Zinc-protoporphyrin formation in nitrite-free Parma Ham and its relationship with intrinsic parameters and red color profile of processed hams.

A total of 134 fresh hams, assayed for Ferrochelatase (FeCH) activity and ultimate pH (pH48), were processed in compliance with the procedures established for PDO Parma ham and finally, analyzed for salt, moisture, Zinc Protoporphyrin IX (ZnPP), heme, iron and zinc contents, and proteolysis index (PI). The variation in ZnPP content was related to the intrinsic parameters of fresh and matured hams by a Partial Least Square Regression model. The most favorable factors on the formation of ZnPP were total iron content (representative of the initial hemoprotein content), and FeCH activity, demonstrating the main role played by these raw matter-specific predictors in the long matured dry-cured hams. To a lesser extent, zinc content and pH48 were involved with a positive and negative role, respectively. Salt content and PI of matured hams showed an inhibitory and a favorable influence, respectively, toward the ZnPP formation. Principal Component Analysis showed the associations between the sensory red color profile and the physicochemical traits of matured hams. The red color intensity increased in agreement with the red-violet and red-pink hues scores. The formation of a high amount of ZnPP was associated with the increased perception of the red-violet shade, with a lower lightness (L*) and Hue angle (h°). Moisture increase contributed to the shift in color perception to red-pink, while marked progress in PI strengthened the perception of the red-brown shade. ZnPP and final heme favored the red color of matured hams, although a high concentration of these pigments increased in particular the red-violet perception.

A simple and portable method for on-line texture measurement of Italian "Speck Alto Adige".

Texture is a relevant parameter for the assessment of cured ham's quality. In this study a rapid on-line instrumental technique for the measurement of the texture of pieces of cured smoked ham intended for sale as "Speck Alto Adige" PGI was developed. Speck samples were subjected to a compression test using a portable Shore A tester, and instrumental data were compared with conventional texture analyses (texture profile analysis and stress relaxation test) and with sensory evaluations. First, a hardness range in which a threshold value could be identified for the eligibility of 120 speck samples for the "Speck Alto Adige" PGI indication was established; afterwards, a Shore A hardness threshold value of 48 was defined based on measurements of more than 1000 samples. These findings may help manufacturers to determine the textural properties of Speck, based on a simple and rapid instrumental analysis.

Replacement of Nitrite in Meat Products by Natural Bioactive Compounds Results in Reduced Exposure to N-Nitroso Compounds: The PHYTOME Project.

SCOPE: It has been proposed that endogenously form N-nitroso compounds (NOCs) are partly responsible for the link between red meat consumption and colorectal cancer (CRC) risk. As nitrite has been indicated as critical factor in the formation of NOCs, the impact of replacing the additive sodium nitrite (E250) by botanical extracts in the PHYTOME project is evaluated. METHOD AND RESULTS: A human dietary intervention study is conducted in which healthy subjects consume 300 g of meat for 2 weeks, in subsequent order: conventional processed red meat, white meat, and processed red meat with standard or reduced levels of nitrite and added phytochemicals. Consumption of red meat products enriched with phytochemicals leads to a significant reduction in the faecal excretion of NOCs, as compared to traditionally processed red meat products. Gene expression changes identify cell proliferation as main affects molecular mechanism. High nitrate levels in drinking water in combination with processed red meat intake further stimulates NOC formation, an effect that could be mitigated by replacement of E250 by natural plant extracts. CONCLUSION: These findings suggest that addition of natural extracts to conventionally processed red meat products may help to reduce CRC risk, which is mechanistically support by gene expression analyses.

Investigating the Features of PDO Green Hams during Salting: Insights for New Markers and Genomic Regions in Commercial Hybrid Pigs.

