Perspective of sodium reduction based on endogenous proteases via the strategy of sodium replacement in conjunction with mediated-curing.

NaCl is the main curing agent in dry-cured meat products, and a large amount of NaCl addition leads to high salt content of final products. Salt content and composition are important factors affecting the activity of endogenous proteases, which in turn could affect proteolysis as well as the quality of dry-cured meat products. With the increasing emphasis on the relationship between diet and health, reducing sodium content without sacrificing quality and safety of products is a great challenge for dry-cured meat industry. In this review, the change of endogenous proteases activity during processing, the potential relationship between sodium reduction strategy, endogenous proteases activity, and quality were summarized and discussed. The results showed that sodium replacement strategy and mediated-curing had a complementary advantage in influencing endogenous proteases activity. In addition, mediated-curing had the potential to salvage the negative effects of sodium substitution by affecting endogenous proteases. Based on the results, a sodium reduction strategy that sodium replacement in conjunction with mediated-curing based on endogenous proteases was proposed for the future perspective.

Effects of Different Levels of Inclusion of Apulo-Calabrese Pig Meat on Microbiological, Physicochemical and Rheological Parameters of Salami during Ripening.

This study focused on the characterization of salami produced with meat from different pig breeds. The aim consisted in evaluating the added value of the inclusion of Apulo-Calabrese meat in the production of salami, which was characterized by production until the end of maturation (1, 30, 60, and 120 days). The experimental design involved three types of salami, two of which were produced by partial inclusion of 50 and 75% of the Italian breed pork meat (S50 and S75, respectively). Physicochemical (pH, aw, fatty acid analysis, and malondialdehyde concentration), rheological parameters (texture analyses and color measurement), and bacterial biodiversity were evaluated. Results showed that the partial inclusion of Apulo-Calabrese meat influences the fatty acid profile of final products, which were characterized by a higher percentage of monounsaturated fatty acids compared to traditional salami; however, due to the high content of unsaturated fatty acids, S50 and S75 showed higher values of secondary lipid oxidation up to the 120th day. The linoleic and palmitic acid content significantly affected hardness and brightness. Overall, the ripening process was able to control the microbiological profile and the S50 formulation appeared as a suitable choice that could satisfy consumers for nutritional expectations and sensory profiles.

Influence of an industrial dry-fermented sausage processing on ochratoxin A production by Penicillium nordicum.

Dry-fermented sausages are prone to be colonised by Penicillium nordicum, which is one of the main ochratoxin A (OTA)-producing species. Its ability to produce this mycotoxin on dry-fermented sausages has been reported. However, the influence of the conditions of a traditional processing of a Spanish dry-fermented sausage and the intrinsic physicochemical parameters of this product such as water activity (aw) and pH on OTA production has not been studied yet. Thus, the aim of this study was to evaluate the influence of traditional processing (interaction of relative humidity (RH) x temperature x ripening days) on the evolution of pH and aw during maturation of dry-fermented sausage "salchichón" and its relationship with OTA synthesis by P. nordicum. The expression of otapks and otanps genes, both involved in the biosynthesis of the mycotoxin, was also assessed. For this, 27 raw sausages were inoculated with P. nordicum and ripened for 26 days in a drying chamber (3 days at 5 °C and 84% RH, 17 days at 12 °C and 84% RH, and 6 days at 12 °C and 80% RH). From results, although it seems that the pH slightly influenced on OTA biosynthesis, the aw had a great impact on this mycotoxin production. In fact, the two highest OTA concentrations found coincided with a dramatic rise of the aw value (0.92 aw) by day 18 of incubation when the RH of the drying chamber was still 84% and at the end of the incubation time when the aw decreased noticeably (0.87 aw). The expression of the otapks and otanps genes correlated with the OTA produced by P. nordicum. Results from this work confirm that the traditional processing of Spanish dry-fermented sausages favours itself OTA synthesis by P. nordicum. Our findings may help in informed decision-making in relation to RH/temperature of drying chambers and shortening of the ripening process. This may be then effectively incorporated into the hygienic production system in the framework of HACCP together with other measures including the use of Penicillium nalgiovense as protective culture or the monitoring of otapks gene expression, and aw during the processing of dry-fermented sausages. All these strategies together may put ochratoxigenic Penicillia at a disadvantage and minimise OTA contamination risks in dry-fermented sausages.

