Colour formation in fermented sausages by meat-associated staphylococci with different nitrite- and nitrate-reductase activities

Three Staphylococcus strains, S. carnosus, S. simulans and S. saprophyticus, selected due to their varying nitrite and/or nitrate-reductase activities, were used to initiate colour formation during sausage fermentation. During fermentation of sausages with either nitrite or nitrate added, colour was followed by L^*a^*b measurements and the content of nitrosylmyoglobin (MbFe^IINO) quantified by electron spin resonance (ESR). MbFe^IINO was rapidly formed in sausages with added nitrite independent of the presence of nitrite reducing bacteria, whereas the rate of MbFe^IINO formation in sausages with added nitrate depended on the specific Staphylococcus strain. Strains with high nitrate-reductase activity showed a significantly faster rate of pigment formation, but other factors were of influence as well. Product stability for the sliced, packaged sausage was evaluated as surface colour and oxidation by autofluorescence and hexanal content, respectively. No significant direct effect of the Staphylococcus addition was observed, however, there was a clear correspondence between high initial amount of MbFe^IINO in the different sausages and the colour stability during storage. Autofluorescence data correlated well with hexanal content, and may be used as predictive tools. Overall, nitrite- and nitrate-reductase activities of Staphylococcus strains in nitrite-cured sausages were of limited importance regarding colour development, while in nitrate-cured sausages strains with higher nitrate reductase activity were crucial for ensuring optimal colour formation during initial fermentation stages.     

The effects of lactate on nitrosylmyoglobin formation from nitrite and metmyoglobin in a cured meat system

Two experiments were conducted to determine the effects of lactate on nitrite during meat curing. In the first experiment, using a model system, eight reaction components including nitrite and lactate, were used to assess the effect of each component on metmyoglobin reducing activity by excluding one component at a time. Excluding lactate, nicotinamide adenine dinucleotide (NAD), l-lactate dehydrogenase (LDH) or phenazine methosulfate (PMS) resulted in no reducing activity. A second experiment, utilising a meat mixture, investigated the effects of lactate (0%, 2%, 4% or 6%), nitrite (0 or 156 ppm), and packaging (oxygen-permeable or vacuum) on residual nitrite, meat colour and pH. Addition of lactate reduced residual nitrite in the meat mixtures. Both experiments support the hypothesis that lactate generates NADH which then reduces metmyoglobin to deoxymyoglobin. The resulting greater concentration of reduced myoglobin subsequently reacted with nitrite to produce more nitric oxide, reducing nitrite concentration and accelerating curing reactions.     

Nitrate and nitrite quantification from cured meat and vegetables and their estimated dietary intake in Australians

High dietary nitrate and nitrite intake may increase the risk of gastro-intestinal cancers due to the in vivo formation of carcinogenic chemicals known as N-nitroso compounds. Water and leafy vegetables are natural sources of dietary nitrate, whereas cured meats are the major sources of dietary nitrite. This paper describes a simple and fast analytical method for determining nitrate and nitrite contents in vegetables and meat, using reversed-phase HPLC-UV. The linearity R² value was >0.998 for the anions. The limits of quantification for nitrite and nitrate were 5.0 and 2.5 mg/kg, respectively. This method is applicable for both leafy vegetable and meat samples. A range of vegetables was tested, which contained <23 mg/kg nitrite, but as much as 5000 mg/kg of nitrate. In cured and fresh meat samples, nitrate content ranged from 3.7 to 139.5 mg/kg, and nitrite content ranged from 3.7 to 86.7 mg/kg. These were below the regulatory limits set by food standards Australia and New Zealand (FSANZ). Based on the average consumption of these vegetables and cured meat in Australia, the estimated dietary intake for nitrate and nitrite for Australians were 267 and 5.3 mg/adult/day, respectively.     

