awareness and perception of milk quality be provided.
    • Prediction of naturally-occurring, industrially-induced and total trans fatty acids in butter, dairy spreads and Cheddar cheese using vibrational spectroscopy and multivariate data analysis

      Zhao, Ming; Beattie, Renwick J.; Fearon, A.M.; O'Donnell, Colm P.; Downey, Gerard; Department of Agriculture, Food and the Marine; Teagasc Walsh Fellowship Programme (Elsevier, 08/08/2015)
      This study investigated the use of vibrational spectroscopy [near infrared (NIR), Fourier-transform mid-infrared (FT-MIR), Raman] and multivariate data analysis for (1) quantifying total trans fatty acids (TT), and (2) separately predicting naturally-occurring (NT; i.e., C16:1 t9; C18:1 trans-n, n = 6 … 9, 10, 11; C18:2 trans) and industrially-induced trans fatty acids (IT = TT – NT) in Irish dairy products, i.e., butter (n = 60), Cheddar cheese (n = 44), and dairy spreads (n = 54). Partial least squares regression models for predicting NT, IT and TT in each type of dairy product were developed using FT-MIR, NIR and Raman spectral data. Models based on NIR, FT-MIR and Raman spectra were used for the prediction of NT and TT content in butter; best prediction performance achieved a coefficient of determination in validation (R2V) ∼ 0.91–0.95, root mean square error of prediction (RMSEP) ∼ 0.07–0.30 for NT; R2V ∼ 0.92–0.95, RMSEP ∼ 0.23–0.29 for TT.
    • Structure and antioxidant activity of Maillard reaction products from α-lactalbumin and β-lactoglobulin with ribose in an aqueous model system

      Jiang, Zhanmei; Brodkorb, Andre; Innovative Research Team of Higher Education of Heilongjiang Province; Program for New Century Excellent Talents in Heilongjiang Province University (Elsevier, 11/02/2012)
      Maillard reaction products (MRPs) were prepared from aqueous model mixtures containing 3% (w/w) ribose and 3% (w/w) of the dairy proteins α-lactalbumin (α-LA) or β-lactoglobulin (β-LG), heated at 95 °C, for up to 5 h. The pH of MRPs decreased significantly during heat treatment of α-LA-Ribose and β-LG-Ribose mixtures from 8.4 to 5.3. The amino group content in MRPs, derived from the α-LA-Ribose and β-LG-Ribose model system, was decreased noticeably during the first hour and did not change thereafter. The loss of free ribose in MRPs was higher for β-LG-Ribose than for α-LA-Ribose. During the Maillard reaction, the concentration of native and non-native α-LA, or β-LG, decreased and the formation of aggregates was observed. Fluorescence intensity of the β-LG-Ribose MRPs reached maximum within 1 h, compared to 2 h for α-LA-Ribose MRPs. Meanwhile, modification of the UV/vis absorption spectra for α-LA and β-LG was mainly due to a condensation reaction with ribose. Dynamic light scattering showed a significant increase in the particle size of the MRPs. Size exclusion chromatography of MRPs revealed the production of both high and low molecular weight material. Electrophoresis of MRPs indicated polymerization of α-LA and β-LG monomers via inter-molecular disulfide bridge, but also via other covelant bonds. MRPs from α-LA-Ribose and β-LG-Ribose exhibited increased antioxidant activities, therefore theses MRPs may be used as natural antioxidants in food products.
    • Feasibility Study on the Use of Visible–Near-Infrared Spectroscopy for the Screening of Individual and Total Glucosinolate Contents in Broccoli

      Hernandez-Hierro, Jose Miguel; Valverde, Juan; Villacreces, Salvador; Reilly, Kim; Gaffney, Michael; Gonzalez-Miret, Maria Lourdes; Heredia, Francisco J.; Downey, Gerard; Spanish MICINN; Junta de Andalucia; et al. (American Chemical Society, 11/07/2012)
      The potential of visible–near-infrared spectroscopy to determine selected individual and total glucosinolates in broccoli has been evaluated. Modified partial least-squares regression was used to develop quantitative models to predict glucosinolate contents. Both the whole spectrum and different spectral regions were separately evaluated to develop the quantitative models; in all cases the best results were obtained using the near-infrared zone between 2000 and 2498 nm. These models have been externally validated for the screening of glucoraphanin, glucobrassicin, 4-methoxyglucobrassicin, neoglucobrassicin, and total glucosinolates contents. In addition, discriminant partial least-squares was used to distinguish between two possible broccoli cultivars and showed a high degree of accuracy. In the case of the qualitative analysis, best results were obtained using the whole spectrum (i.e., 400–2498 nm) with a correct classification rate of 100% in external validation being obtained.
    • Isolation and characterisation of κ-casein/whey protein particles from heated milk protein concentrate and role of κ-casein in whey protein aggregation