Effect of production process and high-pressure processing on viability of Salmonella spp. in traditional Italian dry-cured coppa.

The aim of the study was to investigate the combined effect of the manufacturing process followed by HPP treatment on the inactivation of Salmonella spp. in artificially contaminated coppa samples, in order to verify the ability of the combined processes to achieve the objective of a 5-log reduction of Salmonella spp. needed for exportation to the U.S. Fresh anatomical cuts intended for coppa production were supplied by four different delicatessen factories located in Northern Italy. Raw meat underwent experimental contamination with Salmonella spp. using a mixture of 3 strains. Surface contamination of the fresh anatomical cuts was carried out by immersion into inoculum containing Salmonella spp. The conditions of the HPP treatment were: pressure 593 MPa, time 290 seconds, water treatment temperature 14°C. Surface and deep samples were performed post contamination (T0), end of the cold phase (T1), end of process (Tend), and after HPP treatment (postHPP) and Salmonella spp. Enumerated. The results of this study show a significant reduction of Salmonella spp. all through the production process (P<0.01) for all companies, followed by an additional reduction of bacterial counts due to HPP treatment (P<0.01), both in superficial and deep contaminations (P<0.01). The superficial overall reduction resulted of 1.58 to 5.04 log CFU/g during the production process. HPP treatment resulted in a significant (P<0.01) superficial and deep decrease in Salmonella spp. enumeration varying from 0.61 to 4.01 log and from 1.49 to 4.13 log. According to the data presented in this study, only the combined approach of coppa manufacturing process followed by HPP treatment always led to a 5-log reduction of Salmonella spp. required by USDA/FSIS guidelines.

Effect of production process and high-pressure processing on viability of Listeria innocua in traditional Italian dry-cured coppa.

In this study the effect of the application of High Pressure Treatment (HPP) combined with four different manufacturing processes on the inactivation of Listeria innocua, used as a surrogate for L. monocytogenes, in artificially contaminated coppa samples was evaluated in order to verify the most suitable strategy to meet the Listeria inactivation requirements needed for the exportation of dry-cured meat in the U.S. Fresh anatomical cuts intended for coppa production were supplied by four different delicatessen factories located in Northern Italy. Raw meat underwent experimental contamination with Listeria innocua using a mixture of 5 strains. Surface contamination of the fresh anatomical cuts was carried out by immersion into inoculum containing Listeria spp. The conditions of the HPP treatment were: pressure 593 MPa, time 290 seconds, water treatment temperature 14°C. Listeria innocua was enumerated on surface and deep samples post contamination, resting, ripening and HPP treatment. The results of this study show how the reduction of the microbial load on coppa during the production process did not vary among three companies (P>0.05) ranging from 3.73 to 4.30 log CFU/g, while it was significantly different (P<0.01) for the fourth company (0.92 log CFU/g). HPP treatment resulted in a significant (P<0.01) deep decrease of L. innocua count with values ranging between 1.63-3.54 log CFU/g with no significant differences between companies. Regarding superficial contamination, HPP treatment resulted significant (P<0.01) only in Coppa produced by two companies. The results highlight that there were processes less effective to inhibit the pathogen; in particular for company D an increase of L. innocua count was shown during processing and HPP alone cannot be able to in reaching the Listeria inactivation requirements needed for exportation of dry-cured meat in the U.S. According to the data reported in this paper, HPP treatment increases the ability of the manufacturing process of coppa in reducing Listeria count with the objective of a lethality treatment.

Effect of Spanish smoked paprika "Pimentón de La Vera" on control of ochratoxin A and aflatoxins production on a dry-cured meat model system.

