Hubert Paelinck

Evaluation of the spoilage lactic acid bacteria in modified-atmosphere-packaged artisan-type cooked ham using culture-dependent and culture-independent approaches

Aims:  To investigate microbial diversity and population dynamics of spoilage‐sensitive modified‐atmosphere‐packaged (MAP) artisan‐type cooked ham in relation to storage temperature.

Effect of dietary antioxidant and fatty acid supply on the oxidative stability of fresh and cooked pork

The effect of dietary oil (linseed or soybean oil) and antioxidant treatment (α-tocopheryl acetate (AT; 40ppm) versus a cocktail (AOC; 200ppm): α-tocopheryl acetate+rosemary+citric acid+gallic acid) on colour, lipid and protein oxidation of fresh and processed pork was investigated. No effect of oil source on different parameters of oxidation was seen. No effect of antioxidant treatment on colour stability of fresh longissimus thoracis (LT) or cooked cured ham (CCH) was observed. For both antioxidant treatments, lipid oxidation in fresh LT and CCH was well controlled during display. However, lipid oxidation increased significantly in pre-frozen uncured cooked meat under aerobic conditions. No unambiguous effect of antioxidant treatment on protein oxidation was observed. There seemed to be no clear link between colour, protein and lipid oxidation. At the dose used in this study, no additional or synergistic effects of the extra components of the AOC on the different oxidation parameters was found.

The application of staphylococci with flavour-generating potential is affected by acidification in fermented dry sausages

Differences in the production of bacterial metabolites with potential impact on fermented sausage flavour were found in meat simulation medium when comparing different strains of Staphylococcus xylosus and Staphylococcus carnosus as starter cultures. Overall, higher levels of 3-methyl-1-butanol and acetoin were found for S. xylosus, with some intraspecies variability. In addition, sausage fermentation parameters affected staphylococcal growth and metabolism. Strong acidification, as in Northern-European types of fermented dry sausage, inhibited S. xylosus 3PA6 but not S. carnosus 833. During a milder, Southern-European type of acidification, both strains displayed survival over time. During in situ sausage trials, variations in the degree of acidification and the choice of starter microorganisms were of importance, whereas modifications in fat and salt contents had no effects. Staphylococcus sciuri alphaSg2, Staphylococcus succinus 4PB1, and S. xylosus 3PA6 were unable to survive the fermentation of a Northern-European type of fermented dry sausage, characterized by low or no 3-methyl-1-butanol and acetoin production. Inoculation with S. sciuri alphaSg2, S. succinus 4PB1, or S. xylosus 3PA6 led to 3-methyl-1-butanol and acetoin production in Southern-European type of fermented dry sausages, which was not observed with S. carnosus 833.

Influence of putrescine, cadaverine, spermidine or spermine on the formation of N-nitrosamine in heated cured pork meat

The influence of biogenic amines (i.e. putrescine, cadaverine, spermidine and spermine) on the N-nitrosamine formation in heated cured lean meat was studied in the presence or absence of sodium nitrite and at different meat processing temperatures. Experimental evidence was produced using gas chromatography with thermal energy analysis detection (GC-TEA). Concentration of N-nitrosamines was modelled as a function of the temperature and the nitrite concentration for two situations, i.e. presence or absence of added biogenic amines to the meat. The significance of the influence of the changing parameters was evaluated by ANOVA (Analysis of Variance). It was found that higher processing temperatures and higher added amounts of sodium nitrite increase the yields of N-nitrosodimethylamine (NDMA) and N-nitrosopiperidine (NPIP). Spermidine and putrescine amplify the formation of NDMA, but spermine and cadeverine do not influence the formation of this N-nitrosamine. Spermidine and cadeverine cause a significant increase of NPIP. Beside N-nitrosopyrrolidine (NPYR) in some rare cases, no other volatile N-nitrosamines are detected.

Effect of rosemary extract dose on lipid oxidation, colour stability and antioxidant concentrations, in reduced nitrite liver pâtés

The oxidative stability of liver pâté was investigated in relation to different doses of rosemary extract (RE) and sodium nitrite. Colour stability, lipid oxidation (TBARS) and concentrations of ascorbic acid, α-tocopherol, carnosic acid and nitrite were measured on the batters before cooking and on the cooked liver pâté before and after exposure to light and air for 48 h at 4°C. Results showed that the use of RE significantly reduced lipid oxidation, whereas it had no effect on colour stability. Ascorbic acid and nitrite concentrations were significantly higher and lower respectively when RE was added. RE dose-dependently increased the concentration of carnosic acid. Lower sodium nitrite doses resulted in significantly lower nitrite concentrations and slightly lower TBARS values. It was concluded that in liver pâté sodium nitrite levels may be lowered to 80 mg/kg without negatively affecting colour and lipid stability and that the use of RE may help in maintaining lipid stability.

The occurrence of N-nitrosamines, residual nitrite and biogenic amines in commercial dry fermented sausages and evaluation of their occasional relation.

Regarding food borne intoxications, the accumulation of biogenic amines must be avoided in all kinds of food products. Moreover, biogenic amines can function as precursors for the formation of carcinogenic N-nitrosamines when nitrite is present. To estimate the food safety of the dry fermented sausages available on the Belgian market, a screening of the residual sodium nitrite and nitrate contents, biogenic amines and volatile N-nitrosamine concentrations was performed on 101 samples. The median concentrations of residual NaNO2 and NaNO3 were each individually lower than 20mg/kg. In general, the biogenic amine accumulation remained low at the end of shelf life. Only in one product the amounts of cadaverine and putrescine reached intoxicating levels. Concerning the occurrence of N-nitrosamines, only N-nitrosopiperidine and N-nitrosomorpholine were detected in a high number of samples (resp. 22% and 28%). No correlation between the presence of N-nitrosamines and the biogenic amines content was observed. Although the N-nitrosamines could not been linked to specific product categories, the occurrence of N-nitrosopiperidine could probably be attributed to the use of pepper.