Formation and identification of nitrosylmyoglobin by Staphylococcus xylosus in raw meat batters: A potential solution for nitrite substitution in meat products

Staphylococcus xylosus and Pediococcus pentosaceus isolated from Chinese dried sausage were assessed for their ability to convert metmyoglobin into nitrosylmyoglobin in Mann–Rogosa–Sharp broth model systems and raw pork meat batters without the addition of nitrite. The results showed that samples in model systems with S. xylosus cultures had an absorption spectra that is typical of nitrosylmyoglobin, an obvious pink colour (judged by visual inspection) and a significantly higher a*-value than the control samples or samples inoculated with P. pentosaceus. In raw meat batters, the a*-values of the S. xylosus samples were almost the same as those for the meat with nitrite added. The complementary analysis of meat batter samples by photochemical information from UV–vis, electron spin resonance and resonance Raman spectroscopy revealed that the existing status of the myoglobin in meat batters inoculated with S. xylosus was mainly pentacoordinate nitrosylmyoglobin. This study provides a potential solution for nitrite substitute in meat products.     

Zn-porphyrin formation in cured meat products: Effect of added salt and nitrite

Zn-porphyrin (Zn-pp) was quantified by fluorescence spectroscopy in the cured and dry cured meat products: Parma ham, Iberian ham, dry-cured ham with added nitrite, cooked ham with added nitrite, raw ham meat, raw bacon and Karree-Speck. The highest amount of Zn-pp was found in dry-cured Parma ham and Iberian ham, while the use of nitrite as curing agent was found to inhibit completely the formation of Zn-pp in meat products. A positive correlation between both Zn content and Fe content and the logarithmic transformed Zn-pp content (measured as fluorescence intensity I_fl) was found for the different cured and dry cured meat products, with correlation coefficients of 0.79 (p < 0.001) and 0.71 (p < 0.01), respectively. Log I_fl correlates best with the Zn content, indicating that the formation of Zn-pp is proportional to the Zn content. A model system with vacuum packed pork in brine with different added levels of sodium chloride with or without nitrite and Zn acetate was investigated in order to further elucidate the mechanism of Zn-pp formation. Zn-pp increased with time (up to 42 days investigated) in non-cured meat and for meat cured solely with NaCl lower than 9%. Addition of nitrite or Zn(II) in the curing brine was found to inhibit formation of Zn-pp confirming the observations from the various cured meat products. It is suggested that a chloride anion assisted dissociation of iron from myoglobin could be rate-determining for Zn-pp formation in meat products.     

The determination of nitrate and nitrite in cured meat by HPLC/UV

A rapid strong anion exchange HPLC/UV procedure has been developed for the determination of nitrate and nitrite in a wide variety of cured meats. The accuracy of this technique has been confirmed by the good agreement achieved with the existing British Standard colorimetric method. The applicability and repeatability of the procedure has been established in a survey of over 200 samples. The agreement between duplicate determinations and their respective means averaged +/- 3.4% for nitrite and +/- 4.3% for nitrate as defined by the term [(a - b)/(a + b)] X 100% where a and b are the repeat determination values.     

茶多酚和维生素C对腌制猪肉亚硝酸盐残留量和脂肪氧化的影响

以腌制猪肉模型为材料,研究不同含量的茶多酚和维生素C对色泽、残留亚硝酸盐含量以及脂肪氧化TBARS值的影响。结果表明,茶多酚和维生素C都在一定程度上降低了腌制模型中亚硝酸钠的残留量,维生素C的作用效果要显著强于茶多酚(p<0.05)。腌制过程中TBARS值呈逐渐增加的趋势,茶多酚和维生素C有降低TBARS值的作用,茶多酚的作用效果要强于维生素C。在腌制过程中,维生素C组的色泽显著高于茶多酚组(p<0.05),而蒸煮后各组之间色泽差异不显著(p>0.05)。      

Use of natural antimicrobials to improve the control of Listeria monocytogenes in a cured cooked meat model system