Environmental conditions during ripening of dry-cured meat products favour growth of fungal population on their surface. Some of these moulds can produce mycotoxins. Paprika is one of the ingredients usually used in the formulation of raw-cured sausages, and its addition could influence the growth and production of mycotoxins of the moulds present in these products. In this work the effect of Spanish smoked paprika "Pimentón de la Vera" on growth of Aspergillus parasiticus and Penicillium nordicum and production of aflatoxins B1 (AFB1), G1 (AFG1) and ochratoxin A (OTA) respectively, was evaluated. Moulds were grown in a culture medium made from lyophilized fresh pork meat added with 4% salt and different concentrations of Spanish smoked paprika (1, 2 and 3%) at several water activity values (0.98, 0.94 and 0.87) and temperature (20-25 °C), to simulate conditions usually found during ripening of dry-cured meat products. Mould growth was evaluated by measuring the diameter of the colony every 24 h, and the production of mycotoxins by UHPLC-MS/MS every 2 days, during 10 days of incubation. Addition of paprika favours growth of the two mould species tested. However, the synthesis of mycotoxins was reduced at 0.94 and 0.98 aw when at least a 2% of paprika was added. Therefore, the addition of Spanish smoked paprika at 2-3% in the formulations may help to minimize AFs and OTA production in dry-cured meat products such as loins or "chorizo" sausages.

Biocontrol of aflatoxigenic Aspergillus parasiticus by native Debaryomyces hansenii in dry-cured meat products.

Dry-cured meat products, such as dry-cured ham or dry-fermented sausages, are characterized by their particular ripening process, where a mould population grows on their surface. Some of these moulds are hazardous to the consumers because of their ability to produce mycotoxins including aflatoxins (AFs). The use of native yeasts could be considered a potential strategy for controlling the presence of AFs in dry-cured meat products. The aim of this work was to evaluate the antagonistic activity of two native Debaryomyces hansenii strains on the relative growth rate and the AFs production in Aspergillus parasiticus. Both D. hansenii strains significantly reduced the growth rates of A. parasiticus when grown in a meat-model system at different water activity (aw) conditions. The presence of D. hansenii strains caused a stimulation of AFs production by A. parasiticus at 0.99 aw. However, at 0.92 aw the yeasts significantly reduced the AFs concentration in the meat-model system. The relative expression levels of the aflR and aflS genes involved in the AFs biosynthetic pathway were also repressed at 0.92 aw in the presence of both D. hansenii strains. These satisfactory results were confirmed in dry-cured ham and dry-fermented sausage slices inoculated with A. parasiticus, since both D. hansenii strains significantly reduced AFs amounts in these matrices. Therefore, both tested D. hansenii strains could be proposed as biocontrol agents within a HACCP framework to minimize the hazard associated with the presence of AFs in dry-cured meat products.

Survival of African swine fever virus (ASFV) in various traditional Italian dry-cured meat products.

A number of animal diseases can be transmitted to pigs via pork meat and pork products imported from infected areas. Therefore, feeding of swill to pigs is regulated or prohibited in many swine-rearing countries. African swine fever is one of the major porcine diseases recognized as significant in this transmission pathway. Assessment of disease risks associated with pork products requires knowledge about the viral load in the original material and for how long infectious virus can be recovered from the resulting product. In this work, we present the effect of the dry curing process on the inactivation of African swine fever virus (ASFV) in three different Italian dry-cured meat products prepared from experimentally infected pigs slaughtered at the peak of viremia. The meat products were processed using commercial methods and industrial procedures currently being followed in Italy. Samples collected at predetermined intervals during processing were analyzed for virus survival by virus isolation and animal inoculation. ASFV was detected by in vivo experiments for up to 18, 60, and 83 days of curing in Italian salami, pork belly, and loin, respectively. These data provide valuable information for the pork processing industry when planning the export of these products.

Stability of Antiradical Activity of Protein Extracts and Hydrolysates from Dry-Cured Pork Loins with Probiotic Strains of LAB.