Protected Designation of Origin (PDO) dry-cured hams production is greatly dependent on raw meat quality. This study was performed to identify genetic markers associated with the quality of dry-cured ham. Carcass traits of 229 heavy pigs belonging to three commercial genetic lines were registered (weight, EUROP classification). Phenotypic traits (Semimembranosus muscle ultimate pH, ham weight and lean meat content, adsorbed salt) of the corresponding thighs, undergone PDO ham process in three different plants, were measured, using a fast and non-invasive technology. Green ham weight and lean meat percentage influenced the estimated salt content and the weight loss during salting, even if the processing plant greatly affected the variability of the measured ham traits. The genomic data were obtained with the GeneSeek Genomic Profiler (GGP) 70k HD Porcine Array, using the slaughter day and the sex of the animals in the statistical analyses. The phenotypic traits were associated with the genotypes through GenAbel software. The results showed that 18 SNPs located on nine porcine chromosomes were found to be associated with nine phenotypic traits, mainly related to ham weight loss during salting. New associations were found between markers in the genes Neural Precursor Cell Expressed Developmentally Down-Regulated 9 (NEDD9, SSC7), T-Cell Lymphoma Invasion and Metastasis 2 (TIAM2, SSC1), and the ham quality traits. After validation, these SNPs may be useful to improve the quality of thighs for the production of PDO dry-cured hams.

Impact of salt levels on the sensory profile and consumer acceptance of Italian dry-cured ham.

BACKGROUND: Reduction of the salt (NaCl) content in typical Italian dry-cured ham is intended to reduce sodium dietary intake to meet World Health Organization (WHO) recommendations for the prevention of serious diseases. This reduction is a complex issue because of the multiple roles that salt plays in the product. Hams that were processed to obtain a final product with target salt levels (hams with reduced salt concentration and controls) were analyzed using chemical parameters (salt, moisture, fat, and proteolysis indexes). Their sensory properties were described by a trained panel and 87 consumers evaluated the impact of salt reduction on their liking for the products. RESULTS: The dry-cured hams under investigation are representative of Italian dry-cured hams, which are considered to be low and medium to high in terms of salt content. Consistent with the role of salt in proteolytic enzymes inhibition, there was an increase in the proteolysis index in fully matured dry-cured hams with reduced salt content. Salt level affected saltiness perception but also texture, color, and, to a lesser extent, flavor intensity in matured dry-cured hams. On average, reduced-salt dry-cured hams resulted in increased consumer liking. However, the resulting three consumer segments (37%, 27%, and 36% of the consumer cohort, respectively), differed in the acceptability ranking given to reduced-salt hams and controls. CONCLUSION: The tested reduction in salt content, besides improving the nutritional profile of dry-cured ham, was effective in increasing consumers' acceptance. These results were useful for exploring the effect of reducing salt concentration on consumer liking for dry-cured hams. © 2020 Society of Chemical Industry.

Proteomics of Parma Dry-Cured Ham: Analysis of Salting Exudates.

The production of Parma dry-cured ham involves the steps of salting, drying, and ripening. Although sea salt is the only preserving agent, there are strategies being developed with the goal of reducing salt content in order to decrease its negative impact on consumer health. A 24 h pressure treatment was applied before salting to reduce thickness and inequalities in shape. To evaluate the potential impact of the pressure step on the process outcome, differential proteomic analyses by complementary 2D-PAGE and LC-MS/MS were carried out on exudates collected at day 1, 5, and 18 of the salting phase for hams treated or untreated with pressure. Specific proteins were found differentially abundant in exudates from pressed vs unpressed hams and as a function of time. These changes include glycolytic enzymes and several myofibrillar proteins. These findings indicate that pressure causes a faster loosening of the myofibrillar structure with the release of specific groups of proteins.

Association study between single nucleotide polymorphisms in porcine genes and pork quality traits for fresh consumption and processing into Italian dry-cured ham.

Single nucleotide polymorphisms (SNPs) of six genes (TTN, PRKAG3, CAST, CTSB, CTSF, and MYPN), known for associations with carcass and meat quality traits, post mortem proteolysis, were screened in a commercial crossed population of 368 heavy pigs (Large White x Landrace)×Duroc, reared according to the rules of Italian Protected Designation of Origin (PDO) dry-cured ham. Carcass, longissimus thoracis et lumborum muscle (LTL), and green ham traits were obtained after slaughtering, main weight losses of dry-cured hams were collected during processing. The results showed the impact of CAST variants on carcass weight, of CTSF on LTL tenderness, ham weight and fatness, of PRKAG3 and TTN on ultimate pH, hamweight. This study, while confirming significant associations between SNPs of genes and qualitative traits of carcass, longissimus and ham, supports CTSF as candidate gene suitable for fresh consumption purpose (tenderness of longissimus at 24h post mortem), and for dry-cured ham processing (higher thickness of ham subcutaneous fat).

