• Antioxidant active packaging systems to extend the shelf life of sliced cooked ham

      Pateiro, Mirian; Domínguez, Rubén; Bermúdez, Roberto; Munekata, Paulo E.S.; Zhang, Wangang; Gagaoua, Mohammed; Lorenzo, José M.; INIA-MINECO; Axencia Galega de Innovación; CYTED; et al. (Elsevier BV, 2019-11)
      The effectiveness of active packaging systems with green tea extract and oregano essential oil was checked for their use in sliced cooked ham. Three packaging systems were evaluated: i) control group without active film, ii) ATGT packed with active film of green tea extract (1%) and iii) ATRX with a mixture of green tea extract and oregano essential oil (1%). The evolution of microbiological, physicochemical (pH, aw, colour and lipid oxidation) and sensory attributes were analysed after 0, 7, 14 and 21 days of refrigerated storage. Microbial populations were below the limits established by the European Regulations (106 UFC/g). The samples packed with ATGT showed the better antimicrobial activity against total viable counts (TVC) and lactic acid bacteria (BAL), while lower counts of Brochothrix thermosphacta was observed in ATRX film (1.48 vs. 1.78 and 2.59 UFC/g for ATRX vs. ATGT and CON, respectively). Regarding colour, low differences were found between the samples packaged with active and control films. Unlike L*, a* and b* parameters showed a progressive diminution throughout the storage in all batches, being the films that contained green tea (ATGT) were the ones that showed the less discolouration at the end of storage (8.86 vs. 8.63 and 7.50 for ATGT vs. CON and ATRX, respectively). The low fat content of this type of product and the use of anaerobic atmosphere for the packaging of cooked ham did not allow to show an antioxidant effect on lipid oxidation (values below 0.15 mg MDA/kg). Finally, the use of ATGT and ATRX did not suppose a modification of the sensorial attributes of the product, being acceptance scores under the acceptance limit during the whole display.
    • A Comprehensive Review on Lipid Oxidation in Meat and Meat Products

      Domínguez, Rubén; Pateiro, Mirian; Gagaoua, Mohammed; Barba, Francisco J.; Zhang, Wangang; Lorenzo, José M. (MDPI AG, 2019-09-25)
      Meat and meat products are a fundamental part of the human diet. The protein and vitamin content, as well as essential fatty acids, gives them an appropriate composition to complete the nutritional requirements. However, meat constituents are susceptible to degradation processes. Among them, the most important, after microbial deterioration, are oxidative processes, which affect lipids, pigments, proteins and vitamins. During these reactions a sensory degradation of the product occurs, causing consumer rejection. In addition, there is a nutritional loss that leads to the formation of toxic substances, so the control of oxidative processes is of vital importance for the meat industry. Nonetheless, despite lipid oxidation being widely investigated for decades, the complex reactions involved in the process, as well as the different pathways and factors that influenced them, make that lipid oxidation mechanisms have not yet been completely understood. Thus, this article reviews the fundamental mechanisms of lipid oxidation, the most important oxidative reactions, the main factors that influence lipid oxidation, and the routine methods to measure compounds derived from lipid oxidation in meat.
    • Current Trends in Proteomic Advances for Food Allergen Analysis

      López-Pedrouso, María; Lorenzo, José M.; Gagaoua, Mohammed; Franco, Daniel; Programa Iberoamericano de Ciencia y Tecnología para el Desarrollo; Axencia Galega de Innovación, Xunta de Galicia, Spain; 119RT0568; IN607A2019/01 (MDPI AG, 2020-08-25)
      Food allergies are a global food challenge. For correct food labelling, the detection and quantification of allergens are necessary. However, novel product formulations and industrial processes produce new scenarios, which require much more technological developments. For this purpose, OMICS technologies, especially proteomics, seemed to be relevant in this context. This review summarises the current knowledge and studies that used proteomics to study food allergens. In the case of the allergenic proteins, a wide variety of isoforms, post-translational modifications and other structural changes during food processing can increase or decrease the allergenicity. Most of the plant-based food allergens are proteins with biological functions involved in storage, structure, and plant defence. The allergenicity of these proteins could be increased by the presence of heavy metals, air pollution, and pesticides. Targeted proteomics like selected/multiple reaction monitoring (SRM/MRM) have been very useful, especially in the case of gluten from wheat, rye and barley, and allergens from lentil, soy, and fruit. Conventional 1D and 2-DE immunoblotting have been further widely used. For animal-based food allergens, the widely used technologies are 1D and 2-DE immunoblotting followed by MALDI-TOF/TOF, and more recently LC-MS/MS, which is becoming useful to assess egg, fish, or milk allergens. The detection and quantification of allergenic proteins using mass spectrometry-based proteomics are promising and would contribute to greater accuracy, therefore improving consumer information.
    • Evaluation of the Antioxidant and Antimicrobial Activities of Porcine Liver Protein Hydrolysates Obtained Using Alcalase, Bromelain, and Papain