The application of starter cultures to improve quality and safety has become a very common practice in the meat industry. Probiotic strains of lactic acid bacteria (LAB) can also bring health benefits by releasing bioactive peptides. The aim of this work was to evaluate the stability of antiradical activity of protein extracts from LAB-inoculated dry-cured pork loins during long-term aging and evaluate their hydrolysates after simulated gastrointestinal digestion. Analyses of hydrolysates by using liquid chromatography-tandem mass spectrometry (LC-MS/MS) were strengthened with in silico analysis. The highest antiradical activity of the protein extracts was observed after 180 days of aging. The influence of the strain used (LOCK, BAUER, or BB12) on the inactivation ability of ABTS radicals varied during long-term aging. The IC50 values indicated the higher antiradical properties of salt-soluble (SSF) compared to water-soluble fraction (WSF) of proteins. The peptides generated by in vitro digestion have MW between 700 and 4232 Da and their length ranged from 5 to 47 amino acids in a sequence where Leu, Pro, Lys, Glu, and His had the largest share. This study demonstrates that the degradation of pork muscle proteins during gastrointestinal digestion may give rise to a wide variety of peptides with antiradical properties.

Potential of yeasts isolated from dry-cured ham to control ochratoxin A production in meat models.

The environmental conditions reached during the ripening of dry-cured meat products favour the proliferation of moulds on their surface. Some of these moulds are hazardous to consumers because of their ability to produce ochratoxin A (OTA). Biocontrol using Debaryomyces hansenii could be a suitable strategy to prevent the growth of ochratoxigenic moulds and OTA accumulation in dry-cured meat products. The aim of this work was to evaluate the ability of two strains of D. hansenii to control the growth and OTA production of Penicillium verrucosum in a meat model under water activities (aw) values commonly reached during the dry-cured meat product ripening. The presence of D. hansenii strains triggered a lengthening of the lag phase and a decrease of the growth rate of P. verrucosum in meat-based media at 0.97 and 0.92 aw. Both D. hansenii strains significantly reduced OTA production (between 85.16 and 92.63%) by P. verrucosum in the meat-based medium at 0.92 aw. Neither absorption nor detoxification of OTA by D. hansenii strains seems to be involved. However, a repression of the expression of the non-ribosomal peptide synthetase (otanpsPN) gene linked to the OTA biosynthetic pathway was observed in the presence of D. hansenii. To confirm the protective role of D. hansenii strains, they were inoculated together with P. verrucosum Pv45 in dry-fermented sausage and dry-cured ham slices. Although P. verrucosum Pv45 counts were not affected by the presence of D. hansenii in both meat matrices, a reduction of OTA amount was observed. Therefore, the effect of D. hansenii strains on OTA accumulation should be attributed to a reduction at transcriptional level. Consequently, native D. hansenii can be useful as biocontrol agent in dry-cured meat products for preventing the hazard associated with the presence of OTA.

Potential of bacteriocins from lab to improve microbial quality of dry-cured and fermented meat products.

Meat and meat products are an important component of the daily diet. Nevertheless, they are perishable goods and are prone to microbial contamination, which leads to an increased risk to the health of consumers as well as economic losses in the meat industry. Fermentation has been used for thousands of years to preserve meat. As a result of extensive biochemical reactions occurring in meat during fermentation and ripening, the condi- tions inhibiting the growth of pathogenic and spoilage bacteria are formed. These changes are catalyzed by endogenous meat enzymes and exogenous enzymes derived from natural contaminating bacteria or starter cultures applied. In dry-cured and fermented meat products they are represented mainly by lactic acid bacte- ria (LAB) that produce a wide range of compounds, such as bacteriocins, directed against other microorgan- isms. The use of bactericidal peptides does not affect the sensory quality of foodstuffs, so that they attract attention as alternative means of preserving the stability and safety of dry-cured products.

Reduction of ochratoxin A in dry-cured meat products using gamma-irradiation.