      Gaspard, Sophie J.; Auty, Mark; Kelly, Alan L.; O'Mahony, James A.; Brodkorb, Andre; Teagasc Walsh Fellowship Programme; Dairy Levy Research Trust; 2012211 (Elsevier, 12/06/2017)
      Milk protein concentrate (79% protein) reconstituted at 13.5% (w/v) protein was heated (90 °C, 25 min, pH 7.2) with or without added calcium chloride. After fractionation of the casein and whey protein aggregates by fast protein liquid chromatography, the heat stability (90 °C, up to 1 h) of the fractions (0.25%, w/v, protein) was assessed. The heat-induced aggregates were composed of whey protein and casein, in whey protein:casein ratios ranging from 1:0.5 to 1:9. The heat stability was positively correlated with the casein concentration in the samples. The samples containing the highest proportion of caseins were the most heat-stable, and close to 100% (w/w) of the aggregates were recovered post-heat treatment in the supernatant of such samples (centrifugation for 30 min at 10,000 × g). κ-Casein appeared to act as a chaperone controlling the aggregation of whey proteins, and this effect was stronger in the presence of αS- and β-casein.
    • New Insights into Cell Encapsulation and the Role of Proteins During Flow Cytometry

      Doherty, Sinead B.; Brodkorb, Andre; Irish National Development Plan 2007 to 2013; Irish Dairy Levy Research Trust; Science Foundation Ireland; NU518 (Intech, 13/06/2012)
      Modern approaches to science tend to follow divergent paths. On one hand, instruments and technologies are developed to capture as much information as possible with the need for complex data analysis to identify problematic issues. On the other hand, formulation focused, minimalistic approaches that gather only the most pertinent data for specific questions also represent a powerful methodology. This chapter will provide many examples of the latter by integrating Flow Cytometry (FACS - Fluorescence-Activated Cell Sorting) technology with high throughput screening (HTS) of encapsulation systems with extensive utility of one-dimensional (1-D) imaging for protein localisation. In this regard, less information is acquired from each cell, data files will be more manageable, easier to analyse and throughput screening will be significantly enhanced beyond traditional HTS analysis, irrespective of the protein concentration present in the background or delivery media.
    • Observations on the water distribution and extractable sugar content in carrot slices after pulsed electric field treatment

      Aguilo-Aguayo, Ingrid; Downey, Gerard; Keenan, Derek F.; Lyng, James G.; Brunton, Nigel; Rai, Dilip K.; Department of Agriculture, Food and the Marine; Generalitat of Catalonia; Lifelong Learning Programme; FIRM 06/TNI/AFRC6; et al. (Elsevier, 13/06/2014)
      The impact of pulsed electric field (PEF) processing conditions on the distribution of water in carrot tissue and extractability of soluble sugars from carrot slices was studied. Time domain NMR relaxometry was used to investigate the water proton mobility in PEF-treated carrot samples. Three distinct transverse relaxation peaks were observed in untreated carrots. After PEF treatment only two slightly-overlapping peaks were found; these were attributed to water present in the cytoplasm and vacuole of carrot xylem and phloem tissues. This post-treatment observation indicated an increase in water permeability of tissues and/or a loss of integrity in the tonoplast. In general, the stronger the electric field applied, the lower the area representing transverse relaxation (T2) values irrespective of treatment duration. Moreover an increase in sucrose, β- and α-glucose and fructose concentrations of carrot slice extracts after PEF treatment suggested increases in both cell wall and vacuole permeability as a result of exposure to pulsed electric fields.
    • Pilot-scale Production of Hydrolysates with Altered Bio-functionalities based on Thermally-denatured Whey Protein Isolate

      O'Loughlin, Ian B.; Murray, Brian A.; Fitzgerald, Richard J.; Brodkorb, Andre; Kelly, Philip; Enterprise Ireland; Teagasc Walsh Fellowship Programme; CC20080001 (Elsevier, 13/08/2013)
      Whey protein isolate (WPI) solutions (100 g L−1 protein) were subjected to a heat-treatment of 80 °C for 10 min. Unheated and heat-treated WPI solutions were hydrolysed with Corolase® PP at pilot-scale to either 5 or 10% degree of hydrolysis (DH). Hydrolysates were subsequently processed via cascade membrane fractionation using 0.14 μm, and 30, 10, 5 and 1 kDa cut-off membranes. The compositional and molecular mass distribution profiles of the substrate hydrolysates and membrane processed fractions were determined. Whole and fractionated hydrolysates were assayed for both angiotensin-I-converting enzyme (ACE) inhibitory activity and ferrous chelating capabilities. A strong positive correlation (P < 0.01) was established between the average molecular mass of the test samples and the concentration needed to chelate 50% of the iron (CC50) in solution. The lowest ACE inhibition concentration (IC50 = 0.23 g L−1 protein) was determined for the 1 kDa permeate of the heat-treated 10% DH hydrolysate
    • Simulated gastrointestinal digestion of nisin and interaction between nisin and bile

      Gough, Ronan; O'Connor, Paula M.; Rea, Mary; 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.