Concern about nitrite in processed meats has increased consumer demand for natural products manufactured without nitrite or nitrate. Studies on commercial meat products labeled as "Uncured" and "No-Nitrite-or-Nitrate-Added" have shown less control of nitrite in these products and greater potential growth of bacterial pathogens. To improve the safety of the "naturally cured" meats, several natural ingredients were studied in a cured cooked meat model system (80:20 pork, 10% water, 2% salt, and 150 or 50 ppm ingoing sodium nitrite) that closely resembled commercial frankfurters to determine their inhibitory effect on Listeria monocytogenes. Results showed that cranberry powder at 1%, 2% and 3% resulted in 2-4 log cfu/g less growth of L. monocytogenes compared to the control with nitrite alone (P<0.05). Other natural compounds, such as cherry powder, lime powder and grape seed extract, also provided measureable inhibition to L. monocytogenes when combined with cranberry powder (P<0.05).     

A reference isotope dilution headspace GC/MS method for the determination of nitrite and nitrate in meat samples

A novel method for the determination of nitrite and nitrate in meat products is presented. The samples were ground and extracted in hot water with the presence of and internal standards. The solution was buffered with sodium bicarbonate and reacted with triethyloxonium tetrafluoroborate to convert nitrite and nitrate into EtNO₂ and EtONO₂. Such derivatives could be detected by headspace GC/MS in positive chemical ionisation mode with 0.05 µg g⁻¹ and 1.0 µg g⁻¹ LOD. The method was used for and quantitation in the 0.5-300 and 2.5-300 µg g⁻¹ ranges. The method was applied for the analysis of fifteen meat products. Despite minimal sample preparation, the headspace sampling ensured a clean chromatography for over 135 analyses (throughput ten samples per hour). The proposed method offers selective GC/MS detection combined with high-precision isotope dilution calibration, it is suitable for metrological applications and can support regulations on meat safety (European Commission, 2011).     

The transnitrosation of N-methylaniline by a protein-bound nitrite model system in relation to N-nitrosamine formation in cured meats

The reactions of sodium nitrite, S-nitrosocysteine hydrochloride, S-nitrosoglutathione and a ‘protein-bound nitrite’ model system with N-methylaniline at pH 5.5 and 37°C have been compared. The results show that the rate of nitrosamine formation from sodium nitrite and S-nitrosocysteine is greater than that from the model system although the latter depends upon the local concentration of nitrosothiol groups on the matrix. Transnitrosation by S-nitrosoglutathione is considerably slower than the other reactions. The implications of the results with respect to nitrosamine formation in cured meat are discussed.     

Prediction of NaCl,nitrate and nitrite contents in minced meat by using a voltammetric electronic tongue and an impedimetric sensor

A method for predicting levels of sodium chloride, sodium nitrite and potassium nitrate in minced meat by using a combination of two different electrochemical methods; namely an electronic tongue (ET) based on pulse voltammetry and electrochemical impedance spectroscopy measurements is proposed here. The measurements with the voltammetric ET were carried out on both saline solutions (brines) and minced meat, whereas the impedimetric sensor was used only in minced meat. The addition of the salts was performed following an experimental design in which a system of three compounds/three levels was established. Multivariate analysis including Cross validation and Partial Least Square (PLS) techniques were applied for data management and prediction models building. A very good prediction of the concentration of chloride was achieved, whereas the prediction for the concentration of nitrate and nitrite can be considered as moderate.     

Characterization and technological properties of Staphylococcus xylosus strains isolated from a Tunisian traditional salted meat