      Borrajo, Paula; Pateiro, Mirian; Gagaoua, Mohammed; Franco, Daniel; Zhang, Wangang; Lorenzo, José Manuel; INIA; RTA 2017-00024-CO4-04 (MDPI AG, 2020-03-27)
      In order to make the by-products generated from the porcine industry more valuable, pig livers were used in this trial to obtain protein hydrolysates. Three proteases (alcalase, bromelain, and papain) were utilized for enzymatic hydrolysis with two different durations, 4 and 8 hours. Ultrafiltration process was used for the recovery of the extracts, employing three different membrane pore sizes (30, 10, and 5 kDa). The porcine livers contained considerable amounts of protein (19.0%), considering they are almost composed of water (74.1%). The antioxidant activity of the obtained hydrolysates was investigated using four antioxidant methods (2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, 2-2′-Azino-di-[3-ethylbenzthiazoline sulfonate] (ABTS) radical scavenging activity, ferric reducing antioxidant power assay (FRAP), and oxygen radical absorbance capacity assay (ORAC)). Antibacterial properties were also measured against Gram-negative and Gram-positive bacteria. Results indicated that the three studied factors (type of enzyme, membrane pore size, and time) significantly affected the parameters evaluated. Hydrolysates obtained at 8 hours with alcalase had the best antioxidant properties. The 30 kDa alcalase extracts exhibited the highest DPPH (562 µg Trolox/g), FRAP (82.9 µmol Fe2+/100 g), and ORAC (53.2 mg Trolox/g) activities, while for ABTS the 10 kDa alcalase showed the higher values (1068 mg ascorbic acid/100 g). Concerning the antibacterial activity, 30 kDa hydrolysates obtained with bromelain for 4 hours exhibited the highest antimicrobial capacity, providing an inhibition of 91.7%.
    • Inclusion of Healthy Oils for Improving the Nutritional Characteristics of Dry-Fermented Deer Sausage

      Vargas-Ramella, Márcio; Munekata, Paulo E. S.; Gagaoua, Mohammed; Franco, Daniel; Campagnol, Paulo C. B.; Pateiro, Mirian; Barretto, Andrea Carla da Silva; Domínguez, Rubén; Lorenzo, José M.; CYTED; et al. (MDPI AG, 2020-10-18)
      The influence of partial replacement of animal fat by healthy oils on composition, physicochemical, volatile, and sensory properties of dry-fermented deer sausage was evaluated. Four different batches were manufactured: the control was formulated with animal fat (18.2%), while in the reformulated batches the 50% of animal fat was substituted by olive, canola, and soy oil emulsions immobilized in Prosella gel. The reformulation resulted in a decrease of moisture and fat contents and an increase of protein and ash amount. Moreover, reformulated sausages were harder, darker, and had higher pH values. This fact is related to the lower moisture content in these samples. As expected, the fatty acid composition was changed by the reformulation. The use of soy and canola oils increased polyunsaturated fatty acids and omega-3 content and decreased n-6/n-3 ratio and saturated fatty acids. Thus, the use of these two oils presented the best nutritional benefits. The changes observed in the fatty acids reflected the fatty acid composition of the oils employed in the emulsions. Regarding volatile compounds (VOC), the replacement of animal fat by healthy emulsion gels increased the content of both total VOC and most of individual VOC. However, the lipid-derived VOC did not show this trend. Generally speaking, the control samples presented similar or higher VOC derived from lipid oxidation processes, which could be related to the natural antioxidant compounds present in the vegetable oils. Finally, all reformulated sausages presented higher consumer acceptability than control samples. In fact, the sausage reformulated with soy oil emulsion gel was the most preferred. Thus, as a general conclusion, the reformulation of deer sausages with soy emulsion gel improves both composition and sensory quality of the final product, which could be an excellent strategy to the elaboration of healthy fermented sausages.
    • Meta-proteomics for the discovery of protein biomarkers of beef tenderness: An overview of integrated studies