Effect of added autochthonous yeasts on the volatile compounds of dry-cured hams.

Three yeast strains belonging to Debaryomyces and Hyphopichia spp., isolated from dry-cured hams and previously tested for biocontrol activity against toxigenic Penicillium nordicum, were investigated for ability in colonising ham surface. Hams were twice yeast-inoculated onto the unskinned muscle surface during ripening and processed up to full maturation in two manufacturing plants. The yeast strains and the manufacturing plants differed (P < 0.05) in surface populations, volatile compounds and sensory descriptors of matured hams. Sensory scores for each of the yeast-inoculated groups were higher or similar to the non-inoculated ones (controls). Debaryomyces strains were regarded as those most fit to colonise the ham surface under the ecological conditions of dry-curing rooms, hence to qualify as biocontrol agents against the growth of undesired mould and preserve the typical sensory properties of dry-cured hams.

Biocontrol of Penicillium nordicum growth and ochratoxin A production by native yeasts of dry cured ham.

Twelve yeast strains isolated from the surface of Italian typical dry-cured hams, belonging to D. hansenii, D. maramus, C. famata, C. zeylanoides and H. burtonii species, and previously selected for their ability to grow in dry-cured ham-like substrates, were screened for antagonistic activity against a toxigenic strain of P. nordicum and inhibition of ochratoxin A (OTA) biosynthesis. On average, yeast inhibitory activity was lowered by increasing fungal inoculum and enhanced by NaCl presence. In the assay conditions, H. burtonii and C. zeylanoides were the most effective, both in inhibiting P. nordicum growth and OTA production. D. hansenii was the species with the lowest inhibitory activity, especially in the absence of salt. OTA production dropped from the range < LOD - 5000 ppb in P. nordicum control plates to the range < LOD - 200 ppb in yeast-added plates. OTA production increased in the presence of NaCl in P. nordicum control plates, while salt enhanced inhibition against OTA production in yeast-added plates.

Sensory and texture properties of Italian typical dry-cured hams as related to maturation time and salt content.

Maturing time and salt were fixed factors and fat was a covariate in a full factorial design study of sensory and texture properties of 36 dry cured hams. Samples were chosen to fit three ageing and salt classes. Differences (P < 0.05) in sensory scores were found between whole slices (with cover fat) and their biceps femoris (BF) counterparts, with sweet taste enhanced in whole samples and conversely unpleasant bitter taste more intense in BF portions. Increased ageing time resulted in better sensory profiles, as documented by greater scores (P < 0.05) for matured odour, matured taste and sweet taste, whereas salty taste was reduced. Decreased salt was associated with greater intensities (P < 0.05) of undesired green odour and taste. Based on chemical composition, the ham consistence appears improved by long-term dehydration, provided that protein breakdown is under control. It is concluded that for salt reduction of up to 25% to be achieved without negative side effects, extended ageing and restrained proteolysis are key factors.

Determination of ochratoxin A in dry-cured meat products by a HPLC-FLD quantitative method.

A fast and sensitive method for the quantification of the mycotoxin ochratoxin A (OTA) in dry-cured meat products has been developed, which does not require a clean-up step, by HPLC with an alkaline mobile phase (pH 9.8). Validation procedures for specificity, trueness, ruggedness, stability, recovery and repeatability were performed. The decision limit (CC alpha) and the decision capability (CC beta) were calculated at 1.10 and 1.23 microg/kg, respectively. The procedure was applied to representative dehydration levels of dry-cured meat samples.

Penicillium populations in dry-cured ham manufacturing plants.