The technological properties of strains of Staphylococcus xylosus were studied to select the most suitable for use as starter cultures for the production of dried fermented meat products. Strains of S. xylosus were isolated from traditional salted Tunisian meat and were identified by biochemical and molecular methods. Thirty strains of S. xylosus were studied to evaluate their catalase, nitrate reductase, lipolytic, proteolytic and antibacterial activities as well as growth ability at different temperatures, pH's and NaCl concentrations. All strains of S. xylosus had catalase activity and were able to reduce nitrates to nitrites. The nitrate reductase activity increased when the strains were kept under anaerobic conditions. Proteolytic activity on milk and on gelatin agar was demonstrated for 100% and 83.3% of the S. xylosus isolates, respectively. However extracellular proteolytic activity as assessed by the azocasein method was poor in all the strains. Lipolytic activity as assessed by the agar method showed that 76.6% of strains of S. xylosus could hydrolyze Tween 20 against 33.3% that could hydrolyze tributyrin. Tween 80 was hydrolyzed by only 10% of strains. Strains of S. xylosus hydrolyzed pork fat better than beef and lamb fat. The majority of strains had antibacterial activity against Salmonella arizonae, Staphylococcus aureus, Pseudomonas aeuroginosa, Escherichia coli and Enterococcus faecalis.     

Time-dependent depletion of nitrite in pork/beef and chicken meat products and its effect on nitrite intake estimation

The food additive nitrite (E249, E250) is commonly used in meat curing as a food preservation method. Because of potential negative health effects of nitrite, its use is strictly regulated. In an earlier study we have shown that the calculated intake of nitrite in children can exceed the acceptable daily intake (ADI) when conversion from dietary nitrate to nitrite is included. This study examined time-dependent changes in nitrite levels in four Swedish meat products frequently eaten by children: pork/beef sausage, liver paté and two types of chicken sausage, and how the production process, storage and also boiling (e.g., simmering in salted water) and frying affect the initial added nitrite level. The results showed a steep decrease in nitrite level between the point of addition to the product and the first sampling of the product 24 h later. After this time, residual nitrite levels continued to decrease, but much more slowly, until the recommended use-by date. Interestingly, this continuing decrease in nitrite was much smaller in the chicken products than in the pork/beef products. In a pilot study on pork/beef sausage, we found no effects of boiling on residual nitrite levels, but frying decreased nitrite levels by 50%. In scenarios of time-dependent depletion of nitrite using the data obtained for sausages to represent all cured meat products and including conversion from dietary nitrate, calculated nitrite intake in 4-year-old children generally exceeded the ADI. Moreover, the actual intake of nitrite from cured meat is dependent on the type of meat source, with a higher residual nitrite levels in chicken products compared with pork/beef products. This may result in increased nitrite exposure among consumers shifting their consumption pattern of processed meats from red to white meat products.     

Effect of concentrate feeding pattern in a grass silage/concentrate beef finishing system on performance, selected carcass and meat quality characteristics

Steers were offered grass silage ad libitum and 6.4 kg concentrates daily for 126 days or silage ad libitum for 35 days, followed by concentrates ad libitum (Experiment 1). Steers were offered grass silage ad libitum and 6 kg concentrates daily for 154 days, concentrates ad libitum or grass silage ad libitum for 112 days followed by concentrates ad libitum (Experiment 2). All treatments received the same total concentrate allowance. In Experiment 1, there was no difference in any measurement of meat quality. In Experiment 2, ad libitum concentrate feeding per se, decreased redness and increased shear force of muscle at 2 days post-mortem. Delaying concentrate feeding decreased fat yellowness, decreased shear force at 7 and 14 days post-mortem and increased muscle redness at 14 days post-mortem. Modifications of the beef production system examined had minor effects on beef quality which are unlikely to be of commercial significance.     

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 480 mg/kg), and addition of proline and hydroxyproline (1000 mg/kg), followed by heating at different temperatures. Volatile nitrosamines were analyzed with GC-TEA.     