Technology-induced selection towards the spoilage microbiota of artisan-type cooked ham packed under modified atmosphere.

The microbiota associated with a highly-perishable Belgian artisan-type cooked ham was analyzed through plating and (GTG)(5)-fingerprinting of isolates throughout its processing chain. The raw tumbled meat was characterized by the presence of a versatile microbiota around 4.8 log(cfu g(-1)), consisting of lactic acid bacteria, staphylococci, Brochothrix thermosphacta, Gram-negative bacteria, and yeasts. Pasteurisation of the ham logs reduced bacterial counts below 2 log(cfu g(-1)) and subsequent manipulations selected for leuconostocs and carnobacteria. Also, B. thermosphacta and several Enterobacteriaceae were found at this stage. During storage in an intermediate high-care area for 2 days, a selection towards certain Enterobacteriaceae (Hafnia alvei, Enterobacter spp., and Pantoea agglomerans) and lactic acid bacteria (mainly vagococci and Streptococcus parauberis) was observed. B. thermosphacta, Leuconostoc carnosum and carnobacteria were also detected, but only after allowing bacterial outgrowth by incubating the meat logs at 7 degrees C for four weeks. After a mild post-pasteurisation process and subsequent handling, incubation of the meat logs at 7 degrees C for four weeks led to outgrowth of Enterobacteriaceae (mainly Enterobacter spp. and Serratia spp.). B. thermosphacta, and lactic acid bacteria (Enterococcus faecalis, Leuc. carnosum, and Carnobacterium maltaromaticum) were also found. After slicing and packaging under modified atmosphere, the microbiota of the refrigerated end-product consisted of leuconostocs, carnobacteria, and B. thermosphacta.

Role of proline and hydroxyproline in N-nitrosamine formation during heating in cured meat.

N-Nitrosamines are formed in a multi-step reaction of nitrite with free amino acids and amines in the meat products. The aim of this study was to determine the role of proline and hydroxyproline in N-nitrosamines formation during heating of cured meat. A lean meat model was used with different nitrite concentrations (0, 120, and 480mg/kg), and addition of proline and hydroxyproline (1000mg/kg), followed by heating at different temperatures. Volatile nitrosamines were analyzed with GC-TEA. The nitrosamine content never exceeded 10μg/kg and stayed <LOQ as long as the nitrite level of 120mg/kg was not surpassed. The importance of proline as a precursor for N-nitrosamine formation was confirmed. In contrast, hydroxyproline inhibited NPYR formation (N-nitrosopyrrolidine) because no traces were found after addition of hydroxyproline. NPYR formation was not related to nitrite, but was significantly influenced by temperature (⩾200°C) and proline. NDMA-presence (N-nitrosodimethylamine) in heated meat products was influenced by nitrite and temperatures >120°C.

Evaluation of the influence of proline, hydroxyproline or pyrrolidine in the presence of sodium nitrite on N-nitrosamine formation when heating cured meat.

N-nitrosamines are meant to be probable or possible carcinogenic components, possibly formed out of a reaction between nitrite and N-containing substances such as amino acids and secondary amines. Nitrite is often used for processing meat products because of its colouring and antimicrobial properties. During this experimental setup, the influence of proline, hydroxyproline or pyrrolidine on N-nitrosamine formation in meat samples was evaluated. The N-nitrosamines concentrations were measured with gas chromatography-thermal energy analyzer. Only the concentrations of N-nitrosodimethylamine and N-nitrosopyrrolidine were found above the limit of detection in a number of tested experimental conditions. The concentration of these two N-nitrosamines was modelled as a function of temperature and nitrite concentration for different situations (presence or absence of added natural N-containing meat components). It could be concluded that proline and pyrrolidine promoted the formation of N-nitrosopyrrolidine. It could also be confirmed that the higher the temperature of the meat processing procedure and the higher the sodium nitrite amounts added, the higher were the yields of the respective N-nitrosamines.

Interactions between bacterial isolates from modified-atmosphere-packaged artisan-type cooked ham in view of the development of a bioprotective culture

Growth and metabolite production of three dominant bacterial isolates (Carnobacterium divergens 3M14, Leuconostoc carnosum 3M42, and Brochothrix thermosphacta RMS6) from modified-atmosphere-packaged (MAP), artisan-type cooked ham were assessed for their interactions in view of the development of a bioprotective culture. During monoculture experiments in cooked ham simulation medium, Leuc. carnosum 3M42 converted the available glucose into lactic acid and ethanol, whereas the two other strains produced additional metabolites such as acetic acid and 3-methyl-1-butanol. When grown in co-culture, Leuc. carnosum 3M42 suppressed the growth and metabolism of B. thermosphacta RMS6. In contrast, a co-culture of the latter bacterium with C. divergens 3M14 led to a variety of spoilage-related metabolic compounds. Subsequently, experiments with a commercial cooked ham product indicated that Leuc. carnosum 3M42 dominated the meat matrix and improved acceptability of the product over time, hence acting as a bioprotective culture for MAP, artisan-type cooked ham.