This study investigated the efficiency of gamma (γ)-irradiation in the reduction of ochratoxin A (OTA) present in dry-cured meat products prepared from intentionally contaminated raw materials from OTA-treated pigs. OTA concentrations determined in the samples (n = 24) ranged from 25.8 µg kg(-1) in bacon to 17.8 µg kg(-1) in smoked ham. After γ-irradiation at doses of 3, 7 and 10 kGy (i.e. the doses used in the food industry), a dose-depended OTA reduction was observed; however, it was not statistically significant. The mean OTA reduction achieved with 3-, 7- and 10-kGy γ-doses was approximated to 8.5%, 13.9% and 22.5%, respectively. The storage of irradiated samples (1 month, 4°C) did not significantly affect OTA levels. Based on the correlation between the OTA reduction level and basic chemical composition of dry-cured meat samples, OTA reduction may be linked to the samples' fat content. The results indicate that γ-irradiation can reduce OTA levels in dry-cured meat products, but only to a limited extent due to the complexity of the matrix.

Salazones y chacinados embutidos secos: detección por electroforesis de especies cárnicas y de proteínas extrínsecas agregadas / Dry-cured meat products: electrophoresis detection of meat species and extrinsic proteins added

En la elaboración de salazones y chacinados embutidos secos se pueden utilizar diferentes especies cárnicas y en algunos de ellos también se pueden agregar proteínas extrínsecas. En elpresente trabajo se plantearon los siguientes objetivos: analizar salazones y chacinados embutidos secos elaborados con especies cárnicas de diferente origen (vacuna, porcina, de ciervo, de jabalí y de cordero) para establecer la utilidad de SDS-PAGE en la identificación de las mismas; comparar esta metodología conun método inmunoquímico (ELISA para especie porcina y ELISA para especie vacuna) en muestras que declaraban carne porcina y/o vacuna; detectar la posible presencia de proteínas extrínsecasdeclaradas o no en los respectivos rótulos de estos productos. Se analizaron 5 salazones y 7 chacinados embutidos secos. En todas las muestras se detectaron por electroforesis la o lasespecies cárnicas declaradas, sólo en una muestra que declaraba carnes vacuna y porcina se detectó sólo carne vacuna. Lametodología inmunoquímica confirmó la detección de las carnes vacuna y/o porcina en las muestras que las declaraban. Con respecto a las proteínas extrínsecas se detectaron por electroforesis proteínas de soja en tres muestras, dos de ellas no las declaraban.Se detectaron proteínas lácteas en muy baja concentración en tres muestras que las declaraban y se detectaron proteínas de trigo en dos muestras que no las declaraban. Las tres proteínas detectadas en estas muestras constituyen alergenos alimentarios. Si bien el método ELISA resulta de elección para la detecciónde alergenos alimentarios, ya que tiene una sensibilidad mucho mayor al SDS-PAGE, estas proteínas alergénicas fueron detectadas con facilidad por electroforesis, lo cual indica que estaban agregadas en concentraciones importantes. Resulta entonces imprescindible que los elaboradores de este tipo de productos declaren la totalidad de los ingredientes proteicos utilizados para evitar reacciones alérgicas...

Different meat species can be used in dry-cured meat products, and extrinsic proteins can also be added in some of them. The objectives of this work are to analyze dry-cured meat productselaborated with different meat species (cow, swine, deer, boar and lamb) in order to evaluate the usefulness of the SDS-PAGE method in the identification of the species used; to compare thismethodology with an immunochemical method (ELISA for porcine and bovine species respectively) in samples that declare porcine and/or bovine meat in their label; and to detect the possiblepresence of extrinsic proteins declared or not in these products’ labels. Twelve dry-cured meat products were analyzed. In allsamples, the meat species declared in the labels were found by electrophoresis, except in one case where only bovine meat was detected while the label declared both bovine and porcine meats. The immunochemical method confirmed the detection of bovine and/or porcine meat in the samples that declared them in their labels. As regards extrinsic proteins, soy proteins were detected by electrophoresis in three samples, while two of them did not declare these proteins. Dairy proteins were detected in very low levels in three samples that declared them and wheat proteins were detected in two samples that did notdeclare them. The three proteins detected in these samples are food allergens. Although the method of choice for the detection of allergens in food is ELISA because it is more sensible than SDS-PAGE, these allergenic proteins were easily detected by electrophoresis; this indicates they were added in relevant concentrations. It is then critical that manufacturers declare all proteiningredients used in this kind of products to prevent allergic reactions in consumers.