      Picard, Brigitte; Gagaoua, Mohammed; Marie Skłodowska-Curie Grant; Enterprise Ireland; Pôle Aquitain Agro-Alimentation et Nutrition; National Institute of Agronomical Research; National Institute of Origin and Quality; FNADT Massif Central; DATAR Massif Central; ANR GenAnimal; et al. (Elsevier BV, 2020-01)
      This meta-proteomics review focused on proteins identified as candidate biomarkers of beef tenderness by comparing extreme groups of tenderness using two-dimensional electrophoresis (2-DE) associated with mass spectrometry (MS). We reviewed in this integromics study the results of 12 experiments that identified protein biomarkers from two muscles, Longissimus thoracis (LT) and Semitendinosus (ST), of different types of cattle: young bulls, steers or cows from beef breeds (Charolais, Limousin, Blond d’Aquitaine), hardy breed (Salers) or mixed breed (PDO Maine-Anjou). Comparative proteomics of groups differing in their tenderness evaluated by instrumental Warner-Bratzler shear force (WBSF) or by sensory analysis using trained panelists, revealed 61 proteins differentially abundant (P < 0.05) between tender and tough groups. A higher number of discriminative proteins was observed for LT (50 proteins) compared to ST muscle (28 proteins). The Gene Ontology annotations showed that the proteins of structure and contraction, protection against oxidative stress and apoptosis, energy metabolism, 70 family HSPs and proteasome subunits are more involved in LT tenderness than in ST. Amongst the list of candidate biomarkers of tenderness some proteins such as HSPB1 are common between the 2 muscles whatever the evaluation method of tenderness, but some relationships with tenderness for others (MYH1, TNNT3, HSPB6) are inversed. Muscle specificities were revealed in this meta-proteomic study. For example, Parvalbumin (PVALB) appeared as a robust biomarker in ST muscle whatever the evaluation method of tenderness. HSPA1B seems to be a robust candidate for LT tenderness (with WBSF) regardless the animal type. Some gender specificities were further identified including similarities between cows and steers (MSRA and HSPA9) in contrast to bulls. The comparison of the 12 proteomic studies revealed strong dissimilarities to identify generic biomarkers of beef tenderness. This integrative analysis allowed better understanding of the biological processes involved in tenderness in two muscles and their variations according to the main factors underlying this quality. It allowed also proposing for the first time a comprehensive list of candidate biomarkers to be evaluated deeply to validate their relationships with tenderness on a large number of cattle and breeds.
    • Preliminary investigation of the antimicrobial and mechanisms of resistance of Enterobacteria isolated from minced meat in the Northeast of Algeria: The case of butchers from Constantine

      Leila Dib, Amira; Chahed, Amina; Lakhdara, Nedjoua; Agabou, Amir; Boussena, Sabrina; Ghougal, Khireddine; Lamri, Melisa; Sana Kerrour, Nessrine; Kadja, Louiza; Bouaziz, Assia; et al. (Open Access Text Pvt, Ltd., 2019)
      Food products of animal origin such as fresh meat are easily contaminated by microorganisms if handling, processing and storage conditions are not fully respected. The present study aimed first to evaluate the bacterial load and microbial contamination rates of ground raw beef to identify the main pathogenic flora that dominate and second, to determine the resistance patterns and extended-spectrum beta-lactamase (ESBL) of isolated Gram-negative strains against certain families of antibiotics. Therefore, 39 samples have been collected from 5 butcher shops located in Constantine province in the North-East of Algeria. The samples were analysed for total bacterial count, presence of total and faecal coliforms, Staphylococci and Salmonella. Furthermore, 23 antibiotics were tested using the diffusion method on Mueller-Hinton agar, towards 22 strains isolates. Bacterial analyses showed a high contamination by total aerobic bacteria, total and faecal coliforms. Escherichia coli, Citrobacter spp., Enterobacter spp., Hafnia alvei, Salmonella pullorum and Staphylococcus spp (except Staphylococcus aureus) were further revealed in some samples. The results of the antibiogram test exhibit multi-resistance to more than eight antibiotics with varied effects. From the whole tested strains isolates, the fully susceptibility effect was for spectinomycin (SPT). This study reveals that the analysed minced meat was found to be highly contaminated with antibiotic resistant bacteria. This study allows concluding that appropriate use of antibiotics in compliance with good hygiene practices is essential to reduce the antibiotic resistance identified in this preliminary study.
    • Protein Array-Based Approach to Evaluate Biomarkers of Beef Tenderness and Marbling in Cows: Understanding of the Underlying Mechanisms and Prediction