浓香型大曲中酶系与白酒品质的关系研究

大曲在白酒酿造中有产酒和生香的作用,而这些风味物质的形成与大曲中的酶系有很大的关系,从大曲生化性能中水解酶类、酯化酶和其他酶类与酒体中微量成分关系的分析,探讨了大曲酶系与白酒品质的关系。     

Effect of water activity and temperature on degradation of 5′-inosine monophosphate in a meat model system

The influence of water activity (a_w) (0.7,0.8 and 0.9) and temperature (80° and 120 °C) on the degradation of meat flavor precursor inosine monophosphate (IMP) was studied in a meat fiber model system. Breast and leg muscle from Indian domesticated layer chicken (Gallus gallus) were washed repeatedly with 0.1 mol/l phosphate buffer of pH 6 to obtain pigment free and with minimum content of natural IMP in muscle fiber. The freeze-dried breast and leg meat fiber had a protein content of 86.5±0.48% and 85.6±0.50%, respectively. The IMP contents (mg/100 g) of leg muscle fiber (7.3±0.60) was higher (P?0.05) than in the breast meat fiber (5.1±1.20). The degradation of IMP was temperature-dependent (P?0.05) in both types of meat fiber systems. In the samples of a_w 0.8, the IMP degradation in breast meat fiber system was lower (P?0.05) than in a_w at 0.7 and 0.9 samples, when treated at 80 °C, whereas, there was no significance difference (P>0.05) in the degradation of IMP at a_w 0.7 and 0.9 when heated at 120 °C. The degradation of IMP in leg meat fiber model system at higher a_w (0.9) was more (P?0.05) as compared to lower a_w (0.7 and 0.8) at 80 °C, while the samples treated at 120 °C, the degradation of IMP at a_w 0.8 and 0.9 was more (P?0.05) than at a_w 0.7.     

Effect of breed on performance and meat quality of first parity sows in a seasonal organic rearing system

BACKGROUND: The objective was to compare the performance and meat quality of two different pig breeds: the modern crossbred Landrace × Yorkshire (LY) and the traditional Danish Black‐Spotted (BS) breed. The LY gilts and four of the BS gilts were inseminated with semen from Duroc (D) boars; the remaining BS gilts were served by BS boars. The experiment was performed on 26 gilts over 2 years. RESULTS: The BS breed in general had smaller litter sizes and weaned on average in terms of kg piglet 35% less than the modern breed combination. Crossbred piglets of BS × D had growth rates from birth to weaning comparable with piglets of LY × D. BS first parity sows were significantly fatter and less meaty, and the meat was significantly redder and darker compared to LY. The sensory profiling revealed increased sweetness, crumbliness and tenderness in BS. Further, the fat of the BS breed seemed to be characterized by a special nutty taste. CONCLUSION: The traditional breed has lower productivity and thereby higher costs of production compared to the modern genotypes. On the other hand, the meat and fat of the traditional purebred have special characteristics that might trigger a market premium. Copyright © 2011 Society of Chemical Industry     

Control of food spoiling bacteria in cooked meat products with nisin,lacticin 3147, and a lacticin 3147-producing starter culture

Nisin, in the form of the commercial product Nisaplin, and lacticin 3147 in whey powdered form were added to minced pork-meat in amounts of 0.15% (w/w) and 1.5% (w/w), respectively. The meat was cooked and inoculated with a Staphylococcus aureus strain of meat origin and a Listeria innocua strain at a level of 10⁷ or 10⁵ CFU g^–1. The batches were stored vacuum-packaged for 21 days at 8 °C. Nisin and lacticin 3147 immediately reduced the L. innocua population at the time of inoculation. Nisin showed higher inhibitory activity than lacticin 3147. During the storage period, a slight L. innocua growth was observed in the batches inoculated with the larger inoculum, and a bacteriostatic effect was observed against Listeria in the batches inoculated with 10⁵ CFU g^–1. Nisin maintained a constant S. aureus population in the cooked batch inoculated with 10⁷ CFU g^–1, although the bacteriocin was capable of reducing the amount of S. aureus by 90% in the batch inoculated with 10⁵ CFU g^–1. On the other hand, lacticin 3147 did not show an inhibitory effect against S. aureus in the cooked meat. The starter culture Lactococcus lactis DPC 303-T4 (containing the conjugative plasmid encoding production of lacticin 3147) was inoculated in a portion of a Longissimus dorsi pork muscle with brine. L. lactis DPC 303-T4 performed a good fermentation, but lacticin 3147 production was not found after 7 days at 12 °C of storage.