Seven ham manufacturing plants were sampled for 1 year to assess the mycoflora present in the air and on hams, with special attention given to potential mycotoxin producers. Temperature and relative humidity were recorded in the ripening rooms. Maturing rooms held hams from 2 to 3 through 6 to 7 ripening months, and aging rooms held hams for the following 6 to 7 months, until the 14-month ripening point, when they were ready for the market. Mean temperatures and relative humidities registered during the study were 14.9 degrees C and 62.4%, respectively, in maturing rooms and 16.3 degrees C and 57.6% in aging rooms. Aspergilli and penicillia, potential mycotoxin producers, were isolated in all the plants from the air and the ham. Aspergilli represented 5% of the isolates, while penicillia were largely dominant, with Penicillium nalgiovense being the most represented species (around 60% of the penicillia), followed by Penicillium nordicum, with 10 and 26% of the penicillia isolated, respectively, from the air or the ham. Ochratoxin A production ability, checked in vitro at 250C, was observed in 50% of the P. nordicum isolates obtained both from the air and the ham. Air and ham surface contamination by penicillia was greater in the ripening rooms, where higher temperatures were registered. A certain correlation was also observed between air and ham surface contamination. On the basis of this study, P. nordicum, the ochratoxin A producer that is notable on proteinaceous substrates, is normally present in ham manufacturing plants in Italy, even though not a dominant species. Further studies are necessary to clarify and ensure if dry-curing conditions minimize the potential risk of ochratoxin A formation in the product.

Effect of extended aging of parma dry-cured ham on the content of oligopeptides and free amino acids.

The effect of the dry-curing processing time on the release of oligopeptides and amino acids was evaluated with 158 Parma hams subdivided into three groups: (1) traditional processing (450 days); (2) extended processing (570 days); and (3) extended aging (690 days). Most of the oligopeptides and free amino acids detected increased up to the last deadline (690 days); a sharp increase of peptides below 400 Da was the main change in most aged hams. In particular, gamma-glutamyl dipeptides showed a remarkable increase during ham extended aging, acting like permanent taste-active compounds, being unsuitable for further enzymatic breakdown. The pH of fresh hams showed negative relationships (P < 0.001) with most peptides. With regard to free amino acids, the pattern was modified by different processing lengths, together with their taste categories, so that the amino acids having monosodium glutamate-like and bitter tastes were enhanced in more aged hams.

Magnetic resonance imaging and relaxation analysis to predict noninvasively and nondestructively salt-to-moisture ratios in dry-cured meat.

The current systems are unable to control and predict the cured meat composition nondestructively and in a reasonable time for production needs. In this work, T1 and T2 maps were obtained, with a monoexponential model, on internal sections of Longissimus dorsi muscle at increasing salting times. The maps allow one to visualize the salting process nondestructively and noninvasively. The method goes beyond the simple qualitative visualization, because, for each section of the sample and in any region of the section, it is possible to obtain quantitative information on the progress of salting and to predict salt-to-moisture ratios. In addition, detailed relaxation measurements were performed on samples cored after imaging in order to define better the relaxation properties of the dry-cured meat.

Identification and significance of the N-terminal part of swine pyruvate kinase in aged Parma hams.

Within a project aimed at studying the peptide fraction in dry-cured Parma hams, a peptide was purified by means of reversed phase-high performance liquid chromatography (RP-HPLC) and identified by its molecular mass and amino acid sequence analysis. The peptide showed a very high degree of homology with the N-terminal part of different mammalian pyruvate kinases reported in databases and was accordingly identified as the N-terminal part of swine pyruvate kinase, whose sequence had never been reported before. The peptide was determined quantitatively by comparison with a suitable internal standard (Phe-Phe) in commercial ham samples with different age degrees. The peptide was found to be ubiquitous in Parma ham and its amount to increase during ageing even if a large variability was found within each assayed maturing time. The correlation found with the ham proteolysis degree (P<0.01) suggests that this peptide is related to the endopeptidasic activity. The peptide did not show a significant relationship with bitterness perception in assayed dry-cured ham, in agreement with the hypothesis that this taste is more related to free amino acids and low molecular weight peptides.