      Gagaoua, Mohammed; Bonnet, Muriel; Picard, Brigitte; Pays de Loire Regional Council (MDPI AG, 2020-08-26)
      This study evaluated the potential of a panel of 20 protein biomarkers, quantified by Reverse Phase Protein Array (RPPA), to explain and predict two important meat quality traits, these being beef tenderness assessed by Warner–Bratzler shear force (WBSF) and the intramuscular fat (IMF) content (also termed marbling), in a large database of 188 Protected Designation of Origin (PDO) Maine-Anjou cows. Thus, the main objective was to move forward in the progression of biomarker-discovery for beef qualities by evaluating, at the same time for the two quality traits, a list of candidate proteins so far identified by proteomics and belonging to five interconnected biological pathways: (i) energy metabolic enzymes, (ii) heat shock proteins (HSPs), (iii) oxidative stress, (iv) structural proteins and (v) cell death and protein binding. Therefore, three statistical approaches were applied, these being Pearson correlations, unsupervised learning for the clustering of WBSF and IMF into quality classes, and Partial Least Squares regressions (PLS-R) to relate the phenotypes with the 20 biomarkers. Irrespective of the statistical method and quality trait, seven biomarkers were related with both WBSF and IMF, including three small HSPs (CRYAB, HSP20 and HSP27), two metabolic enzymes from the oxidative pathway (MDH1: Malate dehydrogenase and ALDH1A1: Retinal dehydrogenase 1), the structural protein MYH1 (Myosin heavy chain-IIx) and the multifunctional protein FHL1 (four and a half LIM domains 1). Further, three more proteins were retained for tenderness whatever the statistical method, among which two were structural proteins (MYL1: Myosin light chain 1/3 and TNNT1: Troponin T, slow skeletal muscle) and one was glycolytic enzyme (ENO3: β-enolase 3). For IMF, two proteins were, in this trial, specific for marbling whatever the statistical method: TRIM72 (Tripartite motif protein 72, negative) and PRDX6 (Peroxiredoxin 6, positive). From the 20 proteins, this trial allowed us to qualify 10 and 9 proteins respectively as strongly related with beef tenderness and marbling in PDO Maine-Anjou cows.
    • Proteomic biomarkers of beef colour

      Gagaoua, Mohammed; Hughes, Joanne; Terlouw, E.M. Claudia; Warner, Robyn D.; Purslow, Peter P.; Lorenzo, José M.; Picard, Brigitte; European Union; Enterprise Ireland; 713654; et al. (Elsevier BV, 2020-07)
      Background Implementation of proteomics over the last decade has been an important step toward a better understanding of the complex biological systems underlying the conversion of muscle to meat. These sophisticated analytical tools have helped to reveal the biochemical pathways involved in fresh meat colour and have identified key protein biomarkers. Scope and approach Until recently, there have been no detailed or critical studies on the role of protein biomarkers in determining meat colour. This review presents an integromics of recent muscle proteomic studies to investigate pathways and mechanisms of beef colour. A database was created from 13 independent proteomic-based studies including data on five muscles and a list of 79 proteins which were significantly correlated with colour traits. The database was subjected to a multistep analysis including Gene Ontology annotations, pathway analysis and literature mining. This report discusses the key protein biomarkers and the biological pathways associated with fresh beef colour. Biomarkers were prioritised by the frequency of identification and the need for future validation experiments is discussed. Key findings and conclusions This review identifies six pathways involved in beef colour including energy metabolism, heat shock and oxidative stress, myofibril structure, signalling, proteolysis and apoptosis. The data-mining of the list of the putative biomarkers showed that certain proteins, such as β-enolase (ENO3), Peroxiredoxin 6 (PRDX6), HSP27 (HSPB1), Phosphoglucomutase 1 (PGM1), Superoxide Dismutase [Cu-Zn] (SOD1) and μ-calpain (CAPN1) were consistently reported by multiple studies as being differentially expressed and having a significant role in beef colour. This integromics work proposes a list of 27 putative biomarkers of beef colour for validation using adapted high-throughput methods.
    • Proteomic biomarkers of beef colour