Salazones y chacinados embutidos secos: detección por electroforesis de especies cárnicas y de proteínas extrínsecas agregadas / Dry-cured meat products: electrophoresis detection of meat species and extrinsic proteins added

En la elaboración de salazones y chacinados embutidos secos se pueden utilizar diferentes especies cárnicas y en algunos de ellos también se pueden agregar proteínas extrínsecas. En elpresente trabajo se plantearon los siguientes objetivos: analizar salazones y chacinados embutidos secos elaborados con especies cárnicas de diferente origen (vacuna, porcina, de ciervo, de jabalí y de cordero) para establecer la utilidad de SDS-PAGE en la identificación de las mismas; comparar esta metodología conun método inmunoquímico (ELISA para especie porcina y ELISA para especie vacuna) en muestras que declaraban carne porcina y/o vacuna; detectar la posible presencia de proteínas extrínsecasdeclaradas o no en los respectivos rótulos de estos productos. Se analizaron 5 salazones y 7 chacinados embutidos secos. En todas las muestras se detectaron por electroforesis la o lasespecies cárnicas declaradas, sólo en una muestra que declaraba carnes vacuna y porcina se detectó sólo carne vacuna. Lametodología inmunoquímica confirmó la detección de las carnes vacuna y/o porcina en las muestras que las declaraban. Con respecto a las proteínas extrínsecas se detectaron por electroforesis proteínas de soja en tres muestras, dos de ellas no las declaraban.Se detectaron proteínas lácteas en muy baja concentración en tres muestras que las declaraban y se detectaron proteínas de trigo en dos muestras que no las declaraban. Las tres proteínas detectadas en estas muestras constituyen alergenos alimentarios. Si bien el método ELISA resulta de elección para la detecciónde alergenos alimentarios, ya que tiene una sensibilidad mucho mayor al SDS-PAGE, estas proteínas alergénicas fueron detectadas con facilidad por electroforesis, lo cual indica que estaban agregadas en concentraciones importantes. Resulta entonces imprescindible que los elaboradores de este tipo de productos declaren la totalidad de los ingredientes proteicos utilizados para evitar reacciones alérgicas...(AU)

Different meat species can be used in dry-cured meat products, and extrinsic proteins can also be added in some of them. The objectives of this work are to analyze dry-cured meat productselaborated with different meat species (cow, swine, deer, boar and lamb) in order to evaluate the usefulness of the SDS-PAGE method in the identification of the species used; to compare thismethodology with an immunochemical method (ELISA for porcine and bovine species respectively) in samples that declare porcine and/or bovine meat in their label; and to detect the possiblepresence of extrinsic proteins declared or not in these products labels. Twelve dry-cured meat products were analyzed. In allsamples, the meat species declared in the labels were found by electrophoresis, except in one case where only bovine meat was detected while the label declared both bovine and porcine meats. The immunochemical method confirmed the detection of bovine and/or porcine meat in the samples that declared them in their labels. As regards extrinsic proteins, soy proteins were detected by electrophoresis in three samples, while two of them did not declare these proteins. Dairy proteins were detected in very low levels in three samples that declared them and wheat proteins were detected in two samples that did notdeclare them. The three proteins detected in these samples are food allergens. Although the method of choice for the detection of allergens in food is ELISA because it is more sensible than SDS-PAGE, these allergenic proteins were easily detected by electrophoresis; this indicates they were added in relevant concentrations. It is then critical that manufacturers declare all proteiningredients used in this kind of products to prevent allergic reactions in consumers.(AU)