      Gagaoua, Mohammed; Hughes, Joanne; Terlouw, E.M. Claudia; Warner, Robyn D.; Purslow, Peter P.; Lorenzo, José M.; Picard, Brigitte; Marie Skłodowska-Curie grant agreement; Meat Technology Ireland; 713654; et al. (Elsevier, 2020-05-28)
      Background Implementation of proteomics over the last decade has been an important step toward a better understanding of the complex biological systems underlying the conversion of muscle to meat. These sophisticated analytical tools have helped to reveal the biochemical pathways involved in fresh meat colour and have identified key protein biomarkers. Scope and approach Until recently, there have been no detailed or critical studies on the role of protein biomarkers in determining meat colour. This review presents an integromics of recent muscle proteomic studies to investigate pathways and mechanisms of beef colour. A database was created from 13 independent proteomic-based studies including data on five muscles and a list of 79 proteins which were significantly correlated with colour traits. The database was subjected to a multistep analysis including Gene Ontology annotations, pathway analysis and literature mining. This report discusses the key protein biomarkers and the biological pathways associated with fresh beef colour. Biomarkers were prioritised by the frequency of identification and the need for future validation experiments is discussed. Key findings and conclusions This review identifies six pathways involved in beef colour including energy metabolism, heat shock and oxidative stress, myofibril structure, signalling, proteolysis and apoptosis. The data-mining of the list of the putative biomarkers showed that certain proteins, such as β-enolase (ENO3), Peroxiredoxin 6 (PRDX6), HSP27 (HSPB1), Phosphoglucomutase 1 (PGM1), Superoxide Dismutase [Cu-Zn] (SOD1) and μ-calpain (CAPN1) were consistently reported by multiple studies as being differentially expressed and having a significant role in beef colour. This integromics work proposes a list of 27 putative biomarkers of beef colour for validation using adapted high-throughput methods.
    • What are the drivers of beef sensory quality using metadata of intramuscular connective tissue, fatty acids and muscle fiber characteristics?

      Listrat, Anne; Gagaoua, Mohammed; Andueza, Donato; Gruffat, Dominique; Normand, Jérome; Mairesse, Guillaume; Picard, Brigitte; Hocquette, Jean-François; Agence de l'Environnement et de la Maîtrise de l'Energie, France; European Union; et al. (Elsevier BV, 2020-10)
      The aim of this integrative study was to investigate the relationships between biochemical traits (total, insoluble and soluble collagens (TCol, ICol, SCol), cross-links (CLs), proteoglycans (TPGs), proportion of fiber types, total lipids (TLips), main fatty acids (FAs) families, the n-6/n-3 polyunsaturated FA (n-6/n-3PUFA) ratio and the sensory attributes scores (tenderness, juiciness, flavor) of two muscles from beef: Rectus abdominis (RA) and Longissimus thoracis (LT). For robust analysis, a database was prepared using samples from three studies from animals raised under different production systems. The analyses were performed either on each study separately or on pooled data per muscle after removing as many experimental effects as possible in each study. The CLs (across the muscles and studies) and, to a lower extent, type IIA muscle fibers (mainly for RA muscles), saturated FAs (SFAs), monounsaturated FAs (MUFAs) (for the LT muscles) were the components the most frequently associated with tenderness. The CLs, type IIA muscle fibers (mainly for the RA muscles), TLips, SFAs, MUFAs, conjugated linoleic acids (CLAs) and n-6/n-3 PUFA ratio (mainly for the LT muscles) were the components the most associated with juiciness. The TLips and CLAs (across the muscles and studies), SFAs, MUFAs (mainly for the LT muscles), CLs (mainly for the RA muscles) and TPGs (mainly for the LT muscles) were the components the most associated with flavor liking. The CLs, CLAs, TLips, SFAs, MUFAs, n-6/n-3 PUFA ratio, type IIA and I muscle fibers were the components the most frequently associated with the 3 sensory scores taken together. The SCol, TPGs and type IIX+B muscle fibers were little associated with the sensory scores taken together. The TCol, ICol and PUFAs were components the least associated with sensory scores. The data of this integrative study highlighted for the first time that the CLs were negatively involved in the determination of the three sensory traits mainly in the RA muscle. The muscle fibers in this integrative study had a weak impact on the variations in beef sensory traits. The type IIA and IIX+B muscle fibers were respectively negatively and positively associated with tenderness, negatively associated with juiciness and flavor. The type I muscle fibers were overall positively associated with juiciness and flavor and negatively or positively with tenderness and these associations were muscle and study-dependent. Overall, the TLips and FAs were positively associated with the sensory scores and the n-6/n-3 PUFA ratio was negatively